miR-125b-5p is A Promising Novel Plasma Biomarker for Alveolar Echinococcosis in Patients from the Southern Province of Qinghai

miR-125b-5p is A Promising Novel Plasma Biomarker for Alveolar Echinococcosis in Patients from the Southern Province of Qinghai | 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 Research article miR-125b-5p is A Promising Novel Plasma Biomarker for Alveolar Echinococcosis in Patients from the Southern Province of Qinghai Deping Cao, Bofan Jiang, Yaogang Zhang, Mingquan Pang This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-68399/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 07 Mar, 2021 Read the published version in BMC Infectious Diseases → Version 1 posted 4 You are reading this latest preprint version Abstract Background: Alveolar echinococcosis is an infectious zoonotic disease caused by Echinococcus multilocularis that is endemic to the vast pastoral areas of southern Qinghai province of China. Alveolar echinococcosis affects the human liver, and its clinical manifestations are similar to those of liver tumors. It is also called “worm cancer,” as it induces irreversible liver fibrosis and brain metastases. Alveolar echinococcosis is a serious threat to human health and is a burden to patients and economies. When detected and treated early, patients with alveolar echinococcosis have a strong chance of recovery. Therefore, it is imperative to identify early diagnostic biomarkers of the disease. MicroRNAs (miRNAs) are short, single-stranded, non-coding RNA molecules that play key roles in a wide range of biological processes, and have been recently implicated in tumorigenesis. However, little is known about the role of miRNAs in alveolar echinococcosis. We have previously reported differentially expressed miRNAs in the plasma of patients with alveolar echinococcosis and healthy individuals, using microarray assay chips. Methods: We screened 10 differentially expressed miRNAs based on their fold change and the available literature. Additionally, quantitative polymerase chain reaction was used to verify their expression in the plasma of alveolar echinococcosis patients. We found that hsa-miR-125b-5p was upregulated in the plasma and the hepatic tissue samples of alveolar echinococcosis patients; therefore, its role as a diagnostic biomarker for alveolar echinococcosis was investigated. Accordingly, hsa-miR-125b-5p was upregulated in plasma and hepatic tissue samples obtained from alveolar echinococcosis patients. Receiver operating characteristic curves showed that hsa-miR-125b-5p was associated with an area under the curve of 99.8% and 98.9% in the plasma and hepatic tissues of alveolar echinococcosis patients, respectively. Transfection of an LV-hsa-miR-125b-5p-inhibitor reduced apoptosis of L-O2 hepatocytes in vitro ( P < 0.05). Thus, hsa-miR-125b-5p may promote liver cell proliferation and inhibit liver cell apoptosis. Conclusions: Taken together, hsa-miR-125b-5p may be a promising diagnostic biomarker for the early non-invasive diagnosis of alveolar echinococcosis. We plan to validate the expression of hsa-miR-125b-5p in the plasma of patients with cystic echinococcosis, alveolar echinococcosis, and other liver diseases to determine the potential of hsa-miR-125-5p as a differential biomarker in future studies. Infectious Diseases alveolar echinococcosis diagnostic biomarker hsa-miR-125b-5p plasma hepatic tissue samples Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Highlights Hsa-miR-125b-5p was upregulated in samples from alveolar echinococcosis patients. L-O2 hepatocytes depleted of hsa-miR-125b-5p show increased cell viability. L-O2 hepatocytes depleted of hsa-miR-125b-5p correlated with decreased apoptosis. Hsa-miR-125b-5p may be a promising novel biomarker for alveolar echinococcosis. 1. Background Echinococcus multilocularis causes alveolar echinococcosis, one of the most lethal parasitic infections in humans [ 1 ] . Alveolar echinococcosis is a zoonotic disease that can be found globally, particularly where animal husbandry is practiced. It is particularly endemic to countries with developed animal husbandry, such as the northwest and Tibetan plateau of China, especially the Qinghai Tibetan plateau, where mixed infections of cystic echinococcosis and alveolar echinococcosis [ 2 ] . Alveolar echinococcosis primarily develops in the liver of intermediate hosts. Humans, primates, and other accidental hosts become infected upon ingestion of parasite eggs present in the feces of canine hosts (e.g., dogs and foxes), exposure to egg-contaminated environments, or consumption of contaminated food or drinking water [ 3 , 4 , 5 ] . When left untreated, alveolar echinococcosis is associated with 5- and 10-years mortality rates of 52% and 96%, respectively [ 6 ] . Owing to the tumor-like growth of metacestodes (also called “worm carcinoma”) in the liver and challenging early diagnosis and treatment [ 7 ] , alveolar echinococcosis is a significant public health concern that poses an economic burden for patients and their families in the southern region of the Qinghai province [ 8 ] . The current protocol for diagnosing alveolar echinococcosis predominantly relies on medical history, imaging, and immunological assays. However, none of these methods is accurate and there are no specific and sensitive molecular markers for the early diagnosis of alveolar echinococcosis [ 9 ] . MicroRNAs (miRNAs) are short, endogenous, non-coding RNAs, that post-transcriptionally regulate gene expression by binding to the 3′-untranslated regions of target mRNAs [ 10 ] . Parasite-resident miRNAs are essential for cellular invasion, development, and ability of the host to respond to environmental and developmental signals [ 11 ] . miR-125b-5p upregulation is associated with the severity of liver damage, and high serum miR-125b-5p levels may serve as a predictor for poor outcomes in hepatitis B virus associated acute-on-chronic liver failure cases [ 12 ] . Consistent with other reports, Wang [ 13 ] et al. have shown that circulating exosomes are promising candidates for diagnostic and prognostic biomarkers of human cancers. Furthermore, miR-125-3p was shown to enhance the diagnostic potential of carcinoembryonic antigen for early stage colon cancer [ 13 ] . 2. Methods 2.1. Plasma from alveolar echinococcosis patients and healthy individuals We collected plasma from 44 alveolar echinococcosis patients and 44 healthy individuals (22 males and 22 females) from the echinococcosis sample bank at the Qinghai University Affiliated Hospital. This study was approved by the ethics review broad of the Hospital Affiliated of Qinghai University (approval number: P-SL-2019054). All patients were diagnosed based on their clinical symptoms and imaging data. Whole blood samples were collected in tubes, allowed to clot at 4 °C for 2 h, and centrifuged at 12,000 × g for 10 min at 4 °C. Plasma was separated from the whole blood samples, placed into fresh tubes, and centrifuged at 12,000 × g for 10 min at 4 °C. Hemolyzed samples were excluded. This step was used to remove cell debris and blood platelets. Subsequently, the supernatant was transferred to fresh tubes and stored at -80 °C for further analysis. 2.2. Collection of liver tissues from alveolar echinococcosis patients and healthy individuals Alveolar echinococcosis and normal liver tissues (3 × 3 mm) were resected from the liver, washed three times in phosphate-buffered saline, and stored in liquid nitrogen until further analysis. Normal tissue samples were collected 2 cm away from the alveolar echinococcosis lesions. 