精液体外处理方法的选择

精液体外处理的目的是要达到符合要求的活动精子的密度和活力 ,减少或去除精浆内前列腺素、免疫活性细胞、抗精子抗体、细菌和碎片 ,减少精液的粘稠度 ,促进精子获能 ,改善精子受精能力 ,使能取得丈夫精液人工授精 (ＡＩＨ)的妊娠。但临床报道的受孕率高低不一[1] ,除了精子本身受精功能障碍或卵子异常外 ,精子体外处理方法的选择和效果也是重要的原因之一 ,本文就体外处理方法的选择作一综述。一、精液体外处理方法的选择丈夫精液人工授精的指征至今仍存在争论 ,一般认为性交障碍或异常、精子在女性生殖道内运行障碍、精液异常等均可进行Ａ…     

不同pH值营养液对精液处理后活力的影响

应用营养液对精液进行体外处理是辅助生殖技术实施中的重要环节 ,其目的是去除精液中部分细胞碎片、细菌、前列腺素、抗精子抗体、免疫活性细胞等影响精子活力的物质 ,提高活动精子的百分率 ,改善精子受精能力 ,取得人工授精的成功。营养液的pH值对处理后精液中精子活力的影响     

不育男性精浆碳酸氢钠含量与精子活力等参数的关系

本文对80例不育症患者测定精浆N_aHCO_3含量,同时与粘液质量相比较。认为,精浆中N_aHCO_3含量与精子活力有着密切的关系,而与精液的PH值、粘稠度、精子密度无关,并就精浆N_aHCO_3含量测定在男子不育症诊治方面的意义进行探讨。     

聚精丸对弱精子症患者精浆一氧化氮、超氧化物歧化酶的作用机制研究

目的:研究聚精丸治疗弱精子症的可能机制,为寻找其作用机制提供新的线索。方法:选择弱精子症患者34例,给予聚精丸治疗3个月,治疗前后分别作精液常规及精浆一氧化氮(NO)含量和超氧化物歧化酶(SOD)活性测定,观察治疗前后精液常规参数以及NO含量、SOD活性的变化。结果:与治疗前相比,治疗后精液常规参数均有显著性改变(P<0.05或P<0.01);精浆NO值无明显变化(P>0.05);而SOD活性由(95.97±20.75)μg/L下降为(76.14±19.99)μg/L(P<0.05)。弱精子症治疗前精浆NO含量和SOD活性之间呈负相关(r=-0.246,P<0.05)。结论:聚精丸可以显著改善弱精子症患者精子活动率,但聚精丸的良好疗效可能并不体现在精浆NO含量和SOD活性值的改变。     

不同剂量中波紫外线照射对角质形成细胞的氧化损伤作用

目的:观察不同剂量中波紫外线(UVB)照射对角质形成细胞(Ha Ca T细胞)的氧化损伤作用。方法:体外培养人永生化角质形成细胞,用5m J/cm~2、10m J/cm~2、20m J/cm~2、30m J/cm~2、40m J/cm~2UVB照射,24h后检测细胞丙二醛(MDA)的含量、超氧化物歧化酶(SOD)的活性、细胞上清乳酸脱氢酶(LDH)的含量及角质形成细胞的增殖活性。结果:UVB照射Hacat细胞后,细胞SOD活性降低、MDA含量增加、LDH漏出量增加及细胞增殖抑制均呈剂量依赖性。当剂量达到10m J/cm~2时,首先出现LDH漏出量增加及细胞增殖活性的抑制,20m J/cm~2时Hacat细胞的SOD活性降低,30~40m J/cm~2MDA含量才开始出现明显增加。结论:30~40m J/cm~2的UVB照射角质形成细胞能诱发氧化损伤作用,为今后体外光老化研究提供参考。     

精子体外处理技术

精子体外处理技术辽宁省计划生育科研所李宏军综述少精子、精子活力低下、畸形精子、精液液化不良：精浆中存在抗精子抗体等引起的不育，在经内外科系统治疗都不能使精子质量改善并达到受精目的，而患者又不愿接受供者精液人工授精时，实验室精子体外处理技术是一个重要的...     

