不明原因自发流产妇女配偶射出精子核碱性蛋白组型的电泳分析

本研究从１５例正常生育力男性和２６例不明原因的自发流产妇女配偶的射出精子中提取核碱性蛋白，在１７％的酸性尿素系统聚丙酞胺凝胶中电泳，经微显像测密仪扫描以测定各蛋向工带（组蛋白和鱼精蛋白ＨＰ１～３）的相对含量。结果表明：与正常生育力男性相比，大部分患者（１９／２例）精子的ＨＰＺ和ＨＰ３含量较低（Ｐ＜０．０１），其中４例同时伴有较高量的组蛋白。本实验结果提示过高的组蛋白／ＨＰ１～３比值和／或过低的ＨＰ２＋３／ＨＰ１比值可能与部分不明原因的自发流产有关。     

胰岛素样生长因子及其结合蛋白对体外精子活力的影响

精浆中含有胰岛素样生长因子（ＩＧＦｓ）、ＩＧＦ结合蛋白（ＩＧＦＢＰｓ）和ＩＧＦＢＰ 蛋白酶已被证实,但对它们的功能尚不了解。本文使用上泳法优选精子,并应用计算机辅助精子分析技术（ＣＡＳＡ）研究ＩＧＦｓ 和ＩＧＦＢＰｓ 对体外精子活动参数的影响,如：曲线速度（ＣＶ）、直线速度（ＰＶ）、直线性（Ｌｎ）、前向性（Ｓｔ）、精子头侧摆幅度（ＡＬＨ）和鞭毛摆动频率（ＢＦ）。用方差分析评价各试验组与对照组间随培育时间精子活动参数的变化。结果显示ＩＧＦⅠ使ＣＶ 和ＡＬＨ 显著减少（Ｐ＜ ０．０５）;而ＩＧＦ Ⅱ试验组精子活动参数没有变化。完整的ＩＧＦＢＰ ３使Ｌｎ、Ｓｔ、ＢＦ显著增加（Ｐ＜ ０．０５）,使ＡＬＨ 显著减少（Ｐ＜ ０．０５）;但ＩＧＦＢＰ ２对精子活动参数却没有显著的影响。当ＩＧＦ Ⅰ和ＩＧＦＢＰ ３同时被加到精子样品中培育时,未发现对精子活动参数有统计意义的改变。我们的结论是ＩＧＦ Ⅰ和完整的ＩＧＦＢＰ ３对体外精子活动参数有着不同、而且是相反的影响,这可能在生育调节方面有一定的功用。     

人精子体外获能过程中活动和运动能力和动态变化与体外受精能力的…

通过６５例人精子在普通和高渗ＢＷＷ增基中获能３．６．１２、２４、３６、４８ｈ，观察其活动和运动能力并进行受精能力测定，试图利用常规参数的动力学变化指标评价其受精能力。结果显示：不同时项指标及不同时项的降低百分率均与受精率明显相关（Ｐ＜０．０５－０．０００１），不同分组下各指标的受精率有显著差异（Ｐ＜０．０５－０．０００５）；进一步分析了每小时平均降低百分率（ＡＤＲ）和递降百分率（ＧＤＲ）。报告了一     

人精子体外获能过程中活动和运动能力的动态变化与体外受精能力的相关性

通过６５例人精子在普通和高渗ＢＷＷ培基中获能３、６、１２、２４、３６、４８ｈ，观察其活动和运动能力并进行受精能力测定，试图利用常规参数的动力学变化指标评价其受精能力。结果显示：不同时项各指标及不同时项的降低百分率均与受精率明显相关（Ｐ＜０．０５～０．０００１），不同分组下各指标的受精率有显著差异（Ｐ＜０．０５～０．０００１）；进一步分析了每小时平均降低百分率（ＡＤＲ）和递降百分率（ＧＤＲ）。报告了一组精子活动和运动指标不同变化下的受精率正常值，提出对获能过程中精子基本性质进行动态观察，特别是其动力学表现对研究精子的功能和质量显得更为重要。     

