Use of Anti-Mullerian Hormone (AMH) for Testing of Ovarian Reserve: A Survey of Fifteen (15) Fertility Centres in Ghana

Anti-Mullerian Hormone (AMH) is a dimeric glycoprotein with a molecular weight of 140 kD, encoded by a gene on the short arm of chromosome and a member of the transforming growth factor-beta (TGF-β) superfamily. The expression of AMH is markedly different in males and females, both in concentration and temporality. In males, Sertoli cells maintain a high concentration of AMH in utero which peaks shortly after birth and then drops precipitously at puberty. In females, granulosa cells produce very low levels of AMH in utero followed by a transient spike in the neonatal period. Concentrations of the hormone then rise steadily through adolescence to a peak in the mid-twenties and subsequently decline until becoming undetectable in menopause. The study aimed to understand how Clinicians and Clinical Embryologists used anti-mullerian hormone (AMH) test to assess ovarian reserve, direct patient selection and treatment regimens and guide in vitro fertilization (IVF) cycle management in all registered fertility hospitals in a West African country, Ghana. A web-based survey (questionnaire) using google forms was performed to solicit responses from all IVF hospitals that are registered with the Fertility Society of Ghana (FERSOG). This questionnaire consisted of fifteen (15) broader questions, ten (10) of which assessed the clinics’ use of AMH. Responses were screened for quality to verify that only one (1) survey was completed by each IVF centre. The study was conducted during May and June 2020 at the In Vitro Fertilization (IVF) Department of the Airport Women’s Hospital (AWH) in Accra, Ghana. Results are reported as the proportion of IVF cycles represented by a particular answer choice. Survey responses were completed from 15 IVF centres, representing 2504 IVF cycles performed annually. A good majority (73.3%) [1835 IVF cycles] of the respondent IVF hospitals reported to use AMH as a first line test and 93.3% reported it as the best test for evaluating ovarian reserve. Another 66.7% reported that AMH results were extremely relevant to clinical practice. However, in contrast, for predicting live birth rate, 60% reported age as the best predictor in their practice. Overall, our results indicate that AMH is considered a first line test for assessing ovarian reserve and is relevant to the clinical practice of majority of Assisted Reproductive Technologies (ART) providers in Ghana.

Clinical Value of Serum Anti-Mullerian Hormone and Inhibin B in Prediction of Ovarian Response in Patients with Polycystic Ovary Syndrome

The present study aimed to investigate the clinical value of serum anti-mullerian hormone（AMH） and inhibin B（INHB） in predicting the ovarian response of patients with polycystic ovary syndrome（PCOS）. A total of 120 PCOS patients were enrolled and divided into three groups in terms of the ovarian response： a low-response group（n=36）, a normal-response group（n=44）, and a high-response group（n=40）. The serum AMH and INHB levels were measured by enzyme-linked immunosorbent assay（ELISA）. The follicle stimulating hormone（FSH）, luteinizing hormone（LH）, and estradiol（E2） levels were determined by chemiluminescence microparticle immunoassay. The correlation of the serum AMH and INHB levels with other indicators was analyzed. A receiver operating characteristic（ROC） curve was established to analyze the prediction of ovarian response by AMH and INHB. The results showed that there were significant differences in age, body mass index（BMI）, FSH, total gonadotropin-releasing hormone（Gn RH）, LH, E2, and antral follicle counts（AFCs） between the groups（P〈0.05）. The serum AMH and INHB levels were increased significantly with the ovarian response of PCOS patients increasing（P〈0.05）. The serum AMH and INHB levels were negatively correlated with the age, BMI, FSH level, Gn, and E2 levels（P〈0.05）. They were positively correlated with the LH levels and AFCs（P〈0.05）. ROC curve analysis of serum AMH and INHB in prediction of a low ovarian response showed that the area under the ROC curve（AUC） value of the serum AMH level was 0.817, with a cut-off value of 1.29 ng/m L. The sensitivity and specificity were 71.2% and 79.6%, respectively. The AUC value of serum INHB was 0.674, with a cut-off value of 38.65 ng/m L, and the sensitivity and specificity were 50.7% and 74.5%, respectively. ROC curve analysis showed when the serum AMH and INHB levels were used to predict a high ovarian response, the AUC value of the serum AMH level was 0.742, with a cut-off value of 2.84 ng/m L, and the sensitivity and specificity were 72.7% and 65.9%, respectively; the AUC value of the serum INHB level was 0.551 with a cut-off of 45.76 ng/m L, and the sensitivity and specificity were 76.3% and 40.2%, respectively. It was suggested the serum AMH and INHB levels have high clinical value in predicting the ovarian response of PCOS patients.

