Age- and sex-dependent reference intervals of inhibin B in Chinese children on chemiluminescence analyzer

Age- and sex-dependent reference intervals of inhibin B in Chinese children on chemiluminescence analyzer | 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 Article Age- and sex-dependent reference intervals of inhibin B in Chinese children on chemiluminescence analyzer Bin Zhou, Jinjun Luo, Xiang Dai, Xuelian He, Hui Yao This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4244728/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 10 You are reading this latest preprint version Abstract Backgroud : Inhibin B (INHB) is an emerging biomarker of sexual function diseases, but its clinical use in children is hindered due to the lacking reference intervals (RIs) based on age sex. Methods : A total of 622 healthy Chinese children were included and serum INHB and six conventional sex hormones were measured. INHB was partitioned by four age groups (<1y, 1~5y, 5~10y, 10~18y) and the corresponding RIs were determined with quantile. The effect of age on INHB was assessed using five models including linear, quadratic, cubic, S, and logarithmic. Spearman correlation was used to examine the associations between INHB and sex hormones. Results : Compared to the boys, girls had lower median concentrations and variability in INHB across four age groups. The optimal models for the effect of age on INHB were cubic (R 2 =0.440 for boys). The INHB concentration in the first and oldest age groups was more high than other groups ( P ≤0.001). Spearman’s correlation analysis showed sex-dependent association between INHB and conventional sex hormones (e.g. a positive association with PRL in boys). Conclusions : INHB level is influenced by age and gender, and our results provide specific RIs to facilitate to screen sexual function diseases in Chinese children. Biological sciences/Biochemistry/Hormones/Gonadal hormones Health sciences/Diseases/Reproductive disorders Inhibin B reference intervals children age sexual function Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Inhibin B (INHB), as a gonadal glycoprotein hormone consisting of alpha and beta β subunit, belongs to transforming growth factor-β superfamily. It is mainly secreted by developing follicular granulosa cells for female and by testicular sertoli cells for male, which plays an important role in the female ovarian and male testicular assessment 1-4 . INHB levels vary greatly from birth to adolescence until steady concentrations in adult 5 , with a peak in the first month of life as activation of hypothalamic-pituitary-gonadal. The increasing INHB will promote the functional follicle maturation, and then there is a trough at 5-10 years. In prepuberty, its levels are independent of the presence of actively proliferating germ cells and rise under the stimulation of FSH 6 . During puberty there is a regular increase of INHB 4 . Over the last decade, INHB has received increasing attention of the pediatric endocrinologist. INHB determination is an essential tool in the assessment of pubertal disorders and the function of Sertoli cells 7 . For example, INHB were characterized its clinical usefulness to evaluate gonadal function in children post-treatment for cancer 8, 9 . Raman reported that INHB was as marker of sertoli cell function in children subjected to surgical correction for undescended tests 1 . Furthermore, INHB concentration/ratio to sex hormones is regarded as a newly predictor of age at first menstruation in girls with idiopathic central precocious puberty 10 , and potential markers of progressive central precocious puberty 11 and Bilateral Cryptorchidism in infants 12 . So, inhibin B is very extensive in clinical application. It is generally believed that accurate interpretation of test results, proper diagnostics and treatment rely on accurate reference ranges. Inappropriate pediatric references intervals (RIs) may impact clinical decisions. However, RIs of serum INHB have not been required due to the effects of age, sex, methodologies and difficulties in collecting ample serum from healthy children especially neonates. A few reports have investigated RIs of INHB based on enzyme-linked immunosorbent assays (ELISA), but they are the rough reference ranges from a small amount of children in European and western countries 5, 6, 13 or are acquired from the specific enrolled subjects such as infertile couples 14 . There little is known about RIs of INHB in Chinese pediatric individuals, which hinders pediatric clinical use as a potential diagnostic marker. Therefore, it is necessary to establish normal RIs for serum INHB in heathy Chinese children with considerations of the effect of age and gender. KAESER 1000 is fully automatized chemiluminescence analyzer for measurement of INHB and sex hormones. The platform is developed recently that is more fast and stable than the manual method. Compared to the commercial ELISA, KAESER 1000 have improved in sensitivity and specificity. Therefore, the first objective of this study was to setup the RIs of INHB for healthy children aged from 0 to 18 years in China based on KAESER 1000 chemiluminescence technology. The secondary aim was to investigate the effects of age and gender on INHB levels to provide insights about the INHB application in the reproductive health. Further, it was also aimed to assess the associations of INHB with conventional markers of gonadal function, including follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (TT), prolactin (PRL), progesterone (PROG) and total estradiol (E2). Results Participant characteristics The overall population comprised 302 girls and 320 boys, with mean (± SD) age of 5.9 y (± 4.8) and BMI of 17.78 kg/m 2 (± 3.51). There was no significant difference in age between the boys and girls ( P = 0.911). Weight and BMI for boys were higher than those for girls ( P = 0.043, P = 0.002) (Table 1 ). Compared to the boys, girls had lower INHB and E2 levels among four age subgroups ( P < 0.001), but had higher FSH and PRL levels (Table 2 ). Moreover, changes in serum LH, PROG and E2 levels were negligible among the first three subgroups (Table 2 ). Table 1 Characteristics of children in four age groups [mean ± SD] Girls Boys groups n height weight BMI n height weight BMI < 1y 82 63.8 ± 7.4 7.1 ± 2.2 16.8 ± 3.0 81 66.9 ± 7.7 8.4 ± 2.5 18.4 ± 4.5 1 ~ 5y 60 95.9 ± 11.4 15.2 ± 3.5 16.5 ± 1.8 85 97.2 ± 10 15.7 ± 3.1 16.6 ± 1.8 5 ~ 10y 77 125.6 ± 11.5 27.1 ± 6.9 17.1 ± 2.9 63 130.0 ± 12.4 31.4 ± 10.2 17.9 ± 3.2 10 ~ 18y 83 154.6 ± 9.0 43.9 ± 10.5 18.2 ± 3.0 91 153.4 ± 17.1 46.4 ± 15.2 19.3 ± 4.5 total 302 112.7 ± 37.6 24.6 ± 16.4 17.3 ± 2.9 320 112.9 ± 37.4 26.4 ± 18.6 * 18.2 ± 4.0 * BMI, body mass index; * P < 0.05, independent samples test between boys and girls Table 2 The estimated age-and gender-specific reference intervals for INHB and six sex hormones in four age groups [median (2.5–97.5 percentiles)/ mean ± SD] groups INHB (pg/mL) FSH (mIU/mL) LH (mIU/mL) PRL(ng/mL) Prog(ng/mL) E2(pg/mL) girls < 1y 67 (8, 206) * 4.5 (0.7, 20.3) 0.45 (0.2, 2.01) 18.0 (3.4, 109.2) 0.3 (0.1, 5.5) 3.2 (3.2, 40.1) † 1 ~ 5y 33 (14, 227) * 2.3 (0.7, 18.0) 0.45 (0.45, 0.45) a 11.8 (3.8, 58.2) 0.3 (0.1, 1.6) 3.2 (3.2, 20.9) † 5 ~ 10y 27 (8, 92) * 1.7 (0.3, 23.6) 0.34 (0.45, 2.40) 8.8 (3.7, 33.3) 0.3 (0.2, 1.2) 3.2 (3.2, 33.8) † 10 ~ 18y 81 (12, 219) * 5.0 (1.2, 11.4) 0.45 (3.69, 27.01) 10.7 (3.6, 31.5) 0.3 (0.1, 5.2) 3.2 (41.0, 174.9) total 46 (12, 182) ‡ 3.5 (0.5, 13.3) 0.45 (0.24, 14.33) 12.0 (3.8, 60.9) 0.3 (0.1, 3.7) 13.0 (3.2, 96.0) boys < 1y 236 (75, 476) 0.8 (0.2, 4.0) 0.5 (0.2, 4.0) 16.6 (3.9, 117.5) 0.3 (0.1, 19.5) 12.0 (3.2, 52.1) 1 ~ 5y 148 (62, 355) 0.8 (0.2, 12.3) 0.45 (0.45, 0.45) a 9.9 (4.1, 38.3) 0.3 (0.2, 2.3) 10.5 (3.2, 35.7) 5 ~ 10y 102 (53, 265) 1.0 (0.12, 16.8) 0.45 (0.27, 2.14) 8.21 (3.16, 46.26) 0.3 (0.2, 2.6) 11.0 (3.2, 26.9) 10 ~ 18y 156 (54, 290) 2.6 (0.31, 8.6) 2.01 (0.31, 5.39) 10.94 (3.51, 32.00) 0.2 (0.1, 1.1) 22.0 (10.0, 74.9) total 159 (55, 418) 1.2 (0.19, 8.3) 0.45 (0.23, 4.04) 11.4 (3.9, 62.5) 0.3 (0.1, 3.2) 12.0 (3.2, 40.5) † low detection rate; * P < 0.001, independent samples test of INHB level in corresponding age subgroups between boys and girls. ‡ P < 0.001, independent samples test of INHB level between sex. FSH, follicle-stimulating hormone; LH, luteinizing hormone; TT, testosterone, PRL prolactin, Prog progesterone and E2, total estradiol. Distribution of INHB The distribution of INHB in girls and boys is shown comparatively in Fig. 1 Children in both sexes had the high median INHB concentrations within the first years old and declined sharply from 1-year-old to 5-year-old, then increased between 10 and 18 years old. Significant differences in INHB levels were observed among the multiple age subgroups ( P ≤ 0.001). For girls highest INHB levels is the first age group, but for boys is the oldest age group. In addition, compared to the boys, girls had reduced variability