锌硒宝治疗男性不育患者精浆锌离子与超氧化物歧化酶含量的研究

研究证明精浆中氧自由基的含量直接影响到精液的质量。微量元素锌 (Zn)直接参与超氧化物歧化酶 (SOD)的结构与活性中心 [1 ] 。精浆中锌的含量比外周血液中高出 10 0倍以上 ,故锌对生育能力有密切的关系。材料和方法一、对象及治疗方案1999年 6月至 2 0 0 0年 12月男性科不育专科门诊就诊的患者。需符合下述标准之一 :婚后未避孕不育 2年以上 ;精液常规检查精子活动度 <2级 ;精子密度 <2 0× 10 6 ;精子活动率 <4 0 % ;精液液化时间 >30 min。共接受患者 78例 ,平均年龄 2 9岁 (2 3～ 38岁 )。治疗组给予锌硒宝 (济南体恒健生物工程有限公…     

抗SPIM自身抗体的ELISA测定与临床应用

用ELISA 法定量检测生育、不育男性血清、精浆抗SPIM 自身抗体(SPIM-Ab)。结果表明:不育组和妻子流产组患者血清、精浆SPIM-Ab 含量均显著高于生育组(P<0.01),两组血清、精浆的SPIM-Ab 阳性率与生育组之间有显著性差异(P<0.01);精浆SPIM-Ab 含量高于5.0U/ml 时,精于密度、精子活率及精浆SPIM 活性均明显降低;SPIM-Ab 与血清、精浆抗精子抗体(AsAb)无明显关系。提示:SPIM-Ab 水平增加可能抑制SPIM 活性、与SPIM一起参与局部免疫反应,并减少精子密度和活率。     

中药生精散对精子DNA完整性的影响及其在IVF-ET治疗中的临床疗效

目的:观察中药生精散对不育患者精子DNA完整性的影响及其在IVF-ET治疗中的临床疗效。方法:因少、弱精子症行IVF/ICSI-ET治疗的患者49例,随机分为中药组(22例,服用生精散治疗)和对照组(27例,未经生精散治疗)。观察比较中药组和对照组精液常规分析(精子密度、活率、活力)、DNA碎片指数(DNA fragmentation index,DFI)、受精率、卵裂率、种植率及临床妊娠率。结果:中药治疗后精子活率及活力显著提高(P<0.05),且中药组DFI值较治疗前及对照组明显下降(P<0.05)。中药组胚胎种植率(31.11%)及临床妊娠率(45.45%)明显高于对照组(13.73%和18.51%)(P<0.05)。结论:中药生精散可通过降低不育患者精子DNA损伤程度,改善精子质量,提高IVF-ET的治疗效果。     

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。     

97例不育男子精浆中微量元素的研究

本文对97例不育男子及13例证实有生育能力男子精浆中的九种微量元素(锌、铁、锰、铜、镍、钾、纳、钙和镁)的浓度用原子吸收分光光度法进行了测定。结果表明:钾、钠和钙在不育与正常男子精浆中的水平几乎一样。不育症患者精浆中的铁、锰和铜浓度高于正常生育力男子精浆中的浓度,其中铁(P<0.05)和锰(P<0.01)的差异具有统计学意义。镍的浓度(P<0.05)显著低于正常男子。具有正常精子密度的不育患者精浆中铁(P<0.05)和锰(P<0.01)显著高于正常男子,锌和铜的浓度也高于正常男子,但无统计学意义。无精患者精浆中的镁(P<0.01)、镍(P<0.05)和铁(P<0.05)浓度明显低于正常男子。镁和镍在不育患者精浆中的浓度随精子密度降低而逐渐降低,但无统计学意义。经迴归处理实验数据,结果表明,锌—钾,锌—钙,锌—镁;钙—钾,钙—镁和镁—钾无论在正常组还是不育组均存在明显的正相关。作者认为结合反映前列腺功能的其他生化指标来测定精浆中的铁、锰、镍及镁的水平有利于对不育症的诊断。     