2.3. Cell culture and transfection L-O2 hepatocytes were cultured in Dulbecco’s modified Eagle medium supplemented with 10% fetal bovine serum and penicillin/streptomycin in a humidified atmosphere containing 5% CO 2 at 37 °C. Approximately 50 nM of hsa-miR-125b-5p or control inhibitor, both purchased from JIKAI (Shanghai Jikai), was transfected into L-O2 cells for 48 h using the riboFECTCP transfection kit (Guangzhou RIBO Biocompany) according to the protocol provided. All the experiments were performed using cells in the log phase. 2.4. RNA extraction Total RNA was isolated from plasma and tissue samples using TRIzol according to the protocol provided. The concentration and integrity of isolated RNAs were evaluated using the Agilent Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA, USA). cDNA was synthesized using PrimeScript Reverse Transcriptase (TIANGEN Biotech, Beijing, Co. Ltd) according to the kit protocol. Primers targeting hsa-miR-125b-5p (reverse transcription (RT): 5′-UCCCUGAGACCCUAACUUGUGA-3′; polymerase chain reaction (PCR) forward primer: 5′-TCCCTGAGACCCTAACTTGTGA-3′ and PCR reverse primer: 5′-GTGCAGGGTCCGAGGT-3′) and U6 (RT primer: 5′-UGAGGUAGGAGGUUGUAUAGUU-3′; PCR forward primer: 5′-TGAGGTAGGAGGTTGTATAGTT-3′; and PCR reverse primer: 5′-GTGCAGGGTCCGAGGT-3′) were purchased from Sangon Biotech (Shanghai) Co., Ltd. 2.5. Quantitative PCR (qPCR) qPCR was performed using the LightCycler® 480Ⅱ and Tiangen SYBR Green PCR Kit. U6 was used as the endogenous control for miRNA amplification. The reaction volume was 25 µl and was set up according to the TB GreenTM Premix Ex TaqTMⅡ protocol. Each sample and blank was analyzed in triplicate. The reaction conditions were as follows: pre-denaturation at 95℃ for 30 s, 40 cycles of 95℃ for 5 s, and 60℃ for 30 s. The melting curves were generated using the following temperatures: 95℃ for 5 s and 60℃ for 1 min. hsa-miR-125b-5p expression was analyzed using the ΔCt method. Relative miRNA expression was calculated using the 2 −△△Ct method and normalized to U6 levels. 2.6. Inhibition of hsa-miR-125b-5p in treated L-O2 hepatocytes L-O2 hepatocytes were transfected with lentiviral vector (LV), LV-hsa-miR-125b-5p-inhibitor, and the negative control virus CON137. The L-O2 cells were transfected with the LV-hsa-miR-125b-5p inhibitor for 5 days. The transfected cells were subsequently incubated with the Cell Counting Kit-8 reagents for 2 h to determine the absorption at a wavelength of 450 nm in a time-dependent manner. OD 450 represents the number of viable cells. 2.7. Fluorescence-activated cell sorter analysis for apoptosis LV-hsa-miR-125b-5p-inhibition transfection solution and L-O2 hepatocytes were cultured in a 6-well culture plate in triplicate. In order to ensure that a detectable number of cells was present, a cell density of ≥ 5 × 10 5 /ml was used. The cell fusion was 85% on the fifth day after transfection. The cells were collected, placed into 5 mL centrifuge tubes, and centrifuged at 250 g for 5 min at 4℃. The cells were washed with cold PBS, suspended in complete medium, and centrifuged at 250 g for 3 min. Cells were suspended in 200 µL 1 × binding buffer, and 10 µL Annexin V-APC was added to stain the cells. The cells were incubated at 10–15 min at room temperature in the dark and analyzed using BD cytoflow. 2.8. Statistical analysis Data are presented as the mean ± standard deviation. The independent sample t- test was used to determine the statistical differences between the groups using GraphPad Prism 8. P < 0.05 was considered to indicate a statistically significant difference. 3. Results 3.1. qPCR We used patient plasma to determine the potential of hsa-miR-125b-5p as an early diagnostic biomarker for alveolar echinococcosis. Our preliminary study using miRNA chips revealed upregulation of hsa-miR-125b-5p. 3.1.1 hsa-miR-125b-5p expression in plasma samples from alveolar echinococcosis patients qPCR showed that hsa-miR-125b-5p was upregulated in the plasma samples of alveolar echinococcosis patients ( P < 0.001) (Table 1 and Fig. 1 ). Table 1 The 2 −∆∆Ct values for hsa-miR-125b-5p expression in plasma in patients with alveolar echinococcosis and controls. Group Sample hsa-miR-125b-5p n 2 −∆∆Ct Alveolar echinococcosis patients Plasma 22 2.440268 ± 0.753355 Healthy controls Plasma 22 1.032328 ± 0.261863 3.1. 2 hsa-miR-125b-5p expression in hepatic tissues from alveolar echinococcosis patients qPCR showed that hsa-miR-125b-5p was upregulated in the liver tissue samples obtained from alveolar echinococcosis patients ( P < 0.001) (Table 2 and Fig. 2 ). Table 2 The hsa-miR-125b-5p level in hepatic tissues was statistically different between alveolar echinococcosis patients and the control group (P < 0.001). Group Sample hsa-miR-125b-5p n 2 −∆∆Ct Alveolar echinococcosis patients Liver tissue 8 1.932 ± 0.682 Healthy controls Normal liver 8 1.001 ± 0.049 3.2. Inhibition of hsa-miR-125b-5p in treated L-O2 hepatocytes 3.2.1 Transfection of LV, LV-hsa-miR-125b-5p-inhibitor, and negative control viruses CON137 in normal L-O2 hepatocytes 3.2.2 Cell Counting Kit-8 The proliferation of L-O2 cells increased in the hsa-miR-125b-5p-inhibitor group compared with the negative virus control group. On the fifth day after treatment, the OD 450 of the experimental group was 3.242 ± 0.032, while that of the control group was 1.738 ± 0.008 ( P < 0.05) (Fig. 3 and Fig. 4 ). 3.2.3 Cellular apoptosis The apoptosis (%) of the experimental group was 4.84 ± 0.09, while that of the control group was 1.91 ± 0.18 ( P < 0.05). Apoptosis in the control and LV-hsa-miR-125b-5p inhibitor-transfected L-O2 cells after 5 days was compared. Apoptosis was decreased in cells depleted of hsa-miR-125b-5p ( P < 0.05). Thus, hsa-miR-125b-5p may promote liver cell proliferation and inhibit apoptosis (Fig. 5 ). The expression of miR-125b-5p was increased in the plasma and liver lesion tissues of patients with alveolar echinococcosis. Transfection of the LV-hsa-miR-125b-5p-inhibitor reduced the apoptosis of L-O2 hepatocytes in vitro. It is inferred that miR-125b-5p plays a role in alveolar echinococcosis, and may serve as a promising molecular biomarker for the diagnosis and treatment of the disease. 4. Discussion miRNAs are small non-coding regulatory RNAs that play important roles in the pathogenesis of parasitic diseases [ 14 ] . Lin [ 15 ] et al. (2018) found that miRNA-221/222 is upregulated in the thyroid of papillary thyroid carcinoma patients, and is crucial in invasion and lymph node metastasis, suggesting its potential as a biomarker for invasive papillary thyroid carcinoma. Mogahed [ 16 ] et al. (2018) reported that miR-712-3p could be used as a biomarker for the early diagnosis of toxoplasmosis using qPCR and the plasma of mice acutely infected with Toxoplasma gondii . We have previously reported differences in the expression of hsa-miR-125b-5p, hsa-let7e-5p, hsa-miR-3165, hsa-miR-4725, and hsa-miR-520e in alveolar echinococcosis plasma samples using microarray data [ 17 , 18 ] . In this study, 22 plasma samples from alveolar echinococcosis patients and 22 healthy controls were tested using qPCR. We observed the upregulation of hsa-miR-125b-5p in the plasma from alveolar echinococcosis patients. We also examined the expression of hsa-miR-125b-5p in liver samples obtained from 8 patients with alveolar echinococcosis and 8 controls. qPCR revealed the upregulation of hsa-miR-125b-5p in plasma and lesion tissues obtained from alveolar echinococcosis patients. We found that the area under the curve for hsa-miR-125b-5p was 0.997 in the blood, which was significantly higher than that of the control group, with a 95% confidence interval of 0.9907–1.000 and P < 0.0001. The area under the curve for hsa-miR-125b-5p was 0.937 in the alveolar echinococcosis liver tissues, with a 95% confidence interval of 