探讨血清卵泡刺激素预测非梗阻性无精子症患者精子发生的切点值

目的：利用受试者工作特征曲线（receiver operator characteristic curve ROC曲线）探讨血清卵泡刺激素（FSH）的切点值,以预测非梗阻性无精子症患者睾丸的精子发生。方法选取104例非梗阻性无精子症患者测定其血清FSH（IU/L）值,行经皮睾丸取精子术（TESA）并根据睾丸活检报告分为有精子组（1组）和无精子组（2组）。结果 FSH≤7有52例（50%）,找到精子51例,其概率为98.08%（51/52）;7＜FSH≤14有20例（19.23%）,找到精子17例,其概率为85%（17/20）;14＜FSH≤21有13例（12.50%）,找到精子3例,其概率为23.08%（3/13）;FSH＞21有19例（18.27%）,找到精子6例,其概率为31.58%（6/19）。利用ROC曲线优选的血清FSH切点值是13.78IU/L,该点其敏感性为85.2%,特异性为88.3%,血清FSH水平的ROC曲线下面积为0.895,表明其诊断准确性较高。结论非梗阻性无精子症患者血清FSH水平对预测睾丸精子发生有重要意义。     

血清抗缪勒管激素在梗阻性和非梗阻性无精子症鉴别诊断中的应用价值

目的:通过测定正常生育男性、梗阻性无精子症(OA)患者、非梗阻性无精子症(NOA)患者血清抗缪勒管激素(AMH)水平,探讨其是否作为血清标志物鉴别诊断OA和NOA。方法:选择2018年9月至2019年4月男性不育门诊就诊的患者作为研究对象,采用化学发光免疫法测定正常生育组(n=43)、OA组(n=14)、NOA组(n=45)患者血清中的AMH、抑制素B、FSH水平,经阴囊B超测量睾丸体积。并对正常生育组、OA组和NOA组均进行AMH、抑制素B、FSH、睾丸体积的统计分析。结果:AMH正常生育组[(8.13±3.95) ng/ml]和OA组[(8.51±4.77) ng/ml]均明显高于NOA组[(5.65±3.13) ng/ml,P0.05],抑制素B正常生育组[(127.38±40.50) pg/ml]和OA组[(131.25±52.30) pg/ml]均明显高于NOA组[(25.98±16.29)pg/ml,P0.01],FSH正常生育组[(4.22±3.23) IU/L]和OA组[(4.54±2.09) IU/L]均明显低于NOA组[(19.87±13.09)IU/L,P0.01];正常生育组与OA组AMH、抑制素B、FSH差异无统计学意义(P0.05)。Pearson相关分析结果显示,抑制素B与AMH呈正相关(r=0.326,P=0.01),与FSH呈负相关(r=-0.662,P0.01);FSH与AMH呈负相关(r=-0.468,P0.01);睾丸体积与AMH呈正相关(r=0.339,P0.01),与抑制素B呈正相关(P0.01,r=0.733),与FSH呈负相关(P0.01,r=0.597);精子浓度与抑制素B呈正相关(r=0.522,P0.01),与FSH呈负相关(r=-0.421,P0.01),与睾丸体积呈正相关性(r=0.605,P0.01)。结论:AMH可作为睾丸精子发生的血清标志物之一,并单独或与抑制素B和FSH三者联合用于OA和NOA的鉴别诊断。     

卵泡刺激素正常的无精子症患者无精子因子微缺失检测

研究表明，Yq11，23区域中多个基因片段即无精子因子（azoospermia factor，AZF）的缺失可导致无精子症和严重少精子症。我们选取与无精子症密切相关的Y染色体连锁的13个序列标记位点（sequence taged site，STS），分析无精子症患者Y染色体上AZFa、AZFb、AZFc、AZFd区微缺失情况，     