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。     

一氧化氮、一氧化氮合酶与伴精索静脉曲张不育患者精液参数的关系

目的:探讨一氧化氮(NO)、一氧化氮合酶(NOS)与伴精索静脉曲张(VC)不育患者精液参数之间的关系。方法:根据体格检查和彩色多普勒超声检查选择伴VC的不育患者(组1,n=53),其中临床型和亚临床型分别为21例和32例;同时选择非VC少弱精子症患者(组2,n=29)和正常生育者(组3,n=28)作对照组。采用硝酸还原法分别测定外周血和精浆中NO含量和NOS活性。用计算机辅助精液分析仪测定VC组患者精子密度、活动精子(a+b级精子)和快速前向运动精子百分率。结果:①组1外周血清NO含量和NOS活性与组2及组3相比差异无显著性(P>0.05),但精浆中NO含量和NOS活性组1明显高于其他两组,差异有显著性(P<0.01和P<0.05)。②组1中,随着曲张的精索静脉内径的增加,外周血清和精浆中NO含量和NOS活性均有所上升,但只有精浆中临床型和亚临床型之间相比差异有显著性(P<0.05)。③组1中,随着精子密度和精子活力的下降,外周血清和精浆中NO含量和NOS活性均有上升趋势,且精子密度≥20×106/ml和≤10×106/ml之间,精子活力≥50%和≤25%之间差异有显著性(P均<0.05)。结论:在VC诊断中精浆中NO含量和NOS活性测定较外周血清中更有意义。早期测定精浆NO含量和NOS活性对VC的诊断和治疗具有重要的临床价值。     

温度对体外精子活力的影响

将42名生育力男子的精液置于17°～37℃间的4组不同温度下检查精子活力,以探讨温度对体外精子活力的影响。结果表明:温度对精子前向运动和活动精子百分率有显著影响,而前向运动级别比活动精子百分率对温度的敏感性更高。36～37℃精子活力最好。24°～25℃组与36°～37℃组之间,精子前向运动级别(0～Ⅲ级制)和活动精子百分率的差异无统计学意义。低于24℃则活力下降。文章认为精液常规和有关精子活力的检查宜在24℃以上进行,需要发挥精子活力的实验宜在37℃左右进行。     

锌剂治疗合并前列腺炎的男性不育患者38例

目的:探讨补充锌剂在治疗合并前列腺炎的男性不育中的作用。　方法:应用补充含有生物态锌的活性 蛋白制剂锌硒宝治疗38例合并前列腺炎的男性不育患者,测定治疗前后患者精浆中锌离子浓度,并观察治疗前后 患者的精液常规参数(包括精液液化时间、精子密度、精子存活率、前向运动精子百分率以及精子形态学检查等)的 变化。　结果:38例患者经补充锌剂治疗后,精浆中锌离子浓度明显提高,精子存活率和前向运动精子百分率明 显改善,其改善幅度均显著大于未补充蛋白锌的对照组(P均<0.05)。　结论:补充锌剂可提高合并前列腺炎的 男性不育患者精浆中的锌离子浓度,有助于患者精液质量(特别是精子存活率、前向运动精子百分率)改善,是合并 前列腺炎的男性不育患者的有效辅助治疗方法之一。     

The Effects of Laser Light on Sperm Motility and Velocity in vitro

The effect of Laser light on the motility and the velocity of human spermatozoa were measured by means of multiple exposure photography. Total sperm motility increased after Laser irradiation at 4 J/cm2, 8 J/cm2 and 32 J/cm2 respectively with respect to control. However, no influence on sperm velocity was demonstrated after Laser irradiation. This observation suggests that Laser light stimulates non-motile live spermatozoa.     

Correlates of human sperm motility assessed by laser Doppler spectroscopy

The sperm motility characteristics of 140 men (percentage motile and average velocity of all sperm in motion; percentage progressive and the average velocity of sperm swimming more than 15 microns/sec) were determined using a laser-Doppler technique and correlated with other aspects of sperm quality, including the concentration and the proportion of abnormal and dead sperm in the ejaculate. In addition, the influence of the length of the period of abstinence, the viscosity of seminal plasma and the volume of the ejaculate were also assessed. The four motility characteristics were all highly correlated with each other. The magnitude of all four parameters increased in an exponential fashion with increasing sperm number up to 400 x 10(6) per ejaculate. At higher numbers, no further improvement in motility was observed. Moreover, increasing sperm number was associated with a decline in the proportion of sperm exhibiting abnormalities in morphology but with an increase in viable sperm 30 min after ejaculation. The relative viscosity of the ejaculates had generally no influence on sperm motility. In contrast, certain of the sperm motility characteristics, including the average velocity, were significantly negatively correlated with the length of the abstinence period.     

