精索静脉曲张严重少精子症患者Y染色体微缺失分析

目的:探讨精索静脉曲张和Y染色体微缺失对生精障碍的影响。方法:对随机挑选的100例左侧精索静脉曲张严重少精子症患者(精子浓度<5×106/ml,组1),100例左侧精索静脉曲张轻度少精子症患者[精子浓度(10～20)×106/ml,组2],100例特发性严重少精子症患者(组3),100例特发性轻度少精子症患者(组4)和30例正常生育男性对照组(组5)采用聚合酶链反应(PCR)技术进行Y染色体微缺失检测,选取无精子因子(AZF)a、b和c区9个序列标签位点(STS)。结果:组1中有19例患者存在AZF微缺失(19%);组3中有11例患者存在AZF微缺失(11%),其余各组均未发现AZF微缺失;组1比组3有较高的缺失率。结论:在治疗精索静脉曲张严重生精障碍患者前应先进行Y染色体微缺失检测,避免不必要的治疗。     

伴及不伴精索静脉曲张的无精子、严重少精子症患者Y染色体微缺失对比研究

目的:比较精索静脉曲张(VC)无精子症和严重少精子症与不伴VC无精子症和严重少精子症患者Y染色体微缺失发生率,探讨他们不育的内在原因。方法:A组为VC无精子症和严重少精子症的患者137例,其中无精子症70例(A1组),严重少精子症67例(A2组);B组为不伴有VC的特发性无精子症和严重少精子症患者135例,其中无精子症69例(B1组),严重少精子症66例(B2组)。C组(对照组)为30例正常生育男性。采用多重PCR技术对受试者进行Y染色体微缺失检测。结果:1 A组137例中有23例检测到Y染色体微缺失,缺失率16.8%。B组135例中有23例检测到Y染色体微缺失,缺失率17.0%;C组未检测到Y染色体微缺失;2 A1组、A2组、B1组和B2组Y染色体微缺失率分别为为22.9%、10.4%、20.3%和13.6%;3严重少精子症A2组和B2组共133例中16例检测出Y染色体微缺失,发生率为12.0%;4A组与B组比较,差异无统计学意义(P0.05)。结论:Y染色体微缺失发生率在伴有及不伴有精索静脉曲张的无精子、严重少精子症患者中无显著差异,Y染色体微缺失是精索静脉曲张伴有的无精子、严重少精子症病因之一。     

特发性无精子症和严重少精子症患者Y染色体基因微缺失分析

目的：研究Y染色体基因微缺失与特发性无精子症和严重少精子症的关系，并建立一个灵敏、操作简便的分子检测方法。方法：应用实时荧光定量聚合酶链反应(PCR)法对65例特发性无精子症患者、27例严重少精子症患者进行Y染色体YRRM1、DAZ、DYS1基因微缺失的检测。结果：65例特发性无精子症患者中，3例发生YRRM1基因微缺失，发生率为4．6％；5例发生DAZ基因微缺失，发生率为7．7％。27例严重少精子症患者中，1例发生YRRM1基因微缺失，发生率为3．7％；2例发生DAZ基因微缺失，发生率为7．4％。92例患者中均未发现DYS1基因微缺失。结论：YRRM1和DAZ基因位点的微缺失与特发性无精子症和严重少精子症有一定的相关性，DYS1基因缺失与男性生精障碍的相关性仍需进一步研究明确。应用荧光定量PCR法检测Y染色体微缺失具有灵敏、快速、操作简便的特点。     

特发性无精子症和严重少精子症患者Y染色体基因微缺失研究

目的:研究Y染色体基因微缺失与特发性无精子症和严重少精子症的关系,及探讨Y染色体基因微缺失的位点、缺失率有无民族间的差异性.方法:应用多重实时荧光定量聚合酶链反应(PCR)法,对40例汉族及维吾尔族特发性无精子和严重少精子症患者进行Y染色体Azoospermia Factor(AZF)因子多位点的微缺失检测.结果:23例特发性无精子患者中,3例发生AZF因子缺失,缺失率为13.04%;17例严重少精子症患者中,2例发生AZF因子缺失,缺失率为11.76%.结论:Y染色体AZF因子微缺失的范围和位置对于胞质内体外受精治疗男性不育具有重要意义,但Y染色体AZF因子的缺失的位点及缺失率有无民族间的差异性,值得进一步研究.     