0.8701–1.000 and P < 0.0001. Thus, hsa-miR-125b-5p may serve as a useful diagnostic indicator and therapeutic target for alveolar echinococcosis. Inhibition of hsa-miR-125b-5p in treated L-O2 hepatocytes increased proliferation and decreased apoptosis. This suggested that the inhibition of hsa-miR-125b-5p may promote hepatocyte proliferation to reverse the effects of liver damage. We wish to investigate this further by determining the utility of hsa-miR-125b-5p in treating mice with secondary echinococcosis. miR-125b-5p is a member of the miR-125 family, which was first reported in 2005 by Lee [ 19 ] et al. (2005), and regulates the proliferation of differentiated tumor cells. miR-125b-5p enhances autophagy and apoptosis in multiple myeloma cells and can be used as a novel therapeutic target and early diagnostic biomarker for multiple myeloma [ 20 ] . Thus, the role of circulating miRNAs in the early diagnosis of alveolar echinococcosis may improve our understanding of the pathogenesis of alveolar echinococcosis. Our findings strongly suggest that hsa-miR-125b-5p may serve as a useful diagnostic and prognostic biomarker for alveolar echinococcosis. However, this study has certain limitations. Firstly, the findings in this study are preliminary and do not attempt to elucidate the mechanism by which hsa-miR-125b-5p is implicated in alveolar echinococcosis. Moreover, we have not addressed the association between hsa-miR-125b-5p expression and the size of the diseased tissue. Thus, the correlation between hsa-miRNA-125b-5p expression, diseased tissue size, the physiological characteristics of alveolar echinococcosis lesion(s), and recovery of patients after surgery will be investigated in future. Additionally, the potential of hsa-miRNA-125b-5p to distinguish between alveolar and cystic echinococcosis, as well as other liver diseases, requires further investigation. Our study findings require further validation using a larger cohort samples to determine whether hsa-miR-125b-5p may serve as a biomarker for the early non-invasive diagnosis of alveolar echinococcosis. 5. Conclusions In this study, we report the upregulation of hsa-miR-125b-5p in alveolar echinococcosis using an independent cohort of 22 patients with alveolar echinococcosis and 22 healthy individuals. While hsa-miR-125b-5p may serve as a promising diagnostic marker for alveolar echinococcosis, additional experiments should be performed to further investigate its diagnostic value. Future studies are required to identify the expression of hsa-miR-125b-5p in the plasma of patients with cystic echinococcosis, alveolar echinococcosis, and other liver diseases to determine the potential of hsa-miR-125-5p as a differential biomarker. Declarations Declarations Ethics approval and consent to participate This study was approved by the ethics review broad of the Hospital Affiliated of Qinghai University (approval number: P-SL-2019054). Availability of data and materials Not applicable. competing interests The author(s) declare that they have no competing interests. Consent for publication Agree to publish Authors' contributions CDP conceived and designed the study and drafted the manuscript. JBF did the research for q-PCR and data analysis. ZYG and PMQ helped design the study and revise the draft manuscript. All authors read and approved the final manuscript. Funding This work was supported by the Qinghai Science and Technology Department (grant numbers 2016-SF-A5 and 2019-SF-131). Acknowledgements We would like to thank Editage (www.editage.cn) for English language editing. References Eckert J, Deplazes P. Biological, epidemiological, and clinical aspects of echinococcosis, a zoonosis of increasing concern. Clin Microbiol Rev. 2004;17:107–35. Wang H, Zhang JX, Schantz PM, et al. Epidemiologic survey and analysis on echinococcosis in humans and animals from 1995 to 2005 in Qinghai province. Zhongguo renshou gonghuanbing xuebao. 2006;22:1129–34. (in Chinese). Nunnari G, Pinzone MR, Gruttadauria S, Celesia BM, Madeddu G, Malaguarnera G, Pavone P, Cappellani A, Cacopardo B. Hepatic echinococcosis: clinical and therapeutic aspects. World J Gastroenterol. 2012;18:1448–58. Mariconti M, Bazzocchi C, Tamarozzi F, Meroni V, Genco F, Maserati R, Brunetti E. Immunoblotting with human native antigen shows stage-related sensitivity in the serodiagnosis of hepatic cystic echinococcosis. Am J Trop Med Hyg. 2014;90:75–9. Gottstein B, Wang JH, Blagosklonov O, Grenouillet F, Millon L, Vuitton DA, Müller N. Echinococcus metacestode: in search of viability markers. Parasite. 2014;21:63. Giraudoux P, Pleydell D, Raoul F, Quéré JP, Wang Q, Yang Y, Vuitton DA, Qiu J, Yang W, Craig PS. Transmission ecology of Echinococcus multilocularis : what are the ranges of parasite stability among various host communities in China. Parasitol Int. 2006;55:237–46. Brunetti E, Kern P, Vuitton DA. Expert consensus for the diagnosis and treatment of cystic and alveolar echinococcosis in humans. Acta Trop. 2010;114:1–16. Wang TP, Cao ZG,. Progress in prevention and control of Chinese hydatid disease and its existing problems. Zhongguo jishengchongxue yu jishengchongbing fangzhi zazhi. 2018;36(3):291–6. (In Chinese). Guo X, Zheng Y. Expression profiling of circulating miRNAs in mouse serum in response to Echinococcus multilocularis infection. Parasitology. 2017;144:1079–87. Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004;116:281–97. Bartel DP. Micro RNAs: target recognition and regulatory functions. Cell. 2009;136:215–33. Tao YH, Wang ML, Wang M, et al. Quantification of circulating miR-125b-5p predicts survival in chronic hepatitis B patients with acute-on-chronic liver failure. Digestive Liver Disease. 2019;51:412–8. Wang J, Yan F, Zhao Q, Zhan F, Wang R, Wang L, Zhang Y, Huang X. Circulating exosomal miR-125a-3p as a novel biomarker for early-stage colon cancer. Sci Rep. 2017;7:4150. Jin X, Guo X, Zhu D, Ayaz M, Zheng Y. miRNA profiling in the mice in response to Echinococcus multilocularis infection. Acta Tropica. 2017;166:39–44. Lin ZK, Zhou K, Lu J, et al. ROC curve analysis of mi RNA-RNA-221/222 as a marker for invasive papillary thyroid carcinoma in clinical value. Aizheng jinzhan. 2018;16(1):49–52. (In Chinese). Mogahed NF, Khedr SI, Ghazala RA, Masoud IM. Can miRNA712-3p be a promising biomarker for early diagnosis of toxoplasmosis? Asian Pac. J Trop Med. 2018;11:688–92. Liu J, Cao DP, Zhang XF, et al. Analysis of serum miRNAs expression spectrum in patients with alveolar echinococcosis. Chinese high altitude medicine biology. 2019;40:51–6. (in Chinese). Jiang BF, Sha CQ, Zhang YG, et al. Expression of hsa-miR-125b-5p in serum of patients with alveolar echinococcosis and screening of its signal transduction pathway and target genes. Chinese high altitude medicine biology. 2020;41:39–45. (in Chinese). Lee YS, Kim HK, Chung S, Kim KS, Dutta A. Depletion of human micro-RNA miR-125b reveals that it is critical for the proliferation of differentiated cells but not for the down-regulation of putative targets during differentiation. J Biol Chem. 2005;280:16635–41. Morelli E, Leone E, Gallo, Cantafio,et al. Selective targeting of IRF4 by synthetic microRNA-125b-5p mimics induces anti-multiple myeloma activity in vitro and in vivo. Leukemia. 2015;29:2173–83. Supplementary Files ethicsreviewletter.jpg Cite Share Download PDF Status: Published Journal Publication published 07 Mar, 2021 Read the published version in BMC Infectious Diseases → Version 1 posted Reviewers invited by journal 18 Sep, 2020 Editor assigned by journal 17 Sep, 2020 Submission checks completed at journal 16 Sep, 2020 Editor invited by journal 15 Sep, 2020 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. 