DIRECT PREPARATION AND MEIOTIC ANALYSIS OF HUMAN SEMEN

A meiotic study of human semen specimens was carried out in 60 cases. Among them, 20 were normal controls, 20 suffered from infertility, and 20 had a history of recurrent spontaneous abortions. Of the cases of infertility and abortion, 50% were found to hare numerical and / or structural chromosomal aberrations in spite of the normal blood karyotype. It is suggested that chromosomal aberrations may be the cause of sterility and abortion. The success rate of semen chromosome G-banding in the infertility, abortion and control groups was 70%, 55% and 25% respectively. The semen samples of three cases of heterozygous carriers with reciprocal chromosomal translocation were also studied. The ring quadrivalents were identified in all cases. The results may explain the recurrent abortions in these patients.

A preliminary study on the rheological properties of human ejaculate and changes during liquefaction

Aim: To study the changes in rheological properties, namely the parameters of the hysteresis loops and yield stress versus time for human semen after ejaculation. Methods: Ejaculates were obtained from volunteers and immediately put into the test cup of a Brookfield Programmable DV-11 Rheometer, by which the hysteresis loops and yield stress were determined. Results: (1) Yield stress values dropped down from more than 3000 mPa to 60 mPa in about 5 minutes after ejaculation; (2) The shape of the hysteresis loops of shear stress versus shear rate was changed from the counter-clockwise direction, that enclosed a large area, into the clockwise direction, that enclosed a very small area. Conclusion: Human ejaculate originally possesses semi-solid or visco-elastic body behavior and in 5 minutes after liquefaction, it becomes a thixotropic fluid or shearing thinning fluid with very low viscosity.

男性不育患者人乳头瘤病毒感染与精液质量的相关性研究

目的探讨人乳头瘤病毒(HPV)精液感染对不育男性精液常规参数、精浆生化及精子DNA损伤的影响。方法收集115例2023年5月至2023年12月就诊于武汉华科生殖专科医院男科门诊患者精液样本,采用荧光PCR法进行HPV基因分型检测以评估精液感染率,所有样本完成精液常规分析、精浆生化、精子DNA碎片指数(DFI)及DNA精子高着色性检测分析,比较感染组与未感染组精液质量各参数之间的差异。结果115例男科门诊患者精液样本中HPV检出11例,阳性率占9.57%;与HPV阴性组相比,HPV阳性组在精子总活力、前向运动精子百分率、精液白细胞浓度、精浆锌总量、精浆弹性蛋白酶浓度参数上有所降低,在精子DFI值上有所升高,且差异有统计学意义(P<0.05)。结论男性HPV精液感染会对精子运动能力、精液感染指标及精子DNA完整性产生影响,对男性专科门诊患者诊疗上应考虑HPV感染因素,同时对高危人群必要时进行HPV基因分型检测。

Characteristics of semen parameters of Malawian men from couples seeking assisted reproduction

AIM: To profile semen parameters of Malawian men seeking fertility testing. METHODS: Semen analysis is a key element in the fertility evaluation of men and permits male reproductive potential to be evaluated. Semen samples were collected from consenting men after 3-5 d of sexual abstinence. The samples were collected from 130 males; 78 were male partners of infertile couples while 52 were healthy semen donors. Seminal volume, motility and morphology were assessed. The results were analyzed on Prism 5. All data are expressed as mean ± SD. Student's t-test was used for statistical analysis. Differences were regarded as statistically signifi cant if P < 0.05.RESULTS: Semen volume, sperm concentration, progressive motility and normal morphology were significantly higher in the control group when compared to the participant group. On the other hand, no statistically signifi cant difference was found between the control group total sperm motility when compared to the participant group. Oligozoospermia was found in 25 cases, teratozoospermia detected in 17 cases and abnormal seminal plasma in 16 cases. Asthenozoospermia and azoosper-mia were found in 12 and 8 participants, respectively. This study has shown that most of the infertile patients seeking fertility testing had oligozoospermia. Teratozoospermia was the second most common abnormality found in the patients seeking fertility testing. CONCLUSION: Our study is in agreement with previous studies which reported that these parameters have been shown to be good predictors for fertilization.