Studies on relationship between semen viscosity and other semen parameters, Ureaplasma urealyticum infection and seminal plasma antisperm antibody

Objective: To study the relationship between semen viscosity and other semen parameters, Ureaplasma urealyticum (UU) infection and seminal plasma antisperm antibody (AsAb) in male infertiles. Methods: Semen parameters, Ureaplasma urealyticum (UU) infection and antisperm antibody (AsAb) were measured and analyzed in 4337 infertile men. Results: The seminal viscosity was higherr than normal in 65.02 % of 4337 male infertiles. The sperm motility and grade (a, b) motile sperm were significantly lower in the high viscosity group than in the normal viscosity group (P<0.05-0.01). The rate of abnormal morphology sperm was higher and duration of semen liquefaction was longer in the high viscosity than in the normal viscosity group (P<0.01). The seminal volume, sperm concentration and semen pH were not significantly different between the two groups. The semen viscosity is significantly higher in subjects with higher seminal WBC (>5/ HP) than in those with lower WBC (<5/HP). The positive AsAb and UU infection rates     

不育男性精浆锌、铜和超氧化物歧化酶与衣原体感染的关系

目的观察沙眼衣原体(CT)感染与精浆锌(Zn)、铜(Cu)和超氧化物歧化酶(SOD)含量的关系。方法应用精子活体染色技术和Zn、Cu,SOD含量检测试剂盒,分别检测52例正常生育男性(正常对照组)和58例CT感染不育男性(CT感染组)的精子存活率与精浆Zn、Cu、SOD活性。结果CT感染组中精子存活率、精浆Zn、Cu、SOD含量较正常对照组显著降低(P<0.01)。结论CT感染可引起精浆Zn、Cu、SOD含量降低,精子膜发生氧化损伤,影响精子的存活率和活动力。     

Metenkephalin in Seminal Plasma of Infertile Men

Background: Enkephalin is one of the opioids, which is expressed widely in reproductive organs. However, the function of enkephalin in male reproduction is not completely understood. The effect of metenkephalin on sperm motility remains especially controversial. In this study we examined the level of metenkephalin in seminal plasma from men with normal sperm production and patients with asthenospermia, oligospermia, and azoospermia to investigate the role of metenkephalin in seminal plasma on sperm function. We also investigated the effect of metenkephalin on sperm motility in vitro.
Methods: Sixty nine infertile patients (31 oligospermic, 21 asthenospermic, and 17 azoospermic) were included in this study. The level of metenkephalin in seminal plasma of these men was measured and the effect of the peptide on the motility of human sperm was examined in vitro. Seventeen men with normal seminograms were a control group.
Results: The level of metenkephalin in the seminal plasma of semen from asthenospermic men was significantly lower than that from the controls ( P < 0.05). No significant correlations between the level of metenkephalin and the mean pathing or progressive velocity of sperm, or serum hormone levels were observed. In the in vitro study, which used semen from the controls, treatment of sperm with metenkephalin (50–200 pg/mL) maintained sperm motility for 4 hours. On the other hand, motility of sperm incubated without metenkephalin began to decrease at 3 hours. Metenkephalin levels of 50 pg/mL in seminal plasma is considered to be necessary for maintaining sperm motility.
Conclusion: These results suggest that metenkephalin in seminal plasma is an important clue in the investigation of decreased sperm motility.     

The effect of reactive oxygen species on equine sperm motility, viability, acrosomal integrity, mitochondrial membrane potential, and membrane lipid peroxidation

The objective of this study was to examine the influence of reactive oxygen species (ROS), generated through the use of the xanthine (X)-xanthine oxidase (XO) system, on equine sperm motility, viability, acrosomal integrity, mitochondrial membrane potential, and membrane lipid peroxidation. Equine spermatozoa were separated from seminal plasma on a discontinuous Percoll gradient, and spermatozoa were incubated with 0.6 mM X and 0.05 U/mL XO for 30 minutes. Catalase (150 U/mL), superoxide dismutase (SOD, 150 U/mL), or glutathione (GSH, 1.5 mM) were evaluated for their ability to preserve sperm function in the presence of the induced oxidative stress. At the end of the 30-minute incubation, sperm motility was determined by computer-assisted semen analysis. Viability and acrosomal integrity were determined by Hoechst-Pisum sativum staining, and mitochondrial membrane potential was determined by staining with JC-1. Incubation with the X-XO system led to a significant (P < .01) increase in hydrogen peroxide production and an associated decrease (P < .01) in motility parameters. Total motility was significantly (P < .01) lower in the presence of X-XO compared with the case of the control (29%+/-9% vs 73%+/-1%, respectively). Catalase, but not SOD, prevented a decline in motility secondary to oxidative stress (71%+/-4% vs 30%+/-3%, respectively). The addition of glutathione had an intermediate effect in preserving sperm motility at the end of the 30-minute incubation (53%+/-3%). No influence of X-XO could be determined on viability, acrosomal integrity, or mitochondrial membrane potential. In order to promote lipid peroxidation, samples were incubated with ferrous sulfate (0.64 mM) and sodium ascorbate (20 mM) for 2 hours after the X-XO incubation. No increase in membrane lipid peroxidation was detected. This study indicates that hydrogen peroxide is the major ROS responsible for damage to equine spermatozoa. The decrease in sperm motility associated with ROS occurs in the absence of any detectable decrease in viability, acrosomal integrity, or mitochondrial membrane potential or of any detectable increase in lipid peroxidation.     

