不育病人精浆前列腺特异性抗原的检测及意义

目的 :探讨不育病人精浆前列腺特异抗原 (PSA)检测的意义。　方法 :随机选择 85例不育病人 ,采用ELISA法检测不育病人精浆PSA水平 ,并分析其与精浆酸性磷酸酶 (ACP)、精子密度、活率之间的关系。　结果 :35例不液化病人、30例液化不全病人精浆PSA水平、ACP浓度、精子活率 (a +b +c级精子百分率 )均显著低于 2 0例液化正常病人 (P <0 .0 1) ;而精子密度则无差异 (P >0 .0 5 )。精浆PSA水平与精浆ACP浓度、精子活率显著正相关 (P<0 .0 1)。　结论 :精浆PSA水平与精液液化密切相关 ,其质和量的异常会使精子活率降低 ,从而导致男性生育力下降     

弱精子症、少弱精子症患者血清、精浆和精子锌含量分析

目的:检测弱精子症和少弱精子症患者血清、精浆和精子锌的含量,分析锌含量的变化与精子密度和精子运动之间的关系。方法:按照WHO《人类精液及精子-宫颈粘液相互作用实验室检验手册》第四版的标准进行精液质量分析,随机筛选出90例弱精子症、60例少弱精子症患者以及20例精液质量正常的生育者作为研究对象,利用原子吸收光谱法检测其血清、精浆、精子的锌含量并进行统计学分析。结果:3组间血清锌含量没有显著差异;弱精子症、少弱精子症患者精浆锌含量均显著低于正常生育者(P<0.05);少弱精子症患者精子锌含量显著高于弱精子症患者和正常生育者(P<0.01)。结论:弱精子症、少弱精子症患者精子的发生及运动功能下降可能与精浆锌含量的低下呈正相关;但过高的精子锌含量与精子的发生和运动功能的关系尚不十分明了,有待进一步研究。     

精浆和血清生殖激素与精液质量的关系

目的为了评估精液质量不同的男性精浆和血清生殖激素的浓度与精子浓度及活动力的关系,探索精浆与血清生殖激素的关系。方法对301名男性进行精液检查,按照精液的质量参数将受试对象分成4组:精液正常组(n=176),弱精子症组(n=66),少精子症组(n=40)和非梗阻性无精子症组(n=19)。采用电化学发光免疫法测定各组受试对象血清卵泡刺激素(FSH)、黄体生成素(LH)、泌乳素(PRL)、孕酮(P)、睾酮(T)和雌二醇(E2)六项生殖激素和精浆PRL、T、P和E2四项生殖激素的浓度,比较组间差异并进行相关性分析。结果精液正常组和弱精子症组血清FSH和E2的浓度显著低于少精子症组和非梗阻性无精子症组(P0.05),精液正常组血清LH和P的浓度显著低于弱精子症、少精子症和非梗阻性无精子症的人群(P0.05);而精液正常、弱精子症和少精子症三组精浆PRL的浓度则高于非梗阻性无精子症组(P0.05)。除了非梗阻性无精子症组,受试者血清FSH的浓度与其精子浓度呈负相关(r分别为-0.350、-0.273和-0.448,P0.05)。精液正常组精浆PRL的浓度和精子的浓度之间呈正相关(r=0.269,P0.05);在少精子症组中,亦有相同趋势的相关性(r=0.432,P0.05)。结论精浆PRL及血清FSH的浓度能够反映精子浓度或活动力,在男性不育的病因分析中具有一定的指导价值。     

精索静脉曲张不育患者精子质量及其超微结构的研究

目的:研究精索静脉曲张不育患者精液质量及精子超微结构的变化。方法:不育伴精索静脉曲张患者118例作为实验组(VC组),正常自愿捐精者76例作为对照组,对其精液常规、精浆生化及外周血内分泌水平进行检测,并应用扫描和透射电镜技术对精子的超微结构进行观察。结果:VC组精液常规检查中,精子浓度、前向运动能力、存活率显著低于正常组(P<0.05),精液量、非前向运动能力无差异(P>0.05),精浆生化各项指标中,果糖浓度无显著差异,中性α-葡糖苷酶、锌离子浓度均显著低于正常组(P<0.05),外周血FSH、LH、T、E2水平均无统计学差异(P>0.05)。扫描电镜检查示正常形态精子比率低于对照组[(56.76±15.32)%vs(12.34±6.58)%,P<0.05],精子异常主要发生在头颈部,透射电镜检查示精子畸变以尖头为主,且伴有分化异常的复杂畸形。结论:VC可导致少-弱-畸精子症,进而引起不育,其原因可能是由于精浆微环境及超微结构的改变。     

