男性不育症患者外周血白细胞雄激素受体的改变及意义

目的：探讨男性不育与外周血白细胞雄激素受体表达的关系。方法：根据精液分析及睾丸活检病理检查结果,将67例男性不育症患者分成少精子症组(n=21)、弱精子症组(n=15)、阻塞性无精子症组(n=14)和非阻塞性无精子症组(n=17),采用放射配体结合分析法检测不育症患者外周血白细胞雄激素受体(AR),同时采用放射免疫法检测血清睾酮(T)和雌二醇(E_2)水平,并以22例正常生育男性为对照。结果：外周血白细胞AR含量少精子症组、弱精子症组、阻塞性无精子症组与对照组比,均无统计学差异(P＞0．05)；非阻塞性无精子症组(782±98)与对照组(913±104)相比,差异有统计学意义(P＜0．01)。男性不育症各组与对照组相比,E_2和T水平差异均无统计学意义(P＞0．05)；而男性不育症患者精子密度与白细胞AR含量呈正相关(r=0．233,P=0．010)。结论：一部分男子不育的发生、发展过程可能与AR表达下调有密切关系。     

不育男性精浆左旋肉碱浓度与精子活力和浓度的关系

目的:探讨精浆左旋肉碱测定在男性不育症诊疗中的临床意义。方法:按照第5版《世界卫生组织人类精液检查与处理实验室手册》的参考值,将不育男性按照精液常规分析结果分为精子活力正常组(前向运动精子百分率≥32%)(n=283)和弱精子症组(前向运动精子百分率32%)(n=892)。通过比色法检测精浆中的游离左旋肉碱含量,分析左旋肉碱浓度与精子活力、精液浓度的相关性。通过受试者操作特征分析曲线(receiver operating characteristic curve,ROC curve)确定左旋肉碱浓度的阈值,以阈值为分界点,将弱精子症组患者分为高于左旋肉碱阈值组和低于左旋肉碱阈值组,分析左旋肉碱与精子活力和精子浓度的相关性。结果:弱精子症组的精浆左旋肉碱浓度(384.14±188.81μmol/L)显著低于精子活力正常组(434.04±171.77μmol/L,P0.05)。精子活力正常组和弱精子症组精浆中的左旋肉碱含量与前向运动精子百分率和精子浓度的相关性极低或不相关(r0.2)。肉毒碱检测的ROC曲线下面积(AUC)为0.592,阈值为380.9,低于左旋肉碱阈值弱精子症组与前向运动精子百分率有较弱的正相关关系(r=0.329,P=0.000)。结论:对于拟行辅助生育的不育男性患者,精浆中左旋肉碱浓度作为精浆生化的指标之一,可能对弱精子症患者有一定的临床参考意义。     

人精子中芳香化酶表达与精子功能的关系

目的:研究精子细胞色素P450芳香化酶(P450arom)的表达及其与精子功能状态和受精力的关系。方法:以人精子穿透去透明带金黄地鼠卵异种体外受精试验(SPA)检测精子受精力;以三色法染色观察精子顶体反应(AR)的发生率。采用RT-PCR,用P450arom/GAPDH光密度值的比值代表P450arom的表达水平。结果:精子P450arom表达水平与受精率有一定的相关性(生育组r=0.5622;不育组r=0.6071)。正常男性与不明原因不育症患者精子P450arom/GAPDH比值分别为0.60±0.29,0.39±0.16,有显著差异(P<0.02)。P450arom表达水平与精子AR的发生率也有一定相关性(生育组r=0.5817;不育组r=0.5535)。结论:人精子中存在着P450arom表达产物;P450arom可能与精子功能有一定关系;P450arom表达异常可能与一些不明原因不育有关。     

