不同Y染色体微缺失类型对生精功能的影响研究

目的:回顾分析Y染色体微缺失患者的精液参数以探究其对男性生精功能的影响。方法:从我院就诊并检测Y染色体微缺失的患者中筛选出Y染色体微缺失阳性的男性不育患者151例,并回顾分析这151例患者的精液参数以及对生精功能的影响。结果:筛查出的151例Y染色体微缺失阳性的男性不育患者中AZFc、AZFb、AZFa、AZFa+c、AZFb+c、AZFc+d、AZFb+c+d、AZFa+b+c+d、sY127、sY134、sY86缺失比例各占66.89%、3.97%、3.31%、0.66%、3.97%、0.66%、8.61%、7.95%、1.99%、0.66%、0.66%;其中有48例无精子症,74例隐匿精子症,28例少弱精子症,1例弱畸精子症。结论:Y染色体微缺失会导致男性精子质量下降以及不同的缺失类型对生精功能影响也不同。     

1052例Y染色体微缺失检测及14例微缺失家系调查

目的:通过对不育患者进行Y染色体微缺失筛查以及部分微缺失患者的家系追踪调查,探讨Y染色体微缺失父子间的自然垂直遗传特点。方法:对1 052例患者进行Y染色体无精子因子(AZF)检测,并对12例AZFc缺失患者,1例AZFb和1例AZFb+c缺失患者进行家系追踪调查,绘制AZF缺失患者男性直系家族成员男性不育家系系谱图。结果:1 052例患者,共发现Y染色体微缺失89例,其中AZFc缺失56例,AZFa缺失6例,AZFb缺失5例,AZFb+c缺失14例,AZFa+b+c缺失8例。在追踪调查的AZF缺失家系中,AZFb和AZFb+c仅先证者存在缺失,12例AZFc缺失患者中5例重度少精子症患者存在家族垂直遗传,另外1例重度少精子症患者和6例无精子症患者家系中除先证者有缺失外,其家系成员未发现缺失。结论:通过对Y染色体微缺失患者进一步的家系调查发现,仅重度少精子症的AZFc缺失患者可能由父亲垂直遗传而来,但与父系表型有差异。对AZF缺失的无精子症患者,无论何种缺失类型,由父亲垂直遗传而来的可能都不大。     

多重聚合酶链反应检测Y染色体无精子因子微缺失

目的:研究原发性无精、严重少精症与Y染色体无精子因子(AZF)微缺失之间的关系.方法:采用多重聚合酶链反应技术对103例原发无精子症、72例原发严重少精症患者及60例正常生育男性进行AZFa、AZFb、AZFc 3个区域微缺失分析.结果:60例正常生育男性未发现Y染色体AZF区域微缺失,175例生精障碍患者中发现AZF微缺失19例,总缺失率为10.9%.其中11例无精症患者和4例少精症患者的缺失发生在AZFc区域,缺失率为8.6%;1例无精症患者和2例少精症患者发生AZFb、AZFc双重缺失,缺失率为1.7%;1例无精症患者发生AZFa、b、c 3个区域同时微缺失,缺失率0.6%.生精障碍组与正常生育男性组比较Y染色体AZF区域微缺失率差异有统计学意义(P<0.01).结论:Y染色体AZF区域微缺失是引起男性无精、少精子症的重要原因之一.采用多重聚合酶链反应技术对原发无精、少精子症患者在单精子注射(ICSI)之前进行微缺失筛查是必要的.     

无精子症和严重少精子症患者AZF/DAZ基因微缺失的分子生物学检测

目的　用分子生物学方法检测无精子症和严重少精子症患者无精子基因 (AZF)AZF/DAZ基因微缺失。　方法　应用聚合酶链反应 (PCR)技术对无精子症 4 7例、严重少精子症 4例进行Y染色体AZFa、AZFb、AZFc/DAZ、SRY的微缺失检测。　结果　 5 1例患者缺失率为 35 .3% (18/ 5 1) ,其中AZFa、AZFb、AZFc的微缺失分别为 4例 (7.8% )、5例 (9.8% )和 4例 (7.8% )。无精子症患者 1例 (1.9% )为AZFa、AZFb的双重缺失 ,2例 (3.9% )为AZFb、AZFc的双重缺失 ;2例 (3.9% )为AZFa、AZFb和AZFc的三重缺失 ;5 1例SRY基因PCR扩增均为阳性。 5例已有生育的正常男性均无AZFa、AZFb、AZFc、SRY的微缺失。　结论　AZF/DAZ(包括AZFa、AZFb、AZFc/DAZ)基因的微缺失是引起无精子和严重少精子导致男性不育的重要原因之一。AZF/DAZ基因微缺失的分子生物学检测对不明原因的不育男性行胞浆内单精子注射 (ICSI)时有指导意义。     

