洗必泰醋酸盐和洗必泰葡萄糖酸盐的杀精子效果及阻抑精子对宫颈粘液穿透力的比较研究

本文比较了洗必泰醋酸盐和洗必泰葡萄糖酸盐的杀精子效力和阻抑人精子穿透动情期牛宫颈粘液的作用。结果表明,两种形式的洗必泰盐类均有良好的杀精子效力和阻止精子穿透牛宫颈粘液的作用。应用Sander-Cramer杀精子试验法测定洗必泰醋酸盐和洗必泰葡萄糖酸盐的最低杀精子浓度分别为2.5mg/ml,5.0mg/ml。应用Cremer′s精子穿透法测定两种洗必泰盐类阻止人精子穿透牛宫颈粘液的作用很相似,当药物浓度是0.25%或0.5%时人精子穿透牛宫颈液的距离是0mm(对照是48mm)。     

宫颈粘液葡萄糖果糖水平与不明原因不孕关系的探讨

本文对２３例不孕患者的２３个周期及对照组１５例２０个周期的宫颈粘液葡萄糖果糖水平进行测定。放免法测定血清促黄体素（ＬＨ）、促卵泡素（ＦＳＨ）、雌二醇（Ｅ２）和孕酮（Ｐ）；阴道Ｂ超监测卵泡发育；宫颈粘液改良Ｉｎｓｌｅｒ评分，尿ＬＨ酶联免疫测定和基础体温测定（ＢＢＴ）综合评价预测和确定排卵日。被研究对象均于围排卵期进行性交后试验（ＰＣＴ）。结果：４３个月经周期均为有排卵周期。不孕组（ＰＣＴ正确组７周期，ＰＣＴ异常组１６周期）宫颈粘液改良Ｉｎｓｌｅｒ评分与对照组无显著差异（Ｐ均＞０．０５）；不孕ＰＣＴ异常组卵泡期、排卵前、排卵后、黄体期宫颈粘液葡萄糖果糖水平均明显低于对照组（Ｐ＜０．０１）。而ＰＣＴ正常组与对照组无明显差异（Ｐ＞０．０５）。结论：宫颈粘液葡萄糖果糖水平低落可影响精子的穿透和存活，是不明原因性不孕的重要因素之一。     

ELISA法检测不育男子精浆中抗精子IgG和IgA

将精子经Tritonx－100处理，冷冻高速离心后，经抗人全血清-SephadexG－75亲和柱层析分离，提取分子量为59KD人精子膜蛋白作为抗原，经ELISA间接法对20例生育男子和50例不育男子精浆中抗精子IgG和IgA进行了测定。结果显示：生育男子精浆中抗精子IgG为阴性；抗精子IgA阳性率为5％。不育男子精浆中抗精子IgG和IgA阳性率分别为10％、30％；抗精子IgG和IgA均为阳性者3例，均为阴性者33例；抗精子IgG阳性而IgA阴性者2例；抗精子IgA阳性而IgG阴性者12例。不育组与生育组间抗精子IgG阳性率无显著性差异（P＞0．05）；而IgA阳性率间则有显著性差异（P＜0．05）。不育组抗精子IgG和IgA阳性率间有极显著性差异（P＜0．01）。本文利用ELISA法对精浆中抗精子抗体的分类及可能来源进行了讨论。     

人宫颈粘液葡萄糖、果糖变化规律的研究

本文对月经周期规律的１５例育龄妇女２０个月经周期宫颈粘液及血清葡萄糖、果糖水平进行测定，同时对血清促黄体激素（ＬＨ）、促卵泡激素（ＦＳＨ）、雌二醇（Ｅ２）和孕酮（Ｐ）放免测定；阴道Ｂ超、宫颈粘液Ｉｎｓｌｅｒ评分；尿ＬＨ酶联免疫测定和基础体温（ＢＢＴ）测定，综合评价预测和确定排卵日。２０个周期均为有排卵周期。结果：宫颈粘液葡萄糖、果糖水平均表现为卵泡期稍高，排卵前最低，排卵后逐渐升高，黄体中期达高峰；卵泡期果糖水平与Ｅ２呈负相关（ｒ＝－０．７３，Ｐ＜０．０１）；黄体期葡萄糖、果糖水平均与Ｐ呈正相关（ｒ＝０．９９，Ｐ＜０．０１；ｒ＝０．９８，Ｐ＜０．０１）；血葡萄糖、果糖水平无周期性变化且与宫颈粘液葡萄糖、果糖水平无明显相关性（ｒ＝０．２３，Ｐ＞０．０５）。结论：宫颈粘液葡萄糖、果糖水平变化具有明显规律性，可能与雌孕激素调节有关。     

