Y-chromosomal microdeletions and partial deletions of the Azoospermia Factor c(AZFc)region in normozoospermic,severe oligozoospermic and azoospermic men in Sri Lanka

Aim： To assess for the first time the occurrence of Y chromosomal microdeletions and partial deletions of the Azoospermia Factor c （AZFc） region in Sri Lankan men and to correlate them with clinical parameters. Methods： In a retrospective study, we analyzed 96 infertile men （78 with non-obstructive azoospermia） and 87 controls with normal spermatogenesis. AZFa, AZFb, AZFc and partial deletions within the AZFc region were analyzed by multiplex polymerase chain reaction （PCR） according to established protocols. Results： No AZFa, AZFb or AZFc deletions were found in the control group. Seven patients in the group of infertile men were found to have deletions as following： one AZFa, two AZFc, two AZFbc and two AZFabc. The relative distribution of these patterns was significantly different compared with that found in the German population. Extension analysis confirmed that the deletions occurred according to the current pathogenic model, gr/gr deletions were found to be equally present both in the patients （n = 4） and in the control group （n = 4）. One b2/b3 deletion was found in the patient group. Conclusion： These results suggest that the frequency and pattern of microdeletions of the Y chromosome in Sri Lankan men are similar to those found in other populations and confirm that gr/gr deletions are not sufficient to cause spermatogenetic failure. （Asian J Androl 2006 Jan; 8： 39-44）

中国汉族人群AZFc微缺失区域序列标签位点的探讨

目的:探讨中国人群无精子因子c(azoosperm ia factor c,AZFc)微缺失的类型及AZF多重PCR筛查时序列标签位点(sequence taged sites,STSs)的选择。方法:采用多重PCR反应测定9个STS位点(sY84、sY86、sY127、sY134、sY152、sY145、sY255、sY254、sY157)筛查164例严重少精子症或非梗阻性无精子症汉族男性Y染色体微缺失,并以精子浓度正常的男性105例作为对照;同时对来自多中心的180例已确诊为sY254、sY255缺失的男性进行sY145、sY152和sY157位点分析。结果:164例严重少精子症或无精子症男性中AZFc缺失者14例(8.5%);共194例sY254、sY255缺失男性的sY145、sY152均未缺失,而sY157未缺失者仅2例。发现1例单sY157缺失的严重少精子症男性为sY1206缺失和DAZ基因中DAZ3/DAZ4基因拷贝缺失。结论:sY254、sY255和sY157缺失是中国人群AZFc缺失的最常见类型;sY145和sY152微缺失在入选的研究对象中未曾出现,因此不建议作为AZFc常规筛查的位点。"sY157的单一缺失"可能是中国人群AZFc部分缺失的一种新类型,其临床意义值得进一步探讨。

Y染色体无精子症因子c区缺失男性不育患者同步显微取精术后行卵胞浆内单精子显微注射的临床结局

目的:评价同步显微镜下睾丸切开取精术结合卵胞浆内单精子显微注射(intracytoplasmic sperm injection,ICSI)对Y染色体长臂上无精子症因子c区(azoospermia factor c region,AZFc)缺失所致男性不育患者的临床治疗效果,指导AZFc缺失男性不育患者的临床治疗。方法:回顾性分析了自2015年1月到2019年12月因AZFc缺失所致男性不育患者在北京大学第三医院生殖医学中心行显微取精术的临床资料,追踪随访了同步显微取精手术中成功获取精子的患者使用睾丸精子行ICSI助孕的临床结局,比较了首次同步显微取精术与第二次同步显微取精术患者的临床结局,包括受精率、临床妊娠率、流产率和活产率等。结果:共有195例AZFc缺失的男性不育患者接受显微取精术,其中14例为隐匿性精子症,术中均成功发现精子,精子获得率为100%(14/14);181例为非梗阻性无精子症(non-obstructive aoospermia,NOA),122例成功发现精子,精子获得率为67.4%(122/181),余59例NOA患者在显微取精术中未发现精子,占32.6%(59/181)。我们随访了手术成功发现精子的患者中,选择夫妻双方同步取精、取卵且不行胚胎植入前遗传学检测患者的临床结局,纳入研究的99例患者共进行了118人次显微取精术和120个ICSI周期助孕,最终有38例患者成功活产,累积活产率38.4%(38/99),总共分娩出22个健康男婴和22个健康女婴,其中在首次和二次同步显微取精术的新鲜精子-ICSI周期中,优质胚胎率、新鲜周期临床妊娠率和取卵周期活产率分别为47.7%vs.50.4%、40.5%vs.50.0%、28.3%vs.41.2%,第二次同步手术组略高于首次同步手术组,但组间差异没有统计学意义。结论:对AZFc缺失导致的男性不育患者,有较大的概率通过显微取精在睾丸中成功获取精子,并结合ICSI生育自己的遗传学后代,对首次同步ICSI周期未成功的患者,有机会通过再次手术取精和ICSI成功孕育后代。

