Immune Dysfunction and Systemic Inflammatory infiltration Exist in Neuronal Intranuclear Inclusion Disease

Immune Dysfunction and Systemic Inflammatory infiltration Exist in Neuronal Intranuclear Inclusion Disease | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Short Report Immune Dysfunction and Systemic Inflammatory infiltration Exist in Neuronal Intranuclear Inclusion Disease Lei Bao, Dandan Zuo, Xiaoying Qu, Yingying Cui, Keke Li, Jing Dong, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3703208/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Neuronal intranuclear inclusion disease (NIID) is primarily recognized as a neurodegenerative disorder due to the production of a neurotoxicity protein, uN2CpolyG. However, evidence suggests its systemic nature, prompting an exploration of the immune and systemic inflammatory aspects of NIID in this study. A cohort of 32 diagnosed NIID patients participated in a comprehensive study involving clinical presentations, and tissue specimen analyses. Peripheral blood monocyte cells (PBMCs) were collected to detect uN2CpolyG expression in NIID patients by immunofluorescence staining and Western blotting. NIID patients showed varied neurological and extra-neurological symptoms alongside systemic inflammatory and autoimmune disorders, including ulcerative colitis, Sjögren's syndrome, Hashimoto's thyroiditis, and IgA nephropathy. 19 previous tissue specimens from these patients displayed evidence of inflammatory cell infiltration. Notably, our observations unveiled the novel presence of eosinophilic inclusions within the nuclei of these infiltrating inflammatory cells, primarily concentrated in mononuclear cells. Additionally, uN2CpolyG aggregates, identified as ubiquitin-positive inclusions, were detected in peripheral blood monocyte cells (PBMCs) from NIID patients for the first time, contrasting with the weak signal observed without inclusions in the control group. The detection of uN2CpolyG as a 30 to 40 kDa protein in the PBMCs from three NIID patients further supports our findings. This study highlights NIID's systemic nature, emphasizing immune dysfunction and systemic inflammatory infiltration. The detection of uN2CpolyG aggregates in the PBMCs of NIID patients suggests that it may have a toxic potential and alter the immune response of these cells. Neuronal intranuclear inclusion disease (NIID) uN2CpolyG Immune Inflammatory Peripheral blood monocyte cell (PBMC). Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Neuronal intranuclear inclusion disease (NIID) is a progressive neurodegenerative disorder due to GGC repeat expansions in the NOTCH2NLC gene ( 1 ). This trinucleotide repeat disorder is typically adult-onset with incomplete penetrance and variable phenotypes. Common clinical manifestations of NIID patients include cognitive dysfunction, movement disorders, autonomic dysfunction, paroxysmal symptoms, and muscle weakness ( 2 ). The pathogenic mechanism of NIID is associated with the expansion of GGC repeats, which leads to the production of a polyglycine protein called uN2CpolyG. Due to its specific structural characteristics, this protein continually aggregates and forms insoluble protein deposits, primarily within the cell nucleus as intranuclear inclusions, which serve a toxic effect on neurons, ultimately giving rise to the clinical symptoms of NIID ( 3 ). However, this protein aggregate is not limited to deposition within the nuclei of neurons, it can also be found in nearly all internal organs, leading to a variety of non-neurological symptoms, including visual impairment, hearing loss, irritant dry cough, arrhythmia, cardiac insufficiency, vomiting, and constipation ( 4 , 5 ). These findings have paved the way for the hypothesis that NIID is not solely a condition associated with CNS dysfunctions but rather a systemic disease. A recent study reported a notable presence of macrophage/microglia infiltration within the brain lesion of a NIID patient who suffered an encephalitic episode ( 6 ). This finding indicates the role of neuroinflammation in the development and progression of NIID, particularly in the context of encephalitic episodes. Given the intricate array of extra-neurological clinical symptoms exhibited by patients with NIID, it remains uncertain whether inflammatory cell infiltration also occurs in the peripheral organs of these individuals. Moreover, the vast majority of studies only focus on lesions of solid organs, with few research exploring the presence of inclusions in peripheral blood inflammatory cells or evaluating the immune and inflammatory status of patients with NIID. Given all this, we aim to establish compelling evidence supporting dysregulated immune responses and systemic inflammatory infiltration in NIID. Materials and methods Study participants This study included 32 NIID-diagnosed patients treated at the Affiliated Hospital of Xuzhou Medical University between January 2018 and June 2023. The diagnosis of NIID was established based on clinical presentations and the detection of abnormal GGC repeats (> 65) within NOTCH2NLC, with or without additional confirmation through biopsy. Clinical assessment The neurological and extra-neurological symptoms of this patient group were meticulously evaluated by two experienced clinicians, focusing on identifying the presence of comorbid immune system diseases. Peripheral blood monocyte cell (PBMC) isolation Whole blood samples were collected from NIID patients and healthy individuals, utilizing EDTA tubes. The extraction of PBMCs was performed using a density gradient with Ficoll-PaqueTM PLUS following the manufacturer's protocol. To achieve this, blood samples were diluted at a 1:1 ratio with Dulbecco’s phosphate-buffered saline and gently layered onto an equal volume of Ficoll-Paque solution. Subsequently, the samples underwent centrifugation at 440× g for 30 minutes at room temperature (approximately 22 ˚C). Carefully isolating the PBMCs at the interface, they were washed twice with 1× PBS and then subjected to subsequent experiments. Immunofluorescence staining PBMCs, freshly obtained, were cultured on poly-L-lysine-coated confocal dishes in 10% fetal bovine serum-supplemented RPMI 1640 medium. After 1 hour incubation at 37˚C for adherence, cells were fixed in 4% formaldehyde for 15 minutes and permeabilized with 0.2% PBST (PBS with Triton X) for 15 more minutes. Subsequently, a 4% BSA block was applied for 1 hour. Primary antibodies, including 4D12 (mouse antibody for uN2CpolyG at 1:200, courtesy of Professor Nicolas Charlet-Berguerand) and rabbit anti-ubiquitin (1:200, Abcam, ab7780), were incubated overnight at 4°C. Following PBS washing, secondary fluorescent antibodies—goat anti-rabbit Alexa Fluor Plus 594 and goat anti-mouse Alexa Fluor Plus 488 (both at 1:1000, Invitrogen)—were incubated for 1 hour at room temperature. The cells were then DAPI-stained for 1 minute before imaging with a Leica Stellaris 5 confocal microscope (Leica, Germany). Western blot Analysis PBMCs were lysed in RIPA buffer for 10 minutes, followed by centrifugation at 4°C, 13,000g for 30 minutes. The resultant supernatant, containing proteins, was collected for western blot analysis. Equal protein amounts were electrophoresed, transferred onto nitrocellulose membranes, and blocked with 3% BSA for equal loading. The target membrane was incubated overnight at 4°C with 4D12 (anti-uN2CpolyG antibody, 1:1000). Then, membranes were incubated with goat anti-mouse IgG-HRP secondary antibodies (1:500) for 2 hours at room temperature. Subsequently, an enhanced chemiluminescence kit was used for visualization, and protein bands were detected using a gel imaging system (ChemiDoc XRS+, Bio-Rad). Results Involvement of immune system as another extra-neurological manifestation in NIID Table 1 presented in the study provides an overview of the general information for the 32 patients with NIID, along with details regarding their neurological and extra-neurological clinical manifestations. Within this study, the neurological clinical manifestations observed in NIID patients were categorized into five primary symptom groups. These encompassed cognitive impairment, movement disorders, muscle weakness, paroxysmal symptoms, and autonomic dysfunction. We have also compiled a summary of non-neurological clinical symptoms observed in these patients, categorizing them by the affected systems. Respiratory system involvement primarily presents as irritant dry cough and wheezing. Gastrointestinal system involvement is characterized by poor appetite, nausea, vomiting, diarrhea, and constipation. Urinary system manifestations include proteinuria, urinary retention, and renal insufficiency. Endocrine system abnormalities include diabetes or impaired glucose tolerance and hypothyroidism. In the current study, we obtained a total of 19 archived surgical specimens derived from the cohort of 32 patients diagnosed with NIID. These specimens were collected from various anatomical sources, representing a diverse array of tissues within the patients exhibiting NIID symptoms. Table 1 outlines the specific sources of surgical specimens, encompassing 6 samples from the gastrointestinal tract, 3 from the skin, 2 from the lung, 2 from the bladder, and 1 each from the brain, kidney, adrenal gland, prostate, and fallopian tube. Subsequent histopathological assessment, utilizing H&E-stained sections (refer to Figure 