放置VCu200 IUD不同年限宫颈粘液含铜量

本文用无火焰原子吸收分光光度仪ＨＩＴＡＣＨＩ－１８０测试置ＶＣｕ２００ＩＵＤ妇女宫颈粘液含铜量２６７例。其结果为自第１年平均含铜量２．６６±０．５８μｇ／ｇ粘液湿重至第１５年降为０．９１±０．６μｇ／ｇ粘液湿重，显示宫颈粘液含铜量随放置时间延长而渐减少。金属圆环组２１４例，各不同年限宫颈粘液含铜量平均为０．３５±０．０８～０．５６±０．１４μｇ／ｇ粘液湿重。未带器组２１例宫颈粘液平均含铜量为０．４７±０．１１μｇ／ｇ粘液湿重，ＶＣｕ２００组与金属圆环组各年限相比均有显著差异（Ｐ＜０．０５～０．００１）。与未带器组比各年ｆＫ亦皆有显著差异（Ｐ＜０．０５～０．００１）。文献报道铜离子浓度达１．０μｇ／ｍｌ有杀死或抑制精子活动的作用，浓度为０．７μｇ／ｍｌ能使孕卵胚泡中毒。本文置ＶＣｕ１２００ＩＵＤ者第１０年时铜离子含量为１．１１μｇ／ｇ粘液湿重，第１５年为０．９１±０．１μｇ／ｇ粘液湿重，依此推测ＶＣｕ２００ＩＵＤ至少可放置１０年以上。     

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。     

白细胞介素8与早产的关系

目的　通过测定宫颈粘液中白细胞介素８（ＩＬ－８）的浓度及中性粒细胞数,探讨早产与感染的关系。　方法　用酶联免疫吸附（ＥＬＩＳＡ）法,对８２例先兆早产孕妇（研究组）和１０４例正常孕妇（对照组）宫颈粘液中ＩＬ－８进行检测,ＩＬ－８≥４５０ μｇ／Ｌ为阳性;在４００倍显微镜下对４９例先兆早产孕妇及１０４例正常孕妇宫颈粘液中中性粒细胞数进行检测。　结果　研究组孕妇宫颈粘液中,ＩＬ－８阳性率及中性粒细胞数明显高于对照组。　结论　通过此研究提示早产与生殖道感染有一定的关系,测定宫颈粘液中ＩＬ－８的含量可望成为预知早产及早产管理的一个有效方法     

整合素亚基α_5、β_1在人精子的表达及其与受精的关系

目的 :研究精子表面整合素亚基 α5、β1的表达及其与精子功能状态和受精力的关系 ;整合素亚基 α5、β1的表达与不明原因不育症的关系。方法 :1 3例生育力正常男性的精液标本 ,9例临床诊断为不明原因不育症患者的精液标本。以精子穿透去透明带的金黄地鼠卵行体外受精试验 (SPA)检测精子受精力 ;对新鲜、获能和孕酮诱导顶体反应后精子行间接免疫荧光染色 ,流式细胞仪检测精子表面整合素亚基α5、β1表达阳性的精子百分率。以三色法染色 ,观察精子顶体反应的发生率。结果 :流式细胞仪检测显示正常组新鲜精子表面α5亚基的表达 (1 3 .3± 5 .4% )与对照组 (1 0 .7± 6 .4% )无显著差异 (P>0 .0 5 )。获能组精子表面 α5亚基的表达率 (6 5 .7± 1 7.6 % )比新鲜组和对照组显著增高 (P<0 .0 5 )。孕酮诱导顶体反应后 ,α5亚基的表达率 (6 5 .3± 1 8.9% )比新鲜组和对照组显著增高 (P<0 .0 5 ) ,但与获能精子组相比无显著差异 (P>0 .0 5 )。β1亚基的表达率分别为新鲜组1 2 .9± 7.4% ,获能组 1 5 .9± 7.9% ,诱导顶体反应组 1 6 .9± 6 .2 % ,与对照组 (分别为 :9.3± 3 .4% ,1 7.3± 7.8% ,1 2 .2± 8.7% )比较均无显著差异 (P>0 .0 5 )。获能精子表面α5亚基的阳性表达率与受精率有一定的线性相关 (r=0 .     

