流式细胞计检测人精子顶体及其与精液其他参数的关系

运用流式细胞计及人精子顶体特异性标记物结合异硫氰酸荧光素的花生凝集素检测了３４例精液精子顶体结构的完整性。其中正常生育精液１２例，弱精症精液１２例，畸形弱精症精液１０例。并且进一步检测了２０例精液在体外诱导下精子发生顶体反应的能力，其中正常生育精液１０例，不育精液１０例。结果表明人精子顶体结构的完整性与精于正常形态百分率，存活率，前向运动百分率显著正相关（Ｐ＜０．００１）。不育组精子在体外诱导下发生顶体反应的能力明显低于生育组（Ｐ＜０．０１），说明流式细胞计能用于预测生育力。     

213例不育男性抗精子抗体与精液有关参数的分析

我们检测了９００多例男性不育症患者的血清、精浆抗精子抗体（ＡｓＡｂ）和精液各项指标，对其中２１３例ＡｓＡｂ阳性患者的精液有关参数与ＡｓＡｂ的关系进行了分析，并与生育男性组作了比较。结果表明，ＡｓＡｂ阳性患者的精子密度、精子活率、精子顶体完整率、精浆免疫抑制物（ＳＰＩＭ）及α－糖苷酶均明显低于生育组（Ｐ＜０．０１），不育男性ＡｓＡｂ阳性组ＳＰＩＭ阳性率也明显高于ＡｓＡｂ阴性组（Ｐ＜０．０１）。此外，血清与精浆ＡｓＡｂ同为阳性组的精子活率明显低于单纯血清或精浆ＡｓＡｂ阳性组（Ｐ＜０．０１）。     

植物凝集素荧光标记法检测精子顶体结构的临床意义

目的　探寻评估人精子授精能力的简便有效方法。　方法　采用络合异硫氰酸荧光素的植物凝集素 (FITC PSA)进行精子顶体荧光标记 ,分别测定 5 8例不育患者及 4 6例正常生育男子的精子顶体完整率 ,并对其中 14例精液标本 (不育组及正常组各 7例 )进行人卵透明带诱发的精子顶体反应实验 ,对精子顶体完整率与发生顶体反应的百分率的相关关系进行分析。　结果　正常生育组顶体完整率及发生精子顶体反应百分率显著高于不育组 (P <0 .0 1) ,精子顶体完整率与发生精子顶体反应百分率呈正相关 (r =0 .74 68)。　结论　FITC PSA荧光标记法操作简便 ,技术结果可靠 ,特异性强 ,可作为评价人精子授精能力的有效指标之一。     

精索静脉曲张不育患者精子形态学分析及其治疗后的初步观察

应用改良巴氏染色法对５４例精索静脉曲张不育患者的精子形态学进行了分析并初步观察治疗后的效果。结果表明，精索静脉曲张Ⅰ度组、Ⅱ度组和Ⅲ度组的精子畸形率均显著高于正常生育力组（Ｐ＜０．０１），而且尖头精子、顶体异常、老化精子和不定形精子数都有较高的比率。经手术治疗后，初步观察到Ⅱ度组的精子畸形率显著减少（Ｐ＜０．０１），但是Ⅲ度组却无显著变化（Ｐ＞０．０５）。提示①精索静脉曲张导致不育的原因之一是正常     

去除精浆对test－yolk缓冲液保存大猩猩精子效果的改善

Ｔｅｓｔ－ｙｏｌｋ缓冲液被应用于精子的短时期冷藏，可以提高精子的体外受精能力。过去的冷藏方法中保留了精浆。本实验比较了在卵黄液短期保存精液方法中去除精浆和保留精浆两种情况下大猩猩精子的体外受精能力和运动能力。体外受精能力以去透明带地鼠卵穿透试验来评价，精子运动能力以计算机辅助分析系统定量分析。精液标本取自５只人工饲养的成年大猩猩，１２例标本用于卵穿透试验，２２例用于运动分析。冰浴过夜保存于ｔｅｓｔ－ｙｏｌｋ缓冲液后活动精子百分率下降２６．９％～３０．９％，去除精浆的精子运动速度显著高于保留了精浆的精子，直线速度（ＶＳＬ、Ｐ＜０．０１），曲线运动速度（ＶＣＬ，Ｐ＜０．０１），头部侧向位移（ＡＬＨ，Ｐ＜０．０５）；并且卵穿透试验结果显示前者精子穿卵率和平均每卵穿入精子数均高于后者（Ｐ＜０．０１）。本实验结果提示∶在ｔｅｓｔ－ｙｏｌｋ缓冲液的精子短期冷藏过程中，去除精浆更有利于精子受精能力和运动速度的保存。     