中国人附睾功能指标—精浆肉毒碱研究

精浆肉毒碱90%左右来源于附睾,是附睾的功能性指标。有关实验证明附睾分泌的肉毒碱是精子在附睾中成熟的重要因素,因此是精子在附睾中成熟的重要功能性指标。本文测定了156例中国正常生育力男子精浆中的肉毒碱,并研讨了精浆肉毒碱与精液其他指标的关系。156例精液的几个主要指标: 精子存活率63.36±18.18%;精子密度111.06±90.83×10~6/ml;精子活动度:均在Ⅱ级及Ⅲ级以上;精液体积3.239±0.532ml; 精浆肉毒碱含量: 每毫升精浆肉毒碱量:239.56±105.59nmol/ml 精浆中肉毒碱总量:757.34±471.72nmol 精浆肉毒碱与精子存活率,精子密度及活动度均有相关性。     

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 .     

男性不育患者精浆NO水平及其与抗精子抗体的关系

目的 :观察男性不育患者精浆中 NO水平变化及其与 As Ab的关系。方法 :采用EL ISA法测定了 1 1 6例男性不育患者及 49例正常生育男性血清和精浆 As Ab,同时采用硝酸还原酶法测定 1 1 6例男性不育患者与 49例正常生育男性精浆中 NO水平。结果 :男性不育患者 As Ab阳性率 3 8.8% ,显著高于正常生育组 4.0 8% ( P<0 .0 1 )。不育组精浆中 NO水平 1 4 2 .8± 3 9.2 μmol/ L,显著高于正常生育组 83 .9± 2 6 .1 μmol/ L( P<0 .0 1 )。As Ab阳性的不育患者 NO水平 1 59.3± 42 .6μmol/ L ,显著高于 As Ab阴性组 1 3 4 .2±3 1 .4μmol/ L ( P<0 .0 1 )。结论 :As Ab与男性不育有着密切相关性 ,且为重要的免疫因素 ,男性不育患者精浆内高水平的 NO与 As Ab的产生可能有着密切的关系。     

附睾降解性不活动精子症的研究

目的 :探讨精子不活动是否由附睾降解引起。方法 :本文选择 5例精液中无活动精子的不育患者 ,应用睾丸精子活力检测、精子头 -尾膜完整性结合试验和透射电镜观察 ,探讨其精子不活动的原因。结果 :5例患者睾丸活检组织所分离的睾丸精子 ,经孵育后的活动率为 2 %～ 1 1 %。睾丸精子组中头膜 -尾膜均完整的精子率显著高于射出精子组 ( P<0 .0 1 )。睾丸精子未见明显的降解 ,但透射电镜显示射出精子的浆膜、核等结构表现显著的降解变化。结论 :本组患者的精子可能经历了病理性附睾降解 ,引致精子丧失活动性 ,对这类患者采用活动的睾丸精子作辅助生育治疗有可能改善成功率。     

人精子和精浆内cAMP含量与获能的关系

cAMP与精子的运动、代谢及获能和顶体反应等密切相关。本文用放射免疫分析法测定了正常男性射出精子内和精浆的cAMP含量及在获能液中培育前后精子内cAMP含量的变化,结果如下:(1)51例正常男性射出精子内cAMP含量为9.03±0.60pmol/10~8精子。(2)51例正常精液中的15例精浆cAMP含量为:14.21±2.33nmol/ml。(3)在获能液中培育30分钟后,9例精子内cAMP含量均较培育前显著提高(P<0.05)。     