精液不同体外处理技术对宫腔内人工授精的临床疗效分析

目的　探讨精液的不同体外处理技术对宫腔内人工授精 (IUI)的疗效。　方法　A组为因女方因素引起的不育 ,采用上游法优选精子。B组为因男性性交和射精障碍 ,精液液化不良 ,免疫学异常 ,精液中有核细胞数目 >5× 10 9/L ,单纯精浆异常等引起的不育 ,采用高速离心法处理精子。C组为少、弱、畸精子症等引起的不育 ,采用双层梯度法处理精子。　结果　妊娠成功率A组 5 0 .5 % ,B组为 4 1.4 % ,C组为 32 .4 %。　结论　对不同的病因采用不同的精液体外处理技术 ,能提高IUI的成功率。     

少精子症患者血清、精浆中游离睾酮水平的测定及意义

目的 :通过测定少精子症患者血清、精浆中游离睾酮 (FT)水平 ,分析血清、精浆FT与少精子症的关系。　方法 :正常对照组 (n =4 4 )、少精子症组 (n =4 4 )男性于上午 8:0 0～ 10 :0 0留取血标本 ;正常对照组 (n =30 )、少精子症组 (n =37)同时留取精液。男性精液常规分析判断精子密度 ,放射免疫分析法测定血清、精浆中FT水平。　结果 :少精子症患者血清中FT浓度为 [(94 .88± 4 2 .0 4 )pmol/L],与正常对照组 [(97.5 0± 4 6 .96 )pmol/L]相比差异无显著性 (P >0 .0 5 ) ,但少精子症患者精浆中FT浓度 [(0 .5 2± 0 .4 4 ) pmol/L]显著低于正常对照组 [(2 .0 1±0 .32 )pmol/L],P <0 .0 1。　结论 :精浆中FT的测定较早反映睾丸的功能 ,有利于少精子症患者的早期诊断和治疗。     

不育患者精子中肺癌肿瘤抑制因子-1的表达及意义探讨

目的 探讨肺癌肿瘤抑制因子1(TSLC1)与男性不育的关系.方法 首先免疫小鼠制备TSLC1单克隆抗体(McAb),然后分别选取30份正常组和40份不育组精液标本,Percoll梯度离心分成精浆和精子,将分离出的精子与制备出的McAb做免疫组织化学染色,对结果进行统计学分析.结果 所有正常牛育组精子的TSLC1蛋白表达均为阳性.在小育组中,30例弱精子症患者,25例TSLC1蛋白表达为阴性,10例少精子症患者,8例TSLC1蛋白表达为阴性.正常组与弱精子症组、少精子症组差片分别有统计学意义(P＜0.001),弱精子症组与少精子症组差异无统计学意义(P＞0.05).结论 精子中TSLC1蛋白的低表达或不表达与男性不育密切相关.     