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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-68399","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research article","associatedPublications":[],"authors":[{"id":2410836,"identity":"476fa462-7c3f-4e86-baef-4fc262975621","order_by":0,"name":"Deping Cao","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAnklEQVRIiWNgGAWjYDACCRDBxiDHxt5+gDQtxnw8ZxJI05I4T8LBgDgdfLe7Ez/8KLuT3ibBkMDwo2IbYS2Sd85uluw59yy3TbrxAGPPmduEtRjcyN3GzNh2OLdN5kACkEGClnQ2iQQD0rQkEK9F8kYu2C+GbcBAPkiUX/hu5G4EhZi8fHv7wQc/KojQwnAAgyRJyygYBaNgFIwCrAAAbidBJP9Lyx8AAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0002-1973-8879","institution":"Guilin Medical University","correspondingAuthor":true,"prefix":"","firstName":"Deping","middleName":"","lastName":"Cao","suffix":""},{"id":2410837,"identity":"cf49ae50-9abe-4b35-abdd-d180ca001cc9","order_by":1,"name":"Bofan Jiang","email":"","orcid":"","institution":"Qinghai university medical college","correspondingAuthor":false,"prefix":"","firstName":"Bofan","middleName":"","lastName":"Jiang","suffix":""},{"id":2410838,"identity":"6798d62f-be2e-47d1-9b66-e0f82f5ba46e","order_by":2,"name":"Yaogang Zhang","email":"","orcid":"","institution":"Affliated hospital of Qinghai unniversity","correspondingAuthor":false,"prefix":"","firstName":"Yaogang","middleName":"","lastName":"Zhang","suffix":""},{"id":2410839,"identity":"276738e5-049d-475b-861e-551d010ff1cf","order_by":3,"name":"Mingquan Pang","email":"","orcid":"","institution":"affiliated hospital of Qinghai university","correspondingAuthor":false,"prefix":"","firstName":"Mingquan","middleName":"","lastName":"Pang","suffix":""}],"badges":[],"createdAt":"2020-08-29 10:47:46","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-68399/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-68399/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s12879-021-05940-z","type":"published","date":"2021-03-07T15:00:27+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":2478032,"identity":"8ae4877a-e1d8-434d-a928-affc19ef8dbf","added_by":"auto","created_at":"2020-09-18 15:13:08","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":32983,"visible":true,"origin":"","legend":"Relative expression of hsa-miR-125b-5p in the plasma of healthy (control) and AE (treated) patients (left). The receiver operating characteristic curve for the relative expression of hsa-miR-125b-5p in plasma in the controls and AE patients. AE, alveolar echinococcosis.","description":"","filename":"fig1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-68399/v1/fig1.jpg"},{"id":2478033,"identity":"0d95aeb3-f511-45e5-a669-7345a94894f2","added_by":"auto","created_at":"2020-09-18 15:13:08","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":27730,"visible":true,"origin":"","legend":"Relative expression of hsa-miR-125b-5p in AE and normal liver tissues. The receiver operating characteristic curve for the relative expression of hsa-miR-125b-5p in AE and normal liver tissues. AE, alveolar echinococcosis.","description":"","filename":"fig2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-68399/v1/fig2.jpg"},{"id":2626915,"identity":"bc9ee067-c135-4f27-ac1b-4697d042b2fe","added_by":"acdc","created_at":"2020-09-25 21:03:17","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":51362,"visible":true,"origin":"acdc-manuscripts-figure","legend":"LV-hsa-miR-125b-5p inhibitor-transfected L-O2 hepatocytes (magnification, 100×).","description":"{\"primaryId\":\"undefined\",\"secondaryId\":\"INFD-D-20-03047\",\"acdcId\":\"undefined\",\"revision\":\"1\",\"timestamp\":\"2020-09-16T12:29:13\",\"document\":\"manuscripts\",\"linkRel\":\"figure\"}","filename":"figure3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-68399/v1/figure_3.jpg"},{"id":2626914,"identity":"e1124b56-9770-4845-a127-a540ec05ac0d","added_by":"acdc","created_at":"2020-09-25 21:03:17","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":20493,"visible":true,"origin":"acdc-manuscripts-figure","legend":"t-test analysis of the cell proliferation of LV-hsa-miR-125b-5p inhibitor-transfected and control cells (NC; P \u003c 0.05). ","description":"{\"primaryId\":\"undefined\",\"secondaryId\":\"INFD-D-20-03047\",\"acdcId\":\"undefined\",\"revision\":\"1\",\"timestamp\":\"2020-09-16T12:29:13\",\"document\":\"manuscripts\",\"linkRel\":\"figure\"}","filename":"figure4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-68399/v1/figure_4.jpg"},{"id":2626913,"identity":"91922255-4aff-4688-9bef-3173d4c85760","added_by":"acdc","created_at":"2020-09-25 21:03:17","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":17181,"visible":true,"origin":"acdc-manuscripts-figure","legend":"t-test analysis of apoptosis in control and LV-hsa-miR-125b-5p inhibitor-transfected L-O2 cells (P\u003c0.05).","description":"{\"primaryId\":\"undefined\",\"secondaryId\":\"INFD-D-20-03047\",\"acdcId\":\"undefined\",\"revision\":\"1\",\"timestamp\":\"2020-09-16T12:29:13\",\"document\":\"manuscripts\",\"linkRel\":\"figure\"}","filename":"figure5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-68399/v1/figure_5.jpg"},{"id":13594893,"identity":"ecd857b4-8674-4dc0-9c77-0bd679f61b84","added_by":"auto","created_at":"2021-09-17 05:22:18","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":569741,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-68399/v1/37b9b851-c07e-4a99-b898-5ddaa8d3d7a7.pdf"},{"id":2478038,"identity":"452d1767-72c4-4f20-9245-d3521e3df256","added_by":"auto","created_at":"2020-09-18 15:13:10","extension":"jpg","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":3111649,"visible":true,"origin":"","legend":"","description":"","filename":"ethicsreviewletter.jpg","url":"https://assets-eu.researchsquare.com/files/rs-68399/v1/ethicsreviewletter.jpg"}],"financialInterests":"","formattedTitle":"\u003cp\u003emiR-125b-5p\u0026nbsp;is A Promising Novel Plasma Biomarker\u0026nbsp;for Alveolar Echinococcosis in Patients from the Southern Province of Qinghai\u003c/p\u003e","fulltext":[{"header":"Highlights","content":"\u003cul\u003e\n\u003cli\u003eHsa-miR-125b-5p was upregulated in samples from alveolar echinococcosis patients.\u003c/li\u003e\n\u003cli\u003eL-O2 hepatocytes depleted of hsa-miR-125b-5p show increased cell viability.\u003c/li\u003e\n\u003cli\u003eL-O2 hepatocytes depleted of hsa-miR-125b-5p correlated with decreased apoptosis.\u003c/li\u003e\n\u003cli\u003eHsa-miR-125b-5p may be a promising novel biomarker for alveolar echinococcosis.\u003c/li\u003e\n\u003c/ul\u003e"},{"header":"1. Background","content":" \u003cp\u003e \u003cem\u003eEchinococcus multilocularis\u003c/em\u003e causes alveolar echinococcosis, one of the most lethal parasitic infections in humans\u003csup\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]\u003c/sup\u003e. Alveolar echinococcosis is a zoonotic disease that can be found globally, particularly where animal husbandry is practiced. It is particularly endemic to countries with developed animal husbandry, such as the northwest and Tibetan plateau of China, especially the Qinghai Tibetan plateau, where mixed infections of cystic echinococcosis and alveolar echinococcosis\u003csup\u003e[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]\u003c/sup\u003e. Alveolar echinococcosis primarily develops in the liver of intermediate hosts. Humans, primates, and other accidental hosts become infected upon ingestion of parasite eggs present in the feces of canine hosts (e.g., dogs and foxes), exposure to egg-contaminated environments, or consumption of contaminated food or drinking water\u003csup\u003e[\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]\u003c/sup\u003e. When left untreated, alveolar echinococcosis is associated with 