无精子症及严重少精子症患者的睾丸活检病理研究

在男性不育的患者中 ,无精子症及严重少精子症占有相当的比例 ,无精子症的病因有睾丸性原因、睾丸前性原因和睾丸后性原因 ,其中睾丸性所致无精子症病因及表现又非常复杂 ,睾丸活检对确定是否睾丸性原因 ,睾丸损害类型和程度 ,以及进一步对病因的判断和治疗选择都有重要的意义。 1997年 1月～ 12月在我院泌尿外科诊疗的 4 8例无精子及严重少精子症患者进行了睾丸活检病理检查 ,现将睾丸活检病理结果报告如下。材料和方法1.病例选择　 1997年 1月～ 2 0 0 1年 12月 ,我院泌尿外科门诊 4 8例男性不育患者 ,禁欲 5～ 7天 ,手淫取精液检查 3次以…     

58例无精子症的睾丸活检与染色体检查结果分析

分析５８例无精子症的染色体检查和睾丸活检结果，染色体异常检出率为３７．９３％，其中４７ＸＸＹ（克氏征）占１７．２４％，绝大部分染色体畸变的无精子症患者，其睾丸组织均有不同程度的损害，无生精功能，对于常染色体的结构畸变是否影响睾丸组织功能尚不能肯定，睾丸组织结构异常检出率为７９．３１％，是无精子症的主要原因，而９例输精管缺如患者未发现与染色体变异之间有何联系，由此认为染色体畸变在地精子症的病因中占重     

154例无精子症睾丸活检组织病理和定量组织学观察

本研究对154例无精子症睾丸活检进行了组织病理观察,并在此基础上应用QTM-720图象分析仪对这些活检组织进行了定量组织学研究,测出(1)曲细精管平均直径,(2)曲细精管上皮平均厚度,(3)曲细精管总截面积与测试面积之比,(4)每一测试面积中曲细精管截面数。各病理组间上述四个参数有显著差异,与光镜观察结果相符。     

无精子症患者睾丸活检的临床意义——附70例报告

无精子症患者的诊治非常棘手，如何科学地估计预后，如何指导病人进一步检查治疗，本文就睾丸活检在此方面的作用进行了探讨。材料与方法无精子症70例，年龄20～41岁，平均28．7岁。婚后不育1～14年，平均4．7年。除一例克氏症外，其他患者性生活正常。查体未见明显异常。采用睾丸模型比拟法估测睾丸体积为4～20ml，平均11．68ml。全部病例均经2次以上精液离心镜检，确诊为无精子症。采用局麻下切口法取下米粒大率丸组织，Bouin氏液固定，常规组织学石腊包埋切片，HE染色，光学显微镜下观察。根据吴氏双重诊断法‘1作出睾丸的一般生殖病理…     

无精子症患者睾丸穿刺中评价支持细胞意义

目的 评价无精子症患者睾丸穿刺中检测支持细胞指数的意义。方法 采用Johnson评分法评价睾丸活检组织生精情况；采用瑞-姬染色，人工判读睾丸穿刺液涂片中各期生精细胞和支持细胞，并对其进行分类计数。结果 睾丸活检组织随着Johnson评分降低，支持细胞形态结构越不规则；睾丸穿刺中生精正常组9例，支持细胞指数范围均在20％-40％之间；生精障碍组15例，其中11例支持细胞指数范围在80％～150％，4例支持细胞指数范围在30％以下；生精阻滞组9例，支持细胞指数范围均在150％～500％之间，1例为15％。结论 生精正常组与生精障碍组、生精阻滞组的支持细胞指数差异明显，无精子症患者睾丸穿刺报告中支持细胞指数具有重要意义。     

活检枪与活检钳在无精子症患者睾丸活检应用中的对比研究

<正>无精子症是男性不育的常见病症,在不育男性中的发病率为15%~20%[1]。其病因复杂,治疗难度大,睾丸活检是其组织学诊断、分类、病因分析和预后判断的重要手段。此外,睾丸活检获得的精子和标本还可以用于卵细胞胞质内单精子注射(intracytoplasmic sperm injection,ICSI)[2]以及进行低温冻存[3]。睾丸活检常用的方法有:切开活检、针吸活检和穿刺活检3种。穿刺活检和开放活检成功率     

