无精症患者Y染色体微缺失的多重PCR筛查

目的 建立一套Y染色体微缺失的多重PCR筛查方法。方法 建立5套稳定和可靠的多重PCR筛查方法，对进行ICSI治疗的26例无精子症患者和进行睾丸活检的30例无精子症患者进行Y染色体微缺失的检测。结果 在56例无精子症患者中发现5例AZFc／DAZ缺失，2例同时具有AZFc／DAZ和AZFb／RBMl的缺失，1例仅具有sY153一个片段的缺失。结论 本研究Y染色体微缺失的多重PCR筛查方法是易行和可靠的，对无精子症患者有必要进行Y染色体微缺失的常规筛查。     

精索静脉曲张男性不育患者Y染色体微缺失检测

目的:探讨精索静脉曲张(varicocele,VC)不育患者Y染色体微缺失特点及其与临床表型的关系,为评价VC不育患者是否行手术治疗或ICSI提供依据。方法:VC不育患者174例,分为3组,A组:无精子症47例;B组:严重少精子症57例;C组:轻度少精子症70例;设立正常生育的健康志愿者男性28例作为对照组(D组)。抽取外周血提取DNA,选取Y染色体上AZFa、AZFb、AZFc区共6个序列标签位点,应用多重PCR进行扩增;已生育女性26例作为阴性对照,分别运用琼脂糖凝胶电泳分离,对照阅读扩增产物,判定有无缺失存在以及缺失类型。结果:174例男性不育患者中有22例检测到Y染色体微缺失,缺失率12.64%;A组11例存在微缺失,B组11例存在微缺失,C组未检测到微缺失。A组与C组、B组与C组比较,差异均有显著性。A组缺失病例中有6例为AZFc区缺失,1例为AZFa缺失,2例为AZFb区缺失,2例为AZFb、AZFc区共同缺失;B组缺失病例中有8例为AZFc缺失,2例为AZFb缺失,1例为AZFb、AZFc区共同缺失。结论:①精液异常VC不育与Y染色体微缺失有关;②VC不育患者特别是无精子症和严重少精子症患者,应该进行Y染色体微缺失的检测。     

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。     

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。     

中国男性不育患者染色体核型及Y染色体微缺失分析

目的 探讨男性不育症与染色体畸变及Y染色体微缺失之间的关系.方法临床诊断男性不育患者1975例,采集外周血淋巴细胞常规培养,Giemsa染色,镜下观察并分析染色体核型;选取Y染色体特异性序列标签点(STS),应用PCR技术对无精子症及少精子症患者进行Y染色体微缺失检测.结果 1975例患者中,染色体核型异常305例(15.44%),其中常染色体异常101例(5.11%),患者主要表现为少精子症、畸形精子症;性染色体异常204例(10.33%),主要表现以克氏征(5.62%)为主.728例无精子症或少精子症患者中,Y染色体微缺失109例(14.97%),其中AZFa区缺失3例(2.75%),均表现为无精子症;AZFb区缺失5例(4.59%),表现为无精子症2例、严重少精子症2例,精液正常1例;AZFc区缺失者68例(62.39%),患者主要表现为无精子症和严重少精子症;AZFa区和AZFc区均缺失者5例(4.59%),均表现为无精子症;AZFb区和AZFc区均缺失者15例(13.76%),患者以无精子症表现为主;AZFa区、AZFb区和AZFc区均缺失者6例(5.50%),均表现为无精子症.结论染色体异常及Y染色体微缺失均为男性不育的重要病因.

Abstract：

Objective To study the relationship between chromosomal abnormality and Y chromosome microdeletions and male infertility. Methods Lymphocytes were cultured from peripheral blood of 1975 male infertility patients and stained with Giemsa. The chromosomes were analyzed under microscope. Y chromosome specific sequence tags (STS) were selected, then the Y chromosome microdeletions in AZF regions were screened by polymerase chain reaction (PCR) in azoospermia and oligozoospermia patients. Results There were 305 cases of detected chromosomal abnormalities (15.44%) in the 1975 cases. There were 101 cases (5.11 %) with autosome abnormalities which clinically manifested as oligozoospermia and teratospermia. There were 204 cases (10. 33%) of sexual chromosome abnormalities and the patients were mainly characterized with Klinefelter's syndrome. Y chromosome microdeletions were detected in 109 (14.97 %) of the 728 cases of azoospermia or oligozoospermia. The most common microdeletion of Y chromosome was AZFc (62.39%) and these patients were characterized with azoospermia and oligozoospermia. Five patients (4. 59%) who suffered Y chromosome microdeletion in AZFa region and AZFb region were characterized with azoospermia. Fifteen cases (13.76%) with microdeletion in AZFb region and AZFc region were mainly characterized with azoospermia. There were 6 cases (5. 50 % ) of microdeletion in AZFa, AZFb and AZFc regions,these patients were all characterized with azoospermia. Conclusions Both Chromosome abnormalities and Y chromosome microdeletions are important causes for male infertility.     