1), revealed the presence of eosinophilic inclusions within the nuclei of these tissues. We additionally observed the occurrence of systemic inflammatory and autoimmune disorders within this patient cohort. Specifically, patient 2 with a history of ulcerative colitis exhibited symptoms, including weight loss, rectal bleeding, and chronic diarrhea. Patient 12 displayed hallmark clinical characteristics of Sjögren's syndrome, notably presenting with Raynaud's phenomenon, dry eyes, and dry mouth. Hashimoto's thyroiditis was incidentally discovered in patient 25 during a routine physical examination, with the levels of anti-TPO and anti-thyroglobulin antibodies being significantly elevated. Patient 30 was admitted to the hospital due to early-onset cognitive dysfunction. Following routine assessments, the presence of hematuria, substantial proteinuria, and renal insufficiency was identified. Subsequent biopsy detected the deposition of immunoglobulin A within the kidney tissue confirming the diagnosis of IgA nephropathy. Infiltration of inflammatory cells in tissue specimens from NIID patients. Figure 2 presented a pervasive presence of inflammatory cell infiltration across various tissues from NIID patients. Briefly, in the temporal lobe brain biopsy of a patient experiencing an encephalitis-like attack, a notable presence of scattered microglia was observed. The identification of small focal inflammatory cell infiltrates in a separate patient's skin biopsy is particularly intriguing, especially considering this patient's prior diagnosis of neurodermatitis attributed to recurrent skin itching and ulceration. Further findings included marked inflammatory cell infiltration in a lung tissue biopsy from a patient with an unidentified pulmonary nodule, and diffuse inflammatory cell infiltration in a tissue specimen obtained from a colon polypectomy. Inflammatory cell infiltration was also evident in various other tissue samples: scattered inflammatory cells were detected in the renal tissue of a patient with IgA nephropathy, and patchy inflammatory cell infiltration was observed in surgical tissue specimens from a patient with hydrosalpinx. Notably, we also observed eosinophilic inclusions within the nuclei of these infiltrated inflammatory cells. Although the majority of specimens lacked immunohistochemical analysis for specific inflammatory cell markers, hindering precise cell phenotype characterization, our observations indicated that these eosinophilic intranuclear inclusions were primarily evident in mononuclear cells (Figure 2, zoom). This intriguing finding prompted further investigation into whether a similar phenomenon occurs in PBMCs. Detection of uN2CpolyG aggregates-formed inclusions in PMBCs from NIID patients To investigate whether the intranuclear inclusions also existed in peripheral inflammatory cells, we obtained fresh PBMCs from three NIID patients. In a significant proportion of PBMC cells, we observed the presence of aggregates resulting from the accumulation of uN2CpolyG protein within the nucleus. Moreover, a subset of these protein aggregates displayed positive immunoreactivity for ubiquitin. In sharp contrast, samples collected from healthy individuals with normal CGG repeat lengths exhibited only weak signals and were devoid of any inclusions (Figure. 3a). Furthermore, we conducted western blot analysis to assess the expression of uN2CpolyG protein in PBMCs from these NIID patients. Our results revealed the presence of a 30-40kD uN2CpolyG protein in the PBMCs of three NIID patients. However, in the PBMCs of control individuals without GGC duplications, we did not detect any protein expression (Figure. 3b). Discussion Despite being labeled as “neuronal” intranuclear inclusion disease, our previous research suggests that the term "systemic" intranuclear inclusion disease may be more appropriate. This is because eosinophilic intranuclear inclusions have been identified in the tissue cells of nearly all organs and their presence can lead to organ-specific lesions, resulting in a wide array of extra-neurological manifestations( 5 ). For instance, individuals with visual dysfunction have exhibited widespread intranuclear inclusions and severe gliosis throughout the retina ( 7 ). Similarly, those with cardiomyopathy have displayed diffusely distributed intranuclear inclusions, along with myocyte hypertrophy and fibrosis ( 8 ). In the case of one NIID patient with unexplained massive proteinuria, renal tissue biopsies revealed intranuclear inclusions in podocytes and mesangial cells ( 9 ). However, previous studies, including our own, have predominantly concentrated on the analysis of lesions and symptoms in solid organs while overlooking the assessment of immune and inflammatory status in patients with NIID. Hence, this study aimed to investigate the presence of comorbid immune system disorders in patients with NIID. An intriguing observation in this study is the occurrence of four distinct inflammatory diseases within the cohort of 34 NIID patients, which included ulcerative colitis, Sjogren's syndrome, Hashimoto's thyroiditis, and IgA nephropathy. Indeed, this is not the first occurrence of NIID being associated with inflammatory conditions. Previous case reports have documented instances of patients with this disease presenting with comorbid conditions such as asthma, crescentic IgA nephropathy, lupus nephritis–like nephropathy, and neuropathy associated with IgM monoclonal gammopathy of undetermined significance (MGUS) ( 9 – 12 ). While neurodegenerative diseases and inflammatory diseases are distinct categories of diseases, some studies have indeed found an increased risk of comorbid inflammatory diseases in patients with neurodegenerative diseases, suggesting a potential link between these two types of conditions. A comprehensive review and meta-analysis of 873,643 individuals with Parkinson's disease (PD) found a significantly increased risk of PD in those with autoimmune diseases, including inflammatory bowel disease, Sjögren's syndrome, and Graves' disease ( 13 ). In a nationwide case-control study of 3,561 amyotrophic lateral sclerosis (ALS) patients and 35,610 controls, a 47% higher likelihood of prior autoimmune disease diagnosis was observed in ALS patients. Specific autoimmune conditions included myasthenia gravis, polymyositis or dermatomyositis, Guillain-Barre syndrome, type 1 diabetes, multiple sclerosis, and hypothyroidism ( 14 ). Multiple theories and hypotheses have been proposed to elucidate the association between neurodegenerative diseases and autoimmune diseases, although the precise underlying mechanisms remain incompletely understood. Neurodegenerative diseases may involve immune dysregulation, potentially leading to an exaggerated or misdirected immune response, thereby increasing the susceptibility to autoimmune diseases. Shared genetic factors are another consideration, with common genetic variants or susceptibility genes possibly influencing the development of both neurodegenerative and autoimmune conditions. Furthermore, chronic inflammation emerges as a common element in both neurodegenerative diseases and autoimmune diseases, contributing to the progression of neurodegenerative conditions and playing a pivotal role in many autoimmune disorders ( 15 , 16 ). Our present study revealed a potential association between NIID patients and inflammatory diseases, indicating an altered innate and adaptive immune system in NIID patients. However, the establishment of a causal relationship is inconclusive due to the limited sample size and the absence of controlled studies. It has been reported that more than half of the NIID patients had ever suffered acute encephalitis-like episodes as the disease progressed, presenting with fever, altered mental status, conscious disturbance, or focal neurological deficit ( 2 ). Further brain tissue biopsy revealed numerous macrophages/microglia infiltration in the lesion area ( 6 ). These findings strongly suggest that a pronounced immune response or inflammatory reaction is occurring within the brain tissue of NIID patients, potentially serving as a vital pathological mechanism in the development of NIID. In this study, we present the first evidence of significant infiltration of inflammatory cells within various lesion tissues of NIID patients, indicating, similar to the central nervous system, inflammatory responses may also be implicated in lesions within other systems. Nonetheless, we cannot conclusively determine whether the infiltration of these inflammatory cells is a primary pathological mechanism of multi-system lesions or merely a secondary reaction activated in response to pathological stimulus. Despite previous autopsy findings have consistently revealed intranuclear inclusions in the cells of brain tissues and various internal organs among NIID patients, our study marks the first to report the detection of these inclusions in both circulating and tissue-based immune cells. Detected from the brain and skin tissues of NIID patients, the main component of these inclusions is a glycine-rich protein, the uN2CpolyG protein, which is translated from the GGC repeat expansion in the 5'UTR of the NOTCH2NLC gene ( 3 ). In this study, we utilized immunofluorescence to confirm the formation of intranuclear inclusions comprised of the uN2CpolyG protein within PBMCs. Additionally, we also assessed the protein expression