体外添加黄芪注射液对人精子膜拮抗脂质过氧化的作用

目的 :观察体外添加黄芪注射液对弱精子症患者精子膜脂质过氧化的拮抗作用。方法 :应用计算机辅助精子分析仪 (CASA)和丙二醛 (MDA)及超氧化物歧化酶 (SOD)检测试剂盒 ,分别分析体外添加黄芪注射液 (A组 )对 30例弱精子症患者精子运动参数的影响 ,检测精子悬液MDA含量及总SOD(TSOD)力 ,并进行相关分析 ,同时设立空白对照组进行对比观察。结果 :A组的精子活率 (Mot)、前向运动精子百分率、曲线速度 (VCL)、平均路径速度 (VAP)和精子头摆幅度 (ALH)较空白对照组显著提高 (P <0 .0 5或P <0 .0 1) ;A组精子悬液的MDA含量显著降低 (P <0 .0 5 ) ,TSOD活力与对照组比较差异无统计学意义 (P >0 .0 5 )。A组MDA含量与Mot、前向运动精子百分率、VAP和ALH间均有显著性负相关 (P <0 .0 5或P <0 .0 1)。结论 :体外添加黄芪注射液对弱精子症患者精子具有拮抗脂质过氧化作用 ,为临床应用黄芪减少递质对精子的损害提供理论基础。     

Immunoreactive relaxin in seminal plasma of fertile boars and its correlation with sperm motility characteristics determined by computer-assisted digital image analysis

Ejaculates from 10 mature fertile large white Yorkshire boars were used to examine the correlation between immunoreactive relaxin levels in seminal plasma and sperm motility characteristics. Seminal plasma levels of immunoreactive relaxin were measured by a time-resolved fluoroimmunoassay (TR-FIA). Motility characteristics were assessed using a CellSoft computer-assisted digital image analysis system. The mean +/- SD level of immunoreactive relaxin in seminal plasma was 2.61 +/- 0.62 ng/mL. When the correlation between seminal plasma levels of immunoreactive relaxin and parameters of sperm movement was examined, it was found that relaxin levels were significantly correlated with the percentage of motile spermatozoa (r=0.687, p < 0.05), curvilinear velocity (r=0.745, p < 0.05), straight line velocity (r=0.651, p < 0.05), mean amplitude of lateral head displacement (mean ALH) (r=0.844, p < 0.01) and the maximum amplitude of lateral head displacement (max ALH) (r=0.830, p < 0.01), but not with linearity, beat-cross frequency, or percentage of circular cells. Among these parameters, seminal plasma levels of immunoreactive relaxin showed the strongest correlation with the ALH parameter related to fertilizing ability. These results indicate that immunoreactive relaxin in boar semen may be necessary not only for normal sperm motility but also for normal fertility, suggesting that determination of the profile of immunoreactive relaxin in ejaculates may have value as a potential marker for predicting sperm fertilizing ability of boars.     

Antisperm antibodies and sperm motility: a study using timed exposure photomicrography

This study examined the effect of antisperm antibodies on sperm motility. Antisperm antibodies present in seminal plasmas with different sperm agglutinating titres were transferred passively to normal donor sperm, and the effects on movement characteristics (velocity of forward progression, amplitude of lateral head displacement, percentage progressive motility and percentage non-progressive motility) were analysed using timed exposure photomicrography. There was no significant association between sperm movement characteristics and the presence of titre of antisperm antibodies in seminal plasma. Furthermore, no differences were detected between those samples that possessed sperm agglutinating versus sperm immobilizing activity. These findings do not support the common belief that antisperm antibodies are a cause of poor sperm motility in semen.     

Transforming growth factor βreceptor Ⅱ expression in experimental cryptorchidism and apoptosis in spermatogenic cells in rats

Objective: To evaluate the sensitivity of sperm motility assay for detecting the endotoxin effect on human sperm in vitro. Methods: Motile human sperm were separately incubated for up to 24 hours with different concentrations of endotoxin (0.5, 1, 10, 1000, 10 000 and 50 000 ng/mL). Then the sperm motility was determined. The effect of endotoxin on the sperm motility in media without albumin was also determined. In addition, at the endotoxin concentrations of 0.5, 1 and 10 ng/mL, the sensitivity of the assay was compared to those of 1-cell and 2-cell mouse embryo bioassays. Results: At levels of 0.5-1 000 ng/mL endotoxin in media with 2 mg/mL albumin, sperm did not show significant change in motility after 24 h of incubation (P>0.05), while it was significantly inhibited at endotoxin levels of 10 000 and 50 000 ng/mL. In media without albumin, endotoxin levels of 50 000 and 1 000 ng/mL, markedly inhibited the sperm motility after 2 or 8 h of incubation (P<0.01). With media containing 0.5 and 1 ng/mL endo