丹参加川芎嗪对妊高征孕妇血栓素A2及前列环素变化的影响

用丹参加川芎嗪对３０例妊高征孕妇进行治疗，并分别测定治疗前后孕妇的平均动脉压（ＭＡＰ）、尿蛋白含量，用放射免疫分析法测定母体外周血血栓素Ｂ＿２（ＴＸＢ＿２）和６－酮基－前列腺素Ｆ＿１α（６－ｋｅｔｏ－ＰＧＦ＿１α）含量。通过分析ＴＸＢ＿２和６－ｋｅｔｏ－ＰＧＦ＿１α含量及其比值的改变，结合ＭＡＰ和尿蛋白变化探讨二种中药合用对妊高征的扩血管作用。结果提示：用药后轻、中、重度妊高征ＭＡＰ和尿蛋白含量均显著下降（Ｐ＜０．０５）；ＴＸＢ＿２变化不大；６－ｋｅｔｏ－ＰＧＦ＿１α显著升高（Ｐ＜０．０５～０．０１）；ＴＸＢ＿２与６－ｋｅｔｏ－ＰＧＦ＿１α比值显著下降（Ｐ＜０．０５）。提示丹参加川芎嗪治疗妊高征有扩张血管、降低外周阻力及增加器官血液灌注作用。     

免不1号治疗79例抗精子抗体阳性不育症的临床研究

采用中药免不１号治疗抗精子抗体引起的免疫性不育症７９ 例，并设西药强的松３０例作对照。治疗结果：免不１号妊娠３１ 例，妊娠率３９．２％ ，强的松组妊娠３例，妊娠率１０％ ，两组比较有显著性差异（Ｐ＜ ０．０５）。治疗后，免不１ 号组精液复查，精子密度、存活率、前向精子活力等有明显改善（Ｐ＜ ０．０１），强的松组改善不明显（Ｐ＞ ０．０５）。免疫性不育症中医辨证分三型，其中脾肾两虚型经免不１ 号治疗后妊娠率最高（４２．１％ ）。     

输卵管结扎术后盆腔静脉瘀血综合征患者腹腔液中内皮素-1和心钠素的变化

目的：测定妇女腹腔液内皮素－１（ＥＴ－１）和心钠素（ＡＮＰ）浓度，探讨其在结扎术后盆腔静脉瘀血综合征（ＰＶＣＳＳ）发病中的作用。方法：应用放射免疫技术对２１例ＰＶＣＳＳ患者（ＰＶＣＳＳ组）腹腔液及血浆中ＥＴ－１和ＡＮＰ浓度、腹腔液量和巨噬细胞密度进行测定，并与１２例绝育术后正常妇女（ＮＷＳ组）、１１例正常妇女（ＮＷ组）进行比较。结果：ＰＶＣＳＳ组腹腔液ＥＴ－１浓度、ＥＴ－１／ＡＮＰ比值、巨噬细胞密度均低于ＮＷＳ组和ＮＷ组（Ｐ＜０．００５，Ｐ＜０．００１，Ｐ＜０．００１），且均与疾病严重性评分呈负相关（Ｐ＜０．０５），但腹腔液量多于ＮＷＳ组和ＮＷ组（Ｐ＜０．００１）；各组巨噬细胞密度均与ＥＴ－１浓度呈正相关（Ｐ＜０．０５）；各组血浆ＥＴ－１和ＡＮＰ浓度间差异无显著性（Ｐ＞０．０５）。结论：妇女腹腔液含有一定浓度ＥＴ－１，腹腔液ＥＴ－１浓度和ＥＴ－１／ＡＮＰ比值降低，与ＰＶＣＳＳ发病有关。     