External quality control program for semen analysis: Spanish experience

PURPOSE: Results from an external quality control programme for semen analysis carried out in Spain are analysed. METHODS: Quality control materials were distributed and the following seminal parameters were determined: concentration, total motility, progressive motility, rapid progressive motility, morphology and sperm vitality. The between-laboratories coefficients of variation were assessed on different types of quality control material. RESULTS: The majority of participating laboratories utilised manual versus computer-assisted semen analysis methods. Some between-laboratories coefficients of variation ranges were: 20.8-33.8% for concentration (semen pool suspension); 13.9-19.2% for total motility (videotapes); 54.2-70.2% for sperm morphology (strict criteria using stained smears); and 9.8-41.1% for sperm vitality (stained smears). There was an inverse relation between mean percentage of sperm and coefficients of variation between laboratories for sperm motility, morphology and vitality. CONCLUSIONS: These data highlight the urgent need for improvement in the overall quality of andrology testing.     

Vitality of Oligozoospermic Semen Samples Is Improved by Both Swim-Up and Density Gradient Centrifugation Before Cryopreservation

PURPOSE: To ascertain whether washing sperm from oligozoospermic and normozoospermic samples before cryopreservation improves post-thaw vitality. METHODS: Normozoospermic (n = 18) and oligozoospermic (n = 16) samples were divided into three aliquots. The first aliquot remained untreated and the second and third aliquots were subjected to the swim-up and discontinuous density gradient sperm washing techniques respectively. Vitality staining was performed, samples mixed with cryopreservation media and frozen. Spermatozoa were thawed, stained, and vitality quantified and expressed as the percentage of live spermatozoa present. RESULTS: Post-thaw vitality in untreated aliquots from normozoospermic samples (24.9% +/- 2.3; mean +/- SEM) was significantly higher (unpaired t-tests; P < 0.01) than untreated oligozoospermic samples (11.9% +/- 2.3). Post-thaw vitality was significantly higher after swim-up in normozoospermic samples (35.6% +/- 2.1; P < 0.001; one-way ANOVA) and oligozoospermic samples (27.7% +/- 1.7; P < 0.01). Density gradient centrifugation significantly improved post-thaw vitality in oligozoospermic (22.4% +/- 1.0; P < 0.01) but not normozoospermic (30.8% +/- 1.8) samples. CONCLUSIONS: Sperm vitality in cryopreserved oligozoospermic samples was improved by both the swim-up and density gradient centrifugation washing techniques prior to freezing.     

Effects of antioxidant treatment in oligozoospermic and asthenozoospermic men

OBJECTIVE: To evaluate the effect of sairei-to, an herbal medicine, as an antioxidant in oligozoospermia and asthenozoospermia (nonnormozoospermia). STUDY DESIGN: Forty-seven nonnormozoospermic and 16 normozoospermic men were the subjects of this prospective clinical study. After sairei-to (9.0 g/d) was administered daily to the 2 groups for 3 months, sperm parameters, serum hormones and superoxide dismutase (SOD) activity in the serum and the seminal plasma was analyzed. The testicular artery was also assessed. RESULTS: After therapy, serum hormones and SOD activity did not change significantly in either group. Although men with normozoospermia did not undergo a significant change in sperm conditions or testicular artery flow, total sperm concentration (17.1 +/- 20.0 versus 28.7 +/- 35.5 x 10(6)/mL, P = .02) and sperm motility (30.1% +/- 21.6 versus 45.8% +/- 24.4, P < .0001) were significantly increased, and the pulsatility index of the testicular artery (2.03 +/- 0.84 versus 1.64 +/- 0.48, P = .04) was significantly decreased in nonnormozoospermia. CONCLUSION: Treatment with the herbal antioxidant sairei-to improves sperm condition and testicular artery flow in nonnormozoospermia.     

Use of Clomiphene Citrate in the Treatment of Men with High Sperm Chromatin Stability

Objective: To determine whether improvements of the seminal vesicle function after a 5-day course with clomiphene citrate (CC) may reduce the prevalence of men with high sperm chromatin stability under conditions of sodium dodecyl sulfate (SDS)-ethylenediaminetetraacetic acid (EDTA).