5- and 10-years mortality rates of 52% and 96%, respectively \u003csup\u003e[\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003c/sup\u003e. Owing to the tumor-like growth of metacestodes (also called \u0026ldquo;worm carcinoma\u0026rdquo;) in the liver and challenging early diagnosis and treatment\u003csup\u003e[\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]\u003c/sup\u003e, alveolar echinococcosis is a significant public health concern that poses an economic burden for patients and their families in the southern region of the Qinghai province\u003csup\u003e[\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]\u003c/sup\u003e. The current protocol for diagnosing alveolar echinococcosis predominantly relies on medical history, imaging, and immunological assays. However, none of these methods is accurate and there are no specific and sensitive molecular markers for the early diagnosis of alveolar echinococcosis\u003csup\u003e[\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eMicroRNAs (miRNAs) are short, endogenous, non-coding RNAs, that post-transcriptionally regulate gene expression by binding to the 3\u0026prime;-untranslated regions of target mRNAs\u003csup\u003e[\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]\u003c/sup\u003e. Parasite-resident miRNAs are essential for cellular invasion, development, and ability of the host to respond to environmental and developmental signals\u003csup\u003e[\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]\u003c/sup\u003e. miR-125b-5p upregulation is associated with the severity of liver damage, and high serum miR-125b-5p levels may serve as a predictor for poor outcomes in hepatitis B virus associated acute-on-chronic liver failure cases\u003csup\u003e[\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]\u003c/sup\u003e. Consistent with other reports, Wang\u003csup\u003e[\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]\u003c/sup\u003eet al. have shown that circulating exosomes are promising candidates for diagnostic and prognostic biomarkers of human cancers. Furthermore, miR-125-3p was shown to enhance the diagnostic potential of carcinoembryonic antigen for early stage colon cancer\u003csup\u003e[\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e "},{"header":"2. Methods","content":" \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. Plasma from alveolar echinococcosis patients and healthy individuals\u003c/h2\u003e \u003cp\u003eWe collected plasma from 44 alveolar echinococcosis patients and 44 healthy individuals (22 males and 22 females) from the echinococcosis sample bank at the Qinghai University Affiliated Hospital. This study was approved by the ethics review broad of the Hospital Affiliated of Qinghai University (approval number: P-SL-2019054). All patients were diagnosed based on their clinical symptoms and imaging data. Whole blood samples were collected in tubes, allowed to clot at 4\u0026nbsp;\u0026deg;C for 2\u0026nbsp;h, and centrifuged at 12,000\u0026thinsp;\u0026times;\u0026thinsp;g for 10\u0026nbsp;min at 4\u0026nbsp;\u0026deg;C. Plasma was separated from the whole blood samples, placed into fresh tubes, and centrifuged at 12,000\u0026thinsp;\u0026times;\u0026thinsp;g for 10\u0026nbsp;min at 4\u0026nbsp;\u0026deg;C. Hemolyzed samples were excluded. This step was used to remove cell debris and blood platelets. Subsequently, the supernatant was transferred to fresh tubes and stored at -80\u0026nbsp;\u0026deg;C for further analysis.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Collection of liver tissues from alveolar echinococcosis patients and healthy individuals\u003c/h2\u003e \u003cp\u003eAlveolar echinococcosis and normal liver tissues (3\u0026thinsp;\u0026times;\u0026thinsp;3\u0026nbsp;mm) were resected from the liver, washed three times in phosphate-buffered saline, and stored in liquid nitrogen until further analysis. Normal tissue samples were collected 2\u0026nbsp;cm away from the alveolar echinococcosis lesions.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3. Cell culture and transfection\u003c/h2\u003e \u003cp\u003eL-O2 hepatocytes were cultured in Dulbecco\u0026rsquo;s modified Eagle medium supplemented with 10% fetal bovine serum and penicillin/streptomycin in a humidified atmosphere containing 5% CO\u003csub\u003e2\u003c/sub\u003e at 37\u0026nbsp;\u0026deg;C. Approximately 50\u0026nbsp;nM of hsa-miR-125b-5p or control inhibitor, both purchased from JIKAI (Shanghai Jikai), was transfected into L-O2 cells for 48\u0026nbsp;h using the riboFECTCP transfection kit (Guangzhou RIBO Biocompany) according to the protocol provided. All the experiments were performed using cells in the log phase.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4. RNA extraction\u003c/h2\u003e \u003cp\u003eTotal RNA was isolated from plasma and tissue samples using TRIzol according to the protocol provided. The concentration and integrity of isolated RNAs were evaluated using the Agilent Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA, USA). cDNA was synthesized using PrimeScript Reverse Transcriptase (TIANGEN Biotech, Beijing, Co. Ltd) according to the kit protocol. Primers targeting hsa-miR-125b-5p (reverse transcription (RT): 5\u0026prime;-UCCCUGAGACCCUAACUUGUGA-3\u0026prime;; polymerase chain reaction (PCR) forward primer: 5\u0026prime;-TCCCTGAGACCCTAACTTGTGA-3\u0026prime; and PCR reverse primer: 5\u0026prime;-GTGCAGGGTCCGAGGT-3\u0026prime;) and U6 (RT primer: 5\u0026prime;-UGAGGUAGGAGGUUGUAUAGUU-3\u0026prime;; PCR forward primer: 5\u0026prime;-TGAGGTAGGAGGTTGTATAGTT-3\u0026prime;; and PCR reverse primer: 5\u0026prime;-GTGCAGGGTCCGAGGT-3\u0026prime;) were purchased from Sangon Biotech (Shanghai) Co., Ltd.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5. Quantitative PCR (qPCR)\u003c/h2\u003e \u003cp\u003eqPCR was performed using the LightCycler\u0026reg; 480Ⅱ and Tiangen SYBR Green PCR Kit. U6 was used as the endogenous control for miRNA amplification. The reaction volume was 25\u0026nbsp;\u0026micro;l and was set up according to the TB GreenTM Premix Ex TaqTMⅡ protocol. Each sample and blank was analyzed in triplicate. The reaction conditions were as follows: pre-denaturation at 95℃ for 30\u0026nbsp;s, 40 cycles of 95℃ for 5\u0026nbsp;s, and 60℃ for 30\u0026nbsp;s. The melting curves were generated using the following temperatures: 95℃ for 5\u0026nbsp;s and 60℃ for 1\u0026nbsp;min. hsa-miR-125b-5p expression was analyzed using the ΔCt method. Relative miRNA expression was calculated using the 2\u003csup\u003e\u0026minus;△△Ct\u003c/sup\u003e method and normalized to U6 levels.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.6. Inhibition of hsa-miR-125b-5p in treated L-O2 hepatocytes\u003c/h2\u003e \u003cp\u003eL-O2 hepatocytes were transfected with lentiviral vector (LV), LV-hsa-miR-125b-5p-inhibitor, and the negative control virus CON137. The L-O2 cells were transfected with the LV-hsa-miR-125b-5p inhibitor for 5 days. The transfected cells were subsequently incubated with the Cell Counting Kit-8 reagents for 2\u0026nbsp;h to determine the absorption at a wavelength of 450\u0026nbsp;nm in a time-dependent manner. OD\u003csub\u003e450\u003c/sub\u003e represents the number of viable cells.