in INHB across the four age groups (Fig. 1 A and B) The effect of age on INHB was also assessed using five models including linear, quadratic, cubic, logarithmic, and S (Fig. 2 ). Linear, quadratic, cubic for all population, logarithmic only for boys were effective ( P = 2.36×10 − 14 , P = 2.00×10 − 32 , P = 1.37×10 − 39 ). The optimal models in girls and boys all were cubic models (R 2 = 0.198 for girls and R 2 = 0.440 for boys, Supplementary Table S2). Reference intervals Due to the volatility of INHB concentration along with age and sex, so the RIs for INHB and six sex hormones including the 2.5th and 97.5th quantiles along with the median in boys and girls are shown in Table 2 by age. The median (2.5%~97.5%) INHB level from the oldest age group in girls was the highest in comparison to other age groups. However, the median level of INHB from the first age group in boys was the highest. Correlations between the INHB level and FSH, LH, PRL, PROG and E2 Because the detection rate of serum TT in both girls and boys were too low, the correlation analysis with INHB was exc1uded. In Spearman’s correlation analysis (Fig. 3 and Fig. 4 ), INHB was weakly positively associated with LH in both girls (Rs = 0.321, P = 1.20×10 − 7 ) and boys (Rs = 0.225, P = 2.85×10 − 4 ), and positively associated with FSH (Rs = 0.403, P = 1.61×10 − 11 ) and E2 (Rs = 0.359, P = 2.52×10 − 9 ), but negatively associated with PROG (Rs=-0.136, P = 0.028) only in girls. There was a positive correlation between INHB and PRL only in boys (Rs = 0.300, P = 8.91×10 − 7 ). Discussion The normal values for serum INHB have been often assessed among adults, especially fertile and infertile adults 14 – 16 . However, few study has established sex- and age-related INHB reference data from birth to adulthood (n = 366, boys) 5 , aged 6-11years (n = 705, girls) 6 and during childhood and puberty (n = 52, girls) 13 in European and western countries. Due to the practical difficult in infant’s recruitment, some studies may contain few infants or participants aged range of 5 ~ 18 years old. In the current study, 163 infants (< 1 y subgroup) were regarded as a key strength of our study. To the best of our knowledge, this is the first one to detect serum INHB concentrations in heathy children aged from 0 to 18 years among a large group of more than 600 populations in China. In this study, we investigated the age- and gender-related effects on INHB concentrations and calculated the pediatric RIs from birth to puberty based on a newly introduced fully automatic chemiluminescence analyzer. Our results suggested that gender and age had effects on INHB levels. The level of INHB for boys and girls was similar U-shaped along with age and the optimal estimated models both were cubic. The distribution of INHB levels as four age groups also confirmed the shaped phenomenon. Our finding suggested that INHB was highly variable during the first year of life and attained gradually stability as the age, then increase to adolescence. INHB for girls was high from born, and decreased gradually to reach a trough at 2 ~ 8 years old, and then increased again to reach a crest around 13 years old. However, there were differences for boys, with step to plateau level around 5.5 ~ 9 years old and a crest around 13 years old, which were similar trends with previous study 5 . Increase of INHB at infants and pubertal period were involved the Sertoli cell proliferation 17 , 18 , which in males within one year old can increase fivefold 17 . We found the post-natal peak exceeded the level of INHB in pubertal period in boys. Andersson et al also observed that the peak in infant boys exceeded in adult men 19 . High level of infancy affords an early opportunity to assess hypothalamic-pituitary-gonadal axis function, which is urgently needed, especially, when newborn happens ambiguous genitalia or suspicion of hypopituitarism or hypogonadism 20 . Postnatal increase of INHB also may predict later testicular function, such as spermatogenesis, but abundant studies are requested to evaluate its effectiveness 21 . Our study also showed that INHB raised from age 8 for girl and 9 for boys, which seem to be early than the clinical onset of puberty with mean age of 9 years in girls and 11.5 years in boys, respectively. This demonstrates that INHB secretion takes places in the prepubertal testis, despite the very low levels of both gonadotrophins and TT. Therefore, INHB is used to assess Sertoli cell health and number in pre-pubertal stage. Interestingly, our finding showed that girls not only had lower concentrations but also reduced variability in INHB across four age groups compared to the boys. These results were in agreement with previous studies at all stages of puberty 22 and in infancy 20 . Besides INHB, anti-Müllerian hormone is also expressed at varying degrees in children 23 . However, no published study investigated detailed the difference variability of INHB for girls and boys with different age groups in healthy pediatric populations. The INHB reference ranges have changed over the years in adults among different laboratories 24 . Further, previous reports showed that INHB level was U-shaped with age from birth to adulthood 5 . So, this prompted us to reconsider the INHB cutoff value in childhood and adolescence. Because INHB concentrations in different sex and age individuals were significant difference, the age-specific and sex-specific RIs were calculated separately. Our RIs for INHB [236(75–476) pg/ml for < 1y; 102(53, 265) pg/ml for 5 ~ 10y] was close to the ranges observed in children of the same age range in Ireland [278(68–630) pg/mL for < 1y, n = 51; 84 (35–167) for 6 ~ 10y, n = 80] 5 . However, our RIs for INHB at 10 ~ 18 group [156 (54, 290)] pg/mL was different from the 12 ~ 17 y group [280 (74 ~ 470) pg/mL in Ireland 5 . The remarkable difference of RIs may result from study populations, the establish criteria of reference range, and detection methodologies 24 . Our study indicated that INHB levels were correlated positively with FSH and LH in girls, which were consistent with previous reports in prepubertal, pubertal girls 22 . Other studies also showed no association between FSH and INHB in prepubertal cryptorchidic boys (0.5 to 13 year) 25 and normal prepubertal boys 5 . Our study showed that serum INHB moderately correlated positively with E2 ( P = 2.52×10 − 9 ) and negatively with PROG ( P = 0.028) in girls. INHB correlated positively with PRL ( P = 8.91×10 − 7 ) levels in boys. However, we did not find any reports on correlations between INHB and E2, PROG, and PRL in children. There are several limitations in our study. Firstly, although the inclusion criterion were standardized to select possibly those healthy individuals, it is possible to include pathologic subjects. Secondly, due to certain practical limitations, the participants in adolescence were not identify as Tanner staging and menstrual cycle. Finally, we do not evaluate INHB level in repeated measures. In conclusion, our study administrated the age- and sex-dependent RIs for serum INHB. Our results suggested that INHB level was lower and reduced variability for girls across four age groups compared to boys. INHB concentration changed along with age. Furthermore, significant difference for correlations between INHB and FSH, LH, E2, PROG, PRL were observed in different sex. Our results may broaden the application of INHB levels, which may be sensitively used as an effective indicator of gonad function assessment and distinguish the patients and normal children. Methods Study populations The study population included boys and girls aged 0 ~ 18 years. Samples were acquired from our routine laboratory at Eugenic genetics laboratory, Wuhan Children’s Hospital between January and December 2022. The subject who had the following conditions were excluded: 1) reproductive diseases such as cryptorchidism, inguinal hernia, varicocele, testicular trauma, hermaphroditism, precocious puberty, sexual development delay, and adolescent polycystic ovary syndrome; 2) chronic diseases such as tumor and tuberculosis; 3) endocrine diseases such as diabetes, hyperthyroidism, and obesity; 4) severe liver and kidney damage; 5) serious trauma, infection, and sepsis; 6) neonatal respiratory distress syndrome, jaundice, and shock; 7) use of chemotherapy drugs. After exclusion of the above mentioned criteria, a total of 622 subjects were included in the current study. Among them, 518 children provided sufficient volume of serum samples for other hormone measurements. The baseline characteristics on age, gender, height and weight were collected from the hospital’s medical records. All study participants provided an informed consent, and the present study was approved by the Ethics Committee of Wuhan Children’s Hospital (No.2022R108-E01). Hormone measurements Blood samples were drawn from an antecubital vein and centrifuged after clotting. Serum was