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。     

无精子症患者Y染色体基因微缺失研究

研究表明 ,一些无精子症患者伴有Ｙｑ11.2 3 中多个基因位点的丢失。我们采用ＰＣＲ方法对 6 5例原因不明无精子症患者的外周血细胞Ｙｑ11.2 3 上ＹＲＲＭ 1及ＤＹＳ2 4 0基因位点进行检测 ,报告如下。材料与方法　门诊不育患者 6 5例 ,染色体核型分析均为 4 6 ,ＸＹ。取患者抗     

54例无精子症、少精子症患者Y染色体AZF微缺失的检测

目的 探讨Y染色体上AZF微缺失与男性无精子症及少精子症之间的关系。方法 采用多重PCR技术，对54例无精子症及少精子症患者AZF4个区的15个序列标签位点(STS)进行了微缺失检测，并同时做了细胞遗传学检查。结果 54例患者中共有4例发现微缺失(7．4％)，其中有2例在17例无精子症患者中发现(11．8％)，另2例在37例少精子症患者中发现(5．4％)。结论 AZF微缺失是导致男性无精子及少精子的重要原因之一，细胞遗传学检查与AZF微缺失无相关性。     

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。     

原发性无精子症与严重少精子症患者AZF微缺失筛查

目的:观察Y染色体AZF微缺失与原发性无精子症和严重少精子症之间的关系。方法:所有筛选入实验组的研究对象均进行外周血生殖内分泌激素卵泡刺激素(FSH)、黄体生成素(LH)、睾酮(T)的检测及染色体核型分析,排除激素水平异常者及染色体结构与数目异常者。将符合纳入标准的实验对象67例分为原发性无精子症组(A组)49例与原发性严重少精子症组(B组)18例,正常生育男性对照(C组)40例。确定了8个实验用序列标签位点(STS),分别是:sY84、sY86、sY127、sY134、sY152、sY153、sY254、sY255,并以X/Y连锁锌指蛋白基因(ZFX/Y)为内对照进行多重PCR筛查AZF微缺失。结果:67例实验组样本中,共检测出AZF微缺失8例,缺失率为11.94%,其中AZFc区缺失的有4例,AZFa+AZFc区缺失的有2例,AZFb+AZFc区缺失的有1例,AZFb区缺失的有1例。对照组未检出AZF基因微缺失。经χ2检验,实验组与对照组AZF区域STS总缺失率有显著性差异,实验组高于对照组。结论:Y染色体长臂AZF微缺失与原发性无精子症和严重少精子症相关,多重PCR是一种快速、有效的筛查方法。     

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，     

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。     

Frequency of Y chromosome microdeletions and chromosomal abnormalities in infertile Thai men with oligozoospermia and azoospermia

Aim： To investigate the possible causes of oligozoospermia and azoospermia in infertile Thai men, and to find the frequencies of Y chromosome microdeletions and cytogenetic abnormalities in this group. Methods： From June 2003 to November 2005, 50 azoospermic and 80 oligozoospermic men were enrolled in the study. A detailed history was taken for each man, followed by general and genital examinations. Y chromosome microdeletions were detected by multiplex polymerase chain reaction （PCR） using 11 gene-specific primers that covered all three regions of the azoospermic factor （AZFa, AZFb and AZFc）. Fifty men with normal semen analysis were also studied. Karyotyping was done with the standard G- and Q-banding. Serum concentrations of follicle stimulating hormone （FSH）, luteinizing hormone （LH）, prolactin （PRL） and testosterone were measured by electrochemiluminescence immunoassays （ECLIA）. Results： Azoospermia and oligozoospermia could be explained by previous orchitis in 22.3%, former bilateral cryptorchidism in 19.2%, abnormal karyotypes in 4.6% and Y chromosome microdeletions in 3.8% of the subjects. The most frequent deletions were in the AZFc region （50%）, followed by AZFb （33%） and AZFbc （17%）. No significant difference was detected in hormonal profiles of infertile men, with or without microdeletions. Conclusion： The frequencies of Y chromosome microdeletions and cytogenetic abnormalities in oligozoospermic and azoospermic Thai men are comparable with similarly infertile men from other Asian and Western countries.     

