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。     

2230对生殖异常夫妇男性染色体异常因素分析

目的探讨生育异常夫妇中男性染色体异常出现的频率和类型及少精子症或无精子症患者Y染色体微缺失出现的频率和类型。方法对2230对生育异常夫妇中男性外周血淋巴细胞培养,G显带,核型分析;对其中432例少精子症或无精子症患者外周血提取DNA,通过实时荧光定量PCR进行Y染色体微缺失检测。结果 2230例男性中异常核型75例,异常检出率3.36%(75/2230),432例少精子症或无精子症男性检测出32例存在Y染色体AZF区域缺失,检出率7.41%(32/432)。结论对生育异常的男性进行核型分析,同时对少精子症或无精子症男性进行Y染色体微缺失的检测有助于病因的诊断及遗传咨询,进行生育指导。     

多重聚合酶链反应检测Y染色体无精子因子微缺失

目的:研究原发性无精、严重少精症与Y染色体无精子因子(AZF)微缺失之间的关系.方法:采用多重聚合酶链反应技术对103例原发无精子症、72例原发严重少精症患者及60例正常生育男性进行AZFa、AZFb、AZFc 3个区域微缺失分析.结果:60例正常生育男性未发现Y染色体AZF区域微缺失,175例生精障碍患者中发现AZF微缺失19例,总缺失率为10.9%.其中11例无精症患者和4例少精症患者的缺失发生在AZFc区域,缺失率为8.6%;1例无精症患者和2例少精症患者发生AZFb、AZFc双重缺失,缺失率为1.7%;1例无精症患者发生AZFa、b、c 3个区域同时微缺失,缺失率0.6%.生精障碍组与正常生育男性组比较Y染色体AZF区域微缺失率差异有统计学意义(P<0.01).结论:Y染色体AZF区域微缺失是引起男性无精、少精子症的重要原因之一.采用多重聚合酶链反应技术对原发无精、少精子症患者在单精子注射(ICSI)之前进行微缺失筛查是必要的.     

大Y染色体与精子质量及ICSI妊娠结局的探讨

目的 探讨大Y染色体核型在男性生殖中的临床效应.方法 对严重男性不育患者行外周血染色体核型分析和ICSI-ET助孕.结果 严重男性不育患者中大Y染色体核型的枪出率为10.91%,但大Y核型的无精子症患者经睾丸穿刺均可获精子;大Y与正常核型患者经ICSI移植后的累计妊娠率差异无统计学意义(P＞0.05);大Y与正常核型患者行ICSI移植后新生儿性别比例无统计学差异.结论 男性精子异常与大Y染色体有关,大Y中DNA序列高度重复基因并非完全抑制精子的发生,且不影响ICSI移植后的受精卵发育及着床;通过睾丸穿刺取精行ICSI助孕并未对大Y精子的受精造成不利影响.     

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.     

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。     

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。     

原发性不育患者染色体核型与AZF基因分析

目的建立针对男性不育症的遗传学筛查的诊断方法.方法采用外周血染色体G显带、PCR 技术以及琼脂糖电泳,对255例无精子症和少精子症患者进行外周血染色体核型分析及无精子因子(AZF)基因的6个位点分析.结果255例患者中31例染色体核型异常,占12.16%,在无精子症中染色体异常发生率为17.36%,少精子症和严重少精子症患者染色体异常发生率为7.46%;36例AZF 基因有微缺失,占14.12%,其中无精子症缺失率为16.53%,少精子症缺失率为9.68%,严重少精子症缺失率为13.89%.对照组30例已生育男性标本未检查出染色体核型异常和AZF缺失.结论无精子症和少精子症的发生与遗传缺陷密切相关,应对男性不育症被诊断为无精子症或严重少精子症的患者进行遗传学筛查.     

Y染色体微缺失分子诊断的意义（附122例报告）

目的建立Y染色体无精子因子（azoospermia factor，AZF）微缺失分子诊断的方法。方法实验组选择122例无精子症及少精子症患者，对照组90例为具有正常生育能力的已婚男性。按照欧洲男科协会（EAA）和欧洲分子遗传实验质控网（EMQN）指南选择Y染色体序列标签点和设计引物。取受试者外周血，抽提DNA，进行多重PCR，筛查AZFa、b、C区域的微缺失。结果采用2组多重PCR，122例生精障碍患者中发现Yq AZF微缺失13例（10．06％），共5种缺失类型，其中AZFa1例，AZFb 1例，AZFc 8例，AZFbc 2例，AZFabc 1例。对照组90例未发现微缺失。两组结果经统计学分析，差异有显著意义（P〈0．01）。结论EAA和EMQN推荐的指南对Y染色体AZF微缺失筛查有指导作用。无精子症及少精子症患者选择辅助生殖技术生育子代之前，进行Y染色体AZF微缺失的分子遗传学筛查应为常规检查项目。     