Design: A prospective study.

Setting: Andrology laboratory at the Instituto de Investigaciones de la Altura, Universidad Peruana Cayetano Heredia, Lima, Peru.

Patient(s): Forty-one male partners of infertile couples attending the andrology laboratory.

Intervention(s): Clomiphene citrate was administered orally twice a day. Men were treated with CC at 100 mg daily for 5 days. Blood and semen samples were collected before treatment and 24 hours after the last administration.

Main Outcome Measure(s): Serum testosterone, seminal fructose, sperm motility, sperm chromatin stability after SDS and EDTA, and prevalence of high sperm chromatin stability.

Result(s): The percentage of stable sperm after SDS-EDTA correlated inversely with the basal corrected concentration of seminal fructose (−1.77 ± 0.89, β ± SE). High sperm chromatin stability was observed in 53.8% of the study population and in 66.7% of patients with hypofunction of the seminal vesicles. In those men whose seminal vesicle function improved after treatment with CC, the prevalence of high sperm chromatin stability was reduced from 67% to 25% (χ² = 5.34). Logistic regression analysis showed that the higher the basal corrected seminal fructose levels and the higher the basal serum testosterone levels, the lower the probability of nonresponse of the sperm chromatin stability to treatment with CC (0.54 ± 0.15, odds ratio ± SE for corrected fructose; and 0.50 ± 0.15, odds ratio ± SE for serum testosterone).

Conclusion(s): Hypofunction of the seminal vesicles was associated with high sperm chromatin stability, and this high sperm chromatin stability under SDS-EDTA conditions may be reduced by treatment with CC.     

Leukocytospermia and function of the seminal vesicles on seminal quality.

OBJECTIVE: To determine possible relationships between number of leukocytes, function of seminal vesicles, and seminal quality. DESIGN: The study was carried out on men who consecutively attended an infertility clinic between June 1989 to June 1991. SETTING: This study was conducted in a private immunological center for infertility, a tertiary care center, The Centro Immunológico-Sección Esterilidad y Reproducción. PATIENTS: Semen samples from 280 infertility patients attending an Immunological Center for Infertility were analyzed. MAIN OUTCOME MEASURE: We evaluated the effect of leukocytospermia in the presence of normal or abnormal function of seminal vesicles on seminal quality. RESULTS: Sperm count, percent of motile sperm, and percent of sperm vitality were significantly reduced when both leukocytospermia and hypofunction of seminal vesicles were present (P less than 0.01). Leukocytospermic subjects with normal function of seminal vesicles showed similar seminal parameters to those nonleukocytspermics. The incidence of subjects with antisperm antibodies measured by direct immunobeads was significantly higher in leukocytospermic men with hypofunction of seminal vesicles. No differences in the incidence of antisperm antibodies with nonleukocytospermic samples were observed in those with both leukocytospermia and normal function of seminal vesicles. CONCLUSIONS: These data provide evidence that white blood cells were deleterious for seminal quality when seminal vesicles were also affected.     

宫内窘迫胎鼠脑及肝组织自由基损伤的研究

目的 探讨宫内窘迫胎鼠自由基损伤情况及对不同脏器的影响。方法 通过钳夹双侧子宫动、静脉造成宫内胎鼠完全缺血缺氧,其中Ａ组钳夹５分钟（胎鼠１３只）;Ｂ组钳夹１０分钟（胎鼠１０只）;Ｃ组钳夹２０分钟（胎鼠１１只）;对照组为假手术组（胎鼠１３只）。另一部分钳夹后再灌注,其中Ｄ组钳夹１０分钟后再灌注５分钟（胎鼠１１只）,Ｅ组钳夹１０分钟后灌注２０分钟（胎鼠８只）;以Ｂ组为对照。以比色法测定脑、肝组织匀浆后     

Passive immunization with antisperm monoclonal antibody ZAP-7 increases preimplantation embryo mortality in the mouse

Objective: To evaluate the effect of VIC-1 and ZAP-7 antihuman sperm monoclonal antibodies on in vivo fertility in the mouse.