干细胞疗法在治疗非梗阻性无精子症中的应用

非梗阻性无精子症(NOA)是男性不育的重大难题。干细胞是一群具有自我更新和多向分化能力的细胞。胚胎干细胞和诱导多能干细胞能够通过分化产生精子,但面临着伦理的制约且有成瘤风险。精原干细胞能够通过分化产生单倍体配子,但人精原干细胞培养仍较困难。间充质干细胞通过旁分泌作用促进精子生成。间质干细胞能够分泌睾酮影响精子生成。多种干细胞二维平面共培养及3D睾丸类器官培养更好地模拟了睾丸生精微环境,促进精子生成。干细胞在NOA中的治疗中取得一定的进展,但仍存在不足与困难。本文主要对目前各种干细胞治疗NOA的研究进行总结,介绍了各种干细胞在无精子症治疗中的应用前景与存在缺陷,为干细胞在NOA中的研究及应用提供参考。     

血清抑制素B在鉴别梗阻性与非梗阻性无精子症中的意义

目的通过测定血清抑制素B（INHB）并与卵泡刺激素（FSH）和精浆中性α-葡糖苷酶（α-Glu）等经典指标比较,评价INHB在鉴别诊断梗阻性（OA）和非梗阻性无精子症（NOA）中的应用价值,并对睾丸精子发生障碍作出预判。方法实验采集健康生育男性组（n=60）,以睾丸活检为金标准确定OA组（n=39）和NOA组（n=77）,留取血液和精液标本,进行精液常规分析,检测血清INHB、FSH和精浆中性α—Glu的水平;采用受试者工作特征（ROC）曲线法,通过计算ROC曲线下面积,确定切点值并分析评价检测指标的敏感性和特异性。结果本实验室健康育龄男性血清INHB的95％参考值范围为：20．37-206．21pg／ml。血清INHB、FSH、精浆中性α—Glu、血清INHB／FSH比值以及INHB＋FSH联合在OA组与NOA组之间均差别显著,具有统计学意义（P〈0．01）。其中血清INHB的曲线下面积最大,为0．985,诊断价值最高,敏感性为97．4％,特异性为92．2％,切点值为49．89pg／ml。结论血清INHB比血清FSH、精浆中性α—Glu、血清INHB／FSH比值或INHB＋FSH联合指标在鉴别OA与NOA方面具有更好的敏感性与特异性。     

预测非梗阻性无精子症精子获取因素的研究进展

非梗阻性无精子症(NOA)目前常通过睾丸取精技术结合卵胞浆内单精子注射技术(ICSI),使其获得生育的可能。而睾丸取精技术在不断的发展,睾丸细针穿刺抽吸术(TFNA)、睾丸切开取精术(TESE)、显微睾丸切开取精术(MD-TESE)等技术得到广泛的应用,但是目前仍然缺少可靠的术前预测因素来判断能否成功获取精子。是否可使用特定标记物或非侵入的检查方法对精子获取进行预测,从而避免不必要手术,许多学者进行了多方面的研究。目前已有的预测指标有血清与精浆激素类指标、睾丸指标、影像学和光学技术、Y染色体微缺失、分子蛋白指标、核酸指标等。本文对NOA精子获取的预测因素作一综述。     