stored at -20 ℃ until analysis. Serum INHB was measured in double-antibody chemiluminescence immunoassay (Kangran Biotech, Guangzhou, China). The detection of limit (LOD) for INHB assay was 7 pg/mL, and the intra and interassay coefficients of variation were 6.25%~8.33% and 4.34%~9.73%, respectively. Serum INHB, FSH, LH, PRL, PROG, E2 and TT levels were determined using the Kaeser 1000 automatic chemiluminescence immune analyzer. The parameters for other serum hormones were described in supplementary table S1 . Statistical analysis All statistical analyses were performed with SPSS18.0. P -value less than 0.05 was defined as statistically significant. Independent samples t-test were used to reveal significant differences in height, weight, and BMI between girls and boys. The concentrations of INHB and sex hormones below their LODs were replaced with the LODs divided by the square root of 2. The serum INHB concentrations were stratified into four subgroups by age (< 1y, 1 ~ 5y, 5 ~ 10y, 10 ~ 18y) and into two subgroups by gender (girls and boys). Serum INHB concentrations were the skewed distribution, and thus were transformed by natural logarithm (ln) to meet the normality for the following analysis. The differences of INHB levels among four age groups stratified by gender were compared by Bonferroni multiple comparison test, which was used to control type 1 error multiple comparison. The RIs of INHB at 2.5th, and 97.5th quantiles were expressed for each age group partitioned by gender. Multiple linear and non-linear mathematical models were then performed in order to describe the variation of INHB along with age. The models of curve estimation included linear, quadratic, cubic, logarithmic, and S. Relationships between INHB and other hormone levels were analyzed using Spearman correlation test. Declarations Ethics declarations The present study was approved by the Ethics Committee of Wuhan Children’s Hospital (No.2022R108-E01). All procedures were in accordance with the ethical standards of the institution and with the 1964 Helsinki declaration and its later amendments. All study participants provided an informed consent. Data availability The datasets use and/or analysed during the current study are provided by the corresponding author on a reasonable request. Acknowlegements We would like to thank participants without whom this work would not be possible. Author contributions Bin Zhou: Investigation, Methodology, Writing-original draft; Jin jun Luo and Xiang Dai: Validation, Writing review & editing; Xuelian He and Hui Yao: Writing review & editing. All authors have accepted responsibility for the entire content of this manuscript and approved its submission. Research funding This research did not receive any funding from public agencies, commercial agencies, or not-for-profit sectors. The study was funded by Wuhan Children’s Hospital. Competing Interests The author(s) declare no competing interests. References Raman, V. S., Khanna, S. & Verma, V. A prospective observational study to evaluate the change in inhibin-B as a marker of sertoli cell function in children subjected to surgical correction for undescended testes. Afr J Paediatr Surg 19, 4233–4237(2022). Li, Y., Nie, M. Y., Liu, Y., Zhang, W. Y., Yang, X. K. The dynamic changes of anti-Mullerian hormone and inhibin B during controlled ovarian hyperstimulation in decreased ovarian reserve women and the effect on clinical outcome. Gynecol endocrino l 6, 450–453(2015). Hansen, A. H. et al . 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The Sertoli cell hormones inhibin-B and anti Mullerian hormone have different patterns of secretion in prepubertal cryptorchid boys. J Pediatr Surg 51, 475–480, (2016). Additional Declarations No competing interests reported. Supplementary Files Supplementarymaterials.docx Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 19 Jul, 2024 Reviews received at journal 18 Jul, 2024 Reviewers agreed at journal 15 Jul, 2024 Reviews received at journal 05 Jun, 2024 Reviewers agreed at journal 01 Jun, 2024 Reviewers invited by journal 01 May, 2024 Editor assigned by journal 28 Apr, 2024 Editor invited by journal 27 Apr, 2024 Submission checks completed at journal 24 Apr, 2024 First submitted to journal 09 Apr, 2024 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. 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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-4244728","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":296899842,"identity":"1a179877-cdf9-46cd-9e2d-771e1d8bc567","order_by":0,"name":"Bin Zhou","email":"","orcid":"","institution":"Wuhan Children's Hospital (Wuhan Maternal and Child Healthcare Hospital, Huazhong University of Science \u0026 Technology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Bin","middleName":"","lastName":"Zhou","suffix":""},{"id":296899843,"identity":"01ebdc1b-a258-4b53-be46-361dbdca9d19","order_by":1,"name":"Jinjun Luo","email":"","orcid":"","institution":"Hubei Provincial Center for Disease Control and Prevention","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jinjun","middleName":"","lastName":"Luo","suffix":""},{"id":296899844,"identity":"39141b41-23b2-47c7-8206-a4e8f7d27e3e","order_by":2,"name":"Xiang Dai","email":"","orcid":"","institution":"Wuhan Children's Hospital (Wuhan Maternal and Child Healthcare Hospital, Huazhong University of Science \u0026 Technology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Xiang","middleName":"","lastName":"Dai","suffix":""},{"id":296899845,"identity":"eb97851d-ea61-4f10-b483-994e42c0facf","order_by":3,"name":"Xuelian He","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABD0lEQVRIiWNgGAWjYBACAzBZIMHAwN7YcPBDxQEw/8ADgloMgFp4Dh88LHHmAAMPSEsCYS1ALJGWfIC3DaKFAZ8Wc/YeM4kPBhby5gw5Bgck592Rsxc7/BBoi52cbgN2LZY9x9IkZxhIGO5sOGNwoHDbM2Me6TQDoJZkY7MDOBx2I/mYNI+BBOOGgz1AW7YdTuyRTgBpOZC4DZeW+w/bpP8YSNhvOMxjcIB3DkhL+gf8Wm4wH5MGhljihmNsCQd4G0BacgjYciYt2bLHQCJ5wxnmA4cljh025rmdU3AgwQCPX46fMbzxo6LOdsP9h80fP9QclmOfnb75w4cKOzlcWnABA9KUj4JRMApGwShABQDq02kLLi5R4gAAAABJRU5ErkJggg==","orcid":"","institution":"Wuhan Children's Hospital (Wuhan Maternal and Child Healthcare Hospital, Huazhong University of Science \u0026 Technology","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Xuelian","middleName":"","lastName":"He","suffix":""},{"id":296899846,"identity":"1d92506c-b9d6-4041-84d2-bfde7009be9f","order_by":4,"name":"Hui Yao","email":"","orcid":"","institution":"Wuhan Children's Hospital (Wuhan Maternal and Child Healthcare Hospital, Huazhong University of Science \u0026 Technology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hui","middleName":"","lastName":"Yao","suffix":""}],"badges":[],"createdAt":"2024-04-10 03:02:50","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4244728/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4244728/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":55634136,"identity":"c43152d4-ea32-4846-9f71-1bb9ec778e41","added_by":"auto","created_at":"2024-04-30 20:09:43","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":224311,"visible":true,"origin":"","legend":"\u003cp\u003eAge-stratified distribution of INHB in children.\u003c/p\u003e\n\u003cp\u003eGraphs illustrate the median and interquartile range.\u003cstrong\u003e a \u003c/strong\u003egirls; \u003cstrong\u003eb\u003c/strong\u003e Boys. Inter-group\u003c/p\u003e\n\u003cp\u003estatistical significance is expressed as implied by Bonferroni multiple comparison test after INHB natural logarithm transformed.\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-4244728/v1/66963e7d49522962a61e2baf.jpeg"},{"id":55634821,"identity":"568b129a-1481-4d58-af47-cc102e6aac44","added_by":"auto","created_at":"2024-04-30 20:17:43","extension":"jpeg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":289774,"visible":true,"origin":"","legend":"\u003cp\u003eCurve estimation of the INHB changes along with age\u003c/p\u003e\n\u003cp\u003ea girls; b boys\u003c/p\u003e","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-4244728/v1/f68e19d33e4b5d565475283a.jpeg"},{"id":55634138,"identity":"1743f116-07ac-4953-8147-80fa30e531d5","added_by":"auto","created_at":"2024-04-30 20:09:44","extension":"jpeg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":520835,"visible":true,"origin":"","legend":"\u003cp\u003eSpearman correlation test in girls. a. serum INHB and FSH levels; b. serum INHB and LH levels; c. serum INHB and E2 levels; d. serum INHB and PROG levels; e. serum INHB and PRL levels.\u003c/p\u003e","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-4244728/v1/56c6fe1c6840f8dc22ff0617.jpeg"},{"id":55634137,"identity":"4404349b-6009-4191-9869-8fd3d4f0d124","added_by":"auto","created_at":"2024-04-30 20:09:44","extension":"jpeg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":529548,"visible":true,"origin":"","legend":"\u003cp\u003eSpearman correlation test in boys. a. serum INHB and FSH levels; b. serum INHB and LH levels; c. serum INHB and E2 levels\u003cem\u003e (P=\u003c/em\u003e0.29\u003cem\u003e)\u003c/em\u003e; d. serum INHB and PROG levels(\u003cem\u003eP\u003c/em\u003e\u0026lt;0.05);e.serum INHB and PRL levels.