非人灵长类动物精子冻存技术的研究进展

随着生物医学技术的发展,应用非人灵长类动物模型进行基础科学研究日益广泛。与此同时,由于栖息地破坏、狩猎和基因隔离,许多非人灵长类动物濒临灭绝。因此,改进非人灵长类动物精子冻存技术对物种遗传资源的保藏具有重要的意义。本文概述了非人灵长类动物的精液特征,介绍了精液液化和冷冻精子质量评估的方法,分析了冷冻保护剂、冷冻稀释液及冷冻方法等因素对精子冻存效果的影响,总结了目前非人灵长类精子冷冻常用的冷冻保存液和冷冻方法,并对相关精子参数进行了比较,同时探讨了非人灵长类动物精子冻存研究面临的困境,并提出了可行的方案。总之,本文综述了近年来非人灵长类动物精子冻存的重要研究成果,对开发新的冷冻保护剂及改进冷冻技术具有一定的参考价值。

Age-related changes in seminal polymorphonuclear elastase in men with asymptomatic inflammation of the genital tract

Aim： To investigate age-related inflammatory events in the male genital tract. Methods： In a total of 4 265 randomly collected patients attending the andrological outpatient clinic of the Center for Dermatology and Andrology, University of Giessen, Germany, ejaculate volume, pH-value, sperm concentration, total and progressive sperm motility, concentration of polymorphonuclear （PMN） elastase, number of peroxidase-positive cells and fructose were measured and correlated with patient＇s age. Results： While ejaculate volume, motility and fructose all correlated negatively with age, sperm concentration, PMN elastase and the pH-value showed a positive correlation. The prevalence of male genital tract inflammation （as defined by PMN elastase 〉 250 ng/mL） and its severity increased significantly. PMN elastase did not correlate with sperm motility. Fructose as a marker of seminal vesicle function showed a significant negative relationship with the PMN elastase levels, the number of peroxidase-positive cells and sperm motility. Conclusion： The significant increases of PMN elastase levels as marker of male genital tract inflammation in older men appear to be indicative of age-related changes in local immunoregulatory mechanisms. Because there is no association of PMN elastase with sperm motility, a direct inhibitory effect of this enzyme can be excluded.

人精浆酸性磷酸酶对小鼠T淋巴细胞产生IL-2的影响

本文研究了从正常人精浆中分离纯化的酸性磷酸酶（ACP）对BALB／c小鼠脾脏细胞来源的T淋巴细胞产生IL-2的影响。本文采用MTT生物学测定法测IL-2活性。实验证明：人糖浆ACP对小鼠T淋巴细胞产生IL-2有明显的抑制作用（P＜0．05），并且抑制作用随ACP剂量的增加而增强。这说明了精浆中大量ACP是构成精浆免疫抑制功能的重要组成成份之一。

人类精液一步冷冻法的研究

本文应用聚丙烯1ml注射器作冻贮容器实验人类精液的一步冷冻法。结果表明,正常生育力组(n=45)和不育症组(n=50)的精子冷冻复苏率分别为69.4±9.7%、58.0±14.7%;一步冷冻组与程控仪冷冻组(n=14)的冷冻精子活率比较、与未经冷冻组(n=10)的去透明带地鼠卵穿透试验比较没有显著性差异(P>0.05);冻后精子体外能存活较长时间;37℃水浴解冻有利于精子存活。提示一步冷冻人类精液可获得良好的冷冻效果。

Defining laboratory reference values and decision limits＇ populations, intervals, and interpretations

This article provides a brief overview of various approaches that may be utilized for the analysis of human semen test results. Reference intervals are the most widely used tool for the interpretation of clinical laboratory results. Reference interval development has classically relied on concepts elaborated by the International Federation of Clinical Chemistry Expert Panel on Reference Values during the 1980s. These guidelines involve obtaining and classifying samples from a healthy population of at least 120 individuals and then identifying the outermost 5% of observations to use in defining limits for two-sided or one-sided reference intervals. More recently, decision limits based on epidemiological outcome analysis have also been introduced to aid in test interpretation. The reference population must be carefully defined on the basis of the intended clinical use of the underlying test. To determine appropriate reference intervals for use in male fertility assessment, a reference population of men with documented time to pregnancy of 〈 12 months would be most suitable. However, for epidemiological assessment of semen testing results, a reference population made up ofunselected healthy men would be preferred. Although reference and decision limits derived for individual semen analysis test results will undoubtedly be the interpretational tools of choice in the near future, in the long term, multivariate methods for the interpretation of semen analysis alone or in combination with information from the female partner seem to represent better means for assessing the likelihood of achieving a successful pregnancy in a subfertile couple.