非梗阻性无精子症患者睾丸体积、生殖激素水平与睾丸穿刺取精结果的相关性研究

目的:探讨非梗阻性无精子症患者睾丸体积、生殖激素水平与睾丸穿刺取精术(TESA)结果的相关性,以及可用于预测TESA结果的睾丸体积、生殖激素水平的切点值,从而为非梗阻性无精子症患者进一步诊疗提供重要资料。方法:121例研究对象均为非梗阻性无精子症患者(NOA),测定其睾丸体积和生殖激素水平,并根据TESA结果分为无精子组和有精子组。结果:无精子组和有精子组的左侧睾丸体积(ml)、右侧睾丸体积(ml)、泌乳素(PRL,ng/ml)、卵泡刺激素(FSH,mIU/ml)、黄体生成素(LH,mIU/ml)、雌二醇(E2,pmol/L)、血清总睾酮(TT,nmol/L)水平分别为7.07±1.06和11.75±1.38、7.37±1.37和11.70±1.98、12.43±11.69和9.60±4.55、15.77±10.84和8.01±7.43、6.12±2.92和8.11±20.11、119.36±43.52和141.12±48.33、11.43±4.05和12.46±4.60。无精子组血清FSH和PRL水平平均值高于有精子组,并且有显著的统计学差异。虽然无精子组的睾丸体积平均数小于有精子组,但两组之间没有统计学差异。对于年龄、血清E2和TT水平,两组之间也没有统计学差异。利用ROC曲线优选的睾丸体积切点值为9 ml,此点其敏感性为93.8%/89.6%(左/右),特异性为100%/94.3%(左/右),睾丸体积ROC曲线的AUC为0.984/0.961(左/右),表明其诊断准确性较高;优选的血清FSH水平切点值为8.18 mIU/ml,此点其敏感性为71.2%,特异性为75.0%,FSH水平ROC曲线的AUC为0.743,表明其诊断准确性中等。结论:睾丸体积和FSH水平对于预测NOA患者TESA结果具有重要意义,并且睾丸体积诊断准确性明显优于FSH。     

多因素分析预测来曲唑提高非梗阻性无精子症和隐匿性无精子症患者精子密度研究

探讨来曲唑对提高睾酮／雌二醇比例（T／E2）〈10的非梗阻性无精子症或隐匿性无精子症患者的精子数量的有效性。46例不育症患者,排除染色体异常后随机分为两组：第1组22例患者,包括6例无精子症患者和16例隐匿性无精子症患者,接受来曲唑治疗,每天2．5毫克,连续治疗6个月;第2组24例患者包括5例无精子症患者和19例隐匿性无精子症患者,使用安慰剂。收集以下数据：精液常规分析、临床病史、阴囊超声检查、体重指数（BMI）、Y染色体微缺失筛查、染色体核型分析、囊性纤维化筛查,以及性激素检查卵泡刺激素（FSH）、黄体生成素（LH）、雌二醇、雄激素和催乳素。所有患者在接受来曲唑或安慰剂前后都进行精液分析和性激素评估。采用Mann—Whitney己,检验进行统计学分析,精子浓度与FSH、T／E2的比例、双侧睾丸体积~IIBMI等参数的关系使用多变量分析评估,不良作用评估使用卡方检验进行分析。结果发现第1组患者精子浓度、活力、FSH、LH和T都显著增加,平均精子浓度从400ml。增加到1．29×10^6 ml^-1（P〈0．01）,平均精子活力a级精子从2％增加至15％（P〈0．01）,而第2组则没有明显变化。第一组中的雌二醇水平有显著下降,第2组无明显变化。第1组中有8名患者有副作用,第2组中没有患者出现副作用。来曲唑治疗后精子浓度与T／E2比值,FSH水平以及体重指数呈负相关,精子浓度和睾丸体积之间有直接相关性。     

超声技术在睾丸生精功能评价中的应用及展望

在过去的几十年中,不育症严重影响着人们的生活质量,并成为一个全球性的健康问题.根据WHO的报道,全球约有(50～80)×106对夫妇被不育问题困扰[1].     

睾丸穿刺取精术对非梗阻性无精子症患者性功能的影响

<正>非梗阻性无精子症(NOA)是排除了梗阻性因素的一类生精功能低下性疾病,这类患者不能产生精子或只产生极少量精子,导致精液中找不到精子。睾丸穿刺取精术(TESA)是诊断和治疗无精子症的重要手段,符合睾丸穿刺指征的NOA患者精子获取率约60%。随着卵细胞胞质内单精子注射(IC-SI)的应用,这部分NOA患者可获取睾丸精子治疗     