\u003c/p\u003e","description":"","filename":"floatimage4.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-4244728/v1/71bd9a0a5eccf095426e2926.jpeg"},{"id":55693780,"identity":"50cfb5b7-5693-4ec3-9fa3-dca5da49b596","added_by":"auto","created_at":"2024-05-02 00:34:14","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":775358,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4244728/v1/d35a0ccb-9a6a-40f1-bad2-cb80c97cc77c.pdf"},{"id":55634133,"identity":"7d784735-9ad5-4e65-864d-1b70ec78ccdd","added_by":"auto","created_at":"2024-04-30 20:09:43","extension":"docx","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":22398,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementarymaterials.docx","url":"https://assets-eu.researchsquare.com/files/rs-4244728/v1/0a7aeb75af4c2fbd13b260b8.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Age- and sex-dependent reference intervals of inhibin B in Chinese children on chemiluminescence analyzer","fulltext":[{"header":"Introduction","content":"\u003cp\u003eInhibin B\u0026nbsp;(INHB), as a gonadal glycoprotein hormone consisting of alpha and beta \u0026beta; subunit, belongs to transforming growth factor-\u0026beta; superfamily. It is mainly secreted by developing follicular granulosa cells for female and by testicular sertoli cells for male, which plays an important role in the female ovarian and\u0026nbsp;male\u0026nbsp;testicular assessment\u003csup\u003e1-4\u003c/sup\u003e. INHB levels vary greatly from birth to adolescence until steady concentrations in adult\u003csup\u003e5\u003c/sup\u003e, with a peak in the first month of life as activation of hypothalamic-pituitary-gonadal. The increasing INHB will promote the functional follicle maturation, and then there is a trough at 5-10 years. In prepuberty, its levels are independent of the presence of actively proliferating germ cells and rise under the stimulation of FSH\u003csup\u003e6\u003c/sup\u003e. During puberty there is a regular increase of INHB\u003csup\u003e4\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eOver the last decade, INHB has received increasing attention of the pediatric endocrinologist. INHB determination is an essential tool in the assessment of pubertal disorders and the function of Sertoli cells\u003csup\u003e7\u003c/sup\u003e. For example, INHB were characterized its clinical usefulness to evaluate gonadal function in children post-treatment for cancer \u003csup\u003e8, 9\u003c/sup\u003e. Raman reported that INHB was as marker of sertoli cell function in children subjected to surgical correction for undescended tests\u003csup\u003e1\u003c/sup\u003e. Furthermore, INHB concentration/ratio to sex hormones is regarded as a newly predictor of age at first menstruation in girls with idiopathic central precocious puberty\u003csup\u003e10\u003c/sup\u003e, and potential markers of progressive central precocious puberty\u003csup\u003e11\u003c/sup\u003e and Bilateral Cryptorchidism in infants\u003csup\u003e12\u003c/sup\u003e. So, inhibin B is very extensive in clinical application.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIt is generally believed that accurate interpretation of test results, proper diagnostics and treatment rely on accurate reference ranges. Inappropriate pediatric references intervals (RIs) may impact clinical decisions. However, RIs of serum INHB have not been required due to the effects of age, sex, methodologies and difficulties in collecting ample serum from healthy children especially neonates.\u0026nbsp;A few reports have investigated RIs of INHB based on enzyme-linked immunosorbent assays (ELISA), but they are the rough reference ranges from a small amount of children in European and western countries\u003csup\u003e5, 6, 13\u003c/sup\u003e or are acquired from the specific enrolled subjects such as infertile couples\u003csup\u003e14\u003c/sup\u003e. There little is known about RIs of INHB in Chinese pediatric individuals, which hinders pediatric clinical use as a potential diagnostic marker. Therefore, it is necessary to establish normal RIs for serum INHB in heathy Chinese children with considerations of the effect of age and gender.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eKAESER 1000\u0026nbsp;is fully automatized chemiluminescence analyzer for measurement of INHB and sex hormones. The platform is developed recently that is more fast and stable than the manual method. Compared to the commercial ELISA, KAESER 1000 have improved in sensitivity and specificity.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTherefore, the first objective of this study was to setup the RIs of INHB for healthy children aged from 0 to 18 years in\u0026nbsp;China based on KAESER 1000 chemiluminescence technology. The secondary aim was to investigate the effects of age and gender on INHB levels to provide insights about the INHB application in the reproductive health. Further, it was also aimed to assess the\u0026nbsp;associations\u0026nbsp;of INHB with conventional markers of gonadal function,\u0026nbsp;including follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (TT), prolactin (PRL), progesterone (PROG) and total estradiol (E2).\u003c/p\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec2\" class=\"Section2\"\u003e \u003ch2\u003eParticipant characteristics\u003c/h2\u003e \u003cp\u003eThe overall population comprised 302 girls and 320 boys, with mean (\u0026plusmn;\u0026thinsp;SD) age of 5.9 y (\u0026plusmn;\u0026thinsp;4.8) and BMI of 17.78 kg/m\u003csup\u003e2\u003c/sup\u003e (\u0026plusmn;\u0026thinsp;3.51). There was no significant difference in age between the boys and girls (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.911). Weight and BMI for boys were higher than those for girls (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.043, \u003cem\u003eP\u0026thinsp;=\u003c/em\u003e\u0026thinsp;0.002) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Compared to the boys, girls had lower INHB and E2 levels among four age subgroups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001), but had higher FSH and PRL levels (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Moreover, changes in serum LH, PROG and E2 levels were negligible among the first three subgroups (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\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\u003eCharacteristics of children in four age groups [mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD]\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"10\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e \u003cp\u003eGirls\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c9\" namest=\"c6\"\u003e \u003cp\u003eBoys\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"1\" nameend=\"c10\" namest=\"c10\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003egroups\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eheight\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eweight\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eBMI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eheight\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eweight\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eBMI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"1\" nameend=\"c10\" namest=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;1y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e63.8\u0026thinsp;\u0026plusmn;\u0026thinsp;7.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e7.1\u0026thinsp;\u0026plusmn;\u0026thinsp;2.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e16.8\u0026thinsp;\u0026plusmn;\u0026thinsp;3.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e81\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e66.9\u0026thinsp;\u0026plusmn;\u0026thinsp;7.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e8.4\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e18.4\u0026thinsp;\u0026plusmn;\u0026thinsp;4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"1\" nameend=\"c10\" namest=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u0026thinsp;~\u0026thinsp;5y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e95.9\u0026thinsp;\u0026plusmn;\u0026thinsp;11.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e15.2\u0026thinsp;\u0026plusmn;\u0026thinsp;3.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e16.5\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e97.2\u0026thinsp;\u0026plusmn;\u0026thinsp;10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e15.7\u0026thinsp;\u0026plusmn;\u0026thinsp;3.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e16.6\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"1\" nameend=\"c10\" namest=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u0026thinsp;~\u0026thinsp;10y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e125.6\u0026thinsp;\u0026plusmn;\u0026thinsp;11.