through WB analysis, revealing robust levels of uN2CpolyG expression in PBMCs derived from individuals diagnosed with NIID. To our current understanding, our research group stands as the initial cohort to effectively identify the uN2CpolyG protein in the peripheral blood of individuals diagnosed with NIID. Our findings also propose that the presence of uN2CpolyG inclusions in PBMCs could potentially act as a biomarker, which could significantly aid in the early identification and subsequent treatment of NIID. Previous studies have demonstrated the neurotoxicity of uN2CpolyG protein based on its reduction of cell viability, disruption of nucleocytoplasmic transport, induction of mitochondrial dysfunction, and inhibition of ribosome biogenesis and translation ( 3 , 17 – 19 ). These findings have offered crucial insights into the protein's detrimental effects on cellular mechanisms. PBMCs are a diverse group of immune cells found in the bloodstream, including lymphocytes, monocytes, and dendritic cells, which are key components of the innate and adaptive immune systems and crucial in response to inflammation ( 20 ). Whether the pathological protein uN2CpolyG could also affect the survival and functionality of PBMCs remains ambiguous at present, however, these findings led us to hypothesize that the increased expression of uN2CpolyG in PBMCs may potentially disrupt their typical immune response functions, potentially triggering an aberrant inflammatory response. This potential disruption in PBMC function could shed light on the pronounced escalation in autoimmune and autoinflammatory conditions observed in individuals diagnosed with NIID. This study exhibits several limitations. First, the study cohort consisted of a relatively small and potentially homogeneous group of patients from a single medical center. Consequently, the findings may have limited applicability to a broader population. Secondly, while the study identified inflammatory cell infiltration, it lacked detailed characterization of these cells, particularly their phenotypes and activation states. This absence restricts a comprehensive understanding of the immune response in NIID. Lastly, despite highlighting associations and correlations, the study did not delve into detailed mechanistic insights into how immune dysregulation and uN2CpolyG aggregates precisely contribute to disease pathogenesis. Conclusion The coexistence of inflammatory and autoimmune diseases alongside systemic inflammatory cell infiltration in NIID patients strongly points towards a dysregulated immune response and a systemic inflammatory condition in this disorder (Fig. 4 ). Moreover, the identification of uN2CpolyG aggregates within PBMCs of NIID patients implies a potential toxic effect, suggesting an alteration in the immune response of these cells. These findings collectively underscore the intricate involvement of immune dysregulation in NIID pathophysiology, potentially serving as crucial indicators for both disease understanding and therapeutic interventions. Abbreviations NIID: Neuronal intranuclear inclusion disease PBMC: Peripheral blood mononuclear cell MGUS: Monoclonal gammopathy of undetermined significance PD: Parkinson's disease ALS: Amyotrophic lateral sclerosis Declarations Ethics approval and consent to participate All research involving human subjects received approval from local review boards and adhered strictly to the principles outlined in the Declaration of Helsinki. The protocol of this study was approved by the Institutional Review Board of the Affiliated Hospital of Xuzhou Medical University. Prior to their participation, all individuals involved in the study provided informed written consent, signifying their voluntary agreement and understanding of the research objectives, procedures, and potential risks. Consent for publication We obtained written consent for publication from all NIID patients Availability of data and material The datasets produced or analyzed during this study are not publicly accessible to safeguard personal information. However, they can be made available from the corresponding author upon reasonable request. Competing interests The authors declare that there is no conflict of interest. Funding This work was supported by Project supported by the Affiliated Hospital of Xuzhou Medical University (2023ZL01). Authors' contributions LB designed the study and drafted the manuscript. DDZ created figures and prepared tables. XYQ collected clinical data and tissue specimens. YYC conducted histopathological analysis of tissue specimens. KKL assessed neuroimages. JD prepared pathological specimens. ZZS assessed neurological findings. GYC assessed neurological findings. HC designed the study and assessed extra-neurological findings. Acknowledgments We would like to express our gratitude to Professor Nicolas Charlet-Berguerand for generously providing us with the antibody for uN2CpolyG protein, which has been an invaluable experimental tool and support for our research. We sincerely thank Dr. Fuxing Dong from the Public Experimental Research Center for his enthusiastic help in the experiment of laser scanning confocal microscopy. References Ishiura H, Shibata S, Yoshimura J, Suzuki Y, Qu W, Doi K, et al. Noncoding CGG repeat expansions in neuronal intranuclear inclusion disease, oculopharyngodistal myopathy and an overlapping disease. Nat Genet. 2019;51(8):1222-32. Tian Y, Zhou L, Gao J, Jiao B, Zhang SZ, Xiao Q, et al. Clinical features of NOTCH2NLC-related neuronal intranuclear inclusion disease. J Neurol Neurosur Ps. 2022. Boivin M, Deng J, Pfister V, Grandgirard E, Oulad-Abdelghani M, Morlet B, et al. Translation of GGC repeat expansions into a toxic polyglycine protein in NIID defines a novel class of human genetic disorders: The polyG diseases. Neuron. 2021;109(11):1825-35 e5. Sone J, Mori K, Inagaki T, Katsumata R, Takagi S, Yokoi S, et al. Clinicopathological features of adult-onset neuronal intranuclear inclusion disease. Brain. 2016;139(Pt 12):3170-86. Chen H, Lu L, Wang B, Cui G, Wang X, Wang Y, et al. Re-defining the clinicopathological spectrum of neuronal intranuclear inclusion disease. Ann Clin Transl Neurol. 2020;7(10):1930-41. Mori K, Yagishita A, Funata N, Yamada R, Takaki Y, Miura Y. Imaging findings and pathological correlations of subacute encephalopathy with neuronal intranuclear inclusion disease-Case report. Radiol Case Rep. 2022;17(12):4481-6. Sone J, Ueno S, Akagi A, Miyahara H, Tamai C, Riku Y, et al. NOTCH2NLC GGC repeat expansion causes retinal pathology with intranuclear inclusions throughout the retina and causes visual impairment. Acta Neuropathol Commun. 2023;11(1):71. Oyer CE, Cortez S, O'Shea P, Popovic M. Cardiomyopathy and myocyte intranuclear inclusions in neuronal intranuclear inclusion disease: a case report. Hum Pathol. 1991;22(7):722-4. Morita K, Shinzato T, Endo Y, Suzuki M, Yoshida H, Sone J, et al. A case of unusual renal manifestation in a patient with neuronal intranuclear inclusion disease treated with steroids. Clin Case Rep. 2023;11(8):e7730. Horino T, Matsumoto T, Inoue K, Ichii O, Terada Y. A case of neuronal intranuclear inclusion disease associated with lupus nephritis-like nephropathy. eNeurologicalSci. 2018;10:28-30. Sugiyama A, Takeda T, Koide M, Yokota H, Mukai H, Kitayama Y, et al. Coexistence of neuronal intranuclear inclusion disease and amyotrophic lateral sclerosis: an autopsy case. BMC Neurol. 2021;21(1):273. Fu J, Zhao C, Hou G, Liu X, Zheng M, Zhang Y, et al. A case report of neuronal intranuclear inclusion disease with paroxysmal peripheral neuropathy-like onset lacking typical signs on diffusion-weighted imaging. Front Neurol. 2023;14:1117243. Li M, Wan J, Xu Z, Tang B. The association between Parkinson's disease and autoimmune diseases: A systematic review and meta-analysis. Front Immunol. 2023;14:1103053. Cui C, Longinetti E, Larsson H, Andersson J, Pawitan Y, Piehl F, et al. Associations between autoimmune diseases and amyotrophic lateral sclerosis: a register-based study. Amyotroph Lateral Scler Frontotemporal Degener. 2021;22(3-4):211-9. Doty KR, Guillot-Sestier MV, Town T. The role of the immune system in neurodegenerative disorders: Adaptive or maladaptive? Brain Res. 2015;1617:155-73. DeMaio A, Mehrotra S, Sambamurti K, Husain S. The role of the adaptive immune system and T cell dysfunction in neurodegenerative diseases. J Neuroinflammation. 2022;19(1):251. Zhong S, Lian Y, Luo W, Luo R, Wu X, Ji J, et al. Upstream open reading frame with NOTCH2NLC GGC expansion generates polyglycine aggregates and disrupts nucleocytoplasmic transport: implications for polyglycine diseases. Acta Neuropathol. 2021;142(6):1003-23. Yu J, Liufu T, Zheng Y, Xu J, Meng L, Zhang W, et al. CGG repeat expansion in NOTCH2NLC causes mitochondrial dysfunction and progressive neurodegeneration in Drosophila model. Proc Natl Acad Sci U S A. 2022;119(41):e2208649119. Fan Y, Li MJ, Yang J, Li SJ, Hao XY, Li JD, et al. GGC repeat expansion in NOTCH2NLC induces dysfunction in ribosome biogenesis and translation. Brain. 2023;146(8):3373-91. Anderson J, Toh ZQ, Reitsma A, Do LAH, Nathanielsz J, Licciardi PV. Effect of peripheral blood mononuclear cell cryopreservation on innate and adaptive immune responses. J Immunol Methods. 