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e2.7. Fluorescence-activated cell sorter analysis for apoptosis\u003c/h2\u003e \u003cp\u003eLV-hsa-miR-125b-5p-inhibition transfection solution and L-O2 hepatocytes were cultured in a 6-well culture plate in triplicate. In order to ensure that a detectable number of cells was present, a cell density of \u0026ge;\u0026thinsp;5\u0026thinsp;\u0026times;\u0026thinsp;10\u003csup\u003e5\u003c/sup\u003e/ml was used. The cell fusion was 85% on the fifth day after transfection. The cells were collected, placed into 5\u0026nbsp;mL centrifuge tubes, and centrifuged at 250\u0026nbsp;g for 5\u0026nbsp;min at 4℃. The cells were washed with cold PBS, suspended in complete medium, and centrifuged at 250\u0026nbsp;g for 3\u0026nbsp;min. Cells were suspended in 200\u0026nbsp;\u0026micro;L 1\u0026thinsp;\u0026times;\u0026thinsp;binding buffer, and 10\u0026nbsp;\u0026micro;L Annexin V-APC was added to stain the cells. The cells were incubated at 10\u0026ndash;15\u0026nbsp;min at room temperature in the dark and analyzed using BD cytoflow.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e2.8. Statistical analysis\u003c/h2\u003e \u003cp\u003eData are presented as the mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation. The independent sample \u003cem\u003et-\u003c/em\u003etest was used to determine the statistical differences between the groups using GraphPad Prism 8. \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered to indicate a statistically significant difference.\u003c/p\u003e \u003c/div\u003e "},{"header":"3. Results","content":" \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e3.1. qPCR\u003c/h2\u003e \u003cp\u003eWe used patient plasma to determine the potential of hsa-miR-125b-5p as an early diagnostic biomarker for alveolar echinococcosis. Our preliminary study using miRNA chips revealed upregulation of hsa-miR-125b-5p.\u003c/p\u003e \u003cdiv id=\"Sec13\" class=\"Section3\"\u003e \u003ch2\u003e3.1.1 hsa-miR-125b-5p expression in plasma samples from alveolar echinococcosis patients\u003c/h2\u003e \u003cp\u003eqPCR showed that hsa-miR-125b-5p was upregulated in the plasma samples of alveolar echinococcosis patients (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe 2\u003csup\u003e\u0026minus;∆∆Ct\u003c/sup\u003e values for hsa-miR-125b-5p expression in plasma in patients with alveolar echinococcosis and controls.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eGroup\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eSample\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003ehsa-miR-125b-5p\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003en\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2\u003csup\u003e\u0026minus;∆∆Ct\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAlveolar echinococcosis patients\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePlasma\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e2.440268\u0026thinsp;\u0026plusmn;\u0026thinsp;0.753355\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHealthy controls\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePlasma\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e1.032328\u0026thinsp;\u0026plusmn;\u0026thinsp;0.261863\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section3\"\u003e \u003ch2\u003e3.1.\u003cem\u003e2 hsa-miR-125b-5p expression in hepatic tissues from alveolar echinococcosis patients\u003c/em\u003e\u003c/h2\u003e \u003cp\u003eqPCR showed that hsa-miR-125b-5p was upregulated in the liver tissue samples obtained from alveolar echinococcosis patients (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe hsa-miR-125b-5p level in hepatic tissues was statistically different between alveolar echinococcosis patients and the control group (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eGroup\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eSample\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003ehsa-miR-125b-5p\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003en\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2\u003csup\u003e\u0026minus;∆∆Ct\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAlveolar echinococcosis patients\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLiver tissue\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e1.932\u0026thinsp;\u0026plusmn;\u0026thinsp;0.682\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHealthy controls\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNormal liver\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e1.001\u0026thinsp;\u0026plusmn;\u0026thinsp;0.049\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003e3.2. Inhibition of hsa-miR-125b-5p in treated L-O2 hepatocytes\u003c/h2\u003e \u003cdiv id=\"Sec16\" class=\"Section3\"\u003e \u003ch2\u003e3.2.1 Transfection of LV, LV-hsa-miR-125b-5p-inhibitor, and negative control viruses CON137 in normal L-O2 hepatocytes\u003c/h2\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section3\"\u003e \u003ch2\u003e3.2.2 Cell Counting Kit-8\u003c/h2\u003e \u003cp\u003eThe proliferation of L-O2 cells increased in the hsa-miR-125b-5p-inhibitor group compared with the negative virus control group. On the fifth day after treatment, the OD\u003csub\u003e450\u003c/sub\u003e of the experimental group was 3.242\u0026thinsp;\u0026plusmn;\u0026thinsp;0.032, while that of the control group was 1.738\u0026thinsp;\u0026plusmn;\u0026thinsp;0.008 (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section3\"\u003e \u003ch2\u003e3.2.3 Cellular apoptosis\u003c/h2\u003e \u003cp\u003eThe apoptosis (%) of the experimental group was 4.84\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09, while that of the control group was 1.91\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18 (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Apoptosis in the control and LV-hsa-miR-125b-5p inhibitor-transfected L-O2 cells after 5 days was compared. Apoptosis was decreased in cells depleted of hsa-miR-125b-5p (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Thus, hsa-miR-125b-5p may promote liver cell proliferation and inhibit apoptosis (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe expression of miR-125b-5p was increased in the plasma and liver lesion tissues of patients with alveolar echinococcosis. Transfection of the LV-hsa-miR-125b-5p-inhibitor reduced the apoptosis of L-O2 hepatocytes in vitro. It is inferred that miR-125b-5p plays a role in alveolar echinococcosis, and may serve as a promising molecular biomarker for the diagnosis and treatment of the disease.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e "},{"header":"4. Discussion","content":" \u003cp\u003emiRNAs are small non-coding regulatory RNAs that play important roles in the pathogenesis of parasitic diseases\u003csup\u003e[\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]\u003c/sup\u003e. Lin\u003csup\u003e[\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]\u003c/sup\u003e et al. (2018) found that miRNA-221/222 is upregulated in the thyroid of papillary thyroid carcinoma patients, and is crucial in invasion and lymph node metastasis, suggesting its potential as a biomarker for invasive papillary thyroid carcinoma. Mogahed\u003csup\u003e[\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]\u003c/sup\u003e et al. (2018) reported that miR-712-3p could be used as a biomarker for the early diagnosis of toxoplasmosis using qPCR and the plasma of mice acutely infected with \u003cem\u003eToxoplasma gondii\u003c/em\u003e.