Quantitative Evaluation of the Human Spermatozoal Motility and Acrosome Reaction in Infertile Ollgozoospermic and Fertile Euspermic Men

Idiopathic oligozoospermia is one of the most important problems in Andrology, but up to now it is poorly understood because the often routine conventional semen parameters, unquestionably are not directly related to the evaluation of the morphological and functional integrity that determines the spermatozoan fertilizing capacity. A non complex strategy was designed to determine the presence of alterations in the functional integrity of the spermatozoa from infertile men with idiopathic oligozoospermia and from euspermic fertile men, by the quantitative analysis of the spermatozoan motility and the acrosome reaction. There was a lower percentage of acrosome reacted spermatozoa within the semen of the infertile men, accompanied with a significant decrease in the motility percentage, sperm velocity and motility index in comparison with semen from fertile men. These data strongly support a possible detrimental structural and functional integrity of the spermatozoa from the oligozoospermic men.     

铅作业工人生殖功能调查及精子超微结构观察

本文调查了25名铅作业男工及24名非铅作业男工的生殖功能状况,并观察了精子超微结构的变化。结果表明,铅作业男工的尿铅浓度、精子畸形率及后代先天性癫痫和先天性心脏病患病率均高于非铅作业男工,而精子密度却低于非铅作业组。电镜检查发现,铅作业男工的精子核、线粒体和精子尾均有异常改变。     

Significance of Adenosinetriphosphate in Human Sperm as Clinical Parameter

Washed human spermatozoa from 21 individuals with an average motility of 60% (quality index 277 ± 16) had an endogenous ATP generation of 7.5 ± 3.4 nmole/10⁸spermatozoa. The ATP concentrations in spermatozoa from 16 patients with severely impaired motility of 26% (quality index 98 ± 13) was 16.9 ± 9.9 nmole/10⁸spermatozoa. There was no correlation between ATP content and motility in either group. Sperm penetration into blood serum type AB, Rh-positive, was evaluated using a capillary tube penetration test. The penetration was graded with a maximum score of 14. Spermatozoa with an initial motility of 60% reached a score of 10 ± 0.7. After addition of 20 mmole of ATP the score was significantly improved to 13 ± 0.3. Compared with these results spermatozoa with an average motility of 26% reached a score of 4 ± 1. Exogenous ATP (20 mmole) increased the score to 8 ± 1.0. In both groups reduced glutathione had a negative effect.

Human spermatozoa with high and low motility are capable to synthesize ATP. A dysfunction of the phosphorylating particles in the mitochondria appears not to be associated with low sperm motility.     

Polybrominated Diphenyl Ethers in Human Serum and Sperm Quality

Polybrominated diphenyl ethers (PBDEs) are widely used flame retardants; currently, they are identified as ubiquitous environmental contaminants. Several studies indicate that PBDEs might affect male fertility. We present the results of a pilot study on the relationship between human serum PBDEs and sperm quality. The PBDE levels in Japan are comparable to those found in European countries. Strong inverse correlations were observed between the serum concentration of 2,2',4,4',5,5'-hexabromodiphenyl ether and sperm concentration (r = -0.841, p = 0.002) and testis size (r = -0.764, p = 0.01). Extensive studies on the relationship between PBDEs and sperm quality are required.     

Sperm Immunofluorescence in Infertile Men: Its Relationship with Sperm Agglutination and Gel Agglutination

The spermatozoa from 10 infertile male patients all with sperm agglutination were subjected to direct immunofluorescence using antisera raised against IgA, IgM, and IgG. Sera from these patients were subjected to the Gel Agglutination Test. Although the Gel Agglutination Test related closely with sperm agglutination, none of the spermatozoa from these patients produced a repeatable positive response to direct immunofluorescence. This study suggests that neither sperm agglutination nor the Gel Agglutination Test demonstrate the presence of sperm specific antibodies in the semen of these infertile men.     

Fine Structure and Cytochemistry of the Morphogenesis of Round-Headed Human Sperm

Ejaculates and testicular biopsies of two infertile men were examined at ultrastructural and cytochemical levels. The two patients presented spermograms in which all the spermatozoa had globular heads. Wide Golgi areas and large masses of annulatae lamellae were evident during spermatogenesis. Abnormal acrosomal vesicles were evident during early spermatid stage. Among the late spermatids, there was a small group characterized by a fibrous sheath showing considerable malformation, and “spindle shaped body.” In Sertoli cells, detached acrosomes undergoing degeneration were noted. Low TPPase activity was found in Golgi complex and in abnormal acrosomal vesicles of early spermatids. As regards acid phosphatase, was localized in spermatocytes and in early spermatids, but not in late spermatids and mature spermatozoa. Leydig cells had high phosphatase activity.     