Investigating structural impact of a valine to isoleucine substitution on anti-Müllerian hormone in silico and genetic association of the variant and AMH expression with egg production in chickens

Anti-Mullerian hormone(AMH)acts in maintaining orderly cyclic recruitment of early follicles,suggesting that it is a promising can didate for in flue ncing an imal reproductive efficiency.This study aimed to elucidate the effect of a missense mutati on of Val566lle on the structure of AMH protein and the genetic association of Val566lle and AMH expression with egg production in chickens.Structural perturbations of Val566lle were predicted by homology modeling.The association of the variant with the number of eggs was tested using a quantitative trait transmission disequilibrium test model.AMH expression in granulosa cells in Lueyang black-boned chickens was compared with that in Nick chickens.The Vai566 of AMH is a non?conservative amino acid among mammals and birds,but its hydrophobicity is completely conservative.The substitution of Vai566 for lie566 pote ntially disrupted hydroge n bonds and solve nt accessibility of 22 residues and created a short a-helix in the C terminus of AMH.Despite having striking structure-disrupting potential,the variant was not statistically associated with the number of eggs(P>0.05)in the Lueyang black-boned chickens.We did not detect differential expression of AMH betwee n Lueya ng black-b oned chicke ns and Nick chickens(P>0.05).These results con firmed the structural impact of Val566lle,but suggested that Val566lle and AMH expression might not be the major genetic determinants for egg production in Lueyang black-boned chickens.

Anti-Mullerian Hormone and Follicle Counts as Predictors of Superovulatory Response and Embryo Production in Beef Cattle

This study evaluated Anti-Mullerian hormone (AMH) and/or follicle counts as predictors of subsequent superovulatory response and embryo production in 79 beef cows. Before superovulation, 3 to 5 mm follicles presented on the ovaries of donor cows were counted, and blood was collected for measure of serum AMH. Across cows, serum AMH ranged from 0.013 to 0.898 ng/mL, with a mean of 0.293 ng/mL. The distribution of both AMH concentrations and follicle counts were divided into quartiles (AMH Q1 through Q4, with Q1 the lowest and Q4 the highest) for analysis. Donor cows in AMH Q4 had a greater (P < 0.001) number of 3 to 5 mm follicles at the start of superovulation than did donors in either Q1 or Q2. At embryo collection, cows in AMH Q3 and 4 had more (P < 0.001) palpable corpora lutea (CL) than cows in AMH Q1. The mean number of embryos recovered from donor cows in AMH Q4 was greater (P < 0.001) than those recovered from cows in either AMH Q1 or 2, but similar to that of AMH Q3. Analysis indicated that AMH was positively correlated (P < 0.001) with mean follicles (r = 0.458), CL (r = 0.452) and embryos recovered (r = 0.430). Donor cows with higher follicle counts (F Q3 and 4) at the start of superovulation had more (P < 0.001) palpable CL at embryo collection than donor cows in F Q1 or 2. More (P < 0.001) embryos were recovered from cows with the highest follicle counts (F Q4) as compared with cows having lower (F Q1 and 2) follicle counts. Results confirm that relative AMH concentration was positively correlated with number of small antral follicles in the ovaries of cows;both AMH measure and antral follicle counts might be used to predict subsequent superovulatory response in beef cows.

Expression of anti-Mullerian hormone receptor on the appendix testis in connection with urological disorders

The female internal sex organs develop from the paramesonephric （Mullerian） duct. In male embryos, the regression of the Mullerian duct is caused by the anti-Mullerian hormone （AMH）, which plays an important role in the process of testicular descent. The physiological remnant of the Mullerian duct in males is the appendix testis （AT）. In our previous study, we presented evidence for the decreased incidence of AT in cryptorchidism with intraoperative surgery. In this report, the expression of the anti-Mullerian hormone receptor type 2 （AMHR2）, the specific receptor of AMH, on the AT was investigated in connection with different urological disorders, such as hernia inguinalis, torsion of AT, cysta epididymis, varicocele, hydrocele testis and various forms of undescended testis. The correlation between the age of the patients and the expression of the AMHR2 was also examined. Reverse transcriptase-polymerase chain reaction （RT-PCR） and immunohistochemistry were used to detect the receptor＇s mRNA and protein levels, respectively. We demonstrate that AMHR2 is expressed in the ATs. Additionally, the presence of this receptor was proven at the mRNA and protein levels. The expression pattern of the receptor correlated with neither the examined urological disorders nor the age of the patients; therefore, the function of the AT remains obscure.