来曲唑在非梗阻性无精子症中应用效果研究

目的探讨来曲唑在治疗非梗阻性无精子症(NOA)患者中的应用效果。方法收集2009年8月至2016年7月在我院接受来曲唑治疗的184例NOA患者为研究对象,根据睾丸体积不同分为3组:A组(81例)(睾丸体积≥12ml),B组(63例)6ml≤睾丸体积12ml,C组(40例)睾丸体积6ml。比较各组患者用药前后的睾丸体积、精液质量、性激素水平变化及睾丸活检组织病理情况。结果 184例NOA患者在规律用药后,所有患者精液标本离心沉淀后镜检未见精子。睾丸体积和体重指数在用药前后无显著性差异(P0.05)。各组用药后睾丸活检未见生精功能活跃的病理表现。各组患者用药前后催乳素(PRL)无显著性差异(P0.05)。用药第7天,3组的FSH和LH显著上升(P0.05),A组和B组的E_2显著下降(P0.05),3组的T值较用药前显著上升(P0.05)。用药第14天,3组的FSH和LH继续上升,A组和B组的T值继续上升,均显著高于用药第7天(P0.05)。最后一次活检前,A、B组的FSH和LH较用药第14天上升减缓(P0.05),C组的FSH和LH则显著下降(P0.05),3组的T值较第14天有所下降(P0.05)。结论来曲唑在治疗NOA患者中未见明显优势,或许需要多中心、大样本数据对这一结论加以验证。     

Preliminary study of letrozole use for improving spermatogenesis in non-obstructive azoospermia patients with normal serum FSH

We investigated whether letrozole (2.5 mg day(-1)) improves sperm count in non-obstructive azoospermia (NOA) patients. Four men were included in this study, and they had folliculo-stimulating hormone and other hormone levels within the normal range and no varicoceles or chromosomal aberrations. These four patients were administered letrozole for 3 months. Sperm count, testicular volume, gonadotropin, testosterone (T) and estradiol (E2) blood levels were assessed before, during and 1 week after the suspension of treatment. All patients showed spermatozoa in their ejaculate, increased gonadotropin and T levels and lower E2 levels (P<0.05 in all cases), when letrozole was administered. This suggests that letrozole treatment might improve sperm count in an NOA sub-population; however, more studies, including the proper controls, are needed to confirm its efficacy.     

非梗阻性无精子症患者睾丸穿刺获精子结局的预测因素研究

目的探讨非梗阻性无精子症(NOA)患者睾丸体积、血清抑制素B(INHB)及性激素水平对预测睾丸穿刺(TESA)获精子结局的意义。方法实验组为162例NOA患者,按TESA获精子结局分为有精子组(TESA⁺,n=74)和无精子组(TESA^-,n=88);正常组为60例同期精液常规参数正常者。比较各组的睾丸体积、血清INHB及性激素水平,筛选有显著性差异的指标,应用ROC曲线评价其对预测TESA获精子结局的意义。结果睾丸体积、血清INHB及FSH三项指标在TESA⁺组和TESA^-组间存在显著性差异(P<0.05),因此为优选的预测指标。三者的ROC曲线下面积(AUC)分别为0.816、0.861、0.777,且后两者间有显著性差异(P<0.05),最佳切点值分别为8.15 ml、70.15 pg/ml、5.52 U/L,对应的诊断灵敏度分别为93.2%、87.8%、74.3%,特异度分别为65.9%、81.8%、77.3%。结论睾丸体积、血清INHB及FSH对预测NOA患者TESA获精子结局...     

Integrative bioinformatics approaches for identifying potential biomarkers and pathways involved in non-obstructive azoospermia