Multiplex sequence-tagged site PCR for efficient screening of microdeletions in Y chromosome in infertile males with azoospermia or severe oligozoospermia

The multiplex STS-PCR method was used to detect microdeletions in the long arm of the Y chromosome (Yq) of cytogenetically normal men. One hundred infertile men with azoospermia or oligozoospermia were screened with the multiplex PCR method using 58 STSs, which are specific to Yq for detecting microdeletions on this chromosome. Correlations between the microdeletions on Yq and phenotypes of spermatogenetic disturbance were also examined. Ten patients (10%) had microdeletions on Yq. Seven of the 60 azoospermic patients (11.7%), and 3 of the 40 oligozoospermic patients (7.5%) had microdeletions on Yq. None of the patients showed microdeletions in the AZFa region, but 2 had deletions in the AZFb region, another 2 in the AZFc region, including DAZ, and 1 had deletions in both the AZFb and in the AZFc, including RBM and DAZ. Single microdeletions were found in 4 patients, all of them in the AZFc around DAZ, and 1 patient had 2 microdeletions in the AZFb. The improved multiplex STS-PCR method efficiently detected microdeletions in 10% of azoospermic or severe oligozoospermic men who were cytogenetically normal. All of these microdeletions were presented in the AZFb and/or AZFc regions. This suggests that these regions contain candidate genes for spermatogenesis.     

多重连接探针在无精子症和严重少精子症患者AZF微缺失检测中的应用

目的:探讨多重连接探针扩增技术(MLPA)在无精子症及严重少精子症不育男性无精子因子(AZF)微缺失筛查中的应用可能。方法:提取147例无精子症或严重少精子症患者及154例正常对照男性外周血DNA,经95℃变性后与设计合成的AZF区域探针特异杂交,杂交产物经连接酶连接后用带有FAM荧光标记的通用引物扩增,毛细管电泳将产物分离生成MLPA图谱。所有样本同时行AZF序列标签位点(STS)的多重PCR分析。结果:病例组STS缺失检出率为15.0%(22/147),对照组中未检出STS缺失者;MLPA法于病例组中检出40例AZF区探针缺失患者,检出率为27.2%,其中包括22例STS缺失型患者。对照组中亦有20例AZF探针缺失者。结论:相比较传统的多重PCR,MLPA技术在AZF微缺失筛查中具有更佳的检测灵敏度,同时MLPA图谱所展现的高分辨的AZF区遗传学信息将有助于男性生精障碍病因学机制的深入探索。     

Detection of chromosomal abnormality and Y chromosome microdeletion in patients with azoospermia and oligozoospermia

Objective:To investigate the chromosomal abnormality and Y chromosome microdeletion in patients with azoospermia and oligozoospermia.Methods:Cytogenetic karyotype analysis and multiplex PCR were used to detect chromosomal abnormality and Y chromosome microdeletion in 99 azoospermic and 57 oligospermic patients(total 156).45 fertile men were includ-ed as controls.Results:31 patients were found with chromosomal abnormalities in 156 cases(31/156,19.9 %),20 cases showed 47,XXY,2 cases showed 46,XY/47,XXY,7 cases had Y chromosome structural abnormalities and 2 had autosomal chromosome abnormalities.There were significant differences between the frequency of AZF microde-letion in 125 cases with normal karyotype and 45 controls(P<0.01).The frequency of AZF microdeletion in 68 azoospermic and 57 oligospermic patients were 14.7%(10/68)and 15.8%(9/57)respectively,the difference was not significant(P>0.05).AZFa,AZFb,AZFa+b,AZFb+c,AZFa+b+d and AZFb+c+d mierodeletions were found in azoospermic patients.AZFb,AZFc,AZFd,AZFb+c+d and AZFc+d microdeletions were found in oligo-spermic patients.Conxlusion:The frequency of chromosomal abnormality was 19.9% and the frequency of Y chromosome mi-crodeletion was 15.2% in patient with azoospermia and oligozoospermia.We should pay close attention to this prob-lem.     

Evaluation of sperm chromatin quality and screening of Y chromosome microdeletions in Greek males with severe oligozoospermia

Normal (n = 20) and abnormal (n = 21) semen samples were explored for possible relationships between conventional semen parameters, chromatin status, and microdeletions in the Y chromosome. DNA fragmentation was detected by the terminal deoxynucleotidyl trasferase-mediated dUTP-nick end labelling (TUNEL) assay, chromatin condensation, and DNA packaging quality were assessed by chromomycin A3 (CMA(3)) staining. All men were investigated for Y chromosome microdeletions using polymerase chain reaction (PCR). No deletions were detected in 21 severely oligozoospermic men for the three screened regions (AZFa, AZFb, and AZFc). Men with normal semen parameters showed better chromatin condensation. Spermatozoa with low motility were more likely to contain loosely packaged chromatin. In the abnormal semen group, DNA fragmentation (TUNEL) correlated significantly with sperm motility, concentration, and chromatin packaging assessed by chromomycin A3. However sperm morphology did not correlate significantly with TUNEL and CMA(3) staining.