显微切割睾丸活检在非梗阻性无精子症中的应用

一直以来,学者们认为非梗阻性无精子症因睾丸生精功能受损,导致精液中无精子,而无法生育自己的后代。但随着卵胞质内单精子注射技术的问世,近十几年来涌现出多种睾丸取精术(包括开放性睾丸活检、细针穿刺抽吸、显微切割睾丸活检等)。之后,大量研究表明非梗阻性无精子症患者睾丸中仍存有局部的生精灶,即使是Klinefelter综合征,也可成功取出精子。2010年欧洲泌尿外科学会(EAU)指南明确推荐非梗阻性无精子症采用开放性睾丸活检或显微切割睾丸活检取精。与开放性睾丸活检相比,显微切割睾丸活检的取精成功率高且并发症少,本文就其取精前预测指标、手术操作方法、取精成功率及术后并发症进行综述。     

紧密连接蛋白11在非梗阻性无精子症患者睾丸组织中的表达

目的:分析紧密连接蛋白11(CLAUDIN-11)在非梗阻性无精子症(NOA)患者不同生精障碍类型的睾丸组织中的表达情况,探讨其临床意义。方法:62例NOA患者按不同病理类型分为精子发生能力低下(HS)组和唯支持细胞综合征(SCO)组,其中HS组30例,SCO组32例。免疫组化法检测CLAUDIN-11在睾丸组织中的表达,实时荧光定量PCR检测CLAUDIN-11 mRNA的表达变化,化学发光法检测生殖激素在两组间的差异。结果:免疫组化结果显示,CLAUDIN-11在HS组的表达主要分布于靠近生精小管管壁的支持细胞胞质,在SCO组的表达定位混乱。实时荧光定量PCR结果显示,CLAUDIN-11 mRNA在SCO组表达高于HS组,分别为0.013±0.002、0.008±0.001(t=10.616,P0.01)。血清LH和FSH在HS组和SCO组患者中差异显著[(3.62±1.34)IU/L vs(4.96±3.10)IU/L,(5.36±2.80)IU/L vs(10.65±9.18)IU/L,P均0.05]。结论:CLAUDIN-11在睾丸支持细胞上的表达上调,可能在NOA患者睾丸生精功能障碍的发生和发展中起重要作用。     

Predictive factors of successful testicular sperm recovery in non-obstructive azoospermia patients

Recovery of testicular spermatozoa from non-obstructive azoospermic patients for intracytoplasmic sperm injection (ICSI) is a recent advance in the treatment of male infertility. The purpose of this study is to identify predictive factors for sperm recovery in non-obstructive azoospermic patients. A total of 178 men with non-obstructive azoospermia had multiple testicular sperm extraction (TESE) procedures to recover spermatozoa for intracytoplasmic sperm injection (ICSI) from June 1996 to February 1999. Testicular volume, serum follicle stimulating hormone (FSH) level and testicular histology were examined as positive predictive factors for sperm recovery. Testis biopsies were categorized as severe hypospermatogenesis, maturation arrest and Sertoli cell-only syndrome based on the most advanced pattern of spermatogenesis seen on histology. Sperm retrieval success rates for the patients in three histopathological categories were compared. Spermatozoa were successfully recovered in 94 of 178 (52.8%) men. Sperm were retrieved in 13 of 80 (16.3%) with Sertoli cell-only syndrome, 15 of 24 (62.5%) with maturation arrest, and 66 out of 74 (89.2%) with severe hypospermatogenesis. Spermatozoa recovery has no correlation with testicular volume or serum FSH level. When compared against Sertoli cell-only syndrome, the odds of sperm retrieval success rate was 44.3 times higher in severe hypospermatogenesis and 8.4 times in maturation arrest. These results demonstrate meaningful correlation between successful testicular sperm recovery and testis histopathology. Only testicular histopathology can be used as a predictor of successful sperm recovery.     

Correlation of genetic results with testicular histology,hormones and sperm retrieval in nonobstructive azoospermia patients with testis biopsy

To investigate the frequency and types of genetic results in different testicular histology of patients with nonobstructive azoospermia (NOA), and correlated with hormones and sperm retrieval (SR), a retrospective study was conducted in 286 Chinese NOA patients who underwent testis biopsy and 100 age‐matched fertile men as the control group. Chromosome karyotype analyses were performed by the peripheral blood chromosome G‐band detection method. Screening of Y chromosome microdeletions of azoospermia factor (AZF) region was performed by polymerase chain reaction (PCR) amplification of 11 sequence‐tagged sites (STS). The serum levels of follicle‐stimulating hormone, luteinising hormone and testosterone (T) and the appearance of scrotal ultrasound were also obtained. In 286 cases of NOA, 14.3% were found to have chromosomal alterations. The incidence of chromosomal abnormality was 2.8%. Sex chromosomal abnormalities were seen in six cases (four cases of Klinefelter's syndrome (47, XXY) and two cases of mosaics). The incidence of polymorphic chromosomal variants was 3% in the normal group and 11.5% in the NOA group. In total, 15.7% of NOA patients were found to have AZF microdeletions and AZF (c + d) was the most frequent one. The results of hormone and SR were found to be significantly different among all testicular histological types, whereas no significant differences were found when it comes to genetic alterations. It is concluded that the rate of cytogenetic alterations was high in NOA patients. So screening for chromosomal alterations and AZF microdeletions would add useful information for genetic counselling in NOA patients with testis biopsy and avoid vertical transmission of genetic defects by assisted reproductive technology.     