\u003c/p\u003e \u003cp\u003eWe have previously reported differences in the expression of hsa-miR-125b-5p, hsa-let7e-5p, hsa-miR-3165, hsa-miR-4725, and hsa-miR-520e in alveolar echinococcosis plasma samples using microarray data\u003csup\u003e[\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]\u003c/sup\u003e. In this study, 22 plasma samples from alveolar echinococcosis patients and 22 healthy controls were tested using qPCR. We observed the upregulation of hsa-miR-125b-5p in the plasma from alveolar echinococcosis patients. We also examined the expression of hsa-miR-125b-5p in liver samples obtained from 8 patients with alveolar echinococcosis and 8 controls. qPCR revealed the upregulation of hsa-miR-125b-5p in plasma and lesion tissues obtained from alveolar echinococcosis patients. We found that the area under the curve for hsa-miR-125b-5p was 0.997 in the blood, which was significantly higher than that of the control group, with a 95% confidence interval of 0.9907\u0026ndash;1.000 and \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001. The area under the curve for hsa-miR-125b-5p was 0.937 in the alveolar echinococcosis liver tissues, with a 95% confidence interval of 0.8701\u0026ndash;1.000 and \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001. Thus, hsa-miR-125b-5p may serve as a useful diagnostic indicator and therapeutic target for alveolar echinococcosis.\u003c/p\u003e \u003cp\u003eInhibition of hsa-miR-125b-5p in treated L-O2 hepatocytes increased proliferation and decreased apoptosis. This suggested that the inhibition of hsa-miR-125b-5p may promote hepatocyte proliferation to reverse the effects of liver damage. We wish to investigate this further by determining the utility of hsa-miR-125b-5p in treating mice with secondary echinococcosis.\u003c/p\u003e \u003cp\u003emiR-125b-5p is a member of the miR-125 family, which was first reported in 2005 by Lee\u003csup\u003e[\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]\u003c/sup\u003e et al. (2005), and regulates the proliferation of differentiated tumor cells. miR-125b-5p enhances autophagy and apoptosis in multiple myeloma cells and can be used as a novel therapeutic target and early diagnostic biomarker for multiple myeloma\u003csup\u003e[\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]\u003c/sup\u003e. Thus, the role of circulating miRNAs in the early diagnosis of alveolar echinococcosis may improve our understanding of the pathogenesis of alveolar echinococcosis. Our findings strongly suggest that hsa-miR-125b-5p may serve as a useful diagnostic and prognostic biomarker for alveolar echinococcosis.\u003c/p\u003e \u003cp\u003eHowever, this study has certain limitations. Firstly, the findings in this study are preliminary and do not attempt to elucidate the mechanism by which hsa-miR-125b-5p is implicated in alveolar echinococcosis. Moreover, we have not addressed the association between hsa-miR-125b-5p expression and the size of the diseased tissue. Thus, the correlation between hsa-miRNA-125b-5p expression, diseased tissue size, the physiological characteristics of alveolar echinococcosis lesion(s), and recovery of patients after surgery will be investigated in future. Additionally, the potential of hsa-miRNA-125b-5p to distinguish between alveolar and cystic echinococcosis, as well as other liver diseases, requires further investigation. Our study findings require further validation using a larger cohort samples to determine whether hsa-miR-125b-5p may serve as a biomarker for the early non-invasive diagnosis of alveolar echinococcosis.\u003c/p\u003e "},{"header":"5. Conclusions","content":" \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eIn this study, we report the upregulation of hsa-miR-125b-5p in alveolar echinococcosis using an independent cohort of 22 patients with alveolar echinococcosis and 22 healthy individuals. While hsa-miR-125b-5p may serve as a promising diagnostic marker for alveolar echinococcosis, additional experiments should be performed to further investigate its diagnostic value. Future studies are required to identify the expression of hsa-miR-125b-5p in the plasma of patients with cystic echinococcosis, alveolar echinococcosis, and other liver diseases to determine the potential of hsa-miR-125-5p as a differential biomarker.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003e \u003cb\u003eDeclarations\u003c/b\u003e \u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e "},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003e Ethics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was approved by the ethics review broad of the Hospital Affiliated of Qinghai University (approval number: P-SL-2019054).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;competing interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe author(s) declare that they have no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAgree to publish\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;Authors' contributions \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCDP conceived and designed the study and drafted the manuscript. JBF did the research for q-PCR and data analysis. ZYG and PMQ helped design the study and revise the draft manuscript. All authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e Funding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by the Qinghai Science and Technology Department (grant numbers 2016-SF-A5 and 2019-SF-131).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe\u0026nbsp;would\u0026nbsp;like\u0026nbsp;to\u0026nbsp;thank\u0026nbsp;Editage\u0026nbsp;(www.editage.cn)\u0026nbsp;for\u0026nbsp;English\u0026nbsp;language\u0026nbsp;editing.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e \u003cspan\u003eEckert J, Deplazes P. Biological, epidemiological, and clinical aspects of echinococcosis, a zoonosis of increasing concern. Clin Microbiol Rev. 2004;17:107\u0026ndash;35.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eWang H, Zhang JX, Schantz PM, et al. Epidemiologic survey and analysis on echinococcosis in humans and animals from 1995 to 2005 in Qinghai province. Zhongguo renshou gonghuanbing xuebao. 2006;22:1129\u0026ndash;34. (in Chinese).\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eNunnari G, Pinzone MR, Gruttadauria S, Celesia BM, Madeddu G, Malaguarnera G, Pavone P, Cappellani A, Cacopardo B. Hepatic echinococcosis: clinical and therapeutic aspects. World J Gastroenterol. 2012;18:1448\u0026ndash;58.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eMariconti M, Bazzocchi C, Tamarozzi F, Meroni V, Genco F, Maserati R, Brunetti E. Immunoblotting with human native antigen shows stage-related sensitivity in the serodiagnosis of hepatic cystic echinococcosis. Am J Trop Med Hyg. 2014;90:75\u0026ndash;9.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eGottstein B, Wang JH, Blagosklonov O, Grenouillet F, Millon L, Vuitton DA, M\u0026uuml;ller N. Echinococcus metacestode: in search of viability markers. Parasite. 2014;21:63.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eGiraudoux P, Pleydell D, Raoul F, Qu\u0026eacute;r\u0026eacute; JP, Wang Q, Yang Y, Vuitton DA, Qiu J, Yang W, Craig PS. Transmission ecology of \u003cem\u003eEchinococcus multilocularis\u003c/em\u003e: what are the ranges of parasite stability among various host communities in China. Parasitol Int. 2006;55:237\u0026ndash;46.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eBrunetti E, Kern P, Vuitton DA. Expert consensus for the diagnosis and treatment of cystic and alveolar echinococcosis in humans. Acta Trop. 2010;114:1\u0026ndash;16.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eWang TP, Cao ZG,. Progress in prevention and control of Chinese hydatid disease and its existing problems. Zhongguo jishengchongxue yu jishengchongbing fangzhi zazhi. 2018;36(3):291\u0026ndash;6. (In Chinese).\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eGuo X, Zheng Y. Expression profiling of circulating miRNAs in mouse serum in response to Echinococcus multilocularis infection. Parasitology. 2017;144:1079\u0026ndash;87.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eBartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004;116:281\u0026ndash;97.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eBartel DP. Micro RNAs: target recognition and regulatory functions. Cell. 2009;136:215\u0026ndash;33.