整合素配体玻连蛋白在人精子的表达及其与受精的关系

目的 :进一步研究整合素配体玻连蛋白 (Vn)在人精子的表达及其与受精的关系。方法 :选用 1 4例生育力正常的成年男性及 8例精液常规分析正常的不明原因不育男性患者的精液标本 ,液化后提取上游精子。精子体外获能后用兔抗人 Vn多克隆抗体及羊抗兔 Ig G-FITC行免疫染色。然后用流式细胞仪计数 Vn表达阳性精子百分数。部分获能精子同时与去透明带金黄地鼠卵行异种体外受精 (SPA)以检测其受精力 ,比较两组获能精子表面Vn表达阳性精子百分率及受精率差异并分析受精率与 Vn表达阳性的获能精子百分数之间的相关性。结果 :生育组与不育组获能精子 Vn表达水平分别为 2 1 .2 4± 1 1 .70 %与3.6 4± 3.2 7% ,不育组明显低于生育组 (P<0 .0 5)。生育组受精率大于 1 0 % ,不育组受精率小于 1 0 % ,符合划分生育力正常与异常的标准。所有标本 Vn表达阳性的获能精子百分率与精子受精率间具有相关性 (r=0 .476 )。结论 :人获能精子表面存在一定水平的整合素配体玻连蛋白表达 ;Vn参与受精过程 ;Vn表达异常可能与一些不明原因的不育有关     

染色体多态性对男性生精能力及生育结局的研究

目的探讨染色体多态性对男性生精能力和生育结局的影响。方法选择3 203例男性不育者(不育组)和4 893例捐精初筛合格者(捐精组)进行回顾性队列研究,其中根据精液精子数量将不孕组患者分为精子正常组、少精子症组、无精子症组。检查并比较各组的精液常规、染色体核型、Y染色体微缺失。对有多态性的捐精者和患者进行随访,了解他们的生育结局。结果不育组和捐精组多态性发生率分别为4.62%和3.78%,差异无统计学意义(P0.05)。不育组的Y异染色质长度减少(Yqh-)发生率(0.59%)高于捐精组(0.27%),差异有统计学意义(P=0.022),且随着精子数量减少Yqh-发生率明显增加(精子数量正常组为0.15%,少精子症组为0.22%,无精子症组为0.99%),差异有统计学意义(精子正常组与无精子症组P=0.033;少精子症组与无精子症组P=0.027)。携带有Yqh-的患者Y染色体微缺失检出率为56.25%(9/16),显著高于其它类型人群(P0.001)。不育组中弱精子者和畸形精子者的多态性发生率分别为3.92%、3.96%,同捐精组(3.78%)相比,差异无统计学意义(P0.05)。随访捐精组和不育组有多态性人群的配偶,结果其自然流产率分别为6.25%(3/48)和6.67%(2/30),差异无统计学意义(P0.05),且无1例发生子代出生缺陷。结论除Yqh-部分伴随无精子因子(AZF)缺失可导致精子发生障碍,使男性生育力下降,其它类型多态对男性生育无明显影响。     

精子核成熟度与精液参数关系研究

目的:研究精子核成熟度与精液参数关系。方法:49例精液标本,其中生育组15例,不育组34例。应用精子质量自动检测系统(CASA)进行精子密度、活力分析,伊红染色进行活率分析,联苯胺染色评价精液白细胞,采用精子形态检测系统下人工修正方法分析精子形态,用苯胺蓝染色评价精子核成熟度。结果:不育组苯胺蓝染色阳性率显著高于生育组(P<0.05)。形态异常精子组中头部异常、颈部异常、尾部异常、无定型、其它畸形精子组苯胺蓝染色阳性率均显著高于形态正常精子组(P<0.05)。苯胺蓝染色阳性精子率与形态正常精子率、活力、活率均呈显著负相关(P<0.05);苯胺蓝染色阳性精子率与精子密度、精液白细胞浓度均无显著相关性。结论:精子核成熟度异常可导致男性生育力下降,精子核成熟度是评价男性生育力重要参考指标。     

不育男性精子及精浆中乳酸脱氢酶C_4同功酶活性的研究

本文测定了97例正常及不育男性精子及精浆中的乳酸脱氢酶C_4同功酶(LDH-C_4)活性。精浆/精子中该同功酶活性的比值在正常对照、精液常规正常而不育(不育Ⅰ组)、精液常规不正常的不育(不育Ⅱ组)和少精症组中依次为0.528,0.880,2.85和4.01(中位数值)。各不育组与对照组间均差异显著(P<0.005及P<0.0005)。以mU/10~6精子表示时,不育Ⅰ和Ⅱ组精浆中的LDH-C_4活性大于正常男性;精子中的活性则小于正常男性(P<0.005及P<0.0005)。此外,精子中LDH总活性与LDH-C_4活性之间有良好的正相关(r=0.9593)。     

