多重聚合酶链反应技术检测严重少精及无精子症患者无精子因子的研究

遗传缺陷引起男性精子发生障碍是男性无精子症、严重少精子症的原因之一。业已证明 ,位于 Y染色体长臂的无精子因子 (azoospermiafactor,AZF)的基因缺失或突变引起精子发生异常 ,为调控精子发生的候选基因之一。本研究采用多重聚合酶链反应技术检测 5 0名正常男性及 36例严重少精及无精子症患者 AZF因子。一、材料与方法1 .对象 :5 0名正常生育男性 ,36例不明原因的严重少精、无精子症患者 (按照 WHO的标准 )。所有患者染色体核型分析正常 ,并排除克氏征及其他因素引起的无精、少精症 ,以及先天性输精管缺如、炎症性输精管梗阻及病毒性…     

特发性无精子症和严重少精子症患者无精子因子的检测及其意义

目的：探讨引起特发性无精子和严重少精子造成男性不育的遗传学原因和检测无精子因子(AZF)的临床意义。方法：对50例特发性男性不育患者(不育组)和50例正常生育者(对照组)的外周血标本．提取基因组DNA,通过多重聚合酶链反应检测Y染色体AZt?微缺失。结果：对照组均可见SRY、SY84、SY86、YRRM1(RBM1)和SY254(DAZ)扩增带。不育组6例(无精子症4例．严重少精子症2例)可见SRY扩增带．但未见SY254扩增带,其中2例同时未见YRRM。扩增带;1例仅未见YRRM。扩增带。结论：Y染色体AZF微缺失是引起无精子和严重少精子并造成男性不育的重要原因之一;AZF微缺失检测对男性不育症患者进行遗传学诊断与筛查有一定意义。     

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.     

特发性无精子症和严重少精子症患者无精子因子检测的临床意义

20 0 1年10月至2 0 0 3年1月,我们采用PCR方法对5 0例正常生育男性和5 0例特发性无精子症和严重少精子症患者进行无精子因子(AZF)检测,现报告如下。材料与方法　5 0例正常生育男性。年龄2 8～38岁,平均33岁。精液常规检查精子数均>4 0×10 6/ml。5 0例特发性不育男性患者年龄2 8～4 2岁,平均34岁。临床检查排除相关的泌尿生殖系疾患。精子计数(2～5×10 6/ml) ,符合WHO诊断标准。患者均有正常4 6XY核型,外周血性激素指标正常。38例无精子症患者睾丸病理检查符合精子发生不完全的诊断。取抗凝血1ml,加入5倍体积重蒸馏水溶解红细胞,离心…     

无精子症和严重少精子症患者Y染色体微缺失、染色体核型和性激素结果分析

目的:研究男性无精子和严重少精子症患者Y染色体微缺失、染色体核型和性激素的相关性。方法:收集无精子症患者63例、严重少精子症患者49例和精液参数正常生育男性60例,抽取外周血分别检测Y染色体微缺失、染色体核型和性激素水平。结果:63例无精子症患者中,7例Y染色体微缺失,微缺失的发生率为11.11%(7/63);49例严重少精子症患者中,4例Y染色体微缺失,微缺失的发生率为8.16%(4/49),与正常精液组(未发现Y染色体微缺失)比较均有统计学差异(P<0.05)。无精子症患者中,染色体核型异常率为9.52%(6/63),而正常生育男性精液组和严重少精子症患者中均未发现异常染色体核型。与正常生育男性精液组[FSH(3.88±2.21)IU/L;LH(4.63±1.51)IU/L]比较,无精子症[FSH(20.41±19.34)IU/L;LH(11.44±9.48)IU/L]和严重少精子症[FSH(8.88±7.04)IU/L;LH(6.78±3.85)IU/L]不育患者FSH和LH水平显著升高(P<0.05)。结论:无精子症和严重少精子症不育患者有必要进行遗传学和性激素检查,便于早期诊断和治疗。     

无精子症和严重少精子症134例遗传学分析

目的：研究无精子症和严重少精子症患者染色体畸变及Y染色体(Yql1区)无精子症因子(azoospermic factor，AZF)缺失情况，建立Y染色体微缺失的临床筛查方法。方法：对134例患者(无精子症97例，严重少精子症37例)经染色体核型分析及AZF、区三个位点8对引物PCR扩增，检测染色体畸变和Y染色体微缺失率。结果：134例中染色体核型异常9例，占6．72％。AZF缺失18例，缺失率为13．43％。无精子症和严重少精子症AZF、缺失率分别为14．43％、10．81％。结论：染色体畸变和Y染色体微缺失是导致无精子症和严重少精子症的主要原因之一。无精子症缺失率高于严重少精子症患者。AZF区三个位点8对引物PCR扩增可作为Y染色体微缺失的临床筛查方法。     

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。     

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。     

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，     

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。     

严重少精子、无精子症患者外周血雄激素受体mRNA的表达研究

目的研究严重少精子或无精子症患者外周血中雄激素受体mRNA的表达。方法采用PT-PCR方法分别检测12例正常男性和31例严重少精子或无精子症患者外周血雄激素受体mRNA的表达。结果 所有正常男性和20例严重少精子或无精子症患者外周血雄激素受体mRNA的表达阳性，11例严重少精子或无精子症患者表达阴性。结论 雄激素受体异常可能是部分严重少精子或无精子症患者的病因。     