Design: A randomized blinded study using a mouse model.

Setting: University-based laboratory.

Animals: B6CBAF1 mice (n = 6 per experimental group).

Intervention(s): Antisperm antibodies were administered intravaginally to female mice before mating. Control mice received no treatment, saline, or nonspecific antibodies. Number and viability of preimplantation embryos were determined by microscopic observation. Mouse sperm, oocytes, and normal preimplantation embryos were used in indirect immunofluorescence assays with antisperm antibodies. The effect of antibody treatment on sperm motility and vitality was evaluated.

Main Outcome Measure(s): Antigen expression, sperm motility and vitality, number and viability of embryos.

Result(s): ZAP-7 antibody recognizes a sperm antigen expressed in zygotes and early preimplantation embryos. Passive immunization with ZAP-7 increases embryo mortality significantly (more than 40% above controls). Passive immunization with VIC-1 has no deleterious effect.

Conclusion(s): ZAP-7 monoclonal antibody disrupts fertilization and embryogenesis in the mouse.     

Effects of stallion seminal plasma on hydrogen peroxide release by leukocytes exposed to spermatozoa and bacteria

The ability of stallion seminal plasma to modify phagocytosis of spermatozoa and Streptococcus zooepidemicus was examined. Phagocytosis was monitored indirectly as the H2O2 produced by peripheral blood leukocytes after addition of spermatozoa or bacteria. Hydrogen peroxide production after addition of ejaculated spermatozoa was greater (P less than 0.01) than after addition of epididymal sperm. Furthermore, pre-incubation of epididymal sperm with 6.25-50% seminal plasma caused a dose-dependent increase in subsequent H2O2 production by leukocytes (P less than 0.05). In addition, equine serum was capable of opsonizing epididymal and ejaculated sperm. Seminal plasma also directly stimulated phagocyte function because leukocytes preincubated with 12.5% or 25% seminal plasma released more H2O2 after addition of S. zooepidemicus than control leukocytes (P less than 0.05). It is suggested that the opsonization of spermatozoa and the direct stimulation of phagocytes by seminal plasma may represent mechanisms for clearing spermatozoa and bacteria from the reproduction tract.     

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.     

Relationship between seminal antioxidant enzymes and the phospholipid and fatty acid composition of spermatozoa

Sperm cell membranes are susceptible to peroxidative damage through an excess of reactive oxygen species. The objective of this study was to determine seminal plasma glutathione peroxidase (GPX) and superoxide dismutase (SOD) activity and relate these to phospholipid profiles and phospholipid-esterified fatty acid composition of spermatozoa. Seminal plasma GPX and SOD activity, phospholipid, phospholipid-esterified fatty acid composition and malondialdehyde (MDA) of spermatozoa were assayed in 10 normozoospermic and 25 asthenozoospermic subjects. Mean seminal GPX and SOD activity in normozoospermic men were not significantly different from asthenozoospermic men. A significant positive correlation was observed between seminal plasma GPX activity and phosphatidylcholine content (r = +0.77, P = 0.037) and there was a significant negative correlation with lysophosphatidylcholine content (r = -0.89, P = 0.02) in normozoospermic sperm samples. Positive correlations were found between SOD activity and polyunsaturated fatty acid composition of spermatozoa. MDA content in the spermatozoa of asthenozoospermic subjects was significantly higher than in normozoospermic males (P < 0.05). Negative correlations were found between MDA content and seminal SOD activity and arachidonic acid content of spermatozoa from normozoospermic samples (r = -0.5; P = 0.046, r = -0.9; P = 0.001 respectively). Seminal plasma GPX and SOD provide protection against lipid peroxidation of phospholipid and phospholipid-bound fatty acids in normozoospermic samples.