轻度子宫内膜异位症伴不育患者腹腔液对精子穿卵及精子膜的影响

本研究对轻度子宫内膜异位症患者（３０例）及正常对照组（１５例）在腹腔镜直视下收集腹腔液,正常人精液按１：１比例稀释,以去透明带地鼠卵穿透试验（ＨＯＰ）、精子尾部低渗肿胀试验（ＨＯＳ）及精子头部ＤＮＡ荧光染色有效精子计数（ＥＳＣ）为指标,探讨轻度子宫内膜异位症（ＥＭ）患者的腹腔液对精子受精能力、精子膜及ＤＮＡ 双螺旋结构的影响。结果显示,ＥＭ组精子穿卵率和受精卵外精子附着数明显低于正常对照组（Ｐ＜０．０１）,ＥＭ组的 ＨＯＳ明显低于正常对照组（Ｐ＜０． ０５）,而 ＥＭ组的 ＥＳＣ虽有所下降,但与正常对照组相比无显著性差异。结果提示：轻度ＥＭ腹腔液对精子受精能力及精子膜结构的完整性有影响。     

雷公藤单体T4对大鼠变态期精子细胞核蛋白转换的影响

大鼠喂服雷公藤单体Ｔ４７周后，取睾丸分离长形精子细胞核，取附睾分离精子核，提取总碱性核蛋白（ＴＮＢＰ），经电泳扫描后分析，发现长形精子细胞核ＴＨ／ＲＰ（总组蛋白／大鼠精核蛋白）比值升高，糖核蛋白含量下降，附睾精子核碱性蛋白也发生类似的改变，表明睾丸变态期精子细胞组蛋白一精核蛋白取代反应受阻，进而导致附睾精子核蛋白异常，这可能是Ｔ４导致大鼠不育的重要原因。文中还对精核蛋白与生育力的关系予以讨论。     

反复自然流产绒毛生化学改变的研究

木研究对３４例反复自然流产（ＲＳＡ）、１０例１次流产及３８例正常妊娠人流绒毛组织，应用生物化学及免疫组化方法，检测绒毛组织中谷胱甘肽硫转移酶（ＧＳＴ）活性、腺苷脱氨酶（ＡＤＡ）活性和还原型谷胱甘肽（ＧＳＨ）含量及γ谷氨酸转肽酶（γ－ＧＴ）免疫组化定位进行对比性研究。结果表明，ＲＳＡ绒毛组织较正常绒毛组织中ＧＳＴ活性和ＡＤＡ活性均有明显改变（Ｐ＜０．０１）。γ－ＧＴ免疫组化定位、阳性颗粒的分布及强度也有明显的差别。自然流产３次以上较２次和１次流产的ＧＳＨ含量和ＧＳＴ活性明显降低，经统计学分析均有非常显著差别（Ｐ＜０．０１）。该结果提示绒毛组织本身某些生化功能的改变，可能是原因不明的ＲＳＡ的原因之一。     

不明原因的不育男性精子核碱性蛋白组型的电泳分析

本研究从15例正常生育力男性和37例不育男性的射出精子中提取核碱性蛋白,在酸性尿素系统聚丙酰胺凝胶中电泳,经微显像测密仪扫描获得各蛋白区带(组蛋白和HP1～3)的相对含量,并计算各蛋白条带的相对比值.实验结果表明:与正常生育力男性相比,不育组男性精子的TH(total histones)/HP1～3比值增高(P<0.01);而HP2+3/HP1比值降低(P<0.01).基于TH/HP1～3和HP2+3/HP1比值的分折,这些不育男性精子的核碱性蛋白组型可分为四种:1.TH/HP1～3比值较高,同时伴有较低的HP2十3/HP1比值;2.TH/HP1～3比值较高,而HP2+3/HP1比值正常;3,TH/HP1～3比值正常,而HP2十3/HP1比值较低;4.正常的TH/HP1～3和HP2十3/HP1比值.作者认为前三种异常的核蛋白组型可能与不育有关.     

Sperm head vacuolization affects clinical outcome in ICSI cycle. A proposal of a cut-off value

Objective

To evaluate the relationship between sperm nuclear vacuoles and sperm morphology and to investigate the influence of the rate of spermatozoa with head vacuolization (SVR) in a seminal sample on the clinical outcomes in couples undergoing intracytoplasmic sperm injection.