Expression of IL-2α and IL-2β Receptors on the Membrane Surface of Human Sperm

Cytokines are secreted proteins that act as local immunological mediators. Increased seminal cytokine concentrations are associated with fertility problems. The purpose of the present study was to investigate the presence of IL-2 &#102, and IL-2 &#103 receptors on fresh and isolated sperm by flow cytometry and transmission electron microscopy. Twenty sperm samples from oligospermic men were incubated with CD25, a mouse monoclonal antibody specific for IL-2 &#102 -chain receptor, and CD122, a mouse monoclonal antibody specific for IL-2 &#103 -chain receptor. The strong initial fluorescence intensity and, subsequently, a labeling index yielded by CD25 and CD122 decreased in sperm centrifuged on a Percoll gradient ( p <. 05). The expression of CD25 and CD122 correlated negatively with fresh sperm concentration, but in sperm centrifuged on a Percoll gradient there was no correlation. Labeling with CD25 and CD122 antibody was evident on the head and the middle piece in fresh sperm, while in sperm centrifuged on a Percoll gradient a weak labeling was observed only on the principal piece. The authors have identified and localized cytokine receptors on human sperm for the first time. Cytokine receptors may be involved in the regulation of pathophysiological events in sperm cell functions and male infertility. The exact pathway involved in modulation of these receptors requires further investigation. These results contribute to the understanding of cytokine-sperm relationships.     

拉米夫定抑制乙肝患者精细胞内HBcAg表达的相关临床研究

[目的]探讨乙肝患者精细胞内HBcAg表达与血清HBVDNA水平的相关性及拉米夫定对精细胞内HBcAg表达的抑制作用。[方法]选择2007年1月至2008年10月在济南市传染病医院就诊的门诊和住院的乙肝患者62例为乙肝组,22名HBV指标全部阴性的健康人为对照组,检测精细胞内的HBcAg;乙肝组同时检测血清HBVDNA。[结果]精细胞HBcAg定性检测阳性率,乙肝组为96．77％（SHBc1≤2．5者7例。2．6～4．9者18例,≥5者37例）。22名健康对照均阴性。乙肝组中,精细胞HBcAg的表达及其高度表达者所占比例,血清HBVDNA高者高于血清HBVDNA低者（P〈0.01）。精细胞HBcAg定量值,乙肝病人拉米夫定治疗的23例治疗后低于治疗前（P〈0.01）,未治疗组10例高于治疗组治疗后（P〈0.01）。[结论]乙肝患者精细胞HBcAg表达与血清HBVDNA水平显著相关．拉米夫定治疗可减少精细胞内HBcAg的表达。     

Sperm Donors in Shanghai,China: A Study of Motivations,Characteristics, and Semen Parameters of Actual Sperm Donors

Objective::To identify the sociodemographic characteristics, motivations, and semen parameters of sperm donors in Shanghai, China.Methods::The participants were...     

Effects of Rosmarinic Acid on DNA Integrity and H19 Differentially Methylated Region Methylation Levels in Human Sperm Preserved by Freeze-Drying

Objective::To investigate the effects of rosmarinic acid (RA) on the DNA integrity and methylation levels of the H19 differentially methylated region (DM...     

优选和冻融对精子顶体反应及顶体反应释放的顶体酶活性影响

目的:研究优选和冻融对人精子顶体反应(acrosome reaction,AR)和AR释放的顶体酶活性影响。方法:10例健康生育男子精液,用Percoll密度梯度分离法和上游法优选精子;用3种冷冻保护剂进行冷冻;用孕酮诱导AR;采用FITC-PSA荧光染色法分析AR;采用分光光度比色法测定顶体酶活性。结果:Percoll法和上游法优选后,顶体反应率和AR释放的顶体酶活性均较优选前显著升高(P<0·05),但Percoll法和上游法优选组间比较差异不显著(P>0·05)。精子冻融后甘油组和GYC组顶体反应率和AR释放的顶体酶活性均下降,与冷冻前比较差异均有显著性(P<0·01),GYCG组与冻融前组比较,差异不显著(P>0·05)。结论:优选后顶体反应率及AR释放的顶体酶活性增加,GYCG对精子顶体反应和AR释放的顶体酶活性有保护作用。     

Impact of Sperm Collection Methods on Sperm Parameters in Spinal Cord Injured Men and Compared to Normal Controls in ICSI Program