Clinical efficacy of anti-Mullerian hormone inspection in supporting diagnosis for climacteric disorders

The diagnosis of climacteric disorders in gynecology is performed on the basis of symptoms rather than hormonal levels. Generally, the follicle-stimulating hormone (FSH) level increases when the 17β-estradiol (E2) level decreases in menopause. However, the problem of these hormones should be determined by strict timing of sampling, and there are individual specificities of decreases of hormone levels. We considered that anti-Mullerian hormone (AMH) can be measured at any time of the menstrual cycle and that it shows ovary functional decline earlier than FSH/E2, and we examined whether AMH would possibly become a good index for climacteric disorders. The subjects were 163 healthy females and 21 patients with climacteric disorders between 20 and 59 years old. We examined AMH, FSH and E2 at the same time. It is understood that in healthy females, AMH decreases with age and a decline in ovary function occurs at a relatively early age. Patients visiting clinics for climacteric disorders often have normal-range serum FSH/E2 levels, and it is clear that these values could not serve as indices of menopause at these inspections. Upon measurement of AMH in patients with climacteric disorders, most showed less than normal range (< 14 pmol/L), suggesting a decline in ovarian function. In addition, AMH was low in females with climacteric disorders compared with those without them. In conclusion, AMH was suggested as an objective index for climacteric disorders and possibly as a new diagnostic marker.

Anti-Mullerian Hormone: Establishing the Ovarian Reserve Range with Age in Rivers State Women, Niger Delta Region of Nigeria

Background: The measurement of circulating anti-Mullerian hormone (AMH) in the plasma of adult women has been used as a tool in the assessment of ovarian reserve. This is based on its ability to reflect the number of growing follicles in the ovaries and knowing the level of AMH in a woman’s blood is generally a good indicator of her ovarian reserve and this can be achieved by making reference to a decision values. Objective: The purpose of this study was to measure the level of this hormone in normal, apparently healthy subjects in Rivers State, Nigeria with respect to age. The percentage of women, at their reproductive age with infertility problems in the state is in the increase and the essence was to establish a reference range for the hormone because of its clinical application in women fertility. Materials/method: A total of one thousand and sixty two apparently healthy women divided into four age groups were recruited for this study;this comprised of 300 each in age group of 20 - 30, 31 - 40, 41 - 50 respectively and 162 in 51 - 60 years age group from May 2014 to June 2017. They were randomly selected after filling a questionnaire form that has the information of age, tribe, last date of menses and whether on medication for any infertility problems. Excluded from the study were subjects identified with different endocrine dysfunction and/or on drugs for different types of anovulatory dysfunction. The Enzyme linked Immunoassay method was used in the measurement of the AMH. Results: The result from the measurement of plasma AMH levels showed a Mean ± SD value of 3.193 ± 0.943, 1.644 ± 0.691, 0.516 ± 0.276 and 0.135 ± 0.081 ng/ml respectively for the 1st, 2nd, 3rd and 4th age groups respectively. The mean value for the AMH decreases with increasing age and was statistically significant at the different age group levels (P &minus;2.250 - 4.136;0.953 - 2.350;0.240 - 0.792 and 0.054 - 0.216 respectively for the age groups. This study summarizes the findings concerning AMH and its role as a marker for the quantitative aspect of ovarian reserve, ovarian aging, as well as ovarian dysfunction in this region of the country.