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eTao YH, Wang ML, Wang M, et al. Quantification of circulating miR-125b-5p predicts survival in chronic hepatitis B patients with acute-on-chronic liver failure. Digestive Liver Disease. 2019;51:412\u0026ndash;8.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eWang J, Yan F, Zhao Q, Zhan F, Wang R, Wang L, Zhang Y, Huang X. Circulating exosomal miR-125a-3p as a novel biomarker for early-stage colon cancer. Sci Rep. 2017;7:4150.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eJin X, Guo X, Zhu D, Ayaz M, Zheng Y. miRNA profiling in the mice in response to \u003cem\u003eEchinococcus multilocularis\u003c/em\u003e infection. Acta Tropica. 2017;166:39\u0026ndash;44.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eLin ZK, Zhou K, Lu J, et al. ROC curve analysis of mi RNA-RNA-221/222 as a marker for invasive papillary thyroid carcinoma in clinical value. Aizheng jinzhan. 2018;16(1):49\u0026ndash;52. (In Chinese).\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eMogahed NF, Khedr SI, Ghazala RA, Masoud IM. Can miRNA712-3p be a promising biomarker for early diagnosis of toxoplasmosis? Asian Pac. J Trop Med. 2018;11:688\u0026ndash;92.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eLiu J, Cao DP, Zhang XF, et al. Analysis of serum miRNAs expression spectrum in patients with alveolar echinococcosis. Chinese high altitude medicine biology. 2019;40:51\u0026ndash;6. (in Chinese).\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eJiang BF, Sha CQ, Zhang YG, et al. Expression of hsa-miR-125b-5p in serum of patients with alveolar echinococcosis and screening of its signal transduction pathway and target genes. Chinese high altitude medicine biology. 2020;41:39\u0026ndash;45. (in Chinese).\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eLee YS, Kim HK, Chung S, Kim KS, Dutta A. Depletion of human micro-RNA miR-125b reveals that it is critical for the proliferation of differentiated cells but not for the down-regulation of putative targets during differentiation. J Biol Chem. 2005;280:16635\u0026ndash;41.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eMorelli E, Leone E, Gallo, Cantafio,et al. Selective targeting of IRF4 by synthetic microRNA-125b-5p mimics induces anti-multiple myeloma activity in vitro and in vivo. Leukemia. 2015;29:2173\u0026ndash;83.\u003c/span\u003e \u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-infectious-diseases","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"infd","sideBox":"Learn more about [BMC Infectious Diseases](http://bmcinfectdis.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/infd","title":"BMC Infectious Diseases","twitterHandle":"#bmcinfectdis","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"alveolar echinococcosis, diagnostic biomarker, hsa-miR-125b-5p, plasma, hepatic tissue samples","lastPublishedDoi":"10.21203/rs.3.rs-68399/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-68399/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground:\u003c/strong\u003e Alveolar echinococcosis is an infectious zoonotic disease caused by \u003cem\u003eEchinococcus multilocularis \u003c/em\u003ethat is endemic to the vast pastoral areas of southern Qinghai province of China. Alveolar echinococcosis affects the human liver, and its clinical manifestations are similar to those of liver tumors. It is also called “worm cancer,” as it induces irreversible liver fibrosis and brain metastases. Alveolar echinococcosis is a serious threat to human health and is a burden to patients and economies. When detected and treated early, patients with alveolar echinococcosis have a strong chance of recovery. Therefore, it is imperative to identify early diagnostic biomarkers of the disease. MicroRNAs (miRNAs) are short, single-stranded, non-coding RNA molecules that play key roles in a wide range of biological processes, and have been recently implicated in tumorigenesis. However, little is known about the role of miRNAs in alveolar echinococcosis. We have previously reported differentially expressed miRNAs in the plasma of patients with alveolar echinococcosis and healthy individuals, using microarray assay chips. \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e We screened 10 differentially expressed miRNAs based on their fold change and the available literature. Additionally, quantitative polymerase chain reaction was used to verify their expression in the plasma of alveolar echinococcosis patients. We found that hsa-miR-125b-5p was upregulated in the plasma and the hepatic tissue samples of alveolar echinococcosis patients; therefore, its role as a diagnostic biomarker for alveolar echinococcosis was investigated. Accordingly, hsa-miR-125b-5p was upregulated in plasma and hepatic tissue samples obtained from alveolar echinococcosis patients. Receiver operating characteristic curves showed that hsa-miR-125b-5p was associated with an area under the curve of 99.8% and 98.9% in the plasma and hepatic tissues of alveolar echinococcosis patients, respectively. Transfection of an LV-hsa-miR-125b-5p-inhibitor reduced apoptosis of L-O2 hepatocytes in vitro (\u003cem\u003eP \u003c/em\u003e\u0026lt; 0.05). Thus, hsa-miR-125b-5p may promote liver cell proliferation and inhibit liver cell apoptosis.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConclusions:\u003c/strong\u003e Taken together, hsa-miR-125b-5p may be a promising diagnostic biomarker for the early non-invasive diagnosis of alveolar echinococcosis. We plan to validate the expression of hsa-miR-125b-5p in the plasma of patients with cystic echinococcosis, alveolar echinococcosis, and other liver diseases to determine the potential of hsa-miR-125-5p as a differential biomarker in future studies.\u0026nbsp;\u003c/p\u003e","manuscriptTitle":"miR-125b-5p\u0026nbsp;is A Promising Novel Plasma Biomarker\u0026nbsp;for Alveolar Echinococcosis in Patients from the Southern Province of Qinghai","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2020-09-18 15:10:35","doi":"10.21203/rs.3.rs-68399/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewersInvited","content":"","date":"2020-09-18T12:00:00+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2020-09-17T12:00:00+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2020-09-16T12:00:00+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2020-09-15T12:00:00+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-infectious-diseases","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"infd","sideBox":"Learn more about [BMC Infectious Diseases](http://bmcinfectdis.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/infd","title":"BMC Infectious Diseases","twitterHandle":"#bmcinfectdis","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"65256e36-e071-411e-966e-b1c105f95188","owner":[],"postedDate":"September 18th, 2020","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":546996,"name":"Infectious Diseases"}],"tags":[],"updatedAt":"2021-03-14T15:00:35+00:00","versionOfRecord":{"articleIdentity":"rs-68399","link":"https://doi.org/10.1186/s12879-021-05940-z","journal":{"identity":"bmc-infectious-diseases","isVorOnly":false,"title":"BMC Infectious Diseases"},"publishedOn":"2021-03-07 15:00:27","publishedOnDateReadable":"March 7th, 2021"},"versionCreatedAt":"2020-09-18 15:10:35","video":"","vorDoi":"10.1186/s12879-021-05940-z","vorDoiUrl":"https://doi.org/10.1186/s12879-021-05940-z","workflowStages":[]},"version":"v1","identity":"rs-68399","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-68399","identity":"rs-68399","version":["v1"]},"buildId":"_2-kVJe1T_tPrBINL-cwx","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}