无卵黄型人类精液冷冻保护剂试验与临床初步应用

不含卵黄的“改良 Tyrode’s液—甘油”型冷冻保护剂(简称 T—G 保护剂)与含卵黄的“甘油—卵黄—柠檬酸钠”型冷冻保护剂的试验比较表明,两者的精子复苏率、膜低渗肿胀率、Kremer’s 精子宫颈粘液穿透试验、去透明带地鼠卵穿透试验没有显著性差异(P>0.05);T-G 保护剂的甘油浓度分别为15%、10%,6%时,精子复苏率没有显著性差异(P>0.05);应用 T-G 保护剂冷冻保护的精液,人工授精已使1名丈夫无精子症的妇女受孕,并足月分娩1男婴,生长发育正常,提示 T-G 保护剂能有效地保护精子的受精能力。此外,对“TES-Tris-甘油-卵黄”型冷冻保护剂去除卵黄后的效果观察,精子的复苏率显著低于对照组(P<0.01)。

五年来我院门诊不育男性精液质量变化情况的研究

目的:分析我院近五年来门诊不育男性精液参数的变化趋势。方法:收集了五年来总计51258名不育男性的精液资料,控制禁欲时间等参数后对精液情况的变化进行统计分析。结果:患者年龄没有明显变化,50%以上是在25～35岁之间。无精症患者比例由6%增加到10%,少精症患者的比例由60%增加到85%,弱精症患者的比例由78%增加到92%,少弱精症患者由40%增加到56%,其中弱精症患者所占的比率最大。随着年龄的增加,精子密度呈现下降趋势,由17×106/mL降到9×106/mL。精子活率由2004年的47.27%降到2008年的32.97%,精子活力由2004的33.60%降到2008年的28.30%。结论:过去五年来我院门诊男性不育患者的年龄主要集中在30岁左右,弱精症患者所占的比例最大,无精症、少精症、弱精症、少弱精症患者的比例都有所增加,精子密度、精子活力和精子活率都表现出下降趋势。

禁欲时间与精液参数的相关性研究

目的:探讨禁欲时间与精液参数的关系。方法:接纳在孕前检查中要求精液检查的620例为对象,分析禁欲时间与精液参数之间的关系。结果:禁欲1～3d精液量无明显增加,为2.2～2.9ml;禁欲4d精液量增加到3.4ml,显著高于禁欲1d;禁欲4d与禁欲7d的精液量无差异。精子密度从禁欲1d的26.3×106/ml上升到3d的64.6×106/ml,增量明显(P<0.05);禁欲4d与6d的精子密度无明显增加;不同禁欲时间a级、b级和(a+b)级精子的比例无明显的变化。结论:在本研究范围内,精液的量、精子密度和总精子数随着禁欲时间延长而增加,精子的活力保持不变。

冻贮人类精液的最适条件

冻贮后的人精液与新鲜精液相比 ,其生殖力明显下降。不同的组织和细胞在冷冻复温过程中要求有其最适冻贮条件。人精液冻贮的最适条件是为了确保冻后精子复苏率达到最佳效果。本文对人精液冻贮过程中影响冻贮效果的关健性条件 (冷冻保护剂类型、冻前冷平衡、降温率及复温率等 )进行了综述 ,提示出一套促进人精液冻贮效果的方法。

人类精液两种冷冻方法的比较研究

对采用程控生物冷冻仪控制冷冻速率冷冻精液的方法,与人工操作逐步降溫冷冻方法进行实验比较,结果表明,两种方法冷冻人类精液后的3组精子复苏率(正常生育力组n=18,精液常规参数正常范围的不育症组n=28,异常精液组n=12);Kremer's精子宫颈粘液穿透试验(n=13);去透明带地鼠卵穿透试验(n=11)没有显著性差异(P>0.05)。在3组实验精液中,以正常生育力组复苏率最高,异常精液组最低。

人精浆免疫抑制因子SF_1对血清补体的影响

SF_1是正常人精浆经Sephadex G 100分离后得到的第一组分。将正常人血清和杂种小鼠混合血清作为补体来源,每份血清均以SF_1处理为实验组,并设不加SF_1的自身对照组,从下列实验中分别观察SF_1对补体系统的影响:1.CH_(50)试验证明,SF_1能抑制人血清中总补体活性(P<0.01);2.SF_1能抑制人血清中补体的溶血作用,并随血清中加入SF_1剂量的增加而抑制作用增强;3.利用YC 花环试验证实SF_1可抑制补体替代途径的激活;4.SF_1不影响用单向琼脂扩散试验测定中C_3、C_4的量,证明SF_1没有影响C_3、C_4的抗原性。上述实验结果提示:SF_1在体外实验中能抑制补体的活性,表现于对补体经典途径和替代途径激活的抑制。

