Y染色体微缺失与无精子症少精子症关系的研究

目的 探讨Y染色体微缺失与无精子症、少精子症的关系.方法 应用多重聚合酶链反应技术(PCR)对127例无精子症(80例)和严重少精子症(47例)的不育患者及60例正常生育男性进行Y染色体AZF基因、DAZ外显子检测.结果 无精子和严重少精子患者Y染色体微缺失7例,缺失率5.51%.其中AZFc缺失2例,DAZ外显子缺失5例.少精子症组缺失率8.51%,无精子症组缺失率3.75%,小睾丸组的缺失率6.54%,正常睾丸组缺失率4.94%,正常生育男性AZF基因和DAZ外显子均未检测到缺失.结论 (1)AZF因子、DAZ外显子微缺失可导致无精子症、严重少精子症:(2)绝大部分无精子、严重少精子患者Y染色体AZF因子、DAZ外显子并没有微缺失,有必要再去寻找新的精子发生基因.     

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。     

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)     

USP26基因突变与精子发生研究进展

泛素特异蛋白酶26(USP26)基因位于Xq26.2,仅有单一外显子,编码由913个氨基酸组成的蛋白质,USP26属于去泛素化酶家族,特异表达于睾丸。USP26基因常见突变类型有插入突变和点突变。目前,该基因与精子发生的关系各研究报道还不一致。本文从USP26基因突变与精子发生障碍之间的关系,USP26基因突变的种族、地域分布差异和USP26基因进化方面,综述了USP26基因与精子发生障碍的关系和研究进展。     

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。     

Y染色体基因微缺失在严重少精子症和无精子症中的应用

目的 利用Y染色体基因微缺失的检测来明确少精子症、无精子症患者病因.方法 采用多重聚合酶链反应技术,针对31例严重少精子症和9例无精子症患者与对照组41名已正常生育的男性,进行AZFa、AZFb、AZFc、3个区域共12个序列标签位点(sequence tag site,STS)的微缺失分析.结果 严重少精子症31例中发现Y染色体微缺失6例,无精子症9例中发现Y染色体微缺失3例,而正常对照组41例均未发现Y染色体微缺失.此研究中发现缺失形式有2种,分别是AZFa+AZFb+AZFc区的全缺失和AZFc区的单独缺失.结论 Y染色体微缺失与精子发生障碍导致的不育有一定的联系.     

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).     

精子发生障碍患者Y染色体AZF区微缺失的筛查及意义

目的　探讨精子发生障碍患者Y染色体AZF区微缺失情况及意义。　方法　选取 6个Y染色体特异性序列标签位点 (STS) ,用PCR技术检测 2 7例精子发生障碍患者AZF区微缺失情况。　结果　 2 7例中AZF区微缺失 2例 ,表现为无精症。缺失均在AZFc区 ,1例为DAZ(sY2 5 4、sY2 5 5 )缺失 ,另 1例为DAZ加sY15 7缺失。　结论　与其他人种一样 ,Y染色体AZF区微缺失也可能是中国人精子发生障碍的原因之一 ,因而精子发生障碍患者在行辅助生育技术前进行AZF区微缺失的筛查是必要的。     

Molecular study on Y chromosome microdeletions in Egyptian males with idiopathic infertility

Aim:To determine the frequency of genetic deletions within the azoospermia factors in Egyptian infertile males.Methods:The Yq microdeletions in 33 infertile males with undetectable chromosomal anomalies were examined by mutiplex polymerase chain reaction (PCR).Deletions were confirmed using single PCR amplifications.Results:Four out of the total 33 (12%) men had Yqll microdeletions,thus supporting the average reported figures in other populations.Three of those 4 cases had single short tandem sequence deletions with discrete histological findings of their testes.Single sY272 deletion within AZFc was associated with Sertoli cell only syndrome,whereas a patient with isolated sY84 deletion within AZFa had immature testicular structure.The remaining case had a large deletion in AZFa-c and short stature.Conclusion:The present study supports the hypothesis that the Yq11 encompasses genetic determinants of stature besides genes controlling spermatogenesis.(Asian J Androl 2004 Mar;6:53-57)     

Y染色体及其微缺失与男性不育:过去、现在与将来

由遗传缺陷所引起的精子发生障碍是男性不育的一个重要病因。一直以来,Y染色体被认为缺乏重要的功能基因。直到睾丸决定基因的发现,Y染色体的研究才重新得到重视。Y染色体的成功测序揭开了Y染色体的真实结构和Y染色体微缺失的分子基础。在Y染色体上的220个基因中,位于AZF区的16个编码基因或基因家族与男性生殖与发育相关。至今,在Y染色体AZF区已发现至少12种缺失。Y染色体上大量的同源序列与回文结构所致非等位的同源性重组是Y染色体微缺失发生的分子基础。Y染色体微缺失是已知的导致男性不育的最主要的分子遗传病因,临床上常使用PCR扩增Y染色体特异的序列标记位点来进行检测。基因组学时代的到来为男性不育的研究带来革命性的工具与方法,有利于加深对Y染色体缺失发生机制的了解,进一步明确Y染色体基因的功能以及相互之间的联系,为基因治疗奠定基础。     

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.     