大、小Y染色体携带者精液参数及辅助生殖结局分析

目的:探讨Y染色体长度异常对精液参数以及对ART结局的影响,以期为临床工作提供指导。方法:963例辅助生育患者按Y染色体长度分为正常对照组(n=854),Y18组(n=24)及Y22组(n=85),并将Y18组和Y22组的精液参数及在ART过程中的胚胎数及优质胚胎数与正常对照组进行比较。用χ~2检验及t检验进行统计学分析。结果:Y18组中弱精子症患者比例明显低于正常对照组(8.33%vs 31.03%),优质胚胎数较对照组多,差异均有显著性(P0.05);Y22组中无精子症比例明显高于正常对照组(16.47%vs 1.87%),在ICSI过程中胚胎数多于正常对照组,差异均有显著性(P0.05),Y18和Y22两组妊娠率与对照组统计学差异均无显著性(P0.05)。结论:小Y染色体可能对精子发生有影响,而携带大、小Y染色体不会降低ART的妊娠率。因此,对于大、小Y染色体携带不育患者,进行辅助生殖技术可获得和正常核型类似的妊娠结局。     

Sperm from Spinal Cord Injured Men Lose Motility Faster than Sperm from Normal Men: The Effect is Exacerbated at Body Compared to Room Temperature

Purpose

We investigated if sperm from spinal cord injured men lose motility and viability faster than sperm from normal men, and if there is an effect of temperature on these parameters.

Materials and Methods

Identical aliquots of fresh antegrade semen from 10 spinal cord injured men and 9 normal controls were stored at room temperature (23C) or body temperature (37C). Sperm motility and viability were measured at 0, 2, 4, 6 and 8 hours after semen collection.

Results

Sperm motility, particularly rapid linear motility, decreased faster in specimens from spinal cord injured men compared to normal controls. This effect was exacerbated at 37C. Heat did not affect the degradation of sperm motility in specimens from normal men. The percent of dead sperm in specimens from spinal cord injured men was significantly greater than that in normal men at all time points. However, the rate of cell death was not faster than normal with time or heat.

Conclusions

When using sperm from spinal cord injured men for purposes of insemination or experimentation the rapid loss of motility must be considered, particularly when stored at body temperature.     

Relation between Semen Quality and Fate of Pregnancy: Retrospective Study on 534 Pregnancies

Full semen analysis including sperm and seminal plasma (SP) was performed on 534 men, within 3 months of their wives conception. 65% of the pregnant wives eventually delivered, while the remainder aborted. The abortions were divided into five groups: early abortion (6%), late abortion (3%), repeated abortion (14%), and habitual abortion (12%). SP quality of the husbands of the various groups were identical. The motility and vitality of spermatozoa of the various groups were in the same range. Sperm concentration was significantly higher in the repeated and habitual abortion groups with a tendency to polyzoospermia. The morphology of spermatozoa was much better in these two groups. Generally, sperm quality of the men whose wives were repeated or habitual aborters was better than in the other groups. We could not find any evidence in the routine semen analysis indicating that sperm quality was responsible for the abortions. The reason for the abortions seems to be a female factor or chromosomal aberrations.     

Sperm chromatin integrity may predict future fertility for unexplained recurrent spontaneous abortion patients

The pathogenesis of recurrent spontaneous abortion (RSA) is multi-factorial, complex and poorly understood. In the present study, semen parameters, including sperm chromatin integrity, sperm concentration, sperm motility and sperm morphology, were compared between 111 men whose partners had a history of unexplained RSA (RSA group) and 30 healthy fertile men (control group). The RSA group was further separated into three subgroups, depending on their reproductive outcome during the 12?months after they were enrolled in the study: the pregnancy subgroup consisted of 43 men whose partners achieved a successful pregnancy up to at least the 24th week of gestation; the abortion subgroup included 31 men whose partners experienced further abortions; and the infertile subgroup had 37 men whose partners did not have any positive pregnancy test after regular, unprotected intercourse. Significantly lower proportion of sperm with normal morphology was found in the abortion subgroup (14.7?±?4.3%) than in the control group (17.5?±?5.0%). Sperm concentrations were significantly lower in the infertile subgroup (55.7?± 24.1%) than in the controls (68.6?±?27.8%). The rates of abnormal sperm chromatin integrity were significantly higher in the abortion (16.7?±?7.7%) and infertile (16.3?±?6.6%) subgroups, compared to the control group (13.0?± 4.4%). Logistic regression analysis showed that the subsequent reproductive outcome of the 111 RSA patients was negatively correlated to the rates of abnormal sperm chromatin integrity. In conclusion, sperm chromatin integrity, sperm morphology, and sperm concentration were associated with future reproductive outcome of RSA patients. The sperm chromatin integrity was a significant predictor for future abortion and infertility.     