Relationship of Anti-Mullerian Hormone to Reproductive Traits in Katahdin Ewes Bred in Late Spring or Fall

Anti-Mullerian hormone (AMH) is an endocrine marker for fertility in many species. This study investigated possible correlations between serum AMH concentrations, mean number of lambs born (out-of-season in spring or fall), and estimated breeding values (EBVs) for number of lambs born (NLB), number of lambs weaned (NLW), maternal weaning weight (MWWT), weaning weight (WWT), and maternal index (determined by Sheep Genetics Australia). Blood was collected at breeding from Katahdin ewes between 0.7 and 7 years of age in 2015 (n = 163) for the analysis of serum concentrations of AMH. Anti-Mullerian hormone concentration was either expressed quantitatively or divided into quartiles (AMH Q1 through Q4, with Q1 the lowest and Q4 the highest, pg/ml). Data were analyzed by PROC CORR, GLM or chi-squared using SAS. Mean serum AMH was 182 ± 11 pg/ml and ranged from 0 to 1112 pg/ml. There was no correlation between serum AMH and EBVs for NLB, NLW, MWWT, WWT and the maternal index. Serum AMH concentration was similar in ewes in different age categories (P = 0.157). There was a correlation between EBV for NLB (0.29;P = 0.0002) and NLW (0.19;P = 0.013) with average number of lambs born. Ewes in the lowest AMH quartile (Q1) had a lower mean number of offspring born from spring breeding compared with ewes in other AMH quartiles (Q2, Q3, and Q4;P < 0.05). Further study is needed to determine the effectiveness of using serum AMH for selecting ewes for out-of-season (spring) breeding.

AMH联合卵巢超声影像学参数对早发性卵巢功能不全的预测价值

目的 探讨育龄期女性不同年龄的性激素、抗苗勒氏管激素(anti-Müllerian hormone, AMH)水平和卵巢超声影像学参数的变化,分析其对早发性卵巢功能不全(premature ovarian insufficiency, POI)的预测价值并建立预测模型。方法 选择2018年2月至2022年6月就诊于西安交通大学第一附属医院的1 968名育龄期女性为研究对象,检测其性激素、AMH、窦卵泡计数(antral follicle count, AFC)、卵巢总体积、卵巢皮质体积、卵巢髓质体积、卵巢总面积、卵巢皮质面积、卵巢髓质面积,分析不同年龄各项指标的变化及其对POI的预测价值,采用AMH联合卵巢超声影像学参数建立预测POI的模型。结果 1 968名研究对象中POI患病率为3.10%,30~39岁年龄组POI的患病率显著高于20~29岁年龄组(P<0.05)。30~34岁组血清AMH水平显著低于20~24岁组和25~29岁组(P<0.05),35~39岁组血清卵泡刺激素(follicle stimulating hormone, FSH)、黄体生成素(luteinizing hormone, LH)水平显著高于30~34岁组(P<0.05),而血清雌二醇(estradiol, E2)和AMH水平、AFC、卵巢体积、卵巢皮质体积、卵巢面积、卵巢皮质面积均显著低于30~34岁组(P<0.05)。POI组患者的血清FSH、LH水平显著高于同龄女性卵巢功能正常者(P<0.05),而血清E2、AMH水平显著低于同龄女性卵巢功能正常者(P<0.05)。POI组患者的AFC、卵巢体积、卵巢皮质体积、卵巢髓质体积、卵巢面积、卵巢皮质面积和卵巢髓质面积均显著低于同龄女性卵巢功能正常者(P<0.05)。对POI的预测价值分析结果显示,激素水平中AMH的预测价值最高(AUC=0.957),其次为E2(AUC=0.840)。超声影像学参数中AFC的预测价值最高(AUC=0.867),其次为卵巢皮质体积(AUC=0.835)。AMH联合卵巢超声影像学参数建立的模型对POI具有较好的预测价值,其灵敏度为94.86%,特异度为91.23%,AUC为0.960,截断值为12分。结论 育龄期女性中30~39岁年龄组POI的患病率较高,AMH联合卵巢超声影像学参数建立的模型对POI具有一定的预测价值,特别对隐匿期女性获得临床关注。