GC／cGMP信号转导途径在雄性生殖作用的研究进展

鸟苷酸环化酶（guanylatecyclases,Gcs）通过催化细胞内第二信使环磷酸鸟苷（cyclicgua—nosinemonophosphate,cGMP）的生成来转导信号;cGMP主要激活其下游效应分子cGMP依赖性蛋白激酶（PKG）、环核苷酸门控离子通道（cNG）和磷酸二酯酶（PDE）,从而介导多种生物学效应。近年来大量研究证实,GC／cGMP信号转导体系与雄性生殖过程密切相关,多种介质通过此信号通路影响精子活力、获能和顶体反应,参与精子发生、运输、成熟及阴茎勃起等过程。     

精子功能检测对选择IVF或ICSI治疗不育症的临床意义

本文评价了精子功能检测对选择体外受精(IVF)或卵胞浆精子注射(ICSI)治疗不育症的临床意义。精子功能缺陷所导致的精子与透明带结合反应(即透明带结合、透明带诱发顶体反应和透明带穿透)异常是IVF受精失败的主要原因。在常规精液分析中,精子形态学测定对预测精子受精能力最有价值,但精子形态学评估则是最难做准确和稳定。IVF受精失败的卵子是很有用的生物材料,可用来测定精子形态和顶体功能。人卵透明带有选择性地与正常形态和完整顶体的精子相结合。透明带诱发顶体反应与精子穿透明带的能力呈显著正相关。在不明原因不育症患者中,约25%的不育原因可能是透明带诱发顶体反应缺陷或低下所引起。在严重畸形精子症(正常形态率≤5%)、少精子症的不育病人中,精子与透明带结合异常比率很高。因此,临床上应把这些有精子缺陷,尤其是会影响精子与透明带结合的患者检测出来,采用ICSI治疗,避免这些病人用IVF治疗而导致很低的受精率。精子形态学分析和精子与透明带结合试验有助于临床选择IVF或ICSI治疗不育患者。     

Ureaplasma urealyticum (T-mycoplasma) in seminal plasma and spermatozoa from infertile and fertile volunteers

138 semen samples from infertile and fertile men were collected, analyzed and examined for the presence of Ureaplasma urealyticum. The organism was present in 29% of samples from infertile men and in 36% of samples from fertile volunteers. Heavy growth was obtained from 32% and 42% of the U. urealyticum-positive semen samples from infertile and fertile men respectively. With few exceptions the organism was isolated from both sperm and seminal plasma fractions of positive semen samples after separation and could be demonstrated on the sperm and between them by electron microscopy. U. urealyticum did not alter the semen characteristics or the sperm penetration and viability in cervical mucus. Tetracycline treatment eradicated the organism from 81% of positive cases but no pregnancies were achieved in the partners of the infertile men during the 6-mth study period.     

Presence of antisperm antibodies in fertile and infertile persons.