精液叶酸、维生素B_(12)、活性氧水平与精子质量关系研究

目的 : 检测人精液叶酸、维生素 B12 ( VB12 ) ,及活性氧 ( ROS)的含量 ,探讨三者与精子质量的关系。方法 : 收集经证实配偶已怀孕的新加坡籍健康成年男性 1 76名的精液 ,根据WHO推荐方法检测精子质量各项指标 ,并测定精液叶酸和 VB12 含量及 ROS水平。结果 : 平均精液量、精子浓度和其活力都在正常范围内。而精子存活率 ( 73.0 6% )略低于 WHO标准 (≥ 75% ) ,形态异常的精子 ( 77.0 6% )高于 WHO标准 ( 70 % )。精液的叶酸含量与 ROS水平呈显著负相关 ( r=- 0 .2 5,P=0 .0 0 3) ;VB12 与 ROS呈负相关 ( r=- 0 .2 5,P=0 .0 0 2 ) ,与精子形态异常也呈负相关( r=- 0 .1 5,P=0 .0 4 8) ,但与精子浓度呈正相关 ( r=0 .31 ,P<0 .0 0 1 )。ROS与精子形态异常呈正相关 ( r=0 .2 5,P=0 .0 0 2 ) ,与精子活力也呈正相关 ( r=0 .1 9,P=0 .0 1 9)。结论 : VB12 对精子生成和维持精子正常功能是必需的 ,ROS水平与精子功能缺陷有关 ,VB12 和叶酸可能通过降低 ROS以维持精子的正常功能。

成年男性精液中19种元素含量的测定及影响因素研究

运用ICP-MS和ICP-AES方法同时分析丹东某硼矿周围工人组及对照组精液中19种元素的含量水平。样品经过双氧水-纯化硝酸体系消解,采用稀释分析及双内标铑(Rh)和铼(Re)进行校正。该法最低检出限、加标回收率及分析精密度能满足精液样品的分析要求。按照95%置信区间进行数据筛选,选出118人的数据统计分析成年男性精液中元素含量水平,并与国内其它研究结果进行比较。分析精液元素含量影响因素表明,硼(B)作业暴露对精液中的B含量具有显著影响;吸烟能显著影响精液中的Cd含量;饮酒对工人组精液中的Cu和Mn的含量具有显著影响。

人精浆免疫抑制因子对B淋巴细胞产生抗体的影响

本文采用体内实验的方法,观察正常人精浆经sephadex G-100分离而得的第一组份(SF_1)对Balb/c小鼠B淋巴细胞的影响。实验采用SF_1腹腔注射为实验组,以生理盐水作对照组,四天后给予同量抗原绵羊红细胞(SRBC)刺激,在SRBC免疫后第三天、第四天,第五天、第六天分别测定这二组小鼠的溶血空斑数和血清中的抗SRBC抗体的凝集效价。实验结果发现:实验组小鼠的溶血空斑数均明显低于对照组(P<0.01),实验组小鼠血清中的特异抗SRBC抗体的凝集效价也均低于对照组(P<0.05)。结果表明:正常人精浆提取物SF_1在异种小鼠体内有明显抑制B淋巴细胞对于抗原刺激转化为浆细胞的功能,同时也抑制了特异性抗体的产生。

薏苡仁脂肪酸类成分对人肝癌细胞株SMMC-7721细胞周期及细胞凋亡的影响

目的研究薏苡仁脂肪酸类成分对人肝癌细胞株SMMC-7721增值的影响及作用机制初探。方法采用MTT法考察不同给药剂量,不同给药时间下薏苡仁脂肪酸类成分对SMMC-7721细胞的增值抑制作用及其与时间、剂量的关系;碘化丙啶PI单染色法检测薏苡仁脂肪酸作用后,SMMC-7721细胞周期DNA水平变化;Annexin V-EGFP/PI双染法研究薏苡仁脂肪酸对SMMC-7721细胞凋亡率的影响。结果不同质量浓度的薏苡仁脂肪酸作用SMMC-7721细胞24、48、72 h后,均呈现出较好的细胞增殖抑制作用,且具有明显的时间-剂量-效应关系,差异有统计学意义(P<0.05);不同质量浓度的薏苡仁脂肪酸处理SMMC-7721细胞22 h后可显著增加肝癌细胞凋亡率,(P<0.001);作用36 h后,可显著降低G2/M期细胞比例,(P<0.001)。结论薏苡仁脂肪酸对人肝癌SMMC-7721细胞具有明显的抑制作用并可诱导其凋亡,其作用是通过抑制G2/M细胞周期实现的。