男性年龄与精子质量和精浆生化的关系

目的探讨不同年龄段男性精子质量与精浆弹性硬蛋白酶和精浆中性α-葡萄糖苷酶的关系。方法回顾性分析在嘉兴市妇幼保健院门诊检测精子质量和精浆生化(精浆弹性硬蛋白酶、精浆中性α-葡萄糖苷酶)的3 262例患者的临床资料。根据年龄分为5组:25岁组、25~29岁组、30~34岁组、35~39岁组和≥40岁组;并按精子质量分为少弱精子症(301例)、少精子症(193例)、弱精子症(1 335例)及正常精子组(1 433例)。统计分析不同精子质量不同年龄段男性的精浆弹性硬蛋白酶、精浆中性α-葡萄糖苷酶资料。结果 (1)3 262例精液分析,精浆中性α-葡萄糖苷酶异常发生率7.14%,精浆弹性硬蛋白酶水平确诊感染率29.25%;(2)各年龄组总体精浆中性α-葡萄糖苷酶异常发生率相似(P0.05),生殖道感染发生率随年龄增加有增高趋势(P0.05);(3)不同精子质量各组总体精浆中性α-葡萄糖苷酶异常发生率不同(P0.05),其中少精子症和少弱精子症组均显著高于弱精子症组及精子正常组(P0.05),但生殖道感染发生率相似(P0.05);(4)随着年龄增加,不同精子质量各组精浆中性α-葡萄糖苷酶异常发生率相似(P0.05),而精浆弹性硬蛋白酶异常发生率有增高趋势(P0.05),其中少弱精子症、少精子症患者各年龄组间比较均有统计学差异(P0.05);(5)各精子质量组精浆弹性硬蛋白酶测定值均随着年龄增加呈增高趋势,且正常精子组的精浆弹性硬蛋白酶测定值在35~39岁组及≥40岁组均显著高于25岁组(P0.05)。结论男科门诊患者中生殖道感染具有普遍性;伴随难以自愈的生殖道感染的少弱精子症、少精子症和弱精子症可能与男性患者年龄增加有关,亦可能与附性腺病变严重程度、病程进展所处不同阶段有关。     

少弱精子症与精浆附性腺标志物的相关性分析

目的分析少弱精子症患者精液参数与精浆附性腺标志物的相关性,探讨附性腺功能对男性生育力的影响。方法采用精液常规、精子形态、精浆附性腺标志物分析方法,检测正常供精者和门诊就诊的少弱精子症患者的精液相关指标。结果少弱精子症组正常形态精子百分数、精子穿透功能、精浆中性α-糖苷酶显著低于正常对照组。相关分析结果显示,少弱精子症患者组,精液量与精子活动率呈负相关(r=-0.415,P<0.05),精子数与形态呈正相关(r=0.393,P<0.05)。结论少弱精子症患者同时存在不同程度的附睾功能障碍,精子功能下降,精子畸形率显著升高,睾丸生精功能越低下,精子畸形发生率越高。     

生育及不育男性血清及精浆抑制素-B水平分析

目的 :探讨生育及不育男性血清及精浆抑制素 B(inhibinB ,INHB)水平是否存在差异 ,了解血清及精浆INHB水平与精子发生的关系。　方法 :生育组 (n =2 0 )、少精子症组 (n =2 0 )、弱精子症组 (n =2 2 )和非阻塞性无精子症 (NOA)组 (n =2 0 )男性于上午 8∶0 0～ 10∶0 0留取精液和血液标本 ,进行精液常规分析 ,血清INHB、FSH、LH、T含量 ,精浆INHB、酸性磷酸酶、果糖、α 葡糖苷酶含量和活性测定。　结果 :血清、精浆INHB水平与血FSH均呈显著负相关 (r =- 0 .5 36 ,P <0 .0 0 1vsr =- 0 .2 88,P =0 .0 1) ,血清、精浆INHB水平与精子密度均呈显著正相关 (r=0 .49,P <0 .0 0 1vsr =0 .48,P <0 .0 0 1) ,血清INHB水平在生育组男性与少精子症组、NOA组男性间(分别为P <0 .0 5和P <0 .0 1)、弱精子症组与NOA组男性间 (P <0 .0 1)及少精子症组与NOA组男性间 (P <0 .0 5 )差异均有显著性 ,而精浆INHB变动范围较大 ,其水平仅在生育组男性与NOA组男性间及弱精子症组与NOA组男性间差异有显著性 (分别为P <0 .0 1和P <0 .0 5 )。精浆INHB水平与精浆α 葡糖苷酶活性呈正相关 (r=0 .377,P =0 .0 0 1)。血清INHB水平与精浆INHB水平间无相关性。　结论 :血清、精浆INHB水平均可反映睾丸的精子发生情况 ,精浆INHB水平还与     

Blood and semen paraoxonase—arylesterase activities in normozoospermic and azoospermic men