不育男性精子DNA碎片化指数与精液解脲脲原体及人型支原体感染关系的研究

目的:探讨解脲脲原体(Uu)和人型支原体(Mh)对不育男性精子浓度、活动力、正常形态率及精子DNA碎片化指数(DNA fragmentation index,DFI)的影响.方法:选取169份精液标本,其中129例特发不育者为不育组,40例正常生育者为对照组,计算机辅助精液分析(CASA)系统分析精子浓度和活动力.精子形态分析采用Shorr染色法,吖啶橙试验(acridine orange test,AOT)检测精子DFI.支原体检测采用培养法.结果:与对照组相比无Uu/Mh、Uu或Uu+Mh感染组精子DFI显著增高(P＜0.05),与无Uu/Mh感染组相比,Uu或Uu+Mh感染组DFI亦显著增高(P＜0.05).与正常生育对照组相比无Uu/Mh、Uu或Uu+Mh感染组正常形态率显著降低(P＜0.05),其余组间无统计学差异(P＞0.05).与正常对照组相比,无Uu/Mh、Uu、Mh和Uu+Mh感染组的精子浓度、a级、a+b级和a+b+c级精子百分比均显著降低(P＜0.05),而除无Uu/Mh感染组外,其余各组间无统计学差异(P＞0.05).结论:特发性不育男性的精子浓度、正常形态率、a级、a+b级和a+b+c级精子百分比较正常男性的均显著下降,Uu/Mh感染并非是最主要原因,可能是其他重要因素或综合因素影响的结果.特发性不育男性精子DFI明显增加,Uu是其中的主要因素之一.Uu可能主要通过对精子DNA,而不是常规参数的影响来造成男性生育力的下降.     

应用高效液相色谱法检测男性不育患者精浆丙二醛水平

目的:探讨高效液相色谱法(HPLC)检测精浆中脂质过氧化产物丙二醛(MDA)水平的意义。方法:93例不育男性分为:阻塞性无精子症组(12例);非阻塞性无精子症组(15例);少精子症组(21例);弱精子症组(19例);少弱精子症组(16例);少弱畸精子症组(10例)。18例生育男性作为对照组。采用HPLC检测生育与不育男性精浆MDA含量。结果:生育组精浆MDA值除与阻塞性无精子症组差异无显著性(P>0.05)外,与其他各组相比均有极显著性差异(P均<0.01)。各不育男性组间精浆MDA值也有统计学差异。结论:应用HPLC检测精浆MDA水平是诊断活性氧产生过高所致男性不育的一个重要指标。     

单侧隐睾史不育患者的精子参数评价

目的:评价患者施行睾丸固定手术时的年龄对其后精子参数的可能影响。方法:47例单侧隐睾史不育患者,按照患者施行单侧睾丸固定术时的年龄分为A组(2-7岁,n=23),B组(8-13岁,n=14)和C组(14-17岁,n=10)。对3组患者的精液作精液分析和精子头-尾膜的完整性检测。结果:3组患者的精液中均可观察到活动精子,精子活动率和精子浓度有很大的个体差异,畸形精子率普遍较高,精子头-尾膜完整率低。少弱畸精子症的发生率在B组(35%)和C组(50%)升高。结论:单侧隐睾史不育患者会有较差的精子参数。单侧隐睾症患者在幼年阶段尽早治疗有利于成年后的精子发生。     

Is there any need for diagnostic laparoscopy in couples undergoing intracytoplasmic sperm injection for severe male-factor infertility?