Background To determine and compare the effect of electro ejaculation (EE), Masturbations, Penile vibratory stimulation (PVS), Percutaneous Epididymal Sperm Aspiration (PESA) on the quality and quantity of sperm parameters in the aspect of assisted reproduction.Methods A retrospective study on collected semen of 89 spinal cords injured (SCI) men (EE = 70, Mast = 3, PVS = 6, PESA = 10 individuals) and 49 neurologically intact men (EE = 5, Mast = 14, PVS = 30) with different methods were included and intracytoplasmic sperm injection (ICSI) with homologue oocytes were performed.Results The quality and quantity of semen in SCI was significantly lower than neurologically intact men (P < 0.01), and showed reduction of sperm parameters in compare with normal group including; 23.19% in motility, 14.56 % in normal morphology, and 41.51%in viability (P < 0.001). Within normal group the EE method has significantly influence on sperm parameters (P < 0.01). In SCI group the morphology was severely impaired. Hyper leukocytes significantly increased (P < 0.01). The fertilization rate of EE/ICSI was 60% which was lower than other methods of semen collection (71.7%).Conclusions The EE method of semen collection compared with PVS and other methods of normal ejaculatory stimulation showed significantly lower grade of sperm parameters. EE/ICSI is effective in treating an ejaculatory infertility. For ejaculatory stimulation it is recommended to Consider PVS as the first line of treatment in SCI men. PESA can be considered when EE and PVS both fail to cause seminal emission.     

长效醋酸甲孕酮用于男性避孕的药代动力学及抑制精子发生的剂量研究

比较两种剂量长效醋酸甲孕酮(DMPA)用于男性避孕的药代动力学特性以及抑制精子发生的效果,推荐进一步临床试验配伍剂量和注射间隔。方法:将筛选合格的正常志愿者30名随机分成3组(每组10名),A组150mgDMPA+1000mg十一酸睾酮酯(TU);B组300mgDMPA+1000mgTU;C组单独注射1000mgTU(为对照组)。A和B组受试者按要求接受指定的配伍剂量注射。注射前后每周静脉取血,直至8周,进行血MPA测定。然后,受试者继续每8周接受指定的配伍剂量或TU单独注射,直至完成24周的治疗期,并按要求定期随访进行精液分析及其它安全性指标监测,最后完成24周的恢复期随访。结果:受试者的各项参数在各组内均匀分布。尽管B组较A组有相对较高的血药浓度,但药代动力学的各项参数在两组之间无显著性差异。A、B和C组抑制精子发生起效时间分别为80±5d、83±8d和92±6d,各组间没有统计学差异。尽管合用组比单用组有更持久地抑制精子发生的趋势,但各组之间没有统计学差异。合用组能更加有效地抑制促性腺激素的分泌,与单用组之间有显著性差异;促性腺激素的抑制程度与配伍中DMPA的剂量无关。结论:A与B组在药代动力学的各项参数与抑制精子发生的疗效上无显著性差异。150mgDMPA与1000mgTU或更长效的雄激素联合应用可能成为2～3个月肌肉注射一针的男性避孕药的最佳剂量组合。推荐使用低剂量DMPA配伍TU进行临床避孕有效性的研究。     

Characterization of Hyperactivated Motility by Human Spermatozoa During Capacitation: Comparison of Fertile and Oligozoospermic Sperm Populations

Suspensions of capacitating human spermatozoa were analyzed for potential hyperactivated movements using videomicrographic methods. Analysis was carried out on aliquots of 22 sperm suspensions, which were proved fertile several hours later during human in vitro fertilization. After approximately 3 h of capacitation, 22.1% of the fertile spermatozoa displayed motility patterns designated as hyperactivated. Over 80% of these hyperactivated spermatozoa moved with a wide-amplitude, two-dimensional whiplash pattern, displaying marked lateral displacement of the head. Only 8.4% of capacitating spermatozoa from oligozoospermic patients showed these hyperactivated movements. The incidence of hyperactivated movements by fertile and oligozoospermic spermatozoa could be significantly increased after exposure to various motility stimulants. The clinical significance of hyperactivation as a functional assay of fertilizing capacity is discussed.     

甲型流感病毒感染致人血管内皮细胞凋亡的初步研究

目的探讨甲型流感病毒感染对血管内皮细胞增殖与凋亡的影响。方法 用纯化的甲型流感病毒体外感染人血管内皮细胞后,用RT-PCR法检测病毒整合细胞并复制状况,用MTT法检测细胞增殖状况,流式细胞术及免疫印迹检测细胞凋亡。结果琼脂糖电泳结果显示,H1N1病毒能感染血管内皮细胞且能复制。MTT结果显示,尽管H1N1病毒不能完全抑制血管内皮细胞的增殖,但能使细胞增殖能力明显下降。流式细胞术结果显示甲型流感病毒感染组的Annein V染色阳性的血管内皮细胞比例明显高于对照组。免疫印迹结果显示病毒感染24、32 h后,细胞出现断裂的PRAP蛋白条带。结论甲型流感病毒能感染人血管内皮细胞及增殖,并能诱导血管内皮细胞凋亡。     