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e27.1\u0026thinsp;\u0026plusmn;\u0026thinsp;6.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e17.1\u0026thinsp;\u0026plusmn;\u0026thinsp;2.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e130.0\u0026thinsp;\u0026plusmn;\u0026thinsp;12.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e31.4\u0026thinsp;\u0026plusmn;\u0026thinsp;10.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e17.9\u0026thinsp;\u0026plusmn;\u0026thinsp;3.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"1\" nameend=\"c10\" namest=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u0026thinsp;~\u0026thinsp;18y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e154.6\u0026thinsp;\u0026plusmn;\u0026thinsp;9.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e43.9\u0026thinsp;\u0026plusmn;\u0026thinsp;10.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e18.2\u0026thinsp;\u0026plusmn;\u0026thinsp;3.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e153.4\u0026thinsp;\u0026plusmn;\u0026thinsp;17.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e46.4\u0026thinsp;\u0026plusmn;\u0026thinsp;15.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e19.3\u0026thinsp;\u0026plusmn;\u0026thinsp;4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"1\" nameend=\"c10\" namest=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003etotal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e302\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e112.7\u0026thinsp;\u0026plusmn;\u0026thinsp;37.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e24.6\u0026thinsp;\u0026plusmn;\u0026thinsp;16.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e17.3\u0026thinsp;\u0026plusmn;\u0026thinsp;2.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e320\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e112.9\u0026thinsp;\u0026plusmn;\u0026thinsp;37.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e26.4\u0026thinsp;\u0026plusmn;\u0026thinsp;18.6\u003cem\u003e*\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e18.2\u0026thinsp;\u0026plusmn;\u0026thinsp;4.0\u003cem\u003e*\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"1\" nameend=\"c10\" namest=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"10\" nameend=\"c10\" namest=\"c1\"\u003e \u003cp\u003eBMI, body mass index; *\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05, independent samples test between boys and girls\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 \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 estimated age-and gender-specific reference intervals for INHB and six sex hormones in four age groups [median (2.5\u0026ndash;97.5 percentiles)/ mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD]\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003egroups\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eINHB (pg/mL)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eFSH (mIU/mL)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eLH (mIU/mL)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003ePRL(ng/mL)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eProg(ng/mL)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eE2(pg/mL)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003egirls\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;1y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e67 (8, 206) \u003cem\u003e*\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.5 (0.7, 20.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.45 (0.2, 2.01)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e18.0 (3.4, 109.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.3 (0.1, 5.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.2 (3.2, 40.1) \u003csup\u003e\u0026dagger;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1\u0026thinsp;~\u0026thinsp;5y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e33 (14, 227) \u003cem\u003e*\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.3 (0.7, 18.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.45 (0.45, 0.45)\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e11.8 (3.8, 58.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.3 (0.1, 1.6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.2 (3.2, 20.9) \u003csup\u003e\u0026dagger;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5\u0026thinsp;~\u0026thinsp;10y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e27 (8, 92) \u003cem\u003e*\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.7 (0.3, 23.6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.34 (0.45, 2.40)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e8.8 (3.7, 33.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.3 (0.2, 1.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.2 (3.2, 33.8) \u003csup\u003e\u0026dagger;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10\u0026thinsp;~\u0026thinsp;18y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e81 (12, 219) \u003cem\u003e*\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.0 (1.2, 11.4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.45 (3.69, 27.01)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e10.7 (3.6, 31.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.3 (0.1, 5.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.2 (41.0, 174.9)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003etotal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e46 (12, 182) \u003csup\u003e\u0026Dagger;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.5 (0.5, 13.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.45 (0.24, 14.33)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e12.0 (3.8, 60.9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.3 (0.1, 3.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e13.0 (3.2, 96.0)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eboys\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;1y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e236 (75, 476)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.8 (0.2, 4.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.5 (0.2, 4.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e16.6 (3.9, 117.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.3 (0.1, 19.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e12.0 (3.2, 52.1)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1\u0026thinsp;~\u0026thinsp;5y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e148 (62, 355)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.8 (0.2, 12.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.45 (0.45, 0.45)\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e9.9 (4.1, 38.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.3 (0.2, 2.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e10.5 (3.2, 35.7)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5\u0026thinsp;~\u0026thinsp;10y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e102 (53, 265)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.0 (0.12, 16.8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.45 (0.27, 2.14)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e8.21 (3.16, 46.26)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.3 (0.2, 2.6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e11.0 (3.2, 26.9)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10\u0026thinsp;~\u0026thinsp;18y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e156 (54, 290)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.6 (0.31, 8.6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.01 (0.31, 5.39)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e10.94 (3.51, 32.00)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.2 (0.1, 1.1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e22.0 (10.0, 74.9)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003etotal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e159 (55, 418)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.2 (0.19, 8.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.45 (0.23, 4.04)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e11.4 (3.9, 62.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.3 (0.1, 3.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e12.0 (3.2, 40.5)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"8\" nameend=\"c8\" namest=\"c1\"\u003e \u003cp\u003e\u003csup\u003e\u0026dagger;\u003c/sup\u003elow detection rate; *\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001, independent samples test of INHB level in corresponding age subgroups between boys and girls. \u003csup\u003e\u003cem\u003e\u0026Dagger;\u003c/em\u003e\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001, independent samples test of INHB level between sex. FSH, follicle-stimulating hormone; LH, luteinizing hormone; TT, testosterone, PRL prolactin, Prog progesterone and E2, total estradiol.