Evidence for association of sex hormone-binding globulin and androgen receptor genes with semen quality

The roles of androgen receptor AR(CAG)n gene polymorphisms and sex hormone-binding globulin SHBG(TAAAA)n gene polymorphisms on semen quality were studied. One hundred fourteen men were included in the study: 85 with normal sperm count and 29 oligospermic. The genotype analysis, on DNA extracted from spermatozoa, revealed five SHBG(TAAAA)n alleles with 6–10 repeats and 18 AR(CAG)n alleles with 12–32 repeats. The SHBG allelic distribution showed that in men with normal sperm count and motility, those with short SHBG alleles had higher sperm concentration than men with long SHBG alleles ( P  = 0.039). As concerns AR(CAG)n polymorphisms, men with short AR alleles had lower sperm motility compared to those with long AR alleles ( P  < 0.001) in both total study population and normal sperm count men. The synergistic effect analysis of the two polymorphisms revealed an association between sperm motility ( P  = 0.036), because of the effect of AR(CAG)n polymorphism on sperm motility. In conclusion, long AR alleles were found to be associated with higher sperm motility, while short SHBG alleles were associated with higher sperm concentration, supporting the significance of these genes in spermatogenesis and semen quality.     

Neutralization of cytokine activity at the receptor level improves sperm motility in men with spinal cord injuries

Men with spinal cord injury (SCI) have a unique semen profile characterized by normal sperm concentrations but abnormally low sperm motility. Previous studies showed that elevated concentrations of cytokines in the seminal plasma of these men contribute to this condition. For example, when elevated concentrations of interlekin-1beta (IL-1beta), IL-6, and tumor necrosis factor-alpha (TNF-alpha) were immunoneutralized in the semen of men with SCI, sperm motility improved. The present study investigated if these cytokines act on sperm cell receptors to inhibit sperm motility. Semen was collected from men with SCI and from healthy non-SCI men. Sperm were separated from the seminal plasma by centrifugation. Eight identical aliquots of 5,000 sperm suspended in 50 microL of seminal plasma were prepared for each subject. Agents were added to the aliquots in order to neutralize IL-1beta, IL-6, and TNF-alpha at the receptor level. In SCI subjects, sperm motility improved in each treatment group compared with the untreated group, but statistical significance was reached only when neutralizing agents to all 3 cytokines were added. Improvement was less pronounced in subjects with close to normal semen cytokine concentrations or close to normal pretreatment sperm motility. In control subjects, IL-1beta, IL-6, and TNF-alpha were within normal values, and addition of receptor blockers to semen had no effect on sperm motility. These data support the hypothesis that cytokines act at the level of the sperm receptor to inhibit sperm motility. These data further support the notion that inactivating semen cytokines leads to improved sperm motility in SCI men. Our goal is to develop this finding into a treatment for low sperm motility in men with SCI.     

Semimal plasma testosterone and dihydrotestosterone levels in men with varicoceles

Testosterone (T) and dihydrotestosterone (DHT) were measured in the seminal plasma of men with varicoceles, azoospermic vasectomized men and men with normal seminal parameters. Testosterone levels were not different significantly in these groups of men. DHT concentrations and the DHT T ratio were significantly lower than normal in men with varicoceles and sperm densities below 30 million/ml and the vasectomized men, but not different from normal in the men with varicoceles whose sperm densities were greater than 30 million/ml. In men whose sperm density and/or motility improved after varicocelectomy, there was an increase in seminal plasma DHT levels and the DHT/T ratio. When seminal parameters were not improved by varicocelectomy, there was no change in DHT levels or in the DHT/T ratio. It is concluded that men with varicoceles and sperm densities below 30 million ml have a deficiency in the epididymal 5α-reduction of testosterone.     