精子尾部卷曲试验及影响因素的探讨

本文介绍了精子尾部卷曲试验及影响因素（时间、温度、ｐＨ）。３０例正常生育男性和４３例不育男性的精液分析表明，精子尾部卷曲率与精子活率具有高度的相关性（ｒ＝０．９６２５），与精子密度无相关性（ｒ＝０．１６２）；男性生育组与不育组之间的精子尾部卷曲率具有显著性差异（ｘ±ｓ，百分率分别为７５．２１±２０．０１和５９．４３±２０．５４，Ｐ＜０．０５）。     

128对不育夫妇血清、精浆抗精浆免疫抑制物抗体与精液检查关系的分析

我们检测了８９对生育夫妇和１２８对不育夫妇血清、精浆中抗精浆免疫抑制物抗体（ＳＰＩ—Ａｂ）的水平。结果表明，不育夫妇血清、精浆ＳＰＩＭ—Ａｂ的水平和检出率均明显高于生育组（Ｐ＜０．０１）；精浆ＳＰＩＭ—Ａｂ水平增高者的精子密度、精子活率、精子运动速度和ＳＰＩＭ活性均明显降低（Ｐ＜０．０１）。提示：ＳＰＩＭ—Ａｂ水平增加可能影响生育。     

128对不育夫妇血清,精浆抗精浆免疫抑制物抗体与精液检查关系的分析

我们检测了８９对生育夫妇和１２８对不育夫妇血清、精浆中抗精浆免疫抑制物抗体（ＳＰＩＭ－Ａｂ）的水平。结果表明，不育夫妇血清、精浆ＳＰＩＭ－Ａｂ的水平和检出率均明显高于生育组（Ｐ＜０．０１）；精浆ＳＰＩＭ－Ａｂ水平增高者的精子密度、精子活率、精子运动速度和ＳＰＩＭ活性均明显降低（Ｐ＜０．０１）。提示：ＳＰＩＭ－Ａｂ水平增加可能影响生育。     

人类精子低渗试验指数分析

目的：探讨精子低渗试验指数（ＳＨＯＴＩ）对评估男性生育力的临床价值。方法：采用两种不同渗量的低渗液结合活体染色技术，对１２９例男性精液进行精子低渗试验。结果：生育组与不育组比较精子卷尾率、ｇ型肿胀精子率和精子低渗试验指数差异均显著（Ｐ＜０．０１），生育组ＳＨＯＴＩ分布范围为：０．０９６～０．３３６；不育组ＳＨＯＴＩ分布范围为０．０２５～０．２１５。结论：ＳＨＯＴＩ综合反映了精子的整体膜功能及受精能力，可作为评估男性生育力的一个新的指标。初步确定临床参考值为ＳＨＯＴＩ＜０．０９６，提示精子功能不良，不能生育；在０．０９６～０．２１５之间精子功能部分正常，有机会生育；在０．２１５～０．３４０之间预示精子功能正常，生育力良好。     

精子与宫颈粘液相互作用在预测宫颈性不育的价值

为了解精子与宫颈粘液相互作用预测生育的价值，对２７例不明原因不孕妇女进行性交后试验（ＰＣＴ）和精子穿透试验（ＳＭＰＴ）。１年后按有无妊娠分为妊娠组（ｎ＝１４）和非妊娠组（ｎ＝１３）。两组宫颈粘液评分无显著性差异（Ｐ＞０．０５）；妊娠组在性交后２和４小时精子数（１９．３±３．３及２２．２±４．４个／Ｈｐ）明显高于非妊娠组（５．０±２２及２．９±１．４个／Ｈｐ）（Ｐ＜０．０１）；妊娠组ＰＣＴ正常率（７１％）明显高于非妊娠组（０％），（Ｐ＜０．０１）；ＳＭＰＴ结果与ＰＣＴ结果基本一致。提示ＰＣＴ是有价值的预测生育指标，ＰＣＴ正常可排除男方因素及宫颈性不孕，ＰＣＴ异常则可选择人工授精提高妊娠率     