替代睾丸活检的新方法——精液细胞学研究

目的：寻找无创伤判断睾丸生精功能及输精管道梗阻的检查方法。方法：对１２２例精子症病人及５０例生育男性同时进行睾丸活检和精液细胞学检查。结果;生育男性精液细胞学及睾丸活检生精细胞发育水平符合率为１００％,无精子症病人总符合率为９１％;经Ｋａｐｐａ检验两法呈高度相关,Ｐ〈０．０１。１４例睾丸活检见精子及各级生精细胞,精液细胞学检查未见生精细胞的病例,经精液生化指标证实１３例为输精管道梗阻,１例为逆行射     

1例罕见的AZFb部分缺失的少精子症

Y染色体上AZF区域的微缺失是临床上最常见的无精子症和少精子症遗传学改变，因此对其检测已成为生殖中心的常规检测项目。到目前为止，基因型一表型虽说不是特别清楚，但基本的轮廓已经勾画出来：AZFa缺失的患者表现为唯支持细胞综合症（Sertoli cell only syndrome，SCOS）；AZFb缺失的患者表现为精子成熟障碍，主要停滞在精母细胞阶段；这两个区域缺失的患者睾丸中都没有精子发现；AZFc的缺失从精子数目正常到精子数目减少直至SCOS。AZFb部分缺失的病人十分罕见。我们报告1例AZFb部分缺失的少精子症的患者。     

替代睾丸活检的新方法——精液细胞学研究

目的：寻找无创伤判断睾丸生精功能及输精管道梗阻的检查方法。方法：对１２２例无精子症病人及５０例生育男性同时进行睾丸活检和精液细胞学检查。结果：生育男性精液细胞学及睾丸活检生精细胞发育水平符合率为１００％,无精子症病人总符合率为９１％;经Ｋａｐｐａ检验两法呈高度相关,Ｐ＜０．０１。１４例睾丸活检见精子及各级生精细胞,精液细胞学检查未见生精细胞的病例,经精液生化指标证实１３例为输精管道梗阻,１例为逆行射精。结论：两种方法所反映的生精细胞水平完全一致,说明精液细胞学检查是一种比较理想的判断睾丸生精功能及输精管道梗阻的无创伤检查方法     

精液细胞学与睾丸活检及针吸细胞学的相关性研究

为了寻找无创伤性判断睾丸生精功能及精道梗阻的检查方法，对６５例无精症患者随机分为两组：第一组６１例无精子症患者，进行精液细胞学与睾丸针吸细胞学检查；第二组１５例正常生育男性及４例无精子症患者，进行睾丸活检与精液细胞学检查。结果：第一组５３例（占８７％），精液中检出生精细胞，检出病例二者的生精细胞发育水平总符合率９１％，经χ２检验两者呈显著相关（Ｐ＜０．００５）；８例（占１３％）精液细胞学检查未见生精细胞的病例，考虑梗阻性无精子症，其中７例经精液生化指标证实，５例睾丸针吸细胞检查见精子。第二组两种方法所反映的生精细胞发育水平完全一致。说明精液细胞学检查既能很好反映睾丸生精状况，又能反映精道梗阻情况，是一种比较理想的判断睾丸生精功能及精道梗阻的无创伤性检查方法。     

无精子因子(AZF)在男性不育症诊治中的临床意义

无精子因子(AZF)的微缺失是无精子和严重少精子男性常见的基因改变之一,大约15%~20%的不育男性发生AZF微缺失。目前已知AZF的微缺失主要分布在Y染色体长臂近、中、远三个非重叠区域,分别称为AZFa、AZFb、AZFc。特异性的AZF缺失类型与特异性的精子生成缺陷似乎存在某种联系,如AZFa缺失通常与精原细胞完全缺失有关(睾丸唯支持细胞综合症);AZFb缺失通常与生殖细胞成熟停滞有关;AZFc缺失较常见,与多种无精子和严重少精子有关等[1]。近年来由于体外受精-配体移植(IVF-ET)、睾丸内穿刺或活检获取精子(TESE)及胞浆内单精子注射(IC…     

严重少精子症及非梗阻性无精子症患者Y染色体微缺失分析

目的探讨严重少精子症及非梗阻性无精子症与Y染色体长臂微缺失之间的关系。方法该病例对照研究包括216例严重少精子症、189例非梗阻性无精子症患者及100例精液参数正常的对照。采用多重PCR对Y染色体AZFa、AZFb、AZFc及AZFd区域进行检测。玷果在严重性少精子症患者中,AZF总缺失率为10．65％（23／216）,其中以AZFc区缺失最常见,占缺失的78．26％（18／23）;在非梗阻性无精子症患者中,AZF总缺失率为13．76％（26／189）,其中也以AZFc区缺失最常见,占缺失的57．69％（15／26）;在正常对照中发现1例AZFb缺失,两病例组AZF区缺失分别与对照组相比较均具有显著差异（X^2=9．066,P=0．003;X^2=10．74,P=0．001）。结论通过对Y染色体微缺失的检查可以从基因水平寻找生精障碍的原因以及为优生优育提供可靠的遗传信息依据。     