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eDistribution of INHB\u003c/h2\u003e \u003cp\u003eThe distribution of INHB in girls and boys is shown comparatively in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003e Children in both sexes had the high median INHB concentrations within the first years old and declined sharply from 1-year-old to 5-year-old, then increased between 10 and 18 years old. Significant differences in INHB levels were observed among the multiple age subgroups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026le;\u0026thinsp;0.001). For girls highest INHB levels is the first age group, but for boys is the oldest age group. In addition, compared to the boys, girls had reduced variability in INHB across the four age groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003eA and B)\u003c/p\u003e \u003cp\u003eThe effect of age on INHB was also assessed using five models including linear, quadratic, cubic, logarithmic, and S (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Linear, quadratic, cubic for all population, logarithmic only for boys were effective (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;2.36\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;14\u003c/sup\u003e, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;2.00\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;32\u003c/sup\u003e, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;1.37\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;39\u003c/sup\u003e). The optimal models in girls and boys all were cubic models (R\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;0.198 for girls and R\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;0.440 for boys, Supplementary Table S2).\u003c/p\u003e \u003cp\u003e \u003cem\u003eReference intervals\u003c/em\u003e \u003c/p\u003e \u003cp\u003eDue to the volatility of INHB concentration along with age and sex, so the RIs for INHB and six sex hormones including the 2.5th and 97.5th quantiles along with the median in boys and girls are shown in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e by age. The median (2.5%~97.5%) INHB level from the oldest age group in girls was the highest in comparison to other age groups. However, the median level of INHB from the first age group in boys was the highest.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eCorrelations between the INHB level and FSH, LH, PRL, PROG and E2\u003c/h2\u003e \u003cp\u003eBecause the detection rate of serum TT in both girls and boys were too low, the correlation analysis with INHB was exc1uded. In Spearman\u0026rsquo;s correlation analysis (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e3\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e4\u003c/span\u003e), INHB was weakly positively associated with LH in both girls (Rs\u0026thinsp;=\u0026thinsp;0.321, \u003cem\u003eP\u0026thinsp;=\u003c/em\u003e\u0026thinsp;1.20\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;7\u003c/sup\u003e) and boys (Rs\u0026thinsp;=\u0026thinsp;0.225, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;2.85\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;4\u003c/sup\u003e), and positively associated with FSH (Rs\u0026thinsp;=\u0026thinsp;0.403, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;1.61\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;11\u003c/sup\u003e) and E2 (Rs\u0026thinsp;=\u0026thinsp;0.359, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;2.52\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;9\u003c/sup\u003e), but negatively associated with PROG (Rs=-0.136, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.028) only in girls. There was a positive correlation between INHB and PRL only in boys (Rs\u0026thinsp;=\u0026thinsp;0.300, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;8.91\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;7\u003c/sup\u003e).\u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe normal values for serum INHB have been often assessed among adults, especially fertile and infertile adults\u003csup\u003e\u003cspan additionalcitationids=\"CR15\" citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e–\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e. However, few study has established sex- and age-related INHB reference data from birth to adulthood (n = 366, boys)\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e, aged 6-11years (n = 705, girls)\u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e and during childhood and puberty (n = 52, girls)\u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e in European and western countries. Due to the practical difficult in infant’s recruitment, some studies may contain few infants or participants aged range of 5 ~ 18 years old. In the current study, 163 infants (\u0026lt; 1 y subgroup) were regarded as a key strength of our study. To the best of our knowledge, this is the first one to detect serum INHB concentrations in heathy children aged from 0 to 18 years among a large group of more than 600 populations in China. In this study, we investigated the age- and gender-related effects on INHB concentrations and calculated the pediatric RIs from birth to puberty based on a newly introduced fully automatic chemiluminescence analyzer.\u003c/p\u003e \u003cp\u003eOur results suggested that gender and age had effects on INHB levels. The level of INHB for boys and girls was similar U-shaped along with age and the optimal estimated models both were cubic. The distribution of INHB levels as four age groups also confirmed the shaped phenomenon. Our finding suggested that INHB was highly variable during the first year of life and attained gradually stability as the age, then increase to adolescence. INHB for girls was high from born, and decreased gradually to reach a trough at 2 ~ 8 years old, and then increased again to reach a crest around 13 years old. However, there were differences for boys, with step to plateau level around 5.5 ~ 9 years old and a crest around 13 years old, which were similar trends with previous study\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e. Increase of INHB at infants and pubertal period were involved the Sertoli cell proliferation\u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e, which in males within one year old can increase fivefold\u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eWe found the post-natal peak exceeded the level of INHB in pubertal period in boys. Andersson et al also observed that the peak in infant boys exceeded in adult men \u003csup\u003e\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e. High level of infancy affords an early opportunity to assess hypothalamic-pituitary-gonadal axis function, which is urgently needed, especially, when newborn happens ambiguous genitalia or suspicion of hypopituitarism or hypogonadism\u003csup\u003e\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e. Postnatal increase of INHB also may predict later testicular function, such as spermatogenesis, but abundant studies are requested to evaluate its effectiveness\u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e. Our study also showed that INHB raised from age 8 for girl and 9 for boys, which seem to be early than the clinical onset of puberty with mean age of 9 years in girls and 11.5 years in boys, respectively. This demonstrates that INHB secretion takes places in the prepubertal testis, despite the very low levels of both gonadotrophins and TT. Therefore, INHB is used to assess Sertoli cell health and number in pre-pubertal stage.\u003c/p\u003e \u003cp\u003eInterestingly, our finding showed that girls not only had lower concentrations but also reduced variability in INHB across four age groups compared to the boys. These results were in agreement with previous studies at all stages of puberty\u003csup\u003e\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e and in infancy\u003csup\u003e\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e. Besides INHB, anti-Müllerian hormone is also expressed at varying degrees in children\u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e. However, no published study investigated detailed the difference variability of INHB for girls and boys with different age groups in healthy pediatric populations.\u003c/p\u003e \u003cp\u003eThe INHB reference ranges have changed over the years in adults among different laboratories\u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e. Further, previous reports showed that INHB level was U-shaped with age from birth to adulthood\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e. So, this prompted us to reconsider the INHB cutoff value in childhood and adolescence. Because INHB concentrations in different sex and age individuals were significant difference, the age-specific and sex-specific RIs were calculated separately.