Purpose: Our purpose was to determine whether there is a need for a preliminary diagnostic laparoscopy in couples undergoing intracytoplasmic sperm injection (ICSI) because of severe male-factor infertility. Methods: In this retrospective study, the results of diagnostic laparoscopy in 342 women with a normal fertility workup undergoing ICSI were evaluated and sperm parameters were correlated with the findings at laparoscopy. Subgroups of patients were defined according to sperm quality, which was expressed as total normal motile count [TNMC = volume (ml) × concentration (10⁶/ml) × percentage progressive motility/100 × percentage normal morphology/100]. Results: When sperm morphology was evaluated according to Kruger’s strict criteria, the probability of finding pathology on laparoscopy in the normal male group (16.7%) was statistically higher than that in the group with severely abnormal sperm (1.8%; P<0.01). Conclusions: There is no need to perform a preliminary diagnostic laparoscopy in the female partner if a full workup is normal in couples with severe male-factor infertility willing to undergo ICSI.     

Sperm parameters in men with suspected infertility. Sperm characteristics, strict criteria sperm morphology analysis and hypoosmotic swelling test

OBJECTIVE: To determine the distribution of sperm abnormalities in a population of suspected infertile men presenting for the initial investigation of male factor infertility. STUDY DESIGN: Results obtained in the analysis of sperm viability, motility, conventional morphology (including 12 sperm anomalies), strict criteria sperm morphology analysis (SCSMA) and hypoosmotic swelling test (HOST) were compared in oligozoospermic (< 5.0, 5.1-10.0 and 10.1-20.0 x 10(6)/mL), normozoospermic (20.1-40.0, 40.1-100.0 and 100.1-250.0 x 10(6)/mL) and polyzoospermic (> 250.0 x 10(6)/mL) semen samples from 233 suspected infertile men. RESULTS: Percentage of sperm viability, category a and categories a plus b of sperm motility, oval-headed sperm, and normal-headed sperm according to SCSMA and HOST had a direct relationship with sperm counts (P < .001). Percentage of amorphous heads, pinheads, tapering and combined defects showed an inverse relationship with sperm counts (P < .001), whereas the percentage of large-headed sperm was highest in semen with > 40.0 x 10(6)/mL (P = .003) and of neck/midpiece defects was lowest in semen with < 10.0 x 10(6)/mL (P = .03). No significant differences were found in the percentage of small heads, double heads, round heads, partially elongated heads, cytoplasmic droplet and tail defects. Based on the cutoff points established previously for the sperm characteristics analyzed, normal values were found in semen with > 250.0 x 10(6)/mL (viability and motility), > 100.0 x 10(6)/mL (conventional morphology) and > 40.0 x 10(6)/mL (SCSMA and HOST). CONCLUSION: The incidence of defective spermatozoa is lowest in semen with the highest sperm count. However, sperm abnormalities that affect male fertility may be detected at any level of sperm density. The data indicate that an increase in any sperm abnormality should be regarded as a possible cause of decreased fertility.     

Observation of spermatozoa by a high-magnification microscope

Semen analyses are the primary tool for evaluating male infertility, as semen parameters are useful for predicting potential fertility. In the field of assisted reproductive technology (ART), the single best motile spermatozoon should be selected, especially when performing intracytoplasmic sperm injection (ICSI). In this context, the motile sperm organelle morphology examination (MSOME) was developed as a method of assessing the detailed morphology of motile spermatozoa in real time at a magnification of up to 6,300× on a video system. The use of ICSI with MSOME-selected sperm is called intracytoplasmic morphologically selected sperm injection (IMSI). IMSI improves the outcomes of ICSI. MSOME can be also applied to evaluate male infertility. Among MSOME parameters, the presence of sperm nuclear vacuoles is the most important finding. Large sperm nuclear vacuoles (LNV) are related not only to poor ART outcomes, but also to poor semen quality and sperm DNA damage, such as DNA fragmentation and chromatin condensation failure. It has been suggested that sperm head vacuoles are produced at earlier stages of sperm maturation. It is possible that the number of LNV can be decreased by surgical or medical treatment for male infertility. Therefore, the level of LNV has the potential to be used as an alternative parameter of semen quality and a new tool for evaluating the therapeutic effects of treatment in male patients with infertility.     