2019;465:61-6. Table Table 1. Overview of clinical information and manifestations in 32 NIID patients Patient Sex Age GGC size Disease duration Neurological symptoms Extra-neurological symptoms Histopathological specimens Cognitive impairment Movement disorders Muscle weakness Paroxysmal symptom Autonomic dysfunction Respiratory system Digestive system Endocrine system Genitourinary system Immune system 1 F 73 122 50 + - + - + - + - + - Kidney; Colon 2 M 70 113 53 + - - - + + + + + + Colon; Lung 3 M 64 139 41 + - - + + - - + + - - 4 F 49 133 30 + - - - - + - - - - Skin 5 M 68 108 32 + + + + + - + - + - - 6 F 74 162 8 + + + + + - + + - - Stomach 7 M 65 79 32 + + - - - - - - + - - 8 F 73 133 30 + + + + - - - - - - - 9 M 64 159 16 + + - + - - - - - - Brain 10 M 68 185 30 + - + + - - - - + - Bladder 11 M 65 121 15 + + + + + - - + + - - 12 F 69 79 32 + - - + - - - + - + - 13 M 67 129 9 - + - + + + + - + - Prostate 14 M 74 172 30 + + + - + - + - + - - 15 F 64 147 52 + + - - + + + - + - Skin 16 F 64 109 30 - - - - + + - - - - - 17 M 56 176 15 + - - - + - + - - - Colon 18 M 66 112 17 + + + - + - - - - - - 19 M 52 109 10 + - - - - + - - + - - 20 M 76 169 9 + + + - - - - - - - Stomach 21 M 58 117 20 + + - - + - + - + - Adrenal gland 22 M 70 78 10 + + - + + - - - + - - 23 M 75 121 30 + + + + + - + - + - Stomach 24 F 68 173 18 + - + - - - - - - - - 25 F 49 182 2 - - - - - - - - - + - 26 M 46 175 10 + - - - - - - - + - Lung 27 F 66 188 24 + - - - - - - - - - Skin 28 M 50 172 15 - - - + - + - - + - - 29 F 59 130 10 + - - + + - + + - - Oviduct 30 F 46 116 3 + + - + + + - - + + Kidney 31 F 57 88 2 + - + - + - - - - - - 32 M 58 142 2 + - - + + + + - - - Bladder Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-3703208","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Short Report","associatedPublications":[],"authors":[{"id":256544363,"identity":"8d721732-a598-43ad-8f1b-5d09e63d418c","order_by":0,"name":"Lei Bao","email":"","orcid":"","institution":"the Affiliated Hospital of Xuzhou Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Lei","middleName":"","lastName":"Bao","suffix":""},{"id":256544364,"identity":"79b3290b-8af4-4eb5-8bfa-8b69c647efbf","order_by":1,"name":"Dandan Zuo","email":"","orcid":"","institution":"the Affiliated Hospital of Xuzhou Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Dandan","middleName":"","lastName":"Zuo","suffix":""},{"id":256544365,"identity":"c61c389a-fcda-40d4-a489-958e16283a7e","order_by":2,"name":"Xiaoying Qu","email":"","orcid":"","institution":"Xuzhou Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Xiaoying","middleName":"","lastName":"Qu","suffix":""},{"id":256544366,"identity":"6e95577c-f325-4b47-b76d-f0a5617d871a","order_by":3,"name":"Yingying Cui","email":"","orcid":"","institution":"the Affiliated Hospital of Xuzhou Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yingying","middleName":"","lastName":"Cui","suffix":""},{"id":256544367,"identity":"c3596abb-1cea-4609-8f9e-5ee59b1691ae","order_by":4,"name":"Keke Li","email":"","orcid":"","institution":"the Affiliated Hospital of Xuzhou Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Keke","middleName":"","lastName":"Li","suffix":""},{"id":256544368,"identity":"d01498fc-b788-480a-81bc-38f489d7f197","order_by":5,"name":"Jing Dong","email":"","orcid":"","institution":"the Affiliated Hospital of Xuzhou Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jing","middleName":"","lastName":"Dong","suffix":""},{"id":256544369,"identity":"da0e316b-9e69-4e0f-a7f5-1c19125a1993","order_by":6,"name":"Renjin Chen","email":"","orcid":"","institution":"Xuzhou Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Renjin","middleName":"","lastName":"Chen","suffix":""},{"id":256544370,"identity":"6705c01d-850b-4e1a-a515-45b0e13c3e5a","order_by":7,"name":"Zunsheng Zhang","email":"","orcid":"","institution":"the Affiliated Hospital of Xuzhou Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Zunsheng","middleName":"","lastName":"Zhang","suffix":""},{"id":256544371,"identity":"2cd50172-5745-427e-ad9b-9967681b685c","order_by":8,"name":"Guiyun Cui","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA6ElEQVRIiWNgGAWjYFACxgYGBgMgLX/44AMgxcNHvBYJtmQQxcNGvG0SPGoSIJqgFv4Zyc2veQoOJ26X7mGr/JpjJ8PGwPzw0Q18Zt9IbLPmMTicuHPO2WO3ZbclAx3GZmycg0eLgURimzFIy4YDeWm3JbcxA7XwsEkTqSXHrFhyWz1RWpofg7XcyDFj/LjtMGEtEmcetjHOMUg33nDmWLI047bjPGzMBPzC357++MObP9ayG443H/z4c1u1PT9788PH+LQAARswOprBLGYeMIlfOVjJBwaGOjCL8Qdh1aNgFIyCUTACAQBI2EoJmWSNIAAAAABJRU5ErkJggg==","orcid":"","institution":"the Affiliated Hospital of Xuzhou Medical University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Guiyun","middleName":"","lastName":"Cui","suffix":""},{"id":256544372,"identity":"97b49b69-18b5-433d-898a-359fececb79d","order_by":9,"name":"Hao Chen","email":"","orcid":"","institution":"the Affiliated Hospital of Xuzhou Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hao","middleName":"","lastName":"Chen","suffix":""}],"badges":[],"createdAt":"2023-12-04 01:44:22","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3703208/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3703208/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":47799346,"identity":"b93200af-7944-4332-991f-7f0d0b61d430","added_by":"auto","created_at":"2023-12-07 15:25:12","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":182197,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEosinophilic inclusions within the nuclei of various tissues.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe tissue specimens obtained from the previous surgery in patients with NIID were stained with Hematoxylin-eosin. a: Inclusions within the nuclei of neurons from brain tissue. b: Inclusions within the nuclei of squamous epithelial cells from skin tissue. c: Inclusions within the nuclei of alveolar epithelial cells from lung tissue. d: Inclusion within the nuclei of mucosal epithelial cells from colon tissue. e: Inclusions within the nuclei of renal tubular epithelial cells from kidney tissue. f: Inclusions within the nuclei of epithelial cells from oviduct tissue.\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3703208/v1/d134d5cde7afbe49476e3765.jpg"},{"id":47799348,"identity":"54550066-b443-4155-98e6-a86246ab9d37","added_by":"auto","created_at":"2023-12-07 15:25:12","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":193114,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eInfiltration of inflammatory cells in various tissue specimens.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eInflammatory cells infiltration of the corresponding tissue specimens in Figure 1. Scattered activated microglia can be identified in brain tissue (a). Inflammatory cell infiltration in skin tissue (b), lung tissue (c), colon tissue (d), kidney tissue (e), and oviduct tissue (f). Eosinophilic intranuclear inclusions can also be identified in these inflammatory cells (zoom).\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3703208/v1/47cf6efb73d977101efb754c.jpg"},{"id":47799347,"identity":"d1bd897e-2fc5-4257-bd47-1e17200bf49e","added_by":"auto","created_at":"2023-12-07 15:25:12","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":66498,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAnalysis of uN2CpolyG protein aggregates in PBMCs from NIID patients. \u003c/strong\u003e(a) Immunofluorescence microscopy of PBMCs obtained from NIID patients and healthy individuals with normal CGG repeat lengths. In NIID patient PBMCs, a substantial proportion of cells exhibited intranuclear aggregates composed of uN2CpolyG protein (green) accumulation within the nucleus. Some aggregates co-localized with ubiquitin (red), indicating a subset with positive ubiquitin immunoreactivity. Conversely, PBMCs from healthy controls displayed weak signals and lacked detectable intranuclear inclusions. (b) Western blot analysis depicting the expression profile of uN2CpolyG protein in PBMCs from three NIID patients and control individuals without GGC duplications. NIID patient PBMCs revealed the presence of a 30-40kD uN2CpolyG protein band, while PBMCs from control individuals did not exhibit any detectable protein expression.\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3703208/v1/e6ebbc67910a5130e4859669.jpg"},{"id":47801400,"identity":"84da6e1e-23cd-4d84-bce3-896a5cf2af03","added_by":"auto","created_at":"2023-12-07 15:33:12","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":63813,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAutoimmune/inflammatory diseases in NIID with inflammatory cell infiltration in affected tissues\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNeuronal intranuclear inclusion disease (NIID) demonstrates a spectrum of inflammatory and autoimmune manifestations. Central nervous system presentations often mirror encephalitis-like episodes, while peripheral involvement may manifest as ulcerative colitis, asthma, IgA nephropathy, neurodermatitis, and other systemic conditions. Histological examination of affected tissues reveals discernible infiltration of inflammatory cells with eosinophilic intranuclear inclusions, indicative of an immune-mediated response.\u003c/p\u003e","description":"","filename":"4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3703208/v1/e5da66df9164f6fdd6597c66.jpg"},{"id":48152276,"identity":"8ab83b02-9710-47f9-80f7-ec202c6f617a","added_by":"auto","created_at":"2023-12-13 19:52:23","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":764407,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3703208/v1/cb324bb0-9ebe-4517-8326-623444095af6.