Materials

26 patients undergoing infertility investigations were included and were divided in two groups according to an SVR ≤ 20,28 % (Group A) or > 20,28 % (Group B), and were investigated to verify the influence of SVR on the fertilization rate, embryo quality, pregnancy and implantation rates.

Results

Abnormal spermatozoa with nuclear vacuoles were significantly higher (p < 0.001) than the percentage of normal spermatozoa with nuclear vacuoles. Patients in group A had a percentage of abnormal sperm with nuclear vacuole significantly lower compared to group B (p < 0,001), but there was no difference in the percentage of normal sperm with nuclear vacuoles. Fertilization rates and the number of top quality embryos did not differ between the two groups. The pregnancy and implantation rates were significantly higher in Group A compared to Group B (respectively p < 0,05 and p < 0.001).

Conclusions

For the first time, we propose a cut off value in the proportion of sperms with nuclear vacuolization on the total of sperm in seminal samples, and demonstrate a relationship between SNV and clinical outcomes after ICSI. The SNV rate could be introduced as an easy diagnostic evaluation prior to perform an ICSI cycle.     

A prospective randomized study to evaluate the effect of hyaluronic acid sperm selection on the intracytoplasmic sperm injection outcome of patients with unexplained infertility having normal semen parameters

Purpose

Sperm quality plays an important role in determining embryo development and intracytoplasmic sperm injection (ICSI) outcome. Selection of competent sperm based on its ability to bind to hyaluronic acid (HA) has been suggested as one of the methods to assess sperm quality. The aim of the present study was to examine whether injection of HA bound sperm helps in improving outcome in patients undergoing ICSI with unexplained infertility having normal semen parameters.

Methods

Patients with unexplained infertility having normal semen parameters in accordance with WHO 2010 criterion, undergoing their first IVF-ICSI cycle were enrolled during the course of the study.156 patients were prospectively randomized after oocyte retrieval and were assigned to either the ICSI group, where sperm selection for injection was based on visual assessment, or the PICSI group, where sperm were selected based on their ability to bind to HA. Only fresh embryo transfers were included in the analysis.

Results

There was no difference in the fertilization rates, number of top quality embryos and clinical pregnancy rates between the ICSI and PICSI groups (65.7 % vs 64.7 %; 45.8 % vs 43.6 % and 35 % vs 35.2 % respectively). However, a higher pregnancy loss rate was observed in the ICSI group (25 % vs 12 %; P = 0.227) as compared to the PICSI group, but the difference was not statistically significant. Implantation rates were 22.03 % and 18.84 % in the PICSI and ICSI groups respectively. There were 22 (31 %) live births in the PICSI group and 21(26.3 %) live births in the ICSI group.

Conclusions

Patients with unexplained infertility having normal semen parameters may constitute a patient group which does not benefit from this sperm selection method. A larger study may be necessary to establish a relationship between PICSI and pregnancy loss rate in patients undergoing IVF with unexplained infertility.     

Sperm DNA and chromatin integrity in semen samples used for intrauterine insemination

Background

Sperm DNA damage is associated with male infertility but whether normozoospermic infertile men also have DNA damage is unknown.

Objective

To evaluate sperm DNA and chromatin integrity in men with mild male factor infertility.

Design, setting and participants

Prospective study of 102 consecutive men (78 normozoospermic, 15 asthenozoospermic, 9 oligozoospermic) enrolled for intrauterine insemination (IUI) and 15 fertile controls.

Outcome measurements and statistical analysis

Standard semen parameters and sperm chromatin and DNA integrity were assessed and compared between groups. Sperm chromatin quality was assessed by (1) aniline blue staining (AB is specific to histone lysines), (2) iodoacetamide fluorescein fluorescence (IAF targets free protamine sulfhydryl groups) and (3) sperm chromatin structure assay (SCSA) with the results expressed as % DNA fragmentation index (%DFI).