Varicocele is associated with abnormal retention of cytoplasmic droplets by human spermatozoa

OBJECTIVE: To determine whether varicocele is associated with retention of sperm cytoplasmic droplets in infertile men. DESIGN: Retrospective study.Setting: University infertility clinic. PATIENT(S): Nonazoospermic men with idiopathic (n = 69) and varicocele-associated infertility (n = 73), and 20 fertile controls presenting for vasectomy. INTERVENTION(S): None. MAIN OUTCOME MEASURES(S): Standard semen parameters and percentage of spermatozoa with cytoplasmic droplets on Papanicolaou smears. RESULT(S): No statistically significant differences were found between the fertile and infertile groups with respect to semen volume. Fertile controls had significantly greater mean percent sperm motility and normal morphology than infertile men. The mean percentage of sperm with residual cytoplasm was statistically significantly different in all three groups. Infertile men with varicocele had the highest percentage of sperm with cytoplasmic droplets, the next highest level being in men with idiopathic infertility and the lowest level in fertile controls (11.7 +/- 1.0, 8.1 +/- 0.9 and 3.2 +/- 0.4%, respectively, P<.0001). CONCLUSION(S): Our data show that idiopathic and even moreso, varicocele-related male infertility are conditions associated with impaired disposal of residual sperm cytoplasm by the testis and/or epididymis. These data provide a possible mechanism for the observed semen abnormalities and reduced fertility potential associated with varicocele and idiopathic male infertility.     

精子线粒体膜电位与男性精液参数和体质量指数的相关性研究

目的探讨精子线粒体膜电位(MMP)与精子常规参数及体质量指数(BMI)的相关性。方法在本院辅助生殖科行精液分析的男性患者,禁欲3~7 d后,以手淫方式获取精液样本。通过计算机辅助精液分析仪检测精液常规,改良巴氏染色检查精子形态,JC-1染色后经流式细胞仪评估MMP。结果与对照组(56.68%±11.13%)相比,弱精子组MMP(41.24%±9.71%)显著降低。MMP与BMI呈显著负相关(r=-0.25,P0.01),与精子总活力(r=0.63,P0.01)、前向运动力(r=0.64,P0.01)及正常精子形态率(r=0.37,P0.01)呈显著正相关,而与年龄、精液量、精子浓度和精子数量其余参数的相关性均无统计学意义。结论精液中精子MMP是评估精子功能的重要指标,对临床综合分析男性不育症因素具有一定的参考意义。     

Leukocytospermia is associated with poor semen quality

Semen samples from 179 infertility patients were analyzed for type and number of white blood cells (WBC) by a combination of immunologic techniques. Forty-one (23%) had more than 10(6) WBC/mL semen (leukocytospermia). Semen parameters in patients with less than 10(6) WBC/mL were similar to those of 15 fertile donors. In contrast, the leukocytospermic group had significant reductions in total sperm number, percent sperm motility, sperm velocity, motility index, and total number of motile sperm. Patients with high concentrations of monocytes/macrophages (greater than 5 X 10(5)/mL; n = 27) had significantly reduced ejaculate volume; patients with high numbers of T lymphocytes (greater than 10(5)/mL; n = 19) had a significant reduction in sperm velocity; and patients with high levels of granulocyte elastase in semen (greater than 1,000 ng/mL; n = 26) had significant reductions in ejaculate volume, total sperm number, and total motile sperm number. There was no correlation between the presence of antisperm antibodies and leukocytospermia. Our data suggest that leukocytospermia may occur frequently in male infertility patients and show that elevated levels of WBC in semen are associated with poor semen quality. This provides further rationale for white blood cell testing in semen of male infertility patients, since antibiotic or anti-inflammatory therapy may be helpful in appropriately selected cases.     

Chlamydia trachomatis and male infertility in Tunisia.