Azoospermia factor microdeletions: occurrence in infertile men with azoospermia and severe oligozoospermia from China

Azoospermia factor (AZF) microdeletions are the most frequent genetic cause of male infertility after Klinefelter's syndrome. Although some assisted reproductive techniques such as intracytoplasmic sperm injection (ICSI) have been successfully introduced to clinical treatment for infertile males, the AZF microdeletions might be transmitted from infertile fathers to their male offspring during these procedures. Thus, it is important to carefully evaluate AZF microdeletions in infertile males before assisted reproductive techniques are performed. In this article, we aimed to investigate the frequencies of AZF microdeletions in 137 infertile males with azoospermia and severe oligozoospermia from Jilin province of China and analyse the relationship between the levels of reproductive hormones and AZF microdeletions. Result analysis showed that AZF microdeletions were present in 8 (8.70%) azoospermic males and 3 (6.67%) severely oligozoospermic males. The most frequent microdeletions were detected in the AZFc region, followed by AZFb + c, AZFb and AZFa. And there was no significant correlation between the AZF microdeletion and the levels of reproductive hormones. These findings reinforce the necessity of AZF microdeletion testing among infertile males prior to employment of assisted reproduction techniques in Jilin province of China.     

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.     

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。     

A follow-up study among 285 men with azoospermia and severe oligozoospermia

The present investigation deals with the answer to a simple questionnaire from 147 men with azoospermia and 138 men with severe oligozoospermia (sperm counts < 10 million/ml), earlier admitted to our clinic as male partners in barren couples. Twenty men in the azoospermia group and 52 men in the oligozoospermia group had received different types of infertility treatment. The conception rates reported were 1.6% and 10% in the azoospermia group, and 19.8% and 17.3% in the oligozoospermia group for the untreated and treated subgroups respectively. Only 21% of the men were interested in adoption, whereas 61% answered that they were favourably inclined towards performance of heterologous insemination of their wives. No significant increase in the conception rate was observed in the couples who had achieved adoption. The marriages appeared to be remarkably stable, only 3.5% of the men were divorced or separated from their wives. These results emphasize the necessity of including control groups in treatment trials of male infertility and would also suggest that female partners in these couples should be treated more actively.     

Relationship of azoospermia to inguinal surgery

Nearly one-third of paediatric surgery consists of operations on the inguinal canal. A late complication of this is male sterility due to damage to the vas deferens. Correction of this preventable cause is problematical and there is poor outlook for the severly oligozoospermic and azoospermic male. The true incidence of unilateral damage to the vas deferens is unknown, as this can be masked by a normal reproductive tract on the contralateral side. However, occasionally more subtle changes other than direct transection and fibrosis of the vas deferens can prevent sperm transport. This may be related to damage to the vas pseudocilia, it's auto-nomic and peptidergic innervation leading to defective contractility. This report presents four cases, which confirm the vunerability of the vas deferens during inguinal surgery and highlight azoospermia as a late complication.     

AZF deletion in patients with azoospermia and Oligozoospermia

AZF deletion in patients with azoospermia and Oligozoospermia     

Does the cause of obstructive azoospermia affect the outcome of intracytoplasmic sperm injection: a meta-analysis

OBJECTIVE: To define whether the outcome of intracytoplasmic sperm injection (ICSI) using sperm surgically retrieved from men with obstructive azoospermia (OA) depends on the cause of obstruction. PATIENTS AND METHODS: We first analysed our data and then used a meta-analysis of published data (including ours) to compare the outcome of ICSI in OA, classified in terms of congenital and acquired causes. The present study comprised 82 couples who underwent 127 ICSI cycles using surgically retrieved sperm. The cause was classified as congenital bilateral absence of vas deferens (CBAVD, in 20), after vasectomy (56), infective/inflammatory (21), noninfective (24) and ejaculatory (five). Five reports (687 cycles) including the present were identified as suitable for meta-analysis. RESULTS: Analysis of the present data showed that fertilization and live-birth rates were highest in men with a previous vasectomy and no infective cause (vasectomy 51% and 23%; not infective 53% and 29%, respectively) and lowest in men with infective or inflammatory causes. There was no difference in outcome if the sperm was fresh or frozen, or whether epididymal or testicular. Meta-analysis comparing congenital (CBAVD) and acquired causes showed a significantly increased fertilization rate (95% confidence interval, 0.84-1) with acquired causes. Meta-analysis of the three papers reporting delivery outcome showed no difference in live-birth rate but a significantly higher miscarriage rate in the congenital group (relative risk 2.67). CONCLUSION: In ICSI cycles in men with OA the cause appears to influence the outcome, but outcome is not affected by whether the retrieved sperm is fresh, frozen, epididymal or testicular. The meta-analysis suggested a higher fertilization rate and lower miscarriage rate in acquired causes of OA.