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Immune Dysfunction and Systemic Inflammatory infiltration Exist in Neuronal Intranuclear Inclusion Disease","fulltext":[{"header":"Introduction","content":"\u003cp\u003eNeuronal intranuclear inclusion disease (NIID) is a progressive neurodegenerative disorder due to GGC repeat expansions in the \u003cem\u003eNOTCH2NLC\u003c/em\u003e gene (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e). This trinucleotide repeat disorder is typically adult-onset with incomplete penetrance and variable phenotypes. Common clinical manifestations of NIID patients include cognitive dysfunction, movement disorders, autonomic dysfunction, paroxysmal symptoms, and muscle weakness (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e). The pathogenic mechanism of NIID is associated with the expansion of GGC repeats, which leads to the production of a polyglycine protein called uN2CpolyG. Due to its specific structural characteristics, this protein continually aggregates and forms insoluble protein deposits, primarily within the cell nucleus as intranuclear inclusions, which serve a toxic effect on neurons, ultimately giving rise to the clinical symptoms of NIID (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e). However, this protein aggregate is not limited to deposition within the nuclei of neurons, it can also be found in nearly all internal organs, leading to a variety of non-neurological symptoms, including visual impairment, hearing loss, irritant dry cough, arrhythmia, cardiac insufficiency, vomiting, and constipation (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e). These findings have paved the way for the hypothesis that NIID is not solely a condition associated with CNS dysfunctions but rather a systemic disease.\u003c/p\u003e \u003cp\u003eA recent study reported a notable presence of macrophage/microglia infiltration within the brain lesion of a NIID patient who suffered an encephalitic episode (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e). This finding indicates the role of neuroinflammation in the development and progression of NIID, particularly in the context of encephalitic episodes. Given the intricate array of extra-neurological clinical symptoms exhibited by patients with NIID, it remains uncertain whether inflammatory cell infiltration also occurs in the peripheral organs of these individuals. Moreover, the vast majority of studies only focus on lesions of solid organs, with few research exploring the presence of inclusions in peripheral blood inflammatory cells or evaluating the immune and inflammatory status of patients with NIID. Given all this, we aim to establish compelling evidence supporting dysregulated immune responses and systemic inflammatory infiltration in NIID.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStudy participants\u003c/h2\u003e \u003cp\u003eThis study included 32 NIID-diagnosed patients treated at the Affiliated Hospital of Xuzhou Medical University between January 2018 and June 2023. The diagnosis of NIID was established based on clinical presentations and the detection of abnormal GGC repeats (\u0026gt;\u0026thinsp;65) within NOTCH2NLC, with or without additional confirmation through biopsy.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eClinical assessment\u003c/h2\u003e \u003cp\u003eThe neurological and extra-neurological symptoms of this patient group were meticulously evaluated by two experienced clinicians, focusing on identifying the presence of comorbid immune system diseases.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003ePeripheral blood monocyte cell (PBMC) isolation\u003c/h2\u003e \u003cp\u003eWhole blood samples were collected from NIID patients and healthy individuals, utilizing EDTA tubes. The extraction of PBMCs was performed using a density gradient with Ficoll-PaqueTM PLUS following the manufacturer's protocol. To achieve this, blood samples were diluted at a 1:1 ratio with Dulbecco\u0026rsquo;s phosphate-buffered saline and gently layered onto an equal volume of Ficoll-Paque solution. Subsequently, the samples underwent centrifugation at 440\u0026times; g for 30 minutes at room temperature (approximately 22 ˚C). Carefully isolating the PBMCs at the interface, they were washed twice with 1\u0026times; PBS and then subjected to subsequent experiments.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eImmunofluorescence staining\u003c/h2\u003e \u003cp\u003ePBMCs, freshly obtained, were cultured on poly-L-lysine-coated confocal dishes in 10% fetal bovine serum-supplemented RPMI 1640 medium. After 1 hour incubation at 37˚C for adherence, cells were fixed in 4% formaldehyde for 15 minutes and permeabilized with 0.2% PBST (PBS with Triton X) for 15 more minutes. Subsequently, a 4% BSA block was applied for 1 hour. Primary antibodies, including 4D12 (mouse antibody for uN2CpolyG at 1:200, courtesy of Professor Nicolas Charlet-Berguerand) and rabbit anti-ubiquitin (1:200, Abcam, ab7780), were incubated overnight at 4\u0026deg;C. Following PBS washing, secondary fluorescent antibodies\u0026mdash;goat anti-rabbit Alexa Fluor Plus 594 and goat anti-mouse Alexa Fluor Plus 488 (both at 1:1000, Invitrogen)\u0026mdash;were incubated for 1 hour at room temperature. The cells were then DAPI-stained for 1 minute before imaging with a Leica Stellaris 5 confocal microscope (Leica, Germany).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eWestern blot Analysis\u003c/h2\u003e \u003cp\u003ePBMCs were lysed in RIPA buffer for 10 minutes, followed by centrifugation at 4\u0026deg;C, 13,000g for 30 minutes. The resultant supernatant, containing proteins, was collected for western blot analysis. Equal protein amounts were electrophoresed, transferred onto nitrocellulose membranes, and blocked with 3% BSA for equal loading. The target membrane was incubated overnight at 4\u0026deg;C with 4D12 (anti-uN2CpolyG antibody, 1:1000). Then, membranes were incubated with goat anti-mouse IgG-HRP secondary antibodies (1:500) for 2 hours at room temperature. Subsequently, an enhanced chemiluminescence kit was used for visualization, and protein bands were detected using a gel imaging system (ChemiDoc XRS+, Bio-Rad).\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eInvolvement of immune system as another extra-neurological manifestation in NIID\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTable 1 presented in the study provides an overview of the general information for the 32 patients with NIID, along with details regarding their neurological and extra-neurological clinical manifestations. Within this study, the neurological clinical manifestations observed in NIID patients were categorized into five primary symptom groups. These encompassed cognitive impairment, movement disorders, muscle weakness, paroxysmal symptoms, and autonomic dysfunction. We have also compiled a summary of non-neurological clinical symptoms observed in these patients, categorizing them by the affected systems. Respiratory system involvement primarily presents as irritant dry cough and wheezing. Gastrointestinal system involvement is characterized by poor appetite, nausea, vomiting, diarrhea, and constipation. Urinary system manifestations include proteinuria, urinary retention, and renal insufficiency. Endocrine system abnormalities include diabetes or impaired glucose tolerance and hypothyroidism.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn the current study, we obtained a total of 19 archived surgical specimens derived from the cohort of 32 patients diagnosed with NIID. These specimens were collected from various anatomical sources, representing a diverse array of tissues within the patients exhibiting NIID symptoms. Table 1 outlines the specific sources of surgical specimens, encompassing 6 samples from the gastrointestinal tract, 3 from the skin, 2 from the lung, 2 from the bladder, and 1 each from the brain, kidney, adrenal gland, prostate, and fallopian tube. Subsequent histopathological assessment, utilizing H\u0026amp;E-stained sections (refer to Figure 1), revealed the presence of eosinophilic inclusions within the nuclei of these tissues.\u003c/p\u003e\n\u003cp\u003eWe additionally observed the occurrence of systemic inflammatory and autoimmune disorders within this patient cohort. Specifically, patient 2 with a history of ulcerative colitis exhibited symptoms, including weight loss, rectal bleeding, and chronic diarrhea. Patient 12 displayed hallmark clinical characteristics of Sj\u0026ouml;gren\u0026apos;s syndrome, notably presenting with Raynaud\u0026apos;s phenomenon, dry eyes, and dry mouth. Hashimoto\u0026apos;s thyroiditis was incidentally discovered in patient 25 during a routine physical examination, with the levels of anti-TPO and anti-thyroglobulin antibodies being significantly elevated. Patient 30 was admitted to the hospital due to early-onset cognitive dysfunction. Following routine assessments, the presence of hematuria, substantial proteinuria, and renal insufficiency was identified. Subsequent biopsy detected the deposition of immunoglobulin A within the kidney tissue confirming the diagnosis of IgA nephropathy.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInfiltration of inflammatory cells in tissue specimens from NIID patients.