BackgroundNon-obstructive azoospermia (NOA) is a disease related to spermatogenic disorders. Currently, the specific etiological mechanism of NOA is unclear. This study aimed to use integrated bioinformatics to screen biomarkers and pathways involved in NOA and reveal their potential molecular mechanisms.Methods{"type":"entrez-geo","attrs":{"text":"GSE145467","term_id":"145467"}}GSE145467 and {"type":"entrez-geo","attrs":{"text":"GSE108886","term_id":"108886"}}GSE108886 gene expression profiles were obtained from the Gene Expression Omnibus (GEO) database. The differentially expressed genes (DEGs) between NOA tissues and matched obstructive azoospermia (OA) tissues were identified using the GEO2R tool. Common DEGs in the two datasets were screened out by the VennDiagram package. For the functional annotation of common DEGs, DAVID v.6.8 was used to perform Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. In accordance with data collected from the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database, a protein–protein interaction (PPI) network was constructed by Cytoscape. Cytohubba in Cytoscape was used to screen the hub genes. Furthermore, the hub genes were validated based on a separate dataset, {"type":"entrez-geo","attrs":{"text":"GSE9210","term_id":"9210"}}GSE9210. Finally, potential micro RNAs (miRNAs) of hub genes were predicted by miRWalk 3.0.ResultsA total of 816 common DEGs, including 52 common upregulated and 764 common downregulated genes in two datasets, were screened out. Some of the more important of these pathways, including focal adhesion, PI3K-Akt signaling pathway, cell cycle, oocyte meiosis, AMP-activated protein kinase (AMPK) signaling pathway, FoxO signaling pathway, and Huntington disease, were involved in spermatogenesis. We further identified the top 20 hub genes from the PPI network, including CCNB2, DYNLL2, HMMR, NEK2, KIF15, DLGAP5, NUF2, TTK, PLK4, PTTG1, PBK, CEP55, CDKN3, CDC25C, MCM4, DNAI1, TYMS, PPP2R1B, DNAI2, and DYNLRB2, which were all downregulated genes. In addition, potential miRNAs of hub genes, including hsa-miR-3666, hsa-miR-130b-3p, hsa-miR-15b-5p, hsa-miR-6838-5p, and hsa-miR-195-5p, were screened out.ConclusionsTaken together, the identification of the above hub genes, miRNAs and pathways will help us better understand the mechanisms associated with NOA, and provide potential biomarkers and therapeutic targets for NOA.     

The value of epididymal protease inhibitor in differential diagnosis between obstructive azoospermia and non-obstructive azoospermia

There are no efficient and noninvasive clinical tests to distinguish between obstructive azoospermia (OA) and non-obstructive azoospermia (NOA). Epididymal protease inhibitor (Eppin) protein is secreted specifically by testes and epididymides in male reproductive system. It does not exist in seminal plasma of patients with OA in theory. The seminal plasma from 40 normal men and 46 azoospermic patients was detected via Western blot for investigating the presence and characteristics of Eppin protein to distinguish between OA and NOA. The cases were diagnosed as NOA whether Eppin in seminal plasma was positive via Western blot analysis. The cases were diagnosed as OA when samples were Eppin-negative. Additionally, percutaneous epididymal sperm aspiration (PESA) and percutaneous testicular sperm aspiration (PTSA) were performed on these patients at the same time as the diagnostic criteria to compare with Western blot analysis. Eppin detection in seminal plasma showed similar effectivity with PESA/PTSA in differential diagnosis between OA and NOA. Compared with PESA/PTSA, Eppin detection is a new, efficient and noninvasive method which has good clinical application.     

Surgical recovery of sperm in non-obstructive azoospermia

The development of intracytoplasmic sperm injection (ICSI) opened a new era in the field of assisted reproduction and revolutionized the assisted reproductive technology protocols for couples with male factor infertility. Fertilisation and pregnancies can be achieved with spermatozoa recovered not only from the ejaculate but also from the seminiferous tubules. The most common methods for retrieving testicular sperm in non-obstructive azoospermia (NOA) are testicular sperm aspiration (TESA: needle/fine needle aspiration) and open testicular biopsy (testicular sperm extraction: TESE). The optimal technique for sperm extraction should be minimally invasive and avoid destruction of testicular function, without compromising the chance to retrieve adequate numbers of spermatozoa to perform ICSI. Microdissection TESE (micro-TESE), performed with an operative microscope, is widely considered to be the best method for sperm retrieval in NOA, as larger and opaque tubules, presumably with active spermatogenesis, can be directly identified, resulting in higher spermatozoa retrieval rates with minimal tissue loss and low postoperative complications. Micro-TESE, in combination with ICSI, is applicable in all cases of NOA, including Klinefelter syndrome (KS). The outcomes of surgical sperm retrieval, primarily in NOA patients with elevated serum follicle-stimulating hormone (FSH) (NOA including KS patients), are reviewed along with the phenotypic features. The predictive factors for surgical sperm retrieval and outcomes of treatment were analysed. Finally, the short- and long-term complications in micro-TESE in both 46XY males with NOA and KS patients are considered.     