The prevalence of azoospermia factor microdeletion on the Y chromosome of Chinese infertile men detected by multi-analyte suspension array technology

Aim： To develop a high-throughput multiplex, fast and simple assay to scan azoospermia factor （AZF） region microdeletions on the Y chromosome and establish the prevalence of Y chromosomal microdeletions in Chinese infertile males with azoospermia or oligozoospermia. Methods： In total, 178 infertile patients with azoospermia （nonobstructed）, 134 infertile patients with oligozoospermia as well as 40 fertile man controls were included in the present study. The samples were screened for AZF microdeletion using optimized multi-analyte suspension array （MASA） technology. Results： Of the 312 patients, 36 （11.5%） were found to have deletions in the AZF region. The rnicrodeletion frequency was 14% （25/178） in the azoospermia group and 8.2% （11/134） in the oligospermia group. Among 36 patients with microdeletions, 19 had deletions in the AZFc region, seven had deletions in AZFa and six had deletions in AZFb. In addition, four patients had both AZFb and AZFc deletions. No deletion in the AZF region was found in the 40 fertile controls. Conclusion： There is a high prevalence of Y chromosomal microdeletions in Chinese infertile males with azoospermia or oligozoospermia. The MASA technology, which has been established in the present study, provides a sensitive and high-throughput method for detecting the deletion of the Y chromosome. And the results suggest that genetic screening should be advised to infertile men before starting assisted reproductive treatments.     

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。     

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。     

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.     

Quantification of seminal germ cells in azoospermia: correlations with testicular histology and TESE outcome

In the treatment of male infertility by intra-cytoplasmic injection of spermatozoa (ICSI) extracted from testicular tissue (TESE), the high incidence of negative TESE outcome calls for non-invasive prognostic methods. Literature suggests that seminal haploid germ cell detection could be one. For this purpose, a multi-parametric stringent flow cytometric method was applied to 50 TESE patients for the quantification of ejaculated germ cells. Cells from 50 ejaculates were identified and quantified as spermatozoa (HC, highly condensed), round spermatids (1N), primary spermatocytes (SPC) (4N) or diploid cells (2N, including somatic and non-testicular cells) by their DNA and mitochondria staining and laser scatter characteristics, and compared with testicular biopsy histology and TESE outcome. Whereas 96% of patients displayed a diploid peak in the distribution histograms, the HC, 1N and 4N peaks were absent from the majority of samples. In 13 ejaculates, either a HC or 1N or 4N peak, or a combination of these, was discernible. Although seminal germ cell numbers bore no overall association with elongated spermatids (ES) in histology or spermatozoa retrieval in TESE outcome, 4N cells per ejaculate were correlated with the percentage of tubule sections showing SPC as the most advanced germ cells. The incidence of HC peaks was higher in patients showing some ES in histology or sperm retrieval than in the sperm-negative groups. In groups with suspected obstruction showing nearly full spermatogenesis and maximal sperm retrieval, there was no incidence of a HC peak. Germ cell peaks were associated with germ cell degeneration noted in testicular histology. In conclusion, seminal germ cells cannot provide good prognosis for TESE, although their presence could indicate the spermatogenic activity in the testis.     

Proteomics of spermatogenesis: from protein lists to understanding the regulation of male fertility and infertility

Proteomic technologies have undergone significant development in recent years, which has led to extensive advances in protein research. Currently, proteomic approaches have been applied to many scientific areas, including basic research, various disease and malignant tumour diagnostics, biomarker discovery and other therapeutic applications. In addition, proteomics-driven research articles examining reproductive biology and medicine are becoming increasingly common. The key challenge for this field is to move from lists of identified proteins to obtaining biological information regarding protein function. The present article reviews the available scientific literature related to spermatogenesis. In addition, this study uses two-dimensional electrophoresis mass spectrometry (2DE-MS) and liquid chromatography (LC)-MS to construct a series of proteome profiles describing spermatogenesis. This large-scale identification of proteins provides a rich resource for elucidating the mechanisms underlying male fertility and infertility.