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFigure 2 presented a pervasive presence of inflammatory cell infiltration across various tissues from NIID patients. Briefly, in the temporal lobe brain biopsy of a patient experiencing an encephalitis-like attack, a notable presence of scattered microglia was observed. The identification of small focal inflammatory cell infiltrates in a separate patient\u0026apos;s skin biopsy is particularly intriguing, especially considering this patient\u0026apos;s prior diagnosis of neurodermatitis attributed to recurrent skin itching and ulceration. Further findings included marked inflammatory cell infiltration in a lung tissue biopsy from a patient with an unidentified pulmonary nodule, and diffuse inflammatory cell infiltration in a tissue specimen obtained from a colon polypectomy. Inflammatory cell infiltration was also evident in various other tissue samples: scattered inflammatory cells were detected in the renal tissue of a patient with IgA nephropathy, and patchy inflammatory cell infiltration was observed in surgical tissue specimens from a patient with hydrosalpinx. Notably, we also observed eosinophilic inclusions within the nuclei of these infiltrated inflammatory cells. Although the majority of specimens lacked immunohistochemical analysis for specific inflammatory cell markers, hindering precise cell phenotype characterization, our observations indicated that these eosinophilic intranuclear inclusions were primarily evident in mononuclear cells (Figure 2, zoom). This intriguing finding prompted further investigation into whether a similar phenomenon occurs in PBMCs.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDetection of uN2CpolyG aggregates-formed inclusions in PMBCs from NIID patients\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTo investigate whether the intranuclear inclusions also existed in peripheral inflammatory cells, we obtained fresh PBMCs from three NIID patients. In a significant proportion of PBMC cells, we observed the presence of aggregates resulting from the accumulation of uN2CpolyG protein within the nucleus. Moreover, a subset of these protein aggregates displayed positive immunoreactivity for ubiquitin. In sharp contrast, samples collected from healthy individuals with normal CGG repeat lengths exhibited only weak signals and were devoid of any inclusions (Figure. 3a). Furthermore, we conducted western blot analysis to assess the expression of uN2CpolyG protein in PBMCs from these NIID patients. Our results revealed the presence of a 30-40kD uN2CpolyG protein in the PBMCs of three NIID patients. However, in the PBMCs of control individuals without GGC duplications, we did not detect any protein expression (Figure. 3b).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eDespite being labeled as \u0026ldquo;neuronal\u0026rdquo; intranuclear inclusion disease, our previous research suggests that the term \"systemic\" intranuclear inclusion disease may be more appropriate. This is because eosinophilic intranuclear inclusions have been identified in the tissue cells of nearly all organs and their presence can lead to organ-specific lesions, resulting in a wide array of extra-neurological manifestations(\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e). For instance, individuals with visual dysfunction have exhibited widespread intranuclear inclusions and severe gliosis throughout the retina (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e). Similarly, those with cardiomyopathy have displayed diffusely distributed intranuclear inclusions, along with myocyte hypertrophy and fibrosis (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e). In the case of one NIID patient with unexplained massive proteinuria, renal tissue biopsies revealed intranuclear inclusions in podocytes and mesangial cells (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e). However, previous studies, including our own, have predominantly concentrated on the analysis of lesions and symptoms in solid organs while overlooking the assessment of immune and inflammatory status in patients with NIID. Hence, this study aimed to investigate the presence of comorbid immune system disorders in patients with NIID. An intriguing observation in this study is the occurrence of four distinct inflammatory diseases within the cohort of 34 NIID patients, which included ulcerative colitis, Sjogren's syndrome, Hashimoto's thyroiditis, and IgA nephropathy. Indeed, this is not the first occurrence of NIID being associated with inflammatory conditions. Previous case reports have documented instances of patients with this disease presenting with comorbid conditions such as asthma, crescentic IgA nephropathy, lupus nephritis\u0026ndash;like nephropathy, and neuropathy associated with IgM monoclonal gammopathy of undetermined significance (MGUS) (\u003cspan additionalcitationids=\"CR10 CR11\" citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eWhile neurodegenerative diseases and inflammatory diseases are distinct categories of diseases, some studies have indeed found an increased risk of comorbid inflammatory diseases in patients with neurodegenerative diseases, suggesting a potential link between these two types of conditions. A comprehensive review and meta-analysis of 873,643 individuals with Parkinson's disease (PD) found a significantly increased risk of PD in those with autoimmune diseases, including inflammatory bowel disease, Sj\u0026ouml;gren's syndrome, and Graves' disease (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e). In a nationwide case-control study of 3,561 amyotrophic lateral sclerosis (ALS) patients and 35,610 controls, a 47% higher likelihood of prior autoimmune disease diagnosis was observed in ALS patients. Specific autoimmune conditions included myasthenia gravis, polymyositis or dermatomyositis, Guillain-Barre syndrome, type 1 diabetes, multiple sclerosis, and hypothyroidism (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e). Multiple theories and hypotheses have been proposed to elucidate the association between neurodegenerative diseases and autoimmune diseases, although the precise underlying mechanisms remain incompletely understood. Neurodegenerative diseases may involve immune dysregulation, potentially leading to an exaggerated or misdirected immune response, thereby increasing the susceptibility to autoimmune diseases. Shared genetic factors are another consideration, with common genetic variants or susceptibility genes possibly influencing the development of both neurodegenerative and autoimmune conditions. Furthermore, chronic inflammation emerges as a common element in both neurodegenerative diseases and autoimmune diseases, contributing to the progression of neurodegenerative conditions and playing a pivotal role in many autoimmune disorders (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e). Our present study revealed a potential association between NIID patients and inflammatory diseases, indicating an altered innate and adaptive immune system in NIID patients. However, the establishment of a causal relationship is inconclusive due to the limited sample size and the absence of controlled studies.\u003c/p\u003e \u003cp\u003eIt has been reported that more than half of the NIID patients had ever suffered acute encephalitis-like episodes as the disease progressed, presenting with fever, altered mental status, conscious disturbance, or focal neurological deficit (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e). Further brain tissue biopsy revealed numerous macrophages/microglia infiltration in the lesion area (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e). These findings strongly suggest that a pronounced immune response or inflammatory reaction is occurring within the brain tissue of NIID patients, potentially serving as a vital pathological mechanism in the development of NIID. In this study, we present the first evidence of significant infiltration of inflammatory cells within various lesion tissues of NIID patients, indicating, similar to the central nervous system, inflammatory responses may also be implicated in lesions within other systems. Nonetheless, we cannot conclusively determine whether the infiltration of these inflammatory cells is a primary pathological mechanism of multi-system lesions or merely a secondary reaction activated in response to pathological stimulus.\u003c/p\u003e \u003cp\u003eDespite previous autopsy findings have consistently revealed intranuclear inclusions in the cells of brain tissues and various internal organs among NIID patients, our study marks the first to report the detection of these inclusions in both circulating and tissue-based immune cells. Detected from the brain and skin tissues of NIID patients, the main component of these inclusions is a glycine-rich protein, the uN2CpolyG protein, which is translated from the GGC repeat expansion in the 5'UTR of the NOTCH2NLC gene (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e). In this study, we utilized immunofluorescence to confirm the formation of intranuclear inclusions comprised of the uN2CpolyG protein within PBMCs. Additionally, we also assessed the protein expression through WB analysis, revealing robust levels of uN2CpolyG expression in PBMCs derived from individuals diagnosed with NIID. To our current understanding, our research group stands as the initial cohort to effectively identify the uN2CpolyG protein in the peripheral blood of individuals diagnosed with NIID. Our findings also propose that the presence of uN2CpolyG inclusions in PBMCs could potentially act as a biomarker, which could significantly aid in the early identification and subsequent treatment of NIID.