The sera of 70 infertile and 30 fertile couples were tested for the presence of sperm agglutinins with Kibrick's macroagglutination technique and Boyden's haemagglutination test. 15.7% of the couples studied showed the presence of sperm agglutinins (by Kibrick's method) of which 5.7% were from males and 10.0% were from females. By the haemagglutination test, 13% of the couples studied were found to possess sperm agglutinins, of which 3% were from males and 10% were from females. 30 fertile men and women studied for sperm agglutinins were found to be negative by both methods. It was also observed that these two tests detected different types of sperm agglutinins. The cervical mucus samples from 45 females (15 fertile and 30 infertile) were tested for sperm agglutinins with a mucus penetration test. 23.1, 16.5, and 57.4% of the samples from infertile females, showed 0-degree, 1-degree and 2-degree penetration respectively. In case of samples from fertile females, 6.6, 13.2 and 79.2% showed 0-degree, 1-degree and 2-degree penetration respectively. Addition of serum from infertile females to cervical mucus from the infertile female increased the 0-degree penetration percent cases to 50%, as compared with 23.1 when only cervical mucus was used. Addition of serum from fertile females, or horse serum or pure albumin or globulins did not increase the 0-degree penetration cases.     

Prognostic value of in vitro sperm penetration into hormonally standardized human cervical mucus

To analyze the prognostic value of the sperm cervical mucus penetration test (SCMPT), fresh semen samples of 99 male patients under infertility investigation were exposed to capillary tubes filled with freshly obtained cervical mucus (CM) of the patients' wives (WCM), fertile donors (DCM), and bovine CM (BCM). The quality of the human CM was standardized by oral administration of estrogens. The overall pregnancy rate after 6 months was 17.2% (17/99), and was significantly different in couples with poor and good SCMPT with WCM (1/44, 2.3% versus 16/55, 29%; P less than 0.001) in a prospective study. Human CM was superior to BCM as a penetration medium in providing more information about sperm function. The results suggest that in vitro sperm penetration testing with hormonally standardized CM of female partners adds an important dimension to sperm analysis with regard to fertility prognosis.     

Penetration of human ejaculated spermatozoa into human and bovine cervical mucus. II. Pattern and velocity of penetration

The pattern of penetration of human ejaculated spermatozoa into human cervical mucus (HCM) and bovine cervical mucus (BCM) was studied in an in vitro penetration system by means of plate capillary tubes. Sperm penetrate the HCM in a unique pattern making their way progressively forwards in a one-way direction in between the filaments. In contrast, sperm move preferably forward in BCM. The different patterns are probably due to different organizations of the glycoproteins. The penetration values (PV) were also different in the two systems when studied after short periods of incubation (15 and 30 min), but incubation for 1 h brought both systems to the same rate of PV. Thus, this period is enough to overcome the differences between the patterns of penetration. Sperm velocity in semen, in HCM, and in BCM were the same. Cervical mucus (CM) did not affect sperm velocity. A strong correlation (r = 0.8) was found between sperm velocity in the semen and its PV rate either in HCM or in BCM. The results of this study recommend the use of BCM as a reference CM in cross penetration tests and suggest its establishment as a routine method for assessment of sperm quality.     

In vitro penetration of human sperm into bovine cervical mucus: effects of sperm washing and exposure to low temperature

Standardized bovine cervical mucus penetration by human sperm in vitro provides information for evaluating male fertility. Normal semen specimens from 20 sperm donors and 17 infertile men were tested for cervical mucus penetration. Rigorous control of test temperature was necessary to guarantee the reliability and reproducibility of cervical mucus penetration. Sperm washing was found to significantly improve cervical mucus penetration for infertile men, from 18 +/- 2.2 to 27 +/- 3.4 mm, P less than .025. Sperm washing for normal donors had no apparent effect on cervical mucus penetration (58 +/- 1.5 mm prewash, 55 +/- 1.7 mm postwash, P greater than .1). The authors conclude that: 1) temperature for cervical mucus penetration testing is critical to reliability, and 2) cervical mucus penetration is a useful screening tool for in vitro procedures proposed to improve sperm function.     

Results of in vitro fertilization in women with antisperm antibodies in serum,cervical mucus,and follicular fluid

This study was undertaken to investigate the presence of antisperm antibodies (ASA) in serum, cervical mucus, and follicular fluid (FF) of women undergoing in vitro fertilization and embryo transfer (IVF-ET). IgG and IgA ASA directed mostly against sperm head were found at similar concentrations in serum, cervical mucus, and FF of 2 of 34 patients. Ninety-one percent fertilization and 100% cleavage rates, respectively, were observed in one of the two patients. No fertilization occurred in the second patient. In both women, in vitro sperm penetration tests revealed hostile mucus and repeated postcoital tests were poor. It is concluded that the sperm-cervical mucus penetration test and mucus ASA measurements are useful in establishing the diagnosis of immunological infertility.     