Hyperactivation of early steps of spermatogenesis compromises meiotic insufficiency in men with hypergonadotropism. A possible quantitative assay for high FSH/low testosterone availabilities

It has been shown previously that blood plasma elevation of FSH is associated with an impaired function of the seminiferous tubules. In the study presented here quantitative analysis of the seminiferous epithelium was performed in testicular biopsies of men with qualitatively complete spermatogenesis and hypergonadotropism (HG group, n = 6) or normogonadotropism (NG group, n = 9). Hormonal determinations were performed also in eight men with Sertoli cells only (SCO group). Plasma levels of gonadotropins found in HG were comparable with those found in SCO, while mean plasma testosterone levels in these patients were significantly lower than in SCO or NG. A significant decrease in the mean number of spermatids was present in HG, while the mean numbers of B spermatogonia (0.6 +/- 0.2) and preleptotene spermatocytes (0.6 +/- 0.1) were significantly elevated in comparison with NG (0.4 +/- 0.1 and 0.3 +/- 0.2, respectively). In all men with HG a GnRH test (100 micrograms i.v.) was performed and the relative increase of plasma FSH (maximum/basal level) correlated positively with the number of A-pale (r = 0.85, P less than 0.05) and B spermatogonia (r = 0.81, P less than 0.05). In NG group basal levels of FSH correlated with A-pale (r = 0.90, P less than 0.001) and B spermatogonia (r = 0.59). It seems that FSH plays a role to maintain the number and the differentiation rate of spermatogonia in men and is responsible for the hyperactivation of spermatogonia when secreted in excess. Hyperactivation of spermatogonia probably develops to compensate quantitative decrease in gamete production.(ABSTRACT TRUNCATED AT 250 WORDS)     

'Value of FSH and inhibin-B measurements in the diagnosis of azoospermia'– A clinician's overview

Azoospermia can be either of obstructive ctiology or due to the testis' failure to initiate or maintain spermatogenesis. FSH acts through its receptor at Sertoli cell level and modulates spermatogenesis initiation and maintenance. Inhibin B is a Sertoli cell product expressing the functional capacity of the cell and in an indirect way the state of seminiferous tubule activity. Both FSH and inhibin B differentiate clearly testicular from extra-testicular pathology of azoospermia while, none of these hormones has been convincingly established as predictory index for the finding of spermatozoa in TESE.     

非梗阻性无精子症患者TSG23基因的单核苷酸变异分析

目的分析TSG23基因在非梗阻性无精子症患者中单核苷酸变异。方法采用第二代高通量测序方法对临床收集的776例非梗阻性无精子症患者和709例已正常生育男性的外周血DNA进行TSG23基因外显子捕获测序;采用传统的Sanger测序技术对第二代高通量测序发现的新的突变位点进行验证。结果经过对测序质量控制和外显子测序结果分析筛选,有757例患者和709例的测序结果纳入生物信息学分析,共获得7个突变位点,其中3个为同义突变,4个为错义突变,有4个位点为新发现突变位点;所有位点中非梗阻性无精子症患者特有的突变位点1个,为同义突变,已育正常男性特有的突变位点2个,均为错义突变。结论现有的检测结果显示,TSG23基因的单核苷酸变异与非梗阻性无精子症的发生无明显的相关性。