The predictive value of parameters of clinical presentations for sperm yield in patients with nonobstructive azoospermia receiving microdissection testicular sperm extraction

ObjectiveWe analyzed a cohort of nonobstructive azoospermia (NOA) patients receiving microdissection testicular sperm extraction (mTESE) to examine the relationship of sperm yield and the parameters of clinical presentations. We aim to identify the parameters that might positively predict a positive sperm yield after mTESE.Materials and methodsA total of 200 patients with NOA who had undergone mTESE were enrolled. Among them, 112 (56%) had received a prior testicular needle biopsy. Clinical data including physical findings, underlying genetic abnormalities, pathologic findings in needle biopsy, and sperm retrieval rate (SRR) during mTESE were reviewed and analyzed.ResultsThe pathological findings of prior needle biopsy demonstrate a predictive value of sperm yield during mTESE. Hypospermatogenesis had SRR of 93.3% during mTESE, early maturation arrest had SRR of 13.3%, late maturation arrest (LMA) had SRR of 66.7%, and Sertoli cell-only syndrome had SRR of 18.1%. Regarding parameters of clinical presentation, we found that SRR during mTESE was 85.7% for patients with hypogonadotropic hypogonadism, 60.0% for men with undescended testes (UDT) history, 50.0% for patients who had been exposed to chemotherapeutics due to malignancy of other organs, 100% for prior mumps infection, 50.0% for AZFc deletion, 50.0% for Klinefelter syndrome, and 33.3% for other sex chromosome-related abnormalities. No sperm was found in patients with AZFa or AZFb microdeletion. The consistency of histopathological findings between initial testis biopsy and mTESE was 77.7%. As much as 17.4% of cases had upgraded on spermatogenesis at later mTESE.ConclusionClinical presentations or phenotypes can be used as predictive factors for successful sperm retrieval during mTESE in patients with NOA. Hypogonadotropic hypogonadism and cases with UDT history have a higher chance of sperm retrieval. Initial testicular needle biopsy, if available, can provide valuable information about chances of sperm retrieval. Hypospermatogenesis predicts high sperm yield rate, and LMA can have best upgrade results of sperm yield after mTESE.     

Probability to retrieve testicular spermatozoa in azoospermic patients

Aim: The degree of probability to retrieve spermatozoa from testicular tissue for intracytoplasmic sperm injection intooocytes is of interest for counselling of infertility patients. We investigated the relation of sperm retrieval to clinical dataand histological pattern in testicular biopsies from azoospermic patients. Methods: In 264 testicular biopsies from142 azoospermic patients, the testicular tissue was shredded to separate the spermatozoa, histological semi-thin sec-tions of which were then evaluated using Johnsen score. Results: The retrieval of spermatozoa correlated signifi-cantly (P < 0.001) with the testicular volume (r = 0.49), the FSH concentration ( r = -0.66), the maximum score(r = 0.85) and the mean Johnsen score (r = 0.81). In the multivariate regression analysis the successful testicularsperm extraction showed the closest relationship to the maximum score. The testicular volume correlated significantlywith the mean Johnsen score ( r = 0.64, P < 0. 001), and the basal serum FSH concen     

Microdissection testicular sperm extraction: effect of prior biopsy on success of sperm retrieval

PURPOSE: We determined the effect of prior biopsies with no sperm seen on the chance of sperm retrieval with microdissection testicular sperm extraction in men with nonobstructive azoospermia. MATERIALS AND METHODS: A total of 311 men with NOA underwent microdissection testicular sperm extraction. Of these patients 135 underwent no prior biopsies, 159 underwent 1 or 2 diagnostic testicular biopsies per testis and 17 underwent 3 or 4. The outcome measure studied was the success of sperm retrieval with microdissection testicular sperm extraction. Serum follicle-stimulating hormone and histopathological diagnosis were examined as predictive factors for sperm recovery. RESULTS: Spermatozoa were retrieved in 150 men by microdissection testicular sperm extraction (48%). The success of sperm retrieval in patients who underwent 3 to 4 biopsies (23%) was lower than the retrieval rate in patients who underwent no prior biopsies (56%) and 1 to 2 biopsies per testis (51%) (p = 0.04). When histopathology was considered, patients with Sertoli-cell-only diagnosis on prior diagnostic biopsy had lower retrieval rates compared to the group with no biopsies (p = 0.02). Men with maturation arrest and hypospermatogenesis had similar microdissection testicular sperm extraction sperm retrieval rates regardless of the number of prior biopsies. Spermatozoa recovery was independent of serum follicle-stimulating hormone. CONCLUSIONS: There is no threshold of prior negative biopsies that precludes the success of sperm retrieval using microdissection testicular sperm extraction. A limited number of testicular biopsies provide limited or no prognostic value for sperm retrieval with microdissection testicular sperm extraction.     