陕西地区不明原因无精子症和少精子症患者Y染色体长臂微缺失分析

目的: 评估陕西地区不明原因无精子症和少精子症不育男性患者Y染色体长臂微缺失的频率,探讨精子密度与Y染色体微缺失发生率的相关性。　方法: 以Y染色体特异性无精子症因子区STS AZFa、AZFb、AZFc和SRY4个基因 5个片段设计引物,采用PCR方法对 64例无精子症和少精子症患者以及 20例正常生育男性进行微缺失检测,并比较不同精子密度患者Y染色体微缺失的发生率。　结果: 20例精子密度正常的生育男性未检出Y染色体微缺失,而 64例特发性无精子症 /少精子症患者AZFc区的缺失率为17. 2% (11 /64),AZFc和AZFb联合缺失 1例,未发现AZFa区缺失,SRY基因均为阳性。其中无精子症组缺失率为21. 43% ( 3 /14 );精子密度 <1×106 /ml组,缺失率为 20. 0% (2 /10);精子密度 (1 ~5)×106 /ml组缺失率为17. 9% (5 /28);精子密度 (5 ~10 )×106 /ml组缺失率为8. 3% (1 /12)。各组缺失率经卡方检验差异有显著性 (χ2 =70. 144,P<0. 005 )。　结论: 无精子症和少精子症不育患者Y染色体AZFc缺失率明显较高,PCR扩增AZF基因是诊断Y染色体微缺失的简单方法。     

Testicular biopsy in azoospermia and severe oligozoospermia

The continued experience of testicular biopsy application in 861 cases of azoospermia and 152 cases of severe oligospermia is reported adding to the previously published cases of 1075 patients with azoospermia. The most common finding in the whole series was that of normal testis denoting obstruction (48%), while among cases of functional azoospermia, Sertoli cell only and spermatogenic arrest were the most frequent (66%).     

无精子症和严重少精子症患者Y染色体微缺失和基因缺失研究

目的研究中国特发性无精子症和少精子症患者Y染色体无精子症因子(AZF)区缺失和其中RBMY1A1、DAZ基因缺失。方法选取AZFa、b和c区6个序列标签位点(STS)对56例少精子症和33例无精子症患者进行外周血Y染色体微缺失分析,对缺失样本进行RBMY1A1和DAZ基因缺失分析。结果共确认6例患者发生Y染色体微缺失和基因缺失、占7%(6/89);其中5例AZFc/DAZ基因缺失,1例AZFb+c/RBMY1A1和DAZ基因缺失。结论AZF部分区域缺失的患者同时伴有与精子生成具有重要作用的基因缺失,并可能由此导致精子生成障碍。     

Molecular genetic analysis of microdeletion on AZF/DAZ gene in patients with idiopathic azoospermia and severe oligozoospermia in Fujian

Objective: To identify microdeletions in azoospermia factor(AZF) gene loci in patients with idiopathic azoospermia and severe oligozoospermia in Fujian. Methods: Molecular genetic detection method was used to detect microdeletion at the AZFa, AZFb, AZFc /DAZ,SRY region of Y chromosome in 47 azoospermia and 4 severe oligozoospermia patients. Genomic DNA was extracted from peripheral blood. The sequence tagged site (STS) primers tested in each cases were sY84(AZFa), sY 143(AZFb) sY254(AZFc).SRY region of Y chromosome for control. The PCR products were analyzed on a 2.0% agarose gel. Results: Microdeletions of the Y-chromosomal AZF loci were revealed in 18(35.3%,18/51) of 51 patients with idiopathic azoospermia and severe oligozoospermia. AZFa deletion was found in four (7.8%) patients, AZF b in five (9.8%) patients, AZF c in four (7.8%) patients. AZF a+b in one(1.9%)patient, AZF b+c in two (3.9%) patients, AZF a+b+c in two (3.9%)patients respectively. No deletion of SRY region was found. No deletion of AZF a, AZF b, AZF c/DAZ,SRY regions was found in five fertile male who had at least one or more children. Conclusions: Microdeletions on AZF/DAZ gene loci were major genetics defects leading to azoospermia and severe oligozoospermia in male idiopathic infertility in Fujian. It is necessary to have genetic counseling and carry out microdeletion detection on AZF/DAZ gene loci before performing intracytoplasmic sperm injection (ICSI).     