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。     

甘肃省不育症患者Y染色体微缺失的临床研究

<正>全球约有15%的夫妻受到不孕不育的困扰,其中约50%归于男性因素~([1-3])。男性不育的原因较为复杂,遗传因素是其中重要而又相对明确的病因,染色体核型和Y染色体微缺失是目前临床上遗传因素最常见的检查指标。Y染色体是人体最短的近端     

无精子症和重度少精子症不育患者Y染色体微缺失率的临床研究

目的对本院男性不育患者的Y染色体长臂上的AZF区进行微缺失检测,研究Y染色体AZF区的微缺失与男性患者不育的相关性。方法回顾性分析2012年4月至2014年4月来我院进行诊治的296例无精子症患者和320例重度少精子症患者,比较Y染色体在无精子症和重度少精子症患者中的缺失率。结果 616例不育男性患者中检出69例AZF区不同程度微缺失,缺失率为11.6%。无精子症患者发生的缺失概率高于重度少精子症患者,无精子症患者中AZF区缺失率为15.2%,重度少精子症患者AZF区缺失率为7.5%。AZF区缺失高频发生于AZFc区,占总缺失的60.9%。结论 Y染色体微缺失在无精子症和重度少精子症患者发生率较高,进行辅助生殖治疗前应进行微缺失检测。     

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、...     

男性不育病人Y染色体AZFc/DAZ微缺失的实时荧光PCR筛查

目的 :建立一套Y染色体微缺失的实时荧光PCR筛查方法 ,对无精子症或少精子症男性不育病人进行Y染色体AZFc/DAZ微缺失的常规筛查。　方法 :采用实时荧光PCR对进行卵细胞胞质内单精子注射 (ICSI)治疗的 87例无精子症和少精子症病人以及睾丸活检的 30例无精子症病人进行Y染色体AZFc/DAZ微缺失的检测 ,同时用多重PCR作为方法学对照。　结果 :AZFc/DAZ缺失 11例 (9.4%) ,其中 6 1例少精子症病人中发现 4例(6 .6 %) ,5 6例无精子症病人中发现 7例 (12 .5 %)。　结论 :Y染色体AZFc/DAZ微缺失的实时荧光PCR筛查方法是易行和可靠的 ,与多重PCR筛查结果完全一致。     

中药生精冲剂对生精障碍小鼠治疗作用的实验研究

目的:研究生精冲剂对环磷酰胺所致生精障碍小鼠的治疗作用。方法:46只雄性昆明小鼠随机分为正常组(n=10)、模型组(n=12)、对照组(n=12)和中药组(n=12),后3组腹腔注射环磷酰胺,建立生精障碍模型。造模完成后,中药组和对照组分别灌胃中药生精冲剂[16 g/(kg.d)]和克罗米芬[21.6 mg/(kg.d)],正常组与模型组每天灌胃等量生理盐水,连续15 d。次日,测量睾丸重量,并计算睾丸指数;放射免疫法测定血清FSH、LH、T水平;对睾丸组织进行组织学观察。结果:中药组小鼠血清T[(7.046±0.291)nmol/L]、FSH[(2.947±0.587)mIU/ml]、LH[(3.254±0.492)mIU/ml]、睾丸指数[(3.958±0.342)g/kg]与模型组各检测值相比[(6.231±0.317)nmol/L、(5.428±0.719)mIU/ml、(5.155±0.460)mIU/ml、(3.525±0.462)g/kg]差异有显著性(P<0.05)。组织学观察中药组睾丸生精小管生精细胞层数及管腔内精子数明显高于模型组。结论:生精冲剂治疗小鼠生精障碍有效。     