血清AMH及性激素水平与大学生体检多囊卵巢综合征的相关性

目的分析血清抗缪勒氏管激素(AMH)及性激素水平与大学生体检多囊卵巢综合征(PCOS)的相关性。方法收集2021年3月-2023年11月我院体检确诊为PCOS的80例大学生临床资料作为观察组,另外收集同时期体检的128例非PCOS人群临床资料为对照组。比较两组一般资料、AMH及性激素水平;受试者操作特征曲线(ROC)下面积(AUC)评价AMH及性激素6项对PCOS的预测效能;皮尔森相关性分析AMH水平与性激素水平的相关关系。结果两组年龄、身高、体重、体质量指数比较差异无统计学意义(P>0.05)。PCOS组促卵泡生成素、促黄体生成素、雌二醇、睾酮、催乳素均高于非PCOS组,孕酮低于非PCOS组,差异显著,P<0.05。将PCOS=1,非PCOS=0作为因变量,AMH、性激素6项作为自变量,进行ROC曲线分析,结果显示,AMH及性激素6项均对PCOS有良好的预测价值,AUC分别为0.946、0.924,灵敏度分别为0.850、0.850,特异度为0.977、0.891。但AMH联合性激素6项的预测效能较高,AUC最大,为0.997,灵敏度为0.988,特异度为0.977。AMH的预测阈值为7.975ng/mL。PCOS患者血清AMH水平与促黄体生成素、雌二醇呈显著正相关关系(P<0.05),与促卵泡生成素、睾酮、孕酮、催乳素的相关关系不显著(P>0.05)。结论AMH及性激素水平在大学生体检确认为PCOS血清中异常表达,血清AMH及性激素水平可能与PCOS具有一定相关性,AMH联合性激素6项目对PCOS有良好的评估价值,AMH与促黄体生成素、雌二醇呈显著正相关关系。

Anti-Müllerian Hormone and Its Utility in Cattle Reproduction

Reproductive biotechnologies offer us greater possibilities to improve animal genetics. However, the success of these depends on different factors such as the proper selection of the donor female. For this reason, endocrine markers have been used to evaluate the ovarian reserve, which allow a successful selection of donors. Recent research has shown, among other things, that concentrations greater than 0.130 ng/mL of anti-Mullerian hormone (also known as Muller-inhibiting substance, which is a member of the transforming growth factor beta superfamily of growth and differentiation factors) are related to donors of more than fifteen transferable embryos. Therefore, this review describes studies showing that the measurement of anti-Müllerian hormone concentrations, before superovulation programs, reduces the costs per embryo produced.

Impact of Systemic Lupus Erythematosus on Ovarian Reserve in Premenopausal Women before Receiving Cyclophosphamide Therapy: Evaluation Using Anti-Müllerian Hormone

Introduction: Anti-Müllerian hormone (AMH) is shown to be a possible indicator of ovarian function. Severe systemic lupus erythematosus (SLE) patients exposed to high-dose cyclophosphamide (CTX) have a much higher risk of developing infertility and premature ovarian failure. Therefore, we performed a prospective case-control study to evaluate the impact of SLE on women’s ovarian reserve using AMH before CTX therapy. Methods: SLE patients before receiving CTX therapy were enrolled in our hospital. Age-matched healthy women were served as controls. Serum AMH level was measured using an enzyme-linked immunosorbent assay. Basal hormone levels were measured including follicle-stimulating hormone, luteinizing hormone, and estradiol on the third day of their menstrual periods. All participants underwent transvaginal ultrasonographic examination for the determination of total antral follicle count on the third day. Results: AMH value in SLE patients was significantly lower compared to healthy control with normal ovarian reserve. No significant difference in AMH levels was found between SLE and healthy control with low ovarian reserve. Conclusions: SLE patients not receiving CTX therapy even with normal menstruation, still had an impaired ovarian reserve. Therefore, early monitoring of AMH levels could better reflect the ovarian function and reproductive outcomes of SLE patients and relative protective strategy needed to reserve fertility.

AMH联合性激素检测在多囊卵巢综合征诊断中的价值

目的疑诊多囊卵巢综合征(PCOS)的病人,探讨联合检测血清抗苗勒激素(AMH)及性激素在诊断中的价值。方法选取我院首次就诊并确诊为多囊卵巢综合征的50例患者,作为观察组;并从我院健康管理中心同期体检的患者中抽取58例体检者作为对照组。两组患者血清中的AMH、卵泡刺激素(FSH)、促黄体生成素(LH)、雌激素(E2)、睾酮(T)的水平均采用电化学发光免疫分析技术进行测定后,再进行相关统计学分析。结果观察组的AMH、LH、T的平均水平高于对照组,而FSH的平均水平低于对照组,组间差异具有统计学意义(P<0.05),E2水平虽低于对照组,但组间差异不具有统计学意义(P>0.05)。与其他性激素相比较,AMH对PCOS诊断的灵敏度(86.00%)较高,且与性激素联合检测后,联合诊断的价值高于单项检测。结论血清AMH对多囊卵巢综合征的诊断有意义,并且联合性激素检测后诊断价值更高。