Paraoxonase and arylesterase enzymes are corner stones of antioxidant defence. We aimed to compare azoospermic infertile men and normozoospermic individuals with respect to total antioxidant status (TAS), total oxidant status (TOS), oxidative stress index (OSI), paraoxonase and arylesterase levels in the blood and seminal plasma. Two‐hundred consecutive infertility patients and voluntarily participated were included. In the normozoospermic group, TAS, PON, arylesterase values were statistically significantly higher when compared with those in the azoospermic group, while lower TOS and OSI levels were observed in the blood and seminal plasma of azoospermic group. In the semen analyses of normozoospermic group, the correlation between semen volume, sperm concentration, sperm motility and morphology and TAS, TOS, OSI, PON and arylesterase values was examined. A negative correlation was determined between semen volume and OSI. Levels of serum oxidative parameters were higher in the azoospermic group relative to normozoospermic group, but antioxidant parameters were lower than those of the normozoospermic group. Oxidative stress performs an essential role in the aetiology of male infertility by negatively influencing sperm quality and function. Assessment of blood and seminal plasma oxidative profiles might be an important tool to better evaluation of sperm reproductive capacity and functional competence.     

生育与不育男性精浆总抗氧化能力分析

目的:分析生育与不育男性精浆中总抗氧化能力(TAC)及其在男性生育中意义。方法:225例男性不育患者分为6组,分别为:梗阻性无精子症组(n=10),非梗阻性无精子症组(n=42),少精子症组(n=20),弱精子症组(n=78),少弱精子症组(n=57),以及正常精子症组(n=18)。28例正常生育男性作为对照(生育组)。分别采用计算机辅助精液分析(CASA)系统进行精液参数分析,采用比色法检测精浆TAC水平。结果:生育组男性精浆TAC为(19.82±6.33)U,梗阻性无精子症组(1.71±1.33)U,非梗阻性无精子症组(12.73±9.44)U,少精子症组(10.85±6.64)U,弱精子症组(13.88±8.24)U,少弱精子症组(11.20±7.02)U,正常精子症组(18.07±8.73)U;与生育组精浆TAC[(19.82±6.33)U]相比,在各不育症组中,除正常精子症组精浆TAC与生育组差异无显著性外,其余各组均显著低于生育组(P<0.01)。精浆TAC与精子密度(r=0.182,P<0.05)和a级精子(r=0.150,P<0.05)呈显著正相关。结论:精浆中TAC水平与男性不育密切相关,精浆中过低的TAC水平可能是引起男性不育的病因之一。     

Quick recovery and characterization of cell-free DNA in seminal plasma of normozoospermia and azoospermia： implications for non-invasive genetic utilities

We established a quick and reliable method for recovering cell-free seminal DNA （cfsDNA）, by using the binding-washing-elution procedure on the DNA purification column. Low variations （below 15%） among the triplicate values of cfsDNA quantity verified the reproducibility of our cfsDNA recovery method. Similar cfsDNA yield and size distribution between seminal plasma acquired by filtration and centrifugation confirmed the presence of cfsDNA. To investigate the general characterization of cfsDNA, the quantitation and size distribution of cfsDNA from normozoospermic and azoospermic semen were analyzed by real-time PCR and electrophoresis, respectively. CfsDNA concentration in semen with normozoospermia （n = 11） was 1.34 ± 0.65 μg ·mL^-1, whereas a higher cfsDNA concentration was observed in azoospermia （2.56 ± 1.43 μg ·mL^-1, n = 9）. The continuous distribution of DNA fragments ranging from -1 kb to 15 kb and a spectrum of multiples of 180-bp fragments were observed in each normozoospermic and azoospermic sample. Distinct characteristic DNA ladder fragmentations in some azoospermic samples implicated that cfsDNA originate partly from apoptotic cells. CfsDNAs of 36 selected azoospermic patients with known information of Y chromosome microdeletion were subjected to the same microdeletion analysis by multiplex PCR and PCR amplification of sY114 （1 450 bp）. All multiplex PCR reactions with cfsDNA amplified successfully and provided the same result as leukocyte DNA. PCR amplification of sY114 gave a 1 450-bp amplicon as expected. Our data suggested the potential use of cfsDNA in search of biomarker or diagnostic procedures.     