精子发生阻滞不育患者睾丸雄激素受体和热休克蛋白90α的表达

目的 :探讨精子发生阻滞与雄激素受体 (AR)和热休克蛋白 90α(HSP90α)表达的关系。　方法 :应用免疫组化二步法 ,检查 5 7例精子发生阻滞引起的不育患者睾丸活检标本AR和HSP90α的表达 ,并以 15例正常健康人睾丸组织作为对照。　结果 :在正常健康人睾丸组织中 ,AR在精原细胞、精母细胞、圆形精子细胞、支持细胞、间质细胞和肌样细胞的胞核均有表达 ,但各类细胞表达AR的强度有差异。在精子发生阻滞的睾丸组织中 ,AR高表达主要在阻滞细胞水平的胞质 ,即在核周形成一环行特异性免疫反应阳性产物带。HSP90α在正常睾丸组织精原细胞、精母细胞、支持细胞、间质细胞和肌样细胞的胞质表达 ;在精子发生阻滞的睾丸组织中 ,HSP90α为高表达 ,正常睾丸组织和精子发生阻滞的睾丸组织 ,其表达强度存在显著性差异 (P <0 .0 5 )。　结论 :AR表达部位异常可能无法介导雄激素进入核内 ,而不能实现对基因转录或翻译的调节 ,致使生精细胞难以进入细胞周期而呈现阻滞。HSP90α的高表达可能加剧AR稳定性降低 ,进而增强了AR的遍在化反应。     

男性不育患者生精细胞HCMV、HSV感染检测及形态学研究

目的:探讨男性不育患者精液中生精细胞人类巨细胞病毒(HCMV)、单纯疱疹病毒Ⅱ型(HSV-Ⅱ)感染状况及其形态学特征。方法:收集83例精液检查中发现较多生精细胞男性不育患者精液,洗涤收集细胞后提取DNA,用PCR方法进行HCMV、HSV-Ⅱ筛查,阳性者精液细胞涂片用免疫细胞化学法(ICC)做相应的病毒标记,并另行细胞学染色作形态学观察。结果:83例精液样本PCR检测HCMV阳性8例,HSV-Ⅱ阳性4例,未发现2种病毒同时阳性病例。8例HCMV阳性病例ICC标记生精细胞HCMV晚期抗原均见阳性表达,HCMV早期抗原均为阴性表达;4例HSV-Ⅱ阳性病例ICC标记生精细胞3例阳性表达,1例弱阳性。12例病毒阳性病例均发现较多未成熟生精细胞,并有不同程度的核染色质固缩、出现空泡、核膜破损、核崩解、凋亡小体等凋亡现象,但未能找到病毒感染的特异性形态学改变(如病毒包涵体)。病毒感染阳性组精子密度明显低于阴性组(P<0.05)。结论:生精细胞HCMV、HSV-Ⅱ感染时可出现不同程度的病理性损害,影响生精功能,感染的生精细胞形态学表现为出现凋亡现象等病理性改变;不育男性患者精液中出现病理性生精细胞,其生精细胞HCMV感染率可能较高。     

精子发生阻滞不育患者睾丸雌激素受体α的表达

目的:探讨雌激素受体α(ERα)在睾丸的表达与精子发生阻滞的关系。方法:用HE染色技术检查120例不育症患者睾丸活检标本,其中精子发生阻滞患者标本应用免疫组化法检查ERα的表达,并以10例健康人睾丸组织作为对照。结果:120例标本中符合精子发生阻滞病理诊断标准者有31例(占25.8%)。正常健康人睾丸组织中,ERα在睾丸支持细胞、类肌细胞和间质细胞的细胞核内表达,生精上皮细胞不表达;而在精子发生阻滞的睾丸组织中,ERα也在睾丸支持细胞、类肌细胞和间质细胞的细胞核内表达,但表达较弱,生精上皮细胞不表达。两者表达强度存在显著性差异(P<0.05)。结论:雄激素与雌激素通过各自的受体发挥协调作用而共同促进精子发生。精子发生阻滞可能与包括雄激素受体、ERα及热休克蛋白90在内的一系列复杂的细胞信号转导障碍有关。     

男性不育患者生精细胞HCMV、HSV感染检测及形态学研究

目的：探讨男性不育患者精液中生精细胞人类巨细胞病毒（HCMV）、单纯疱疹病毒Ⅱ型（HSV-Ⅱ）感染状况及其形态学特征。方法：收集83例精液检查中发现较多生精细胞男性不育患者精液，洗涤收集细胞后提取DNA，用PCR方法进行HCMV、HSV-Ⅱ筛查，阳性者精液细胞涂片用免疫细胞化学法（ICC）做相应的病毒标记，并另行细胞学染色作形态学观察。结果：83例精液样本PCR检测HCMV阳性8例，HSV-Ⅱ阳性4例，未发现2种病毒同时阳性病例。8例HCMV阳性病例ICC标记生精细胞HCMV晚期抗原均见阳性表达，HCMV早期抗原均为阴性表达；4例HSV-Ⅱ阳性病例ICC标记生精细胞3例阳性表达，1例弱阳性。12例病毒阳性病例均发现较多未成熟生精细胞，并有不同程度的核染色质固缩、出现空泡、核膜破损、核崩解、凋亡小体等凋亡现象，但未能找到病毒感染的特异性形态学改变（如病毒包涵体）。病毒感染阳性组精子密度明显低于阴性组（P〈0．05）。结论：生精细胞HCMV、HSV-Ⅱ感染时可出现不同程度的病理性损害，影响生精功能，感染的生精细胞形态学表现为出现凋亡现象等病理性改变；不育男性患者精液中出现病理性生精细胞，其生精细胞HCMV感染率可能较高。     