Clinical and pathological correlation of the microdeletion of Y chromosome for the 30 patients with azoospermia and severe oligoasthenospermia

Aim: To review the accumulated 30 patients with different area of Y chromosome microdeletions, focus-ing on their correlation with the clinical and pathological findings. Methods: A total of 334 consecutive infertile men with azoospermia (218 patients) and severe oligoasthenospermia (116 patients) were screened. Complete physical and endocrinological examinations, general chromosome study and multiplex polymerase chain reaction assay to evaluate the Y chromosome microdeletion were performed. Ten patients received testicular biopsy. Then the clinical and pathological findings were analyzed with reference to the areas of Y chromosome microdeletion. Results: There is a decline of the percentage of sperm appearing in semen in the group that the gene deletion region from AZFc to AZFb. The clinical evidence of the impairment (decreased testicular size and elevated serum FSH) is also relevantly aggravated in this group. However, the pathology of testicular biopsy specimen was poorly correlated with the different deletion areas of the Y chromosome, which may be due to the limited number of specimens. Conclusion:The clinical correlation of spermatogenic impairment to the different AZF deletion regions may provide the information for the infertile couples in pre-treatment counseling. (Asian JAndrol 2004 Dec; 6:369-375)     

Fas和FasL 系统在非梗阻性无精子症睾丸中的表达

目的：为探讨睾丸生精功能障碍与细胞凋亡的关系，研究Fas和FasL系统在非梗阻性无精子症睾丸支持、间质和生精细胞中的表达。方法：对20例非梗阻性无精子症患者行睾丸开放性活检，常规病理检查，按Johnson评分法评价精子发生和发生障碍的程度；采用免疫组化SABC法对睾丸支持、间质和生精细胞进行Fas和FasL表达的检测。结果：睾丸活检生精功能评为8分有14例，3分有2例，6、5、4和2分各有1例。在20例非梗阻性无精子症睾丸的间质、支持和生精细胞均有Fas和FasI。的表达；而支持细胞Fas和FasL的阳性和强阳性表达率明显高于间质和生精细胞。结论：非梗阻性无精子症的睾丸支持、间质及生精细胞Fas和FasL的高表达与精子生成障碍是一致的，非梗阻性无精子症可能与生殖细胞过度凋亡密切相关。     

新疆地区汉族维吾尔族不育症患者Y染色体长臂微缺失分析

目的评估新疆地区汉族、维吾尔族不明原因无精子症和严重少精子症男性患者Y染色体长臂微缺失的频率,探讨不同民族间Y染色体长臂微缺失发生率的差异。方法以Y染色体无精子因子(AZF)区STS- AZFa、AZFb、AZFc和AZFd 4个基因8片段设计引物,采用聚合酶链反应(PCR)方法对123例(汉族61例,维吾尔族62例)无精子症和少精予症不育男性患者进行Y染色体微缺失检测,并比较不同民族的患者Y染色体微缺失发生率的差异。结果61例汉族患者中有27例(44.26%)存在Y染色体微缺失,62例维吾尔族患者检出13例(20.97%)存在Y染色体微缺失,在所有被检出有Y染色体长臂微缺失的患者中AZF区联合缺失23例(58%)。汉族患者与维吾尔族患者Y染色体微缺失率及AZF多位点联合缺失发生率差异有统计学意义(P<0.05)。结论无精子症和严重少精子症不育男性患者中Y染色体长臂微缺失发生率及AZF多位点联合缺失发生率存在民族差异,PCR检测AZF基因是诊断Y染色体长臂微缺失的较好的方法。     

Expression of beta-catenin in human testes with spermatogenic defects

beta-catenin is a multifunctional molecule that functions in intercellular adhesion and signal transduction during assembly of AJs between Sertoli cells as well as between Sertoli cells and germ cells. To assess changes in the testicular beta-catenin in male infertility conditions, testicular tissues from obstructive azoospermia with normal spermatogenesis, spermatogenic arrest (SA) and Sertoli cell-only syndrome (SCO) patients were examined for immunohistochemical localization of beta-catenin. In normal spermatogenic tissue, expression of beta-catenin was largely found in the Sertoli cell-germ cell (primarily spermatocytes) contact areas. Interestingly, perinuclear localization of beta-catenin was found in spermatocytes and spermatids. In spermatogenic arrest, beta-catenin in cell contact areas between Sertoli cells and germ cells was greatly decreased, but perinuclear beta-catenin in spermatocytes was not. In SCO, weak or negligible immunoreactivity of beta-catenin was found in cell contacts between Sertoli cells. Nuclear localization of beta-catenin was found in myotubular cells in all samples. Taken together, altered expression of beta-catenin in cell contacts within the seminiferous epithelia in spermatogenic arrest and SCO suggests that interactions between Sertoli cells and germ cell are crucial for expression of beta-catenin, and thus functional development of AJs in seminiferous epithelia in human testis. It should be also emphasized that perinuclear beta-catenin in germ cells may play a specific role in spermatogenesis.