Quantification of seminal plasma motility inhibitor/semenogelin in human seminal plasma

Semenogelin I and II (Sg I and II) are the major components of human semen coagulum. The protein is rapidly cleaved after ejaculation by the chymotrypsin-like protease prostate-specific antigen (PSA), which results in the liquefaction of the semen coagulum and the progressive release of motile spermatozoa. One of the cleavage products of the protein, a 14-kDa protein, is a sperm motility inhibitor (seminal plasma motility inhibitor [SPMI]). We developed a monoclonal antibody (mAb) that is specific to the fragment of Sgs, SPMI, and a sandwich enzyme-linked immunosorbent assay (ELISA) system for the quantification of Sgs using this mAb. Then, we measured SPMI/Sg levels in human seminal plasma from healthy male volunteers (n = 100, aged 18-24 years). The mean level of SPMI/Sg in seminal plasma was 19 +/- 13 mg/mL (range, 4-68 mg/mL). Log-transformed SPMI/Sg levels were negatively correlated with the sperm motility (r = -0.229, P =.0220) and positively correlated with the total protein concentration (r = 0.793, P <.0001). This result supports that SPMI, one of the fragments of Sg, has its inhibitory effect on ejaculated spermatozoa in liquefied semen under physiological conditions.     

Lack of correlation of selenium level in human semen with sperm count/motility

Considering the importance of selenium (Se) in male fertility, its concentration was measured in 211 semen samples from 211 normozoospermic, oligozoospermic, asthenozoospermic, and azoospermic men using the hydride generation atomic absorption spectrophotometry. No significant correlation of any kind existed between Se level in the seminal plasma and sperm count or motility. In view of the known poor correlation of these two frequently used semen parameters with the incidence of pregnancy, the assessment of the fertilizing potential of normozoospermic ejaculates with low Se levels is warranted.     

Assessment of seminal plasma laminin in fertile and infertile men

Aim： To assess laminin levels in the seminal plasma of infertile and fertile men, and to analyze the correlation of laminin levels with sperm count, age, sperm motility and semen volume. Methods： One hundred and twenty-five recruited men were equally divided into five groups according to their sperm concentration and clinical examination： fertile normozoospermia, oligoasthenozoospermia, non-obstructive azoospermia （NOA）, obstructive azoospermia （OA） and congenital bilateral absent vas deferens （CBAVD）. The patients＇ medical history was investigated and patients underwent clinical examination, conventional semen analysis and estimation of seminal plasma laminin by radioimmunoassay. Results： Seminal plasma laminin levels of successive groups were： 2.82 ± 0.62, 2.49 ± 0.44, 1.77 ± 0.56, 1.72 ± 0.76, 1.35 ± 0.63 U/mL, respectively. The fertile normozoospermic group showed the highest concentration compared to all infertile groups with significant differences compared to azoospermic groups （P 〈 0.05）. Testicular contribution was estimated to be approximately one-third of the seminal laminin. Seminal plasma laminin demonstrated significant correlation with sperm concentration （r = 0.460, P 〈 0.001） and nonsignificant correlation with age （r = 0.021, P = 0.940）, sperm motility percentage （r = 0.142, P = 0.615） and semen volume （r = 0.035, P = 0.087）. Conelusion： Seminal plasma laminin is derived mostly from prostatic and testicular portions and minimally from the seminal vesicle and vas deferens. Estimating seminal laminin alone is not conclusive in diagnosing different cases of male infertility.     