AZFc区部分缺失与男性不育的关系

Y染色体长臂无精子症因子(AZF)的微缺失是导致男性不育的一个重要原因。在AZF微缺失中以AZFc区的全部缺失(b2/b4缺失)最为常见,近年来的研究发现AZFc区的缺失种类繁多,不仅有AZFc区的全部缺失,还存在AZFc区部分缺失(gr/gr缺失,b1/b3缺失,b2/b3缺失),并且部分缺失也可能与男性不育有关。本文旨在对AZFc区部分缺失的研究进展及与男性不育的关系做一综述。     

Testis histopathology in boys with cryptorchidism correlates with future fertility potential

PURPOSE: We studied the ability of testis biopsy to predict eventual fertility potential. MATERIALS AND METHODS: A total of 25 patients with unilateral undescended testis and 11 with bilateral undescended testes who previously underwent orchiopexy at a median age of 9 years were followed until after age 18 years. Semen analysis, testicular volume, follicle-stimulating hormone, luteinizing hormone, testosterone and inhibin B were compared to previously obtained biopsies of each testis. Biopsies were graded as mild, moderate or severe histology based on published data for normal and undescended testis histology. RESULTS: Patients with unilateral undescended testis and moderate histology were more likely to have a sperm density of greater than 20 million per cc than those with severe histology (p = 0.006), although no difference in hormonal parameters was noted. Patients with bilateral undescended testes and moderate histology were more likely to have normal follicle-stimulating hormone and inhibin B than those with severe histology (p = 0.05 and 0.002, respectively), although no difference in semen analysis was observed. Overall 7 of the 9 patients with bilateral undescended testes had less than 20 million sperm per cc. CONCLUSIONS: Testis biopsy provides useful prognostic information about semen analysis in patients with unilateral undescended testis. In the 9 patients with bilateral undescended testes testis biopsy divided them into normal and abnormal groups in terms of hormonal parameters. Testis biopsy was not able to provide additional prognostic information on semen analysis in patients with bilateral undescended testes since they were uniformly poor.     

原癌基因在生精细胞中的表达及其作用

睾丸生精细胞中有许多原癌基因特异表达。在正常睾丸生精过程中,原癌基因的正常表达对精原细胞的有丝分裂、精母细胞的减数分裂及精子变形、成熟有重要的调控作用。     

Mast cells in testicular biopsies of infertile men with 'mixed atrophy' of seminiferous tubules

Mast cells in the bilateral testicular biopsies of 30 patients with a 'mixed atrophy' of seminiferous tubules were analysed. Seven biopsies from vasectomized patients served as controls. With regard to their characteristic location within testicular tissue, two groups of mast cells could be distinguished, in both control and infertile patients: 'interstitial' mast cells (located between Leydig and other interstitial cells as well as in the vicinity of blood vessels) and 'peritubular' mast cells (located in the close proximity of the tubular lamina propria or incorporated in the lamina propria itself). Morphometric data indicated a significant increase in the number and volume of mast cells in infertile patients when compared with controls. In the biopsies of infertile patients that were analysed both 'interstitial' and 'peritubular' mast cells showed a significant increase in their number and volume, although it appeared that 'peritubular' mast cells increased at a higher rate than 'interstitial' mast cells. A significant negative correlation was found between the following variables: volume and number of mast cells, testis volume and the status of spermatogenesis evaluated by Johnsen's scoring. It was concluded that the increased presence of mast cells is closely associated with an impairment of spermatogenesis.     

AZF与男性不育研究进展

AZF微缺失的发生率在1%～55%之间不等。AZF基因缺失与严重的生精功能障碍密切相关,是最常见的导致严重少弱精子症和无精子症的分子遗传学因素。对AZF及其相关基因的研究,可从分子水平阐明精子发生障碍的机制,对男性不育症的诊断、治疗及预后判断具有非常重要的意义。本文综述了AZF基因结构和功能特点,及其与男性不育、隐睾、精索静脉曲张、Klinefelter综合征、精原细胞瘤、习惯性流产等之间的关系。