INHB、AMH及IL-21水平与卵巢良性肿瘤患者卵巢储备功能的相关性分析

目的分析血清抑制素B(Inhibin B,INHB)、抗苗勒管激素(Anti-mullerian hormone,AMH)及白细胞介素-21(Interleukin-21,IL-21)水平与卵巢良性肿瘤患者卵巢储备功能的相关性。方法收集2020年1月-2023年12月黑龙江中医药大学附属第二医院妇科收治的104例卵巢良性肿瘤患者为研究对象,根据患者术后3个月时窦卵泡计数(Antral gollicle count,AFC)分为卵巢储备低功能组(n=46)与正常组(n=58)。比较两组患者的一般资料,术前及术后3个月血清INHB、AMH及IL-21水平。采用多因素Logistic回归模型分析卵巢良性肿瘤患者卵巢储备功能的影响因素,采用受试者工作特征(Receiver operator characteristic curve,ROC)曲线分析INHB、AMH及IL-21水平对卵巢良性肿瘤患者卵巢储备功能的评估价值,采用Pearson分析AFC值与INHB、AMH及IL-21水平的相关性。结果与本组术前相比,术后3个月两组患者血清INHB、AMH水平均降低,IL-21水平均升高(P<0.05)。与正常组术后3个月相比,低功能组血清IL-21水平升高,INHB、AMH水平降低;卵泡刺激素(Follicle stimulating hormone,FSH)、黄体生成激素(Luteinizing hormone,LH)水平升高,雌二醇(Estradiol,E2)水平较低(P<0.05)。Logistic回归模型分析显示,术后3个月时血清IL-21水平是卵巢良性肿瘤患者卵巢储备功能低下的独立危险因素,血清INHB、AMH水平是其保护因素(P<0.05)。ROC曲线分析显示,术后3个月时患者血清INHB、AMH及IL-21水平诊断卵巢储备功能的AUC(95%CI)分别为0.707(0.610~0.792)、0.731(0.635~0.813)、0.802(0.712~0.873),联合评估的AUC(95%CI)为0.866(0.785~0.925),联合检测对卵巢储备功能低下的评估价值高于单项指标应用(P<0.05)。Pearson相关性分析显示卵巢良性肿瘤患者中血清INHB、AMH水平与AFC呈正相关(r=0.554、0.617,P均<0.05),血清IL-21水平与AFC呈负相关(r=-0.466,P<0.05)。结论术后3个月的血清INHB、AMH及IL-21水平与卵巢良性肿瘤患者卵巢储备功能具有密切相关性,三者联合应用对卵巢良性肿瘤患者术后卵巢储备能力的评估有重要参考价值。

多囊卵巢综合征患者血清AMH,LH和FSH水平检测及临床应用研究

目的分析抗缪勒氏管激素(AMH)、黄体生成素(LH)和卵泡刺激素(FSH)在常见妇科疾病患者血清中的表达水平,探讨AMH对多囊卵巢综合征(PCOS)的诊断价值。方法采用化学发光法检测1089例常见妇科疾病患者血清AMH,LH和FSH,比较不同年龄组和不同疾病组中的血清AMH水平差异,分析PCOS患者中AMH与LH/FSH的相关性,绘制ROC曲线,计算出诊断PCOS的最佳截断值。结果①AMH水平随着年龄的增加而下降;各个年龄组PCOS患者的血清AMH水平均明显高于同年龄组的健康女性(P<0.05),其他妇科疾病患者的血清AMH水平与同年龄组的健康女性相比,差异无统计学意义(P>0.05);②PCOS患者和健康女性的血清AMH水平与LH/FSH呈中度正相关(r=0.304,0.464,P<0.00);③AMH=6.895ng/ml为PCOS诊断界值时,敏感度为82.8%,特异度为79.6%,曲线下面积为0.886,具有最高诊断效能。在LH/FSH=0.960时,敏感度为75.5%,特异度为80.2%,ROC曲线下面积为0.832,具有最高诊断效能。结论血清AMH水平升高是PCOS患者生殖内分泌代谢紊乱的重要特征,采用ROC曲线分析确立AMH的cutoff值为6.895ng/ml时,对PCOS诊断具有较大临床价值。