Oocyte Penetration and Acrosome Reactions of Human Sperms II: Correlation with other Seminal Parameters

Washed sperm suspensions of fertile men and men consulting for infertility were evaluated for their ability to penetrate zona-free hamster eggs and for their ability to exhibit an acrosome reaction in vitro. Furthermore, the swelling of the spermatozoa under hypoosmotic conditions as indication of their membrane integrity was determined. In the group of fertile men and in the group of patients with normal spermiogram, significantly more acrosome reactions were observed than in the group of infertile men with abnormal spermiogram parameters (p less than 0.05). This difference was still more significant when men with a positive hamster penetration test (H.O.P. test) and men with a negative H.O.P. test were compared (p less than 0.005). However, within the groups the level of acrosome reactions after incubation appeared to be highly variable. In a second series of experiments, working with semen obtained during our in vitro fertilization program, we found that the fertilization of human ova does not seem to be dependent on a strong progress of the acrosome reaction. Finally, swelling of the spermatozoa in a hypoosmotic medium was weakly correlated (r = 0.70, p less than 0.01; n = 73) with trypan blue exclusion. No significant correlations with other semen parameters, hamster ova test included, were found.     

用有效活精指数评价精液质量

同时应用有效活精指数及精子尾部低渗肿胀试验、精子前向运动级别、毛细管穿透试验，精子顶体染色，对９０例正常男性和１２９例不育男性精液进行检测。结果显示：有效活精指数与精子总肿胀率（Ｔ），ｇ形精子率（Ｇ），前向运动级别（Ｐ），穿透力评分（Ｓ），顶体完整率（Ａ）之间，生育组呈显著正相关，不育组呈非常显著正相关。以上各项试验参数在两组间均有非常显著差异。不育男性结果表明，有效活精指数与Ｔ、Ｇ、Ｐ、Ａ的异常检出率之间均无显著差异，而仅与（Ｓ）的异常检出率有显著差异，提示有效活精指数作为传统的检查手段对精液质量评价仍有重要意义     

男性生育力的研究：三种精子功能测定方法的比较

应用精液常规分析(SFA),精子尾部低渗肿胀试验(HOS)和去透明带地鼠卵穿透试验(HOP)对15名能宵男性和15名不育患者的精液样本进行了综合检测和相关分析。能育组SFA参数异常者占20％,不育组SFA参数正常者占27％;精子尾部低渗肿胀率在能育组与不育组间无显著性差异(P>0.05);精子穿透率和受精指数在两组间差异非常显著(P<0.005,P<0.001)这表明HOP试验用于男性生育力的评价是一种较为准确,可靠的手段。本文还对各实验参数间的相关关系进行了分析和讨论。     

Studies on male infertility--acrosome reaction of the human sperm incubated in vitro

In 1981, P. Talbot developed a triple-stain technique to estimate the number of human sperm undergoing normal acrosome reaction in fixed smears. In this method, live and dead sperm are first differentiated using the vital stain trypan blue. Sperm are then fixed in glutaraldehyde, dried onto slides, and the postacrosomal region and acrosome are differentiated using Bismark brown and Rose Bengal. Slides are examined at 1,000x with a bright-field microscope and assessed for the percentage of sperm that have undergone the normal acrosome reaction. Using this method we examined the time course of the acrosome reaction of human sperm incubated in mBWW the influence of human serum albumin, the calcium concentration of incubation media, Ca ionophore A 23187, and trypsin on the acrosome reaction of human sperm and the difference between the percentage of sperm undergoing normal acrosome reaction of fertile and infertile males. We got the following results: The percentage of human sperm undergoing acrosome reaction increased for the first six hours of incubation. Without human serum albumin the acrosome reaction did not occur. Ca ion could be one of the triggers of the acrosome reaction. Ca ionophore A23187 and trypsin induced the acrosome reaction in vitro. Sperm from oligozoospermic males, especially poorly motile sperm, could not undergo acrosome reaction so easily as sperm from fertile males could.     