Pathological changes in Sertoli cells and dysregulation of divalent metal transporter 1 with iron responsive element in the testes of idiopathic azoospermia patients

Iron is essential for rapidly dividing spermatocytes during normal mammalian spermatogenesis. Decreased transferrin and transferrin receptor levels were observed in seminal plasma from idiopathic azoospermia (IA) patients, suggesting disturbed iron metabolism in IA testes. However, how Sertoli cells (SCs) contribute to the iron homoeostasis in IA is still unclear. In this study, we analysed 30 IA and 30 age‐matched obstructive azoospermia (OA) patients undergoing testicular sperm aspiration (TESA). SCs hyperplasia was indicated by higher SC density and Ki‐67 labelling index in the IA TESA specimens. The attenuated expression of superoxide dismutase (SOD) suggested an impaired antioxidative capacity in IA testes. We further detected increased levels of iron importer divalent metal transporter 1 with iron responsive element (DMT1 + IRE) in IA testes, whereas the increasing trend of iron exporter ferroportin 1 (FPN1) was not statistically significant. Next, we demonstrated that iron regulatory protein 1 (IRP1) and hypoxia‐inducible factor‐1α (HIF‐1α), which can potentially bind to the IRE and hypoxia‐responsive element in the DMT1 + IRE mRNA, were both up‐regulated in IA testes. Unexpectedly, HIF‐2α was down‐regulated in IA testes. These results indicate that there is a dysregulation of DMT1 + IRE in IA testes, which might due to the up‐regulation of IRP1 and HIF‐1α.     

睾丸组织Johnsen评分对非梗阻性无精子症患者取精成功率的预测

目的探讨睾丸组织病理学Johnsen评分是否可以作为非梗阻性无精子症患者取精成功的预测因素.方法选取2008年6月至2011年6月来院就诊的非梗阻性无精子症患者513例.所有患者均行详细的病史采集、体格检查、实验室检查、睾丸活检及活检组织病理学检查等.患者随访1~3年,其主要包括患者是否进一步行睾丸取精术以及取精的结果.结果总计有399例患者接受了活检同侧的睾丸取精术,其中睾丸穿刺取精成功112例,而穿刺取精失败的患者通过进一步接受睾丸显微取精术,成功取精44例.通过分析睾丸取精成功的预测因素后发现,当Johnsen评分≥7时,非梗阻性无精子症患者睾丸取精的成功率将显著提升.结论睾丸组织病理学Johnsen评分≥7可能是睾丸取精成功的预测因素.     

Freezing of testicular tissue as a minced suspension preserves sperm quality better than whole-biopsy freezing when glycerol is used as cryoprotectant

Frozen-thawed testicular spermatozoa have been used successfully for ICSI, especially in cases of obstructive azoospermia with normal spermatogenesis. Fewer attempts, however, have been made to check whether these rather immature spermatozoa, in a different environment with several other cell types present, have cryobiological requirements other than those of ejaculated spermatozoa. This is the reason why the freezing protocols and cryoprotectants (glycerol) used for freezing testicular tissue are based on experience with semen freezing. This study aimed to assess whether cryosurvival and/or motility was influenced by freezing of testicular tissue either as an intact biopsy or as a shredded tissue suspension, when glycerol was used as cryoprotectant. Freezing of testicular tissue as a suspension preserved motility (type B + C) significantly better than freezing of whole biopsies (9.2% vs. 4.0%). Similar observations have been made for vitality (39.3% vs. 25.4%). Centrifugation on 50% Percoll in order to remove the cryoprotectant resulted in a huge loss of spermatozoa (or late spermatids) and should therefore be especially avoided in cases of testicular failure. On the basis of these observations, mincing of the testicular biopsies before freezing may be advocated. Testicular spermatozoa seem to be better preserved when frozen in suspension, at least when slowly permeating glycerol is used as a cryoprotectant.     

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。