\u003c/p\u003e \u003cp\u003eOur RIs for INHB [236(75–476) pg/ml for \u0026lt; 1y; 102(53, 265) pg/ml for 5 ~ 10y] was close to the ranges observed in children of the same age range in Ireland [278(68–630) pg/mL for \u0026lt; 1y, n = 51; 84 (35–167) for 6 ~ 10y, n = 80]\u003csup\u003e5\u003c/sup\u003e. However, our RIs for INHB at 10 ~ 18 group [156 (54, 290)] pg/mL was different from the 12 ~ 17 y group [280 (74 ~ 470) pg/mL in Ireland\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e. The remarkable difference of RIs may result from study populations, the establish criteria of reference range, and detection methodologies\u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eOur study indicated that INHB levels were correlated positively with FSH and LH in girls, which were consistent with previous reports in prepubertal, pubertal girls \u003csup\u003e\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e. Other studies also showed no association between FSH and INHB in prepubertal cryptorchidic boys (0.5 to 13 year)\u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e and normal prepubertal boys\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e. Our study showed that serum INHB moderately correlated positively with E2 (\u003cem\u003eP\u003c/em\u003e = 2.52×10\u003csup\u003e− 9\u003c/sup\u003e) and negatively with PROG (\u003cem\u003eP\u003c/em\u003e = 0.028) in girls. INHB correlated positively with PRL (\u003cem\u003eP\u003c/em\u003e = 8.91×10\u003csup\u003e− 7\u003c/sup\u003e) levels in boys. However, we did not find any reports on correlations between INHB and E2, PROG, and PRL in children.\u003c/p\u003e \u003cp\u003eThere are several limitations in our study. Firstly, although the inclusion criterion were standardized to select possibly those healthy individuals, it is possible to include pathologic subjects. Secondly, due to certain practical limitations, the participants in adolescence were not identify as Tanner staging and menstrual cycle. Finally, we do not evaluate INHB level in repeated measures.\u003c/p\u003e \u003cp\u003eIn conclusion, our study administrated the age- and sex-dependent RIs for serum INHB. Our results suggested that INHB level was lower and reduced variability for girls across four age groups compared to boys. INHB concentration changed along with age. Furthermore, significant difference for correlations between INHB and FSH, LH, E2, PROG, PRL were observed in different sex. Our results may broaden the application of INHB levels, which may be sensitively used as an effective indicator of gonad function assessment and distinguish the patients and normal children.\u003c/p\u003e"},{"header":"Methods","content":"\u003ch2\u003eStudy populations\u003c/h2\u003e\u003cp\u003eThe study population included boys and girls aged 0 ~ 18 years. Samples were acquired from our routine laboratory at Eugenic genetics laboratory, Wuhan Children’s Hospital between January and December 2022. The subject who had the following conditions were excluded: 1) reproductive diseases such as cryptorchidism, inguinal hernia, varicocele, testicular trauma, hermaphroditism, precocious puberty, sexual development delay, and adolescent polycystic ovary syndrome; 2) chronic diseases such as tumor and tuberculosis; 3) endocrine diseases such as diabetes, hyperthyroidism, and obesity; 4) severe liver and kidney damage; 5) serious trauma, infection, and sepsis; 6) neonatal respiratory distress syndrome, jaundice, and shock; 7) use of chemotherapy drugs. After exclusion of the above mentioned criteria, a total of 622 subjects were included in the current study. Among them, 518 children provided sufficient volume of serum samples for other hormone measurements. The baseline characteristics on age, gender, height and weight were collected from the hospital’s medical records. All study participants provided an informed consent, and the present study was approved by the Ethics Committee of Wuhan Children’s Hospital (No.2022R108-E01).\u003c/p\u003e\u003ch3\u003eHormone measurements\u003c/h3\u003e\u003cp\u003eBlood samples were drawn from an antecubital vein and centrifuged after clotting. Serum was stored at -20 ℃ until analysis. Serum INHB was measured in double-antibody chemiluminescence immunoassay (Kangran Biotech, Guangzhou, China). The detection of limit (LOD) for INHB assay was 7 pg/mL, and the intra and interassay coefficients of variation were 6.25%~8.33% and 4.34%~9.73%, respectively. Serum INHB, FSH, LH, PRL, PROG, E2 and TT levels were determined using the Kaeser 1000 automatic chemiluminescence immune analyzer. The parameters for other serum hormones were described in supplementary table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e.\u003c/p\u003e\u003ch2\u003eStatistical analysis\u003c/h2\u003e\u003cp\u003eAll statistical analyses were performed with SPSS18.0. \u003cem\u003eP\u003c/em\u003e-value less than 0.05 was defined as statistically significant. Independent samples t-test were used to reveal significant differences in height, weight, and BMI between girls and boys. The concentrations of INHB and sex hormones below their LODs were replaced with the LODs divided by the square root of 2. The serum INHB concentrations were stratified into four subgroups by age (\u0026lt; 1y, 1 ~ 5y, 5 ~ 10y, 10 ~ 18y) and into two subgroups by gender (girls and boys). Serum INHB concentrations were the skewed distribution, and thus were transformed by natural logarithm (ln) to meet the normality for the following analysis.\u003c/p\u003e\u003cp\u003eThe differences of INHB levels among four age groups stratified by gender were compared by Bonferroni multiple comparison test, which was used to control type 1 error multiple comparison. The RIs of INHB at 2.5th, and 97.5th quantiles were expressed for each age group partitioned by gender. Multiple linear and non-linear mathematical models were then performed in order to describe the variation of INHB along with age. The models of curve estimation included linear, quadratic, cubic, logarithmic, and S. Relationships between INHB and other hormone levels were analyzed using Spearman correlation test.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics declarations\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe present study was approved by the Ethics Committee of Wuhan Children\u0026rsquo;s Hospital (No.2022R108-E01). \u0026nbsp;All procedures were in accordance with the ethical standards of the institution and with the 1964 Helsinki declaration and its later amendments. All study participants provided an informed consent.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets use and/or analysed during the current study are provided by the corresponding author on a reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowlegements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe would like to thank participants without whom this work would not be possible.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBin Zhou: Investigation, Methodology, Writing-original draft; Jin jun Luo and Xiang Dai: Validation, Writing review \u0026amp; editing; Xuelian He and Hui Yao: Writing review \u0026amp; editing. All authors have accepted responsibility for the entire content of this manuscript and approved its submission.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResearch funding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research did not receive any funding from public agencies, commercial agencies, or not-for-profit sectors. The study was funded by Wuhan Children\u0026rsquo;s Hospital.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe author(s) declare no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eRaman, V. S., Khanna, S. \u0026amp; Verma, V. A prospective observational study to evaluate the change in inhibin-B as a marker of sertoli cell function in children subjected to surgical correction for undescended testes. Afr J Paediatr Surg 19, 4233\u0026ndash;4237(2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLi, Y., Nie, M. Y., Liu, Y., Zhang, W. Y., Yang, X. K. The dynamic changes of anti-Mullerian hormone and inhibin B during controlled ovarian hyperstimulation in decreased ovarian reserve women and the effect on clinical outcome. Gynecol endocrino l 6, 450\u0026ndash;453(2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHansen, A. H. \u003cem\u003eet al\u003c/em\u003e. Testicular torsion and subsequent testicular function in young men from the general population. Hum Reprod 38, 216\u0026ndash;224(2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRadicioni, A. F. \u003cem\u003eet al.\u003c/em\u003e Changes in serum inhibin B during normal male puberty. Eur J Endocrinol 152, 403\u0026ndash;409(2005).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCrofton, P. M. \u003cem\u003eet al\u003c/em\u003e. Inhibin B in boys from birth to adulthood: relationship with age, pubertal stage, FSH and testosterone. Clin Endocrinol (Oxf) 56, 215\u0026ndash;221(2002).