Ureaplasma urealyticum (T-mycoplasma) in vaginal fluid and cervical mucus from fertile and infertile women

Ureaplasma urealyticum (T-mycoplasma) was isolated more frequently and in heavier growth from cervical mucus (49%) than from vaginal fluid (34%). It was isolated in 24% of vaginal fluid samples and in 35% of cervical mucus samples from fertile women, and in 29% of vaginal fluid samples and in 47% of cervical mucus samples from infertile women. The incidence of infection was high following abortion or total hysterectomy and during pregnancy or oral contraceptive use. T-mycoplasma was also isolated from the vaginal fluid and cervical mucus of a woman with tubo-ovarian abscess, but was not present in women with Trichomonas vaginalis infection. U. urealyticum did not alter the physiophysiologic characteristics of vaginal fluid and cervical mucus or the sperm penetration and sperm viability in cervical mucus. Treatment with tetracycline eradicated the organism in 88% of the infected women. Pregnancies were recorded during a 6-month follow-up in 1 of 19 infertile women who were treated with tetracycline.     

宫颈糜烂不孕患者应用射频自凝刀治疗术后宫颈黏液和妊娠率的改善

目的:探讨宫颈糜烂不孕患者应用射频自凝刀治疗后宫颈黏液和妊娠率的改变。方法:宫颈糜烂不孕患者62例,依其宫颈糜烂程度分为3组:轻度糜烂组(n=8)、中度糜烂组(n=38)、重度糜烂组(n=16)。应用射频自凝刀治疗术治疗,术前行宫颈黏液评分。治疗后随访1年,评价宫颈黏液和妊娠的情况。结果:射频自凝刀治疗总有效率为100%。轻度、中度、重度组的痊愈率分别为100%、94.7%和87.5%。62例患者治疗后宫颈黏液评分与治疗前的比较有显著提高(P<0.01),宫颈黏液性状明显改善。在随访期内,3例妊娠早期流产,9例在孕,4例分娩,妊娠率为21%。结论:射频自凝刀治疗术能改善宫颈糜烂不孕患者的宫颈黏液性状和妊娠率。     

Cytotoxic sperm antibodies inhibit sperm penetration of zona-free hamster eggs

Zona-free hamster egg sperm penetration assay was used to study the effects of cytotoxic sperm antibodies on egg penetration by the sperm of fertile and infertile men. Twenty-nine fertile and 9 infertile men did not have significant cytotoxic sperm antibodies in their serum and seminal plasma; 7 infertile men were positive for these antibodies in serum and seminal plasma. Two others were positive in sera, and 14 were positive in seminal plasma. Sperm from 18 of 23 (78%) infertile men with sperm antibodies had poor egg penetration (less than or equal to 20%) compared with only 6 of 38 (16%) nonautoimmune men (P less than 0.0001). Sperm from nonautoimmune fertile men were coated with seminal plasma and serum of autoimmune men and serum of isoimmune women, resulting in a significant decrease in hamster egg penetration. Sixteen of 21 (76%) seminal plasma samples with cytotoxic sperm antibodies reduced the control sperm penetration of hamster eggs by greater than or equal to 50%. Coating of sperm from fertile men with serum and seminal plasma samples from non-sperm-immune fertile and infertile subjects did not alter their penetration of hamster eggs. Coating of sperm from autoimmune men with cytotoxic antibody-positive autologous seminal plasma samples resulted in a significant decrease of egg penetration. The inhibitory effect of antibody-positive seminal plasma samples on egg penetration by control sperm was abrogated when the samples were preabsorbed with sperm. It is concluded that cytotoxic sperm antibodies, especially those in seminal plasma, inhibit hamster egg penetration by autologous and control sperm. This may explain in part the incidence of infertility associated with sperm antibodies.