半滑舌鳎抗缪勒氏管激素(AMH)基因的克隆及组织表达分析

本研究克隆了半滑舌鳎(Cynoglossus semilaevis)AMH基因并对其进行表达分析。该基因cDNA序列开放阅读框长为1 563 bp,编码蛋白含520个氨基酸。该蛋白含有TGF-β超家族的特征序列,包含1个AMH-N区域和TGF-β结构域,并在C端生物活性区含有9个保守的半胱氨酸残基。系统进化分析表明,半滑舌鳎AMH和所有鱼类AMH聚为一簇,与鲽形目相似性最高。实时定量结果表明,AMH基因在半滑舌鳎不同组织中有差异表达,在正常雄鱼和伪雄鱼性腺中表达量最高。胚后不同时期性腺的实时定量结果表明,性腺分化之前AMH在精巢中相对表达量较低,70 dph(days post hatching)达到最高峰,而在卵巢中呈现先升高后下降的趋势,预示AMH基因可能在半滑舌鳎性腺发育中起重要作用。较正常雄鱼后代而言,AMH基因在伪雄鱼后代的雄鱼和伪雄鱼性腺中的表达都有升高的趋势,而在雌鱼中无明显差异,也预示其可能参与半滑舌鳎的性反转过程。

奥利亚罗非鱼AMH基因结构及其表达

抗缪勒氏管激素（antiMullerian hormone, AMH），也称缪勒氏管抑制物质（mullerian inhibiting substance, MIS），为肽类生长因子，属于TGFβ生长和分化因子家族。应用RTPCR和RACE法，从奥利亚罗非鱼精巢组织中分离出AMH的cDNA。分离到3′UTR长为621 bp和168 bp的两条cDNA，除了长度不一，两者之间只存在两个碱基的差别。5′UTR为 27 bp，阅读框为1 545 bp，翻译成514个氨基酸，其中，AMH-N域237个氨基酸，TGFβ域93个氨基酸。同源性分析显示，AMH保守性偏低，奥利亚罗非鱼与其它鱼类的相似性为32%~60%，与哺乳类动物只有19%~21%，而TGF-β域的保守性相对较高，与其它鱼类的相似性为54%~72%。基因结构分析显示，奥利亚罗非鱼AMH存在6个内含子，和斑马鱼AMH结构相似，和人、鼠AMH有较大差别，它们缺少奥利亚罗非鱼和斑马鱼AMH基因共有的内含子Ⅰ和Ⅵ。然而，启动子转录因子分析结果表明，奥利亚罗非鱼AMH启动子中也存在SRY, SF-1, GATA-4, Sox5, Sox9等结合位点。使用Taqman探针分析奥利亚罗非鱼AMH在成鱼不同组织中的表达以及鱼苗、鱼种、成鱼性腺中的表达量，结果显示AMH在成鱼的卵巢、精巢中均有表达，而在脑、肝、肌肉以及心脏中不表达。AMH在鱼苗性腺中不表达，在鱼种、成鱼中雄鱼AMH表达量显著高于雌鱼，而鱼种雄鱼AMH表达量是成鱼雄鱼的10倍。

AMH蛋白在性成熟小鼠卵泡发育不同阶段及自然衰老过程卵巢组织中的表达变化

目的:通过检测抗苗勒氏管激素(AMH)蛋白在成年小鼠卵泡发育不同阶段和自然衰老过程卵巢中的表达变化,为卵泡的生长发育调控及卵巢衰老发生发展的机制研究奠定理论基础。方法:选用12,24,36,48周龄自然衰老C57BL/6小鼠为模型。收集12周龄C57BL/6小鼠不同发育阶段的卵巢和24,36,48周龄自然衰老C57BL/6小鼠卵巢。采用免疫组织化学方法和蛋白印迹技术检测各组卵巢中AMH蛋白的表达。结果:免疫组织化学显示,AMH蛋白在始基卵泡、初级卵泡,次级卵泡、窦状卵泡的颗粒细胞中均有表达,但在黄体和闭锁卵泡的颗粒细胞中未见表达。始基卵泡颗粒细胞中AMH蛋白表达明显低于初级卵泡、次级卵泡和窦状卵泡(P<0.05);窦状卵泡颗粒细胞中AMH蛋白表达明显低于初级卵泡和次级卵泡(P<0.05);初级卵泡和次级卵泡的AMH蛋白表达未见明显差异(P>0.05)。不同鼠龄的小鼠同级别卵泡颗粒细胞AMH的染色程度未见明显差异;但是24周龄以后间质中开始出现AMH蛋白表达,并且AMH的表达随鼠龄增加而上升;48周龄小鼠卵巢间质中AMH蛋白表达明显高于24周龄和36周龄(P<0.05);24周龄与36周龄比较差异无统计学意义(P>0.05)。蛋白印迹技术结果显示,AMH蛋白在小鼠卵巢组织中的表达随鼠龄增加而降低,差异有统计学意义(P<0.05)。结论:AMH蛋白主要表达于卵泡发育早期阶段的颗粒细胞,并且其在间质中随鼠龄增加表达增强,但是在整个卵巢组织中其表达量随鼠龄增加而降低。提示AMH可能在卵泡发育以及卵巢衰老中起重要的调控作用。