Effects of clinical stage and immunological status on semen analysis results in human immunodeficiency virus type 1-seropositive men

Summary. Complete semen analyses including computer-assisted sperm motility and morphology assessments were performed to determine if semen and sperm differed between HIV-seropositive men and fertile controls, or differed with symptoms, or CD4+ peripheral cell count categories. Previous studies included small numbers of men and presented conflicting conclusions. Two hundred and fifty non-vasectomized HIV-seropositive men and 38 fertile controls each provided one semen sample. Non-parametric statistics were used to analyse both continuous and nominal data. Fertile men had significantly greater semen volume, sperm concentration, percent motility, percent rapid and linear motility and total strictly normal spermatozoa than HIV seropositive men. Neither total number nor subtypes of leukocytes in semen differed between the two groups. Among the HIV seropositive men, significant differences in semen analyses were found between CD4+ cell count, clinical, and AIDS categories. Lower CD4+ cell counts (<200 mm⁻³) were associated with significantly lower percent motility, percent normal sperm morphology by strict criteria, significantly more spermatids in semen, and higher percentages of teratozoospermia, oligoasthenoteratozoospermia and leukocytospermia. Healthier men, based on clinical categories, had significantly more normal shaped spermatozoa and fewer had azoospermia, oligoasthenoteratozoospermia or leukocytospermia. Many HIV-seropositive men have normal semen analyses, but as the disease progresses more defects are found, particularly in strict criteria sperm morphology.     

附睾和睾丸精子活力对卵胞浆内单精子注射结局的影响

目的比较附睾或睾丸来源及其不同活力精子行卵胞浆内单精子注射（ICSI）的结局。方法回顾性分析2005年1月至2008年5月在本生殖中心经皮附睾精子抽吸术（PESA）、睾丸精子抽吸术（TESA）助孕的218例无精子症患者的资料,比较附睾和睾丸及其不同活力精子的正常受精率、卵裂率、优质胚胎率、临床妊娠率、种植率和早期流产率。结果附睾精子组与睾丸精子组比较,正常受精率、卵裂率、临床妊娠率、种植率和早期流产率无显著性差异（P〉0．05）;优质胚胎率附睾精子组显著高于睾丸精子组（P〈0．05）。附睾活动精子、睾丸活动精子和睾丸不活动精子的正常受精率显著高于附睾不活动精子（P〈0．01）;睾丸活动精子的正常受精率显著高于睾丸不活动精子组（P〈0．05）;以上各组间的优质胚胎率、临床妊娠率、种植率、流产率均无显著性差异（P〉0．05）。结论PESA操作简单且不影响妊娠率,无精子症患者行ICSI治疗时可首选附睾精子;附睾或睾丸不活动精子影响ICSI的受精率,应优先选择活力较好的精子,若无活动精子则选择睾丸不活动精子。     

Semen quality changes among 2343 healthy Slovenian men included in an IVF-ET programme from 1983 to 1996

To determine whether semen quality in Slovenians has changed over 14 years (1983-96), we analysed retrospectively the semen of 2343 healthy men with a normal spermiogram, who were partners of women with tubal infertility included in the IVF-ET programme. Age at semen collection, duration of sexual abstinence, semen volume, sperm concentration, total sperm count, percentage of spermatozoa with progressive motility, and normal morphology were determined. Multiple regression analysis was used to assess the changes in sperm characteristics according to the year of semen collection, year of the man's birth and the duration of sexual abstinence. Semen volume, sperm concentration, sperm count and total sperm motility did not change between 1983 and 1996, whereas between 1988 and 1996 rapid progressive sperm motility decreased by 0.95% per year (p < 0.0001). Semen volume, sperm concentration, and sperm count increased with duration of sexual abstinence. After adjustment for the year of semen collection and duration of sexual abstinence, multiple regression analysis showed that sperm concentration decreased by 0.67% per each successive year of birth (p = 0.03). Thus the sperm concentration decreased from 87.6 x 10(6)/mL in men born in the 1940s to 77.3 x 10(6)/mL in those born between 1956 and 1960. After 1960, sperm concentration was found to increase. In 2343 healthy men, no decline in semen quality, except in rapid progressive motility, was observed in the study period. Lower sperm concentration was found among men born between 1950 and 1960. This could be related to worse socio-economic status, stress or negative environmental factors in this time period.     

人精子透明质酸酶活性测定的临床意义

目的 :测定人精子透明质酸酶 (HYD)活性并分析其与精液常规参数之间的相关性。　方法 :用改良Singer法测定 14 6例男性精子HYD活性 ,常规检测精子密度、活动率和正常形态百分率。　结果 :精子HYD活性与密度、活动率及与正常形态百分率存在显著相关性 (r值分别为 0 .65、0 .63和 0 .72 ,P均 <0 .0 1)。不育男性精子密度在 4 0× 10 6 /ml以上组的HYD活性明显高于精子密度少于 2 0× 10 6 /ml以下组 (P <0 .0 1)。不育男性精子活动率 >60 %组的HYD活性明显高于 <3 0 %组 (P <0 .0 1)。　结论 :精子HYD活性测定是评价精子功能的有效指标之一