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSims, E. K. \u003cem\u003eet al\u003c/em\u003e. Inhibin B and luteinizing hormone levels in girls aged 6\u0026ndash;11 years from NHANES III, 1988\u0026ndash;1994. Clin Endocrinol (Oxf) 77, 555\u0026ndash;563(2012).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNajiba Lahlou, M. D. Marc Roger, M. D. Inhibin B in Pubertal Development and Pubertal Disorders. Semin Reprod Med 22, 165\u0026ndash;175(2004).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVan der Kooi, A. L. F. \u003cem\u003eet al\u003c/em\u003e. Changes in Anti-Mullerian Hormone and Inhibin B in Children Treated for Cancer. J Adolesc Young Adult Oncol 8, 281\u0026ndash;290(2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDrechsel, K. C. E. \u003cem\u003eet al.\u003c/em\u003e Reproductive ability in survivors of childhood, adolescent, and young adult Hodgkin lymphoma: a review. Hum Reprod Update 29, 486\u0026ndash;517(2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTencer, J., Lemaire, P. \u0026amp; Brailly-Tabard, S. Serum inhibin B concentration as a predictor of age at first menstruation in girls with idiopathic central precocious puberty. PLoS One 13(2018)e0205810.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVuralli, D., Ciftci, N. \u0026amp; Demirbilek, H. Serum kisspeptin, neurokinin B and inhibin B levels can be used as alternative parameters to distinguish idiopathic CPP from premature thelarche in the early stages of puberty. Clin Endocrinol (Oxf). 98, 788\u0026ndash;795(2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKato, T. \u003cem\u003eet al\u003c/em\u003e. Low Serum Inhibin B/Follicle-Stimulating Hormones and Anti-M\u0026uuml;llerian Hormone/Follicle-Stimulating Hormones Ratios as Markers of Decreased Germ Cells in Infants with Bilateral Cryptorchidism. J Urol. 207, 701\u0026ndash;709(2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChada, M. \u003cem\u003eet al\u003c/em\u003e. Inhibin B, Follicle Stimulating Hormone, Luteinizing Hormone, and Estradiol and Their Relationship to the Regulation of Follicle Development in Girls during Childhood And Puberty. Physiol Res 52, 341\u0026ndash;346(2003).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMyers, G. M., Lambert-Messerlian, G. M. \u0026amp; Sigman. M. Inhibin B reference data for fertile and infertile men in Northeast America. Fertil Steril 92, 1920\u0026ndash;1923(2009).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAndersson, A. M. \u003cem\u003eet al\u003c/em\u003e. Serum inhibin B and follicle- stimulating hormone levels as tools in the evaluation of infertile men: significance of adequate reference values from proven fertile men. J Clin Endocrinol Metab 89, 2873\u0026ndash;2879(2004).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGrunewald, S. \u003cem\u003eet al\u003c/em\u003e. Age-dependent inhibin B concentration in relation to FSH and semen sample qualities: a study in 2448 men. Reproduction 145, 237\u0026ndash;244(2013).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCortes, D., M\u0026uuml;ller, J. \u0026amp; Skakkebaek. N. E. Proliferation of Sertoli cells during development of the human testis assessed by stereological methods. Int J Androl 10, 589\u0026ndash;596(1987).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYou, X. \u003cem\u003eet al\u003c/em\u003e. Common markers of testicular Sertoli cells. Expert Rev Mol Diagn 21, 613\u0026ndash;626(2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAndersson, A. M. \u003cem\u003eet al\u003c/em\u003e. Longitudinal Reproductive Hormone Profiles in Infants: Peak of Inhibin B Levels in Infant Boys Exceeds Levels in Adult Men. J Cli Endocrinol Metab 83, 675\u0026ndash;681(1998).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKuiri-Hanninen, T., Dunkel, L., \u0026amp; Sankilampi. U. Sexual dimorphism in postnatal gonadotrophin levels in infancy reflects diverse maturation of the ovarian and testicular hormone synthesis. Clin Endocrinol (Oxf) 89, 85\u0026ndash;92(2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKelsey, T. \u003cem\u003eet al\u003c/em\u003e. A normative model of Inhibin B in young males. Plos one 11(2016) e0153843.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSehested, A. \u003cem\u003eet al\u003c/em\u003e. Serum Inhibin A and Inhibin B in Healthy Prepubertal, Pubertal, and Adolescent Girls and Adult Women: Relation to Age, Stage of Puberty, Menstrual Cycle, Follicle-Stimulating Hormone, Luteinizing Hormone, and Estradiol Levels. J Clin Endocrinol Metab 85, 1634\u0026ndash;1640(2000).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang, J. \u003cem\u003eet al\u003c/em\u003e. Age-specific reference intervals for anti-Mullerian hormone in Chinese boys: A population-based study. Clin Chim Acta 506, 154\u0026ndash;159(2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang, J. \u003cem\u003eet al\u003c/em\u003e. Reference range and cutoff value of serum inhibin B to predict successful sperm retrieval: A cross-sectional study of 30613 Chinese men. Clin Endocrinol (Oxf) 92, 232\u0026ndash;240(2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCortes, D. \u003cem\u003eet al\u003c/em\u003e. The Sertoli cell hormones inhibin-B and anti Mullerian hormone have different patterns of secretion in prepubertal cryptorchid boys. J Pediatr Surg 51, 475\u0026ndash;480, (2016).\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":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Inhibin B, reference intervals, children, age, sexual function","lastPublishedDoi":"10.21203/rs.3.rs-4244728/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4244728/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackgroud\u003c/strong\u003e: Inhibin B (INHB) is an emerging biomarker of sexual function diseases, but its clinical use in children is hindered due to the lacking reference intervals (RIs) based on age sex.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e: A total of 622 healthy Chinese children were included and serum INHB and six conventional sex hormones were measured. INHB was partitioned by four age groups (\u0026lt;1y, 1~5y, 5~10y, 10~18y) and the corresponding RIs were determined with quantile. The effect of age on INHB was assessed using five models including linear, quadratic, cubic, S, and logarithmic. Spearman correlation was used to examine the associations between INHB and sex hormones.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e: Compared to the boys, girls had lower median concentrations and variability in INHB across four age groups. The optimal models for the effect of age on INHB were cubic (R\u003csup\u003e2\u003c/sup\u003e=0.440 for boys). The INHB concentration in the first and oldest age groups was more high than other groups (\u003cem\u003eP\u003c/em\u003e≤0.001). Spearman’s correlation analysis showed sex-dependent association between INHB and conventional sex hormones (e.g. a positive association with PRL in boys).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions\u003c/strong\u003e: INHB level is influenced by age and gender, and our results provide specific RIs to facilitate to screen sexual function diseases in Chinese children.\u003c/p\u003e","manuscriptTitle":"Age- and sex-dependent reference intervals of inhibin B in Chinese children on chemiluminescence analyzer","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-04-30 20:09:39","doi":"10.21203/rs.3.rs-4244728/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-07-19T05:47:47+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-07-18T10:40:54+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"192808992836433520832242663377990615880","date":"2024-07-15T06:33:54+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-06-05T14:42:22+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"141226712253606969749921014828558920213","date":"2024-06-01T15:10:01+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-05-01T13:14:37+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-04-28T19:27:58+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2024-04-27T17:59:43+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-04-25T03:29:36+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2024-04-10T03:01:28+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"8cd0e74b-9122-4aac-811a-408e75b8a966","owner":[],"postedDate":"April 30th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":31316851,"name":"Biological sciences/Biochemistry/Hormones/Gonadal hormones"},{"id":31316852,"name":"Health sciences/Diseases/Reproductive disorders"}],"tags":[],"updatedAt":"2024-10-07T03:53:21+00:00","versionOfRecord":[],"versionCreatedAt":"2024-04-30 20:09:39","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4244728","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4244728","identity":"rs-4244728","version":["v1"]},"buildId":"rHA-KDH7Qsr4HCuvH75dn","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}