\u003c/p\u003e \u003cp\u003ePrevious studies have demonstrated the neurotoxicity of uN2CpolyG protein based on its reduction of cell viability, disruption of nucleocytoplasmic transport, induction of mitochondrial dysfunction, and inhibition of ribosome biogenesis and translation (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan additionalcitationids=\"CR18\" citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e). These findings have offered crucial insights into the protein's detrimental effects on cellular mechanisms. PBMCs are a diverse group of immune cells found in the bloodstream, including lymphocytes, monocytes, and dendritic cells, which are key components of the innate and adaptive immune systems and crucial in response to inflammation (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e). Whether the pathological protein uN2CpolyG could also affect the survival and functionality of PBMCs remains ambiguous at present, however, these findings led us to hypothesize that the increased expression of uN2CpolyG in PBMCs may potentially disrupt their typical immune response functions, potentially triggering an aberrant inflammatory response. This potential disruption in PBMC function could shed light on the pronounced escalation in autoimmune and autoinflammatory conditions observed in individuals diagnosed with NIID.\u003c/p\u003e \u003cp\u003eThis study exhibits several limitations. First, the study cohort consisted of a relatively small and potentially homogeneous group of patients from a single medical center. Consequently, the findings may have limited applicability to a broader population. Secondly, while the study identified inflammatory cell infiltration, it lacked detailed characterization of these cells, particularly their phenotypes and activation states. This absence restricts a comprehensive understanding of the immune response in NIID. Lastly, despite highlighting associations and correlations, the study did not delve into detailed mechanistic insights into how immune dysregulation and uN2CpolyG aggregates precisely contribute to disease pathogenesis.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe coexistence of inflammatory and autoimmune diseases alongside systemic inflammatory cell infiltration in NIID patients strongly points towards a dysregulated immune response and a systemic inflammatory condition in this disorder (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Moreover, the identification of uN2CpolyG aggregates within PBMCs of NIID patients implies a potential toxic effect, suggesting an alteration in the immune response of these cells. These findings collectively underscore the intricate involvement of immune dysregulation in NIID pathophysiology, potentially serving as crucial indicators for both disease understanding and therapeutic interventions.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eNIID: Neuronal intranuclear inclusion disease\u0026nbsp;\u003c/p\u003e\n\u003cp\u003ePBMC: Peripheral blood mononuclear cell\u003c/p\u003e\n\u003cp\u003eMGUS: Monoclonal gammopathy of undetermined significance\u003c/p\u003e\n\u003cp\u003ePD: Parkinson\u0026apos;s disease\u003c/p\u003e\n\u003cp\u003eALS: Amyotrophic lateral sclerosis\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll research involving human subjects received approval from local review boards and adhered strictly to the principles outlined in the Declaration of Helsinki. The protocol of this study was approved by the Institutional Review Board of the Affiliated Hospital of Xuzhou Medical University. Prior to their participation, all individuals involved in the study provided informed written consent, signifying their voluntary agreement and understanding of the research objectives, procedures, and potential risks.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe obtained written consent for publication from all NIID patients\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and material\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets produced or analyzed during this study are not publicly accessible to safeguard personal information. However, they can be made available from the corresponding author upon reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that there is no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by\u0026nbsp;Project supported by the Affiliated Hospital of Xuzhou Medical University (2023ZL01).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eLB designed the study and drafted the manuscript. DDZ created figures and prepared tables. XYQ collected clinical data and tissue specimens. YYC conducted histopathological analysis of tissue specimens. KKL assessed neuroimages. JD prepared pathological specimens. ZZS assessed neurological findings. GYC assessed neurological findings. HC designed the study and assessed extra-neurological findings.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe would like to express our gratitude to Professor Nicolas Charlet-Berguerand for generously providing us with the antibody for uN2CpolyG protein, which has been an invaluable experimental tool and support for our research. We sincerely thank Dr. Fuxing Dong from the Public Experimental Research Center for his enthusiastic help in the experiment of laser scanning confocal microscopy.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eIshiura H, Shibata S, Yoshimura J, Suzuki Y, Qu W, Doi K, et al. Noncoding CGG repeat expansions in neuronal intranuclear inclusion disease, oculopharyngodistal myopathy and an overlapping disease. Nat Genet. 2019;51(8):1222-32.\u003c/li\u003e\n\u003cli\u003eTian Y, Zhou L, Gao J, Jiao B, Zhang SZ, Xiao Q, et al. Clinical features of NOTCH2NLC-related neuronal intranuclear inclusion disease. J Neurol Neurosur Ps. 2022.\u003c/li\u003e\n\u003cli\u003eBoivin M, Deng J, Pfister V, Grandgirard E, Oulad-Abdelghani M, Morlet B, et al. Translation of GGC repeat expansions into a toxic polyglycine protein in NIID defines a novel class of human genetic disorders: The polyG diseases. Neuron. 2021;109(11):1825-35 e5.\u003c/li\u003e\n\u003cli\u003eSone J, Mori K, Inagaki T, Katsumata R, Takagi S, Yokoi S, et al. Clinicopathological features of adult-onset neuronal intranuclear inclusion disease. Brain. 2016;139(Pt 12):3170-86.\u003c/li\u003e\n\u003cli\u003eChen H, Lu L, Wang B, Cui G, Wang X, Wang Y, et al. Re-defining the clinicopathological spectrum of neuronal intranuclear inclusion disease. Ann Clin Transl Neurol. 2020;7(10):1930-41.\u003c/li\u003e\n\u003cli\u003eMori K, Yagishita A, Funata N, Yamada R, Takaki Y, Miura Y. Imaging findings and pathological correlations of subacute encephalopathy with neuronal intranuclear inclusion disease-Case report. Radiol Case Rep. 2022;17(12):4481-6.\u003c/li\u003e\n\u003cli\u003eSone J, Ueno S, Akagi A, Miyahara H, Tamai C, Riku Y, et al. NOTCH2NLC GGC repeat expansion causes retinal pathology with intranuclear inclusions throughout the retina and causes visual impairment. Acta Neuropathol Commun. 2023;11(1):71.\u003c/li\u003e\n\u003cli\u003eOyer CE, Cortez S, O\u0026apos;Shea P, Popovic M. Cardiomyopathy and myocyte intranuclear inclusions in neuronal intranuclear inclusion disease: a case report. Hum Pathol. 1991;22(7):722-4.\u003c/li\u003e\n\u003cli\u003eMorita K, Shinzato T, Endo Y, Suzuki M, Yoshida H, Sone J, et al. A case of unusual renal manifestation in a patient with neuronal intranuclear inclusion disease treated with steroids. Clin Case Rep. 2023;11(8):e7730.\u003c/li\u003e\n\u003cli\u003eHorino T, Matsumoto T, Inoue K, Ichii O, Terada Y. A case of neuronal intranuclear inclusion disease associated with lupus nephritis-like nephropathy. eNeurologicalSci. 2018;10:28-30.\u003c/li\u003e\n\u003cli\u003eSugiyama A, Takeda T, Koide M, Yokota H, Mukai H, Kitayama Y, et al. Coexistence of neuronal intranuclear inclusion disease and amyotrophic lateral sclerosis: an autopsy case. BMC Neurol. 2021;21(1):273.\u003c/li\u003e\n\u003cli\u003eFu J, Zhao C, Hou G, Liu X, Zheng M, Zhang Y, et al. A case report of neuronal intranuclear inclusion disease with paroxysmal peripheral neuropathy-like onset lacking typical signs on diffusion-weighted imaging. Front Neurol. 2023;14:1117243.\u003c/li\u003e\n\u003cli\u003eLi M, Wan J, Xu Z, Tang B. The association between Parkinson\u0026apos;s disease and autoimmune diseases: A systematic review and meta-analysis. Front Immunol. 2023;14:1103053.\u003c/li\u003e\n\u003cli\u003eCui C, Longinetti E, Larsson H, Andersson J, Pawitan Y, Piehl F, et al. Associations between autoimmune diseases and amyotrophic lateral sclerosis: a register-based study. Amyotroph Lateral Scler Frontotemporal Degener. 2021;22(3-4):211-9.\u003c/li\u003e\n\u003cli\u003eDoty KR, Guillot-Sestier MV, Town T. The role of the immune system in neurodegenerative disorders: Adaptive or maladaptive? Brain Res. 2015;1617:155-73.\u003c/li\u003e\n\u003cli\u003eDeMaio A, Mehrotra S, Sambamurti K, Husain S. The role of the adaptive immune system and T cell dysfunction in neurodegenerative diseases. J Neuroinflammation. 2022;19(1):251.\u003c/li\u003e\n\u003cli\u003eZhong S, Lian Y, Luo W, Luo R, Wu X, Ji J, et al. Upstream open reading frame with NOTCH2NLC GGC expansion generates polyglycine aggregates and disrupts nucleocytoplasmic transport: implications for polyglycine diseases. Acta Neuropathol. 