Results and limitations

The mean (±SD) percentage of spermatozoa with positive IAF fluorescence was significantly higher in the IUI population compared to fertile controls (17 % ± 10 % vs. 8 % ± 6 %, P = 0.0011) and also in the normozoospermic subset (n = 78) compared to controls (16 % ± 9 % vs. 8 % ± 6 %, P < 0.0001, ANOVA). We also observed a trend toward lower %progressive motility, and higher %AB staining and %DFI in the IUI group compared to controls. We observed significant relationships between sperm %DFI and progressive motility (r = −0.40, P < 0.0001) and between positive AB staining and IAF fluorescence (r = 0.58, P < 0.0001).

Conclusions

The data indicate that sperm chromatin integrity may be abnormal in men enrolled in IUI treatment cycles, despite the fact that most of these men are normozoospermic.     

溶脲脲原体感染与精子顶体反应

本实验对精液溶脲脲原体培养阳性的不育男性和其它不明原因不育男性及正常生育男性分别进行了精子明胶膜顶体反应和透射电镜观察。结果发现：在精液溶脲脲原体培养阳性的不育男性组中，严重感染者精子顶体反应率比其它不明原因不育组和正常生育男性组低，经统计学处理有显著意义（Ｐ１＜０．０１，Ｐ２＜０．０１）。电镜观察显示：溶脲脲原体感染者精子膜上有溶脲脲原体颗粒附着，出现精子膜和顶体膜缺损及破坏现象。本实验证实：溶脲脲原体感染可引起精子膜包括顶体膜的破损，精子顶体反应降低或消失。     

棉酚对体外家兔精子顶体酶类的抑制作用

本文对棉酚体外抑制１０种家兔精子顶体酶活性进行了测定。当醋酸棉酚浓度为１２～７６μｍｏｌ／Ｌ时，可完全或明显抑制顶体酶（ａｃｒｏｓｉｎ）、Ａｚｏｃｏｌｌ蛋白酶、芳香基硫酸酯酶和神经氨酸苷酶活性；而当浓度高达３８０μｍｏｌ／Ｌ时才能抑制透明质酸酶、β－葡糖苷酸酶和酸性磷酸酶，但不抑制磷酸酯酶Ｃ、碱性磷酸；酶和β－Ｎ－乙酰氨基葡糖苷酶。棉酚对芳香基硫酸酯酶的抑制作用是一种非竞争性抑制作用，Ｋｉ为１２０μｍｏｌ／Ｌ。其抑制作用是可逆的，呈剂量依赖关系。由于抑制家兔精子顶体酶所需棉酚浓度比精子糖酵解或能量代谢酶要低，因此，这些敏感酶似可作为监护棉酚引起不育的指标。     

不同源性及不同参数精子对ICSI结局的影响

目的:研究不同源性精子、不同参数精子及冻融精子对ICSI结局的影响。方法:接受ICSI治疗的510对不育夫妇共进行了517个周期,分为6组。射出精液少弱畸精子症组(A组)82例,共进行85个周期;射出精液严重少弱精子症组(B组)170例,共进行174个周期;附睾细针穿刺抽吸精子组(C组)108例,共进行108个周期;睾丸细针穿刺抽吸精子组(D组)71例,共进行71个周期;冻融射出精子组(E组)34例,共进行34个周期;冻融附睾睾丸精子组(F组)45例,共进行45个周期;比较其妊娠结局。结果:6组患者一般情况比较无统计学差异(P>0.05),6组患者的受精率、优胚率、临床妊娠率、早期流产率无统计学差异(P>0.05)。结论:不同源性精子、不同参数精子及冻融精子对ICSI结局无明显影响。     

人精子膜上凝集素受体的研究

从糖专一性相同或不同的24种凝集素中筛选出SML、MDL、CKL和PHA-S四种能与人精子发生强烈反应的凝集素,浓度为0.98μg/ml时就能使精子凝集成块,6 h后,凝集团块也不分散,用机械方法分散团块所得精子已完全失去活动能力;其凝集作用可被凝集素各自的抑制糖所抑制;用FITC标记的SML、CKL、MDL与正常人精子作用后,在荧光显微镜下可观察到各凝集素受体的分布部位各不相同;用去垢剂增溶精子,Brij 58的增溶效果最好,增溶的膜蛋白可强烈地抑制凝集素对兔红细胞的凝集作用。