Use of theophylline to enhance sperm function.

Poor sperm motility is an important factor in male infertility. Preliminary results in our laboratory on a group of 19 men (10 suspected infertile men and 9 fertile donors) showed stimulation of sperm fertilizing ability after sperm washing with theophylline as demonstrated by zona free hamster egg penetration test. The egg penetration rate for the control spermatozoa samples from subfertile men was 16%. Incubation with theophylline (10 mM) increased the penetration rate to 46%, whereas semen incubation with theophylline (20 mM) increased the penetration rate to 51%. A similar twofold increase in egg penetration was observed in the semen of fertile men incubated with theophylline of similar concentrations. Subfertile patients with ejaculate volumes of less than or equal to 1 ml or total motile sperm count of less than or equal to 10 x 10(6)/mL or increased semen viscosity did not exhibit beneficial effects with theophylline washing as measured by hamster egg penetration test score. The increase in percentage of penetrated eggs with theophylline use in both fertile and subfertile men was significant at 10 mM concentration (p less than .001) and 20 mM (p less than .001) when compared to control (untreated) samples. No significant difference in penetration rate was seen between 10 and 20 mM theophylline concentrations. It appears that theophylline may be useful in improving the fertilizing capacity of selected human semen samples with poor motility and poor penetration ability under artificial insemination conditions.     

Heterologous ovum penetration test and seminal parameters in fertile and infertile men

Sperm penetration rates in the heterologous ovum penetration test were correlated with results of routine semen analysis in 30 fertile and 50 infertile men. There was no difference in penetration rates when comparing infertile men with normal and abnormal seminal parameters, nor was any difference seen between fertile (15-83%) and infertile men (8-83%). Of the 22 infertile men with normal seminal characteristics, seven had partners with no discernible reproductive dysfunction. The penetration rates of these men (38-81%) did not differ from those of fertile men. Stepwise regression analysis of seminal characteristics, with penetration as the dependent variable, indicated that sperm count and morphology are the most important seminal parameters for fertility assessment. Discrimination analysis revealed that sperm numbers and morphologic variables provide significant information for discriminating between fertile men and infertile men with normal or abnormal seminal characteristics. Sperm penetration and motility were not indicated as important factors. The present data suggest that, in unselected male patients seeking reproductive evaluation, the sperm penetration assay did not yield any additional information on the cause of infertility.     

精浆游离L-肉毒碱水平及其与精子密度、活动率及活力的相关性研究

目的:研究正常生育及不育男性精浆中游离L-肉毒碱水平差异及其与精子密度、活动率(a+b+c级精子百分率)及活力(a+b级精子百分率)之间的相关性,探讨精浆中游离L-肉毒碱水平对男性生育力的影响及其在不育症检查和治疗中的作用。方法:分别采用高效液相色谱法和计算机辅助精液分析系统,测定了230例不育症患者(精子密度正常117例,少精子症81例,无精子症32例)和30例正常生育男性精浆中游离L-肉毒碱水平及精子密度、活动率、活力等参数。根据检查结果对不育症患者分组后,以SPSS12.0软件包进行统计学分析,比较各组间游离L-肉毒碱水平的差异以及游离L-肉毒碱水平与精子密度、活动率、活力之间的相关性。结果:正常生育组精浆游离L-肉毒碱水平明显高于不育组(P<0.01)。精液中精子密度越低、活力越弱,这种差异性越显著。相关性分析结果显示,精浆游离L-肉毒碱水平与精子密度呈显著正相关关系(r=0.521,P<0.01),与精子活动率和活力之间也具有正相关关系(r=0.319,P<0.01;r=0.251,P<0.01)。结论:精浆L-肉毒碱水平与精子密度、活动率和活力之间密切相关,其含量测定作为一项有用的生化指标,可为男性不育症检查及临床诊治和进行有关男性生殖功能机制研究提供参考。