诱导人胚胎干细胞向子宫内膜样细胞分化的初步研究

目的:探讨无饲养层培养的人胚胎干细胞诱导分化为子宫内膜样细胞的方法。方法:将无饲养层培养的人胚胎干细胞分别通过共培养诱导法和细胞因子诱导法向子宫内膜样细胞诱导,流式细胞术检测分化的细胞中子宫内膜上皮细胞标志物细胞角蛋白18和间质细胞标志物波形蛋白的表达情况。结果:共培养诱导法和细胞因子诱导法在分化第7天均出现长梭状细胞样改变。分化第8天通过流式细胞术检测发现细胞波形蛋白表达均为阴性,部分细胞角蛋白18表达阳性,共培养诱导法分化率高于细胞因子诱导法[(55.63±10.29)%vs.(13.90±0.26)%,P<0.05]。结论:共培养诱导法和细胞因子诱导法均可使人胚胎干细胞分化为子宫内膜上皮样细胞,但前者的分化效率更高。     

铅诱导人脐静脉内皮细胞损伤及机制研究

目的探讨铅诱导人脐静脉内皮细胞损伤效应及机制。方法用不同浓度的铅染毒人脐静脉内皮细胞,观察细胞形态和活性,通过检测细胞上清液丙二醛(MDA)、非对称性二甲基精氨酸(ADMA)、一氧化氮(NO)的含量和一氧化氮合酶(NOS)的活性等指标,观察铅对人脐静脉内皮细胞损伤的量-效关系(终浓度分别为1、2、4、8mg/L的铅处理细胞24h)和时-效关系(终浓度为4mg/L的铅分别于处理0、3、6、12、24、48h后观察检测指标)。实验分为正常对照组(加入与处理组等体积的D-Hank’s液,铅浓度为0mg/L)、铅处理组(加终浓度分别为1、2、4、8mg/L的铅)、铅VitC组(铅终浓度为4mg/L,VitC终浓度为100mg/L)和阳性对照组(加终浓度为500μg/L的过氧化氢)。结果 2～8mg/L的铅能显著改变细胞形态、降低细胞活性;能显著性诱导内皮细胞MDA升高。2～8mg/L的铅还能显著性诱导内皮细胞NO含量降低,具有显著的时间依赖性,各处理组间两两比较差异有统计学意义(P0.05或P0.01)。4mg/L铅处理时细胞ADMA含量最高,且分别在处理后24h到达最高峰。结论 2～8mg/L铅能显著诱导人脐静脉内皮细胞损伤;ADMA在铅诱导人脐静脉内皮细胞损伤中有着重要作用。     

Spermiogenesis and DNA Repair: A Possible Etiology of Human Infertility and Genetic Disorders

This paper reviews the possible origin of sperm DNA fragmentation and focuses on the nuclear events associated with spermiogenesis as a potential source of genetic instability and reduced fertilizing potential of the mature male gamete. Recent findings suggest a programmed DNA fragmentation and DNA damage response during the chromatin remodeling steps in spermatids. We also discuss the spermatid DNA repair mechanisms and the possible involvement of condensing proteins, such as transition proteins and protamines, in the process, as this DNA fragmentation is normally not found in late spermatids. We propose that alterations in the chromatin remodeling steps or DNA repair in elongating spermatids may lead to persistent DNA breaks. This vulnerable step of spermiogenesis may provide a clue to the etiology of sperm DNA fragmentation associated with infertility in humans. This vulnerability is further emphasized given the haploid character of spermatids that must resolve programmed double-stranded breaks by an error-prone DNA repair mechanism. Therefore, spermiogenesis has probably been overlooked as an important source of genetic instability.     

氢醌对人支气管上皮细胞DNA损伤及细胞周期的影响

目的探讨氢醌(HQ)对体外培养人支气管上皮细胞DNA损伤及细胞周期的影响。方法将HBE细胞用不同浓度(0、5、10、20、40、80、160、320μmo/lL)的氢醌作用24 h,采用噻唑蓝(MTT)比色法测定16HBE细胞的相对存活率,用单细胞凝胶电泳(SCGE)检测细胞DNA的损伤情况,流式细胞术检测细胞周期分布。结果在0～40μmo/lL范围内HQ作用24 h后,16HBE细胞的存活率未见明显变化(P>0.05);当染毒剂量超过80μmo/lL时,细胞存活率明显下降(P<0.01)。SCGE显示随着HQ浓度的升高,16HBE细胞的DNA断裂程度加重。HQ作用浓度在10～320μmo/lL范围内,16HBE细胞的细胞周期表现为G2期阻滞,G1期比例下降。结论 HQ会对16HBE细胞的DNA产生损害,并且引起G2期细胞阻滞。