OBJECTIVE: Chlamydia trachomatis is a common sexually transmitted micro-organism. The impact of chlamydial infection on semen parameters and male fertility is controversial. Our purpose was to determine the prevalence of C. trachomatis in the male partners of infertile couples in Tunisia and to assess the relationship between chlamydial infection markers and male infertility. METHODS: Chlamydial DNA in urethral and in semen specimens was determined by using the Cobas Amplicor polymerase chain reaction (PCR) assay and chlamydial immunoglobulin G (IgG) antibodies were measured by micro-immunofluorescence in serum samples in 92 male partners, with or without pathological standard semen parameters, according to the guidelines of the World Health Organization (sperm count, progressive sperm motility, sperm morphology and sperm viability). In parallel, chlamydial infection markers in endocervical material were determined by PCR and chlamydial IgG antibodies were measured by micro-immunofluorescence in serum samples from the female partners of the patients. RESULTS: C. trachomatis was found in 35.9% (33/92) of the male partners of the infertile couples and in 38% (35/92) of their female partners. There was a significant correlation between the detection of C. trachomatis in both partners (p = 0.004). C. trachomatis DNA was detected in 18.5% (17/92) of urethral specimens and in 16.3% (15/92) of semen specimens. Chlamydial IgG antibodies were present in 9.8% (9/92) of the serum samples. A standard semen analysis showed that 88% (81/92) were pathological. Sperm viability, progressive sperm motility, morphology and sperm concentration were abnormal in 73.8%, 70.2%, 34.5% and 13%, respectively, of the 92 evaluated semen specimens. Comparison of the parameters of the standard semen analysis between the male partners of the infertile couples with or without chlamydial infection markers showed that only the presence of C. trachomatis DNA in semen samples can affect sperm motility. Parameters of the standard semen analysis were not significantly related either to the detection of chlamydial DNA in urethral samples or to the presence of serum chlamydial antibodies. CONCLUSION: Our results show that C. trachomatis seems to be widespread among the male partners of infertile couples in Tunisia and show that this organism can affect sperm motility and, thus, can play an important role in the etiology of male infertility.     

Sperm DNA fragmentation and male fertility: a retrospective study of 5114 men attending a reproductive center

PurposeThe sperm DNA fragmentation index (DFI) was quantitatively measured and its relationship with age, semen quality, and infertility conditions was investigated.MethodsSemen routine test and sperm DFI were performed in 2760 infertile male and 2354 male whose spouse experienced at least one unexplained miscarriage to analyze the correlation between sperm DNA damage, semen routine parameters, and age.ResultsSperm DFI was significantly lower from patients whose wife experienced unexplained miscarriage compared to infertility males (p = 0.000). An inverse correlation between sperm DFI and sperm progressive motility was observed (r_s = − 0.465, p = 0.000) and sperm DFI was positively correlated with age (r_s = 0.255, p = 0.000). However, the correlation between sperm DFI and sperm concentration, semen volume, total sperm count, and motile sperm count were not proved.ConclusionsSperm DFI is an important indicator for evaluating the quality of semen. Sperm DNA integrity testing is preferentially recommended to those who have decreased sperm progressive motility, especially older men. An integrative analysis of sperm DFI, sperm progressive motility, age, and infertility conditions can provide a more comprehensive assessment of male fertility.     

The clinical value of conventional semen analysis

The objectives of this study were, firstly, to relate semen variables to treatment independent conception rates by life-table analysis after having accounted for known female factors; secondly, to assess the relationship between the length of involuntary infertility before investigation and the predictive value of semen parameters; and thirdly, to examine the relationship between the type of progressive spermatozoal motility and fertility outcome. Laboratory error in the assessment of semen variables was minimized by using one consistent observer. Seven hundred thirty-nine subjects were recruited to the study over a 34-month period, and a 96.5% follow-up rate was achieved. Where the female partner had regular spontaneous ovulation, no pelvic pathology, and more than 48 months' preceding infertility, the Grade 2 motile sperm density (the concentration of spermatozoa exhibiting slow or sluggish linear or nonlinear motility) was the variable that best predicted fertility outcome (X1(2) = 20.24, P less than 0.0001). Where the Grade 2 motile sperm density was below 5 X 10(6)/ml in the latter group (19%), no conceptions were reported at 32 months' follow-up. Semen variables were not of predictive value where there were fewer than 48 months' infertility before investigation, or where the female partner had ovulatory dysfunction or pelvic pathology.