羔羊卵巢AMH的免疫组织化学分析

为探讨AMH在羔羊和成年羊卵巢中时间和空间表达变化,揭示卵泡生长发育调控的机制。收集1月龄羔羊和正常成年羊卵巢,采用HE染色观察卵巢的组织学特征,免疫组化法检测绵羊卵巢AMH蛋白表达分布,RT-PCR技术分析卵巢卵丘细胞中AMH基因的表达变化。结果显示,卵巢HE染色后,对外源激素处理有反应的卵巢存在大量的大直径卵泡,且无中等卵泡和小卵泡,也没有黄体化和排卵;对外源激素没有反应的卵巢仅存在原始卵泡和少量的初级卵泡,无有腔卵泡。免疫组化结果显示,AMH存在于生长期的腔前卵泡和小的有腔卵泡,以及卵丘细胞,大多原始卵泡无AMH的表达,而在原始卵泡向初级卵泡转变,外层的颗粒细胞由扁平向柱状转变时AMH有表达,颗粒细胞存在表达AMH和不表达AMH 2种类型。RT-PCR结果显示,羔羊卵丘细胞的表达量高于成年羊。AMH可能参与卵泡发育的启动、向促性腺激素依赖期的转变等过程的调控,其功能作用还有待于深入研究。

卵巢子宫内膜异位囊肿患者血清及卵巢组织中AMH的表达

目的 检测卵巢子宫内膜异位囊肿患者血清及卵巢组织的抗苗勒管激素(AMH)表达水平,并与健康人及其他卵巢良性囊肿患者进行对比,以判断异位囊肿本身对患者卵巢储备的影响.方法 选择2016年7月至2017年8月在海南省人民医院妇科住院行手术治疗的单侧卵巢子宫内膜异位囊肿患者20例(实验组),因单侧其他卵巢良性囊肿住院行手术治疗的患者20例(对照组A)和健康女性20例(对照组B).抽取所有受检者的外周静脉血,测定AMH水平.卵巢囊肿患者在行卵巢囊肿剔除术中,取囊肿旁的一小部分正常卵巢组织,采用免疫组化的方法 检测卵巢组织中AMH抗原表达,采用荧光定量RT-PCR的方法 检测卵巢组织中AMH mRNA相对表达量;对照组B仅检测血清AMH水平.结果 三组受检者间的年龄比较差异无统计学意义(P>0.05);实验组和对照组A患者的囊肿大小比较差异无统计学意义(P>0.05);实验组患者的血清AMH水平为(2.34±0.98)ng/mL,明显低于对照组A的(3.14±0.73)ng/mL及对照组B的(3.46±1.02)ng/mL,差异均有统计学意义(P<0.05),而对照组A与对照组B间的血清AMH水平比较差异无统计学意义(P>0.05);实验组患者卵巢组织中的AMH阳性表达率为53.3%,明显低于对照组A的80.0%,差异有统计学意义(P<0.05);荧光定量RT-PCR显示,实验组患者的AMH mRNA的相对表达量为0.48±0.16,明显低于对照组A的0.63±0.21,差异有统计学意义(P<0.05).结论 异位囊肿患者的血清和卵巢组织中的AMH的水平均明显低于健康人和其他卵巢良性囊肿患者,异位囊肿对周边卵巢组织存在损伤,进而影响了卵巢储备功能,对异位囊肿患者早诊断早治疗或许可保护患者的卵巢功能.