Effect of smoking on seminal plasma ascorbic acid in infertile and fertile males

This work aimed to assess the relationship of seminal ascorbic acid levels with smoking in infertile males. One hundred and seventy men were divided into four groups: nonobstructive azoospermia [NOA: smokers (n = 20), nonsmokers (n = 20)]; oligoasthenozoospermia [smokers (n = 30), nonsmokers (n = 20)]; asthenozoospermia [smokers (n = 20), nonsmokers (n = 20)] and normozoospermic fertile men [smokers (n = 20), nonsmokers (n = 20)]. The patients underwent medical history, clinical examination, conventional semen analysis and estimation of ascorbic acid in the seminal plasma calorimetrically. There was a significant decrease in the mean seminal plasma ascorbic acid levels in smokers versus nonsmokers in all groups (mean +/- SD; 6.03 +/- 2.18 versus 6.62 +/- 1.29, 7.81 +/- 1.98 versus 9.44 +/- 2.15, 8.09 +/- 1.98 versus 9.95 +/- 2.03, 11.32 +/- 2.15 versus 12.98 +/- 12.19 mg dl(-1) respectively). Fertile subjects, smokers or not, demonstrated significant higher seminal ascorbic acid levels than any infertile group. Seminal plasma ascorbic acid in smokers and nonsmokers was correlated significantly with sperm concentration (r = 0.59, 0.60, P < 0.001), sperm motility (r = 0.65, 0.55, P < 0.001) and negatively with sperm abnormal forms per cent (r = -0.53, -0.50, P < 0.001). Nonsignificant correlations were elicited with semen volume (r = 0.2, 0.09) or liquefaction time (r = 0.03, 0.06). It is concluded that seminal plasma ascorbic acid decreased significantly in smokers and infertile men versus nonsmokers and fertile men, and is significantly correlated with the main sperm parameters: count, motility and normal morphology. Also, cigarette smoking is associated with reduced semen main parameters that could worsen the male fertilizing potential, especially in borderline cases.     

Proteomic analysis of seminal plasma in men with different spermatogenic impairment

Seminal plasma is a potential source of biomarkers for many disorders of the male reproductive system including male infertility. The identification and characterisation of differentially expressed proteins in seminal plasma of man with normal and impaired spermatogenesis can help in the elucidation of the molecular basis of male infertility. We compared the protein expression profiles of seminal plasma from four different groups of men as follows: normozoospermic, asthenozoospermic, oligozoospermic and azoospermic groups, using two-dimensional differential gel electrophoresis (2-D DIGE). We found eight proteins with statistically significant increased expression in azoospermia compared with at least one of the other studied groups. The differentially expressed spots were fibronectin, prostatic acid phosphatase (PAP), proteasome subunit alpha type-3, beta-2-microglobulin, galectin-3-binding protein, prolactin-inducible protein and cytosolic nonspecific dipeptidase. Notably, PAP was increased in patients with azoospermia compared with that of all other groups. We have observed no statistically significant differences in protein expression between three of the groups: normozoospermic, oligozoospermic and asthenozoospermic. We suggest that the identified panel of proteins in our study especially PAP have a strong potential to be used as azoospermia markers. However, further investigations will be necessary to validate these markers in samples of larger and independent patient cohorts and to clarify their role in the pathogenesis of male infertility.     

Heme oxygenase enzyme activity in human seminal plasma of fertile and infertile males

This work aimed to assess heme oxygenase (HO) enzyme activity relationship with different human semen parameters. One hundred and twenty men were divided according to their sperm count and clinical examination into: obstructive azoospermia (n = 20), nonobstructive azoospermia (NOA) (n = 25), oligozoospermia (n = 35) and normozoospermia (n = 40). Semen analysis, western blot for HO-1 and HO-2, and estimation of seminal plasma HO enzyme activity chemically in the form of bilirubin concentration were carried out. Seminal plasma HO enzyme activity was very low in OA specimens, low in NOA, moderate in oligozoospermia while higher in normozoospermia (mean +/- SD; 6.26 +/- 2.2, 81.4 +/- 35.5, 283.8 +/- 90.1, 657.4 +/- 227.6 pmol ml(-1) min(-1)) with significant differences. Western blot analysis demonstrated HO-2 expression in all studied groups whereas HO-1 was highly expressed in fertile normozoospermic group compared with other groups. There was positive correlation between seminal plasma HO enzyme activity and sperm concentration, sperm motility percentage, motile spermatozoa ml(-1) and sperm normal morphology per cent. It is concluded that HO enzyme activity in the human seminal plasma is related to spermatogenesis and sperm-motility processes.