Y染色体AZFc微缺失规律的研究

目的:探讨中国汉族人群Y染色体AZFc微缺失的规律。方法:采用9个STS位点(sY84、sY86、sY127、sY134、sY152、sY145、sY255、sY254、sY157)的多重PCR,排除sY84、sY86、sY127、sY134缺失的患者,共有194例严重少弱精子症或非梗阻性无精子症汉族男性发生AZFc微缺失;对其中192例sY254、sY255、sY157缺失的患者,再通过sY1191、sY1197、sY1054、sY1125及sY1206 STS位点检测,分析其缺失规律。结果:192例sY254、sY255、sY157缺失患者存在5种缺失模式,其中常见的为sY1197(+)、sY1191(-)、sY1206(-)、sY1054(-)、sY1125(+),占54.17%(104/192);sY1197(+)、sY1191(+)、sY1206(-)、sY1054(-)、sY1125(+),占28.12%(54/192);sY1197(+)、sY1191(-)、sY1206(-)、sY1054(+)、sY1125(+),占14.58%(28/192)。70.83%的AZFc微缺失患者,近端缺失区域发生在sY1197和sY1191之间(属于b2区域);82.29%的AZFc微缺失患者,远端缺失区域发生在sY1054和sY1125之间(属于b4区域)。结论:中国汉族人群AZFc微缺失有5种缺失模式,AZFc微缺失近、远端缺失位点基本集中于复制子b2和b4。

DAZ1/DAZ2 cluster deletion mediated by gr / gr recombination per se may not be sufficient for spermatogenesis impairment： a study of Chinese normozoospermic men

Aim： To explore the possible effect of the deleted in azoospermia （DAZ） copy cluster deletion on spermatogenesis in the Chinese population, the deletion of the azoospermia factor c （AZFc） region was analyzed in 346 normozoospermic men. Methods： Three DAZ single nucleotide variant loci and seven AZFc-specific sequence-tagged sites were examined with polymerase chain reaction （PCR）-restriction fragment length polymorphism and routine PCR. Results： Five （1.4%） of the normozoospermic men were found to have deletion of grlgr-DAZ1/DAZ2. None of the men were found to have b2/b4--entire DAZ deletion. Conclusion： The presence of grlgr-DAZ1/DAZ2 deletion in five men with normozoospermia suggests that this deletion per se may not be sufficient for spermatogenic impairment in Chinese men. （Asian J Androl 2006 Mar; 8： 183-187）

Partial AZFc duplications not deletions are associated with male infertility in the Yi population of Yunnan Province,China

Objective:There are many reports on associations between spermatogenesis and partial azoospermia factor c(AZFc) deletions as well as duplications;however,results are conflicting,possibly due to differences in methodology and ethnic background.The purpose of this study is to investigate the association of AZFc polymorphisms and male infertility in the Yi ethnic population,residents within Yunnan Province,ChinaMethods:A total of 224 infertile patients and 153 fertile subjects were selected in the Yi ethnic population.The study was performed by sequence-tagged site plus/minus(STS+/) analysis followed by gene dosage and gene copy definition analysis.Y haplotypes of 215 cases and 115 controls were defined by 12 binary markers using single nucleotide polymorphism on Y chromosome(Y-SNP) multiplex assays based on single base primer extension technology.Results:The distribution of Y haplotypes was not significantly different between the case and control groups.The frequencies of both gr/gr(7.6% vs.8.5%) and b2/b3(6.3% vs.8.5%) deletions do not show significant differences.Similarly,single nucleotide variant(SNV) analysis shows no significant difference of gene copy definition between the cases and controls.However,the frequency of partial duplications in the infertile group(4.0%) is significantly higher than that in the control group(0.7%).Further,we found a case with sY1206 deletion which had two CDY1 copies but removed half of DAZ genes.Conclusions:Our results show that male infertility is associated with partial AZFc duplications,but neither gr/gr nor b2/b3 deletions,suggesting that partial AZFc duplications rather than deletions are risk factors for male infertility in Chinese-Yi population.