2021;142(6):1003-23.\u003c/li\u003e\n\u003cli\u003eYu J, Liufu T, Zheng Y, Xu J, Meng L, Zhang W, et al. CGG repeat expansion in NOTCH2NLC causes mitochondrial dysfunction and progressive neurodegeneration in Drosophila model. Proc Natl Acad Sci U S A. 2022;119(41):e2208649119.\u003c/li\u003e\n\u003cli\u003eFan Y, Li MJ, Yang J, Li SJ, Hao XY, Li JD, et al. GGC repeat expansion in NOTCH2NLC induces dysfunction in ribosome biogenesis and translation. Brain. 2023;146(8):3373-91.\u003c/li\u003e\n\u003cli\u003eAnderson J, Toh ZQ, Reitsma A, Do LAH, Nathanielsz J, Licciardi PV. Effect of peripheral blood mononuclear cell cryopreservation on innate and adaptive immune responses. J Immunol Methods. 2019;465:61-6.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Table","content":"\u003cp\u003e\u003cstrong\u003eTable 1. Overview of clinical information and manifestations in 32 NIID patients\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"929\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.41334768568353%\" valign=\"top\"\u003e\n \u003cp\u003ePatient\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9063509149623252%\" valign=\"top\"\u003e\n \u003cp\u003eSex\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1216361679224973%\" valign=\"top\"\u003e\n \u003cp\u003eAge\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.6598493003229278%\" valign=\"top\"\u003e\n \u003cp\u003eGGC size\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.180839612486544%\" valign=\"top\"\u003e\n \u003cp\u003eDisease duration\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"31.646932185145317%\" colspan=\"5\" valign=\"top\"\u003e\n \u003cp\u003eNeurological symptoms\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"37.56727664155005%\" colspan=\"5\" valign=\"top\"\u003e\n \u003cp\u003eExtra-neurological symptoms\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.503767491926803%\" valign=\"top\"\u003e\n \u003cp\u003eHistopathological specimens\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\" valign=\"top\"\u003e\n \u003cp\u003eCognitive impairment\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\" valign=\"top\"\u003e\n \u003cp\u003eMovement disorders\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\" valign=\"top\"\u003e\n \u003cp\u003eMuscle weakness\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\" valign=\"top\"\u003e\n \u003cp\u003eParoxysmal symptom\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\" valign=\"top\"\u003e\n \u003cp\u003eAutonomic dysfunction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\" valign=\"top\"\u003e\n \u003cp\u003eRespiratory system\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\" valign=\"top\"\u003e\n \u003cp\u003eDigestive system\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\" valign=\"top\"\u003e\n \u003cp\u003eEndocrine system\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\" valign=\"top\"\u003e\n \u003cp\u003eGenitourinary system\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\" valign=\"top\"\u003e\n \u003cp\u003eImmune system\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e122\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003eKidney; Colon\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e113\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003eColon; Lung\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e139\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e133\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003eSkin\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e108\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e162\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003eStomach\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e133\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e159\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003eBrain\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e185\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003eBladder\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e121\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e69\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e129\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003eProstate\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e172\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e147\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003eSkin\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e109\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e176\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003eColon\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e112\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e109\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e169\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003eStomach\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e117\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003eAdrenal gland\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e121\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003eStomach\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e173\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e182\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e175\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003eLung\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e188\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003eSkin\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e172\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e59\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e130\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003eOviduct\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e116\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003eKidney\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.403866809881848%\"\u003e\n \u003cp\u003e32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"2.9001074113856067%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.1149301825993554%\"\u003e\n \u003cp\u003e58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.651987110633727%\"\u003e\n \u003cp\u003e142\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.16326530612245%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.337271750805585%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"7.518796992481203%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"8.485499462943071%\"\u003e\n \u003cp\u003eBladder\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Neuronal intranuclear inclusion disease (NIID), uN2CpolyG, Immune, Inflammatory, Peripheral blood monocyte cell (PBMC).","lastPublishedDoi":"10.21203/rs.3.rs-3703208/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3703208/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eNeuronal intranuclear inclusion disease (NIID) is primarily recognized as a neurodegenerative disorder due to the production of a neurotoxicity protein, uN2CpolyG. However, evidence suggests its systemic nature, prompting an exploration of the immune and systemic inflammatory aspects of NIID in this study. A cohort of 32 diagnosed NIID patients participated in a comprehensive study involving clinical presentations, and tissue specimen analyses. Peripheral blood monocyte cells (PBMCs) were collected to detect uN2CpolyG expression in NIID patients by immunofluorescence staining and Western blotting. NIID patients showed varied neurological and extra-neurological symptoms alongside systemic inflammatory and autoimmune disorders, including ulcerative colitis, Sj\u0026ouml;gren's syndrome, Hashimoto's thyroiditis, and IgA nephropathy. 19 previous tissue specimens from these patients displayed evidence of inflammatory cell infiltration. Notably, our observations unveiled the novel presence of eosinophilic inclusions within the nuclei of these infiltrating inflammatory cells, primarily concentrated in mononuclear cells. Additionally, uN2CpolyG aggregates, identified as ubiquitin-positive inclusions, were detected in peripheral blood monocyte cells (PBMCs) from NIID patients for the first time, contrasting with the weak signal observed without inclusions in the control group. The detection of uN2CpolyG as a 30 to 40 kDa protein in the PBMCs from three NIID patients further supports our findings. This study highlights NIID's systemic nature, emphasizing immune dysfunction and systemic inflammatory infiltration. The detection of uN2CpolyG aggregates in the PBMCs of NIID patients suggests that it may have a toxic potential and alter the immune response of these cells.\u003c/p\u003e","manuscriptTitle":"Immune Dysfunction and Systemic Inflammatory infiltration Exist in Neuronal Intranuclear Inclusion Disease","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-12-07 15:25:08","doi":"10.21203/rs.3.rs-3703208/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"d652f844-c69e-4103-af7b-057c2c67cf21","owner":[],"postedDate":"December 7th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2023-12-13T19:44:16+00:00","versionOfRecord":[],"versionCreatedAt":"2023-12-07 15:25:08","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-3703208","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3703208","identity":"rs-3703208","version":["v1"]},"buildId":"cBFmMYwuxLRRLfASyISRj","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}