Reproductive outcomes of intracytoplasmic sperm injection using testicular sperm and ejaculated sperm in patients with AZFc microdeletions:a systematic review and meta-analysis

Studies have explored the assisted reproductive technology(ART)outcomes of Y-chromosome azoospermia factor c(AZFc)microdeletions,but the effect of sperm source on intracytoplasmic sperm injection(ICSI)remains unknown.To determine the ART results of ICSI using testicular sperm and ejaculated sperm from males with AZFc microdeletions,we searched Embase,Web of Science,and PubMed to conduct a systematic review and meta-analysis.The first meta-analysis results for 106 cycles in five studies showed no significant differences in the live birth rate between the testicular sperm group and the ejaculated sperm group(risk ratio:0.97,95%confidence interval[CI]:0.73-1.28,P=0.82).The second meta-analysis of 106 cycles in five studies showed no difference in the abortion rate between the testicular sperm group and ejaculated sperm group(risk ratio:1.06,95%Cl:0.54-2.06,P=0.87).The third meta-analysis of 386 cycles in seven studies showed no significant difference in clinical pregnancy rates between the testicular sperm group and the ejaculated sperm group(risk ratio:1.24,95%Cl:0.66-2.34,P=0.50).Inevitable heterogeneity weakened our results.However,our results indicated that testicular sperm and ejaculated sperm yield similar ART outcomes,representing a meaningful result for clinical treatment.More properly designed studies are needed to further confirm our conclusions.

Y染色体AZFc区不同部分缺失对严重少弱精子症患者ICSI结局的影响

目的探讨Y染色体无精子因子(azoospermia factor, AZF)c区不同部分缺失对严重少弱精子症患者卵胞质内单精子显微注射(intracytoplasmic sperm injection, ICSI)临床结局的影响。方法本研究为回顾性队列研究, 分析在河南省人民医院生殖医学中心2017年12月至2020年7月期间经Y染色体AZF区高通量测序筛查、精子浓度<5×106/mL且行射精精子ICSI助孕的严重少弱精子症患者的临床资料, 根据筛查结果将携带AZFc缺失的患者纳入缺失组(n=108), 无AZFc缺失的患者纳入对照组(n=445), 同时按照缺失位点类型将AZFc缺失组分为b2/b4缺失(n=19)、b2/b3缺失(n=30)、gr/gr缺失(n=43)3个亚组, 分别比较各组间及亚组间胚胎发育及妊娠结局情况。结果存在AZFc缺失的患者第3日(day 3, D3)可利用胚胎率、优质胚胎率以及囊胚形成率均低于对照组, 差异均有统计学意义[(70.4%(556/790)比78.5%(2867/3651), P<0.001;24.7%(199/807)比34.3%(1284/3747), P<0.001;51.7%(277/536)比58.0%(1540/2592), P=0.007];种植率、临床妊娠率、移植周期活产率两组间比较差异均无统计学意义(均P>0.05)。AZFc区域b2/b3缺失亚组D3可利用胚胎率、优质胚胎率、囊胚形成率、种植率、临床妊娠率及活产率与对照组比较, 差异均无统计学意义(均P>0.05);b2/b4缺失亚组优质胚胎率[23.2%(32/138)]低于对照组(P=0.004), 而D3可利用胚胎率、囊胚形成率、种植率、临床妊娠率及活产率与对照组相比, 差异均无统计学意义(均P>0.05);gr/gr缺失亚组D3可利用胚胎率[71.6%(280/391)]、优质胚胎率[20.8%(84/403)]以及囊胚形成率[48.7%(133/273)]均低于对照组(P=0.002、P<0.001、P<0.001), 而种植率、临床妊娠率及活产率与对照组比较, 差异均无统计学意义(均P>0.05)。结论 Y染色体AZFc区域b2/b3缺失和b2/b4缺失对严重少弱精子症患者ICSI胚胎发育及妊娠结局均无显著影响, gr/gr缺失对胚胎发育不利影响最大, 但对妊娠结局无影响。

63例成人无精子症AZFc缺失患者的临床特点及其与性激素水平相关性分析

目的探讨无精子症无精子因子C区域(Azoospermia factor C region,AZFc)缺失患者的临床特点及其与性激素的相关性。方法回顾性分析63例无精子症并AZFc缺失患者的临床资料,包括年龄、身高、体重、体重指数(BMI)、睾丸体积和性激素。结果无精子症AZFc缺失组患者身高、体重、睾丸体积与精液正常组差异有统计学意义(P<0.05);2组FSH、LH、睾酮、催乳素水平差异也有统计学意义(P<0.05)。BMI与雌二醇呈正相关(r=0.327,P=0.009),与FSH呈负相关(r=-0.251,P=0.04);年龄与睾酮呈负相关(r=-0.256,P=0.04);睾丸体积与FSH呈负相关(r=-0.289,P=0.02);身高与睾酮、雌二醇、FSH、LH无相关性(P>0.05)。结论无精子症AZFc缺失患者更容易出现体重超标现象,容易更早期出现雄激素水平的下降,生活方式的调整和补充睾酮可能对该类患者的长期生活质量有益。