解脲支原体感染对男性不育的影响

解脲支原体(UU)是人类泌尿生殖系统感染的常见病原体，可引起男性生殖道炎症。许多研究表明男性生殖道炎症是造成男性不育的原因之一，可导致精液质量异常、男性生育力降低。本文用培养法对774例受检男性的精液作UU培养检测，并同时进行精液常规分析，从精液体积、精子活动率及活动力、精子畸形率和精子密度几个方面来探讨男性精液UU感染对不育的影响，现报告如下。     

精子DNA损伤的机制及检测方法研究进展

DNA是精子的遗传物质,其完整性易受各种因素影响而受损,继而影响精子质量、最终导致不孕不育.其损伤机制涉及染色质组装异常、氧化应激、凋亡异常等.精子DNA完整性受损的患者,用传统的精液检查往往找不到不育原因,因此,对这部分患者进行精子DNA完整性检查具有更重要的意义.目前已经建立了很多检测方法.现就精子DNA完整性受损的机制、常用检测方法及其临床应用作一综述.     

精子DNA损伤与男性不育的关系

近年来,不育症日益受到社会的广泛关注.发病率约为15％,其中男性因素约占40％[1].在临床实践中,男性不育的评估主要依靠精液分析.然而常规的精液分析(精子密度,精子活动力,精子形态)往往不能全面评估男性的生育力,即部分精液分析正常的人仍无生育能力,而某些精液分析异常的人却能够生育.这无疑给不育症的诊断和治疗带来了困难.有研究表明,男性不育可能与精子DNA的损伤紧密相关[2],不育男性的精子DNA碎片率显著高于正常生育男性的精子DNA碎片率[3].精子DNA损伤形式包括精子DNA碎片、异常的染色质包装和鱼精蛋白缺乏,其中最主要的损伤形式是精子DNA碎片,包括精子DNA双链或单链的断裂.精子DNA的完整性是父系遗传信息传递给子代的前提.精子DNA损伤对受精、胚胎的形态和植入有负影响.如果精子的遗传物质发生严重的损伤,胚胎的发育就会停滞,并导致流产的发生.这里面似乎存在一个阈值,如果精子DNA损伤超过了这个阈值,胚胎发育和妊娠就会受到损害[4].因而,评估精子DNA损伤程度有望成为诊断男性不育的一个指标,并对辅助生殖技术(artificial reproductive technology,ART)有指导意义.目前,已有许多方法用来检测精子DNA损伤,精子DNA损伤的评估在临床上的应用也日益受到重视.本综述的目的就是介绍在这个领域的新进展.     

周细胞在脊髓损伤中的作用

脊髓损伤(SCI)会导致患者自主神经功能、运动和感觉功能丧失,对患者的生活质量产生极大影响[1].SCI的病理变化包括损伤部位缺血、缺氧、神经元受损、瘢痕组织形成和炎性反应等[2].近年来, SCI的病理变化机制研究日趋深入,周细胞作为脊髓微环境的重要组成部位,在SCI的病理过程中发挥着重要作用.SCI发生后神经元轴突遭到破坏,这一过程伴随着血-脊髓屏障的破坏,神经胶质细胞和神经元细胞活化,并分泌多种副产物(包括基质金属蛋白酶、游离氧自由基、趋化因子和细胞因子)[3].各种免疫细胞渗入损伤部位[4],受损区域周围还会产生星形胶质细胞,小胶质细胞同样会被激活[5-6].     

不育症患者精子头部及尾部超微结构的研究

目的研究不育男性精子超微结构的形态特征。方法利用透射电镜对8例不育男性新鲜精液标本中的精子头及尾部超微结构进行观察。结果在电镜下不育男性精子存在多种形态超微结构异常,有以下几种类型：(1)顶体异常精子,包括顶体膜受损,顶体发育不良、缺失,顶体内形成包涵体精子。(2)头部异常精子,包括尖头精子、圆头精子、头部含空泡精子。(3)尾部异常精子,①尾部形态异常精子,包括无尾精子、短尾精子、卷尾精子、体尾胞质残余。②尾部结构异常精子,包括线粒体缺失精子、尾部线粒体多种形态和结构异常。结论不育男性精子存在顶体、头部、尾部线粒体、微管多种形态和结构异常。     

司机职业不育男性的精液分析

目的 :探讨司机职业与男性精液质量有无相关性。　方法 :对 12 2 3例不育男性 (司机 78例、非司机 114 5例 )和 10 0例生育男性精液从液化、精子密度、精子活力、精子活率、精子形态等方面进行全面分析。　结果 :从事司机职业的不育男性精液质量异常率显著高于非司机职业不育男性 (P <0 .0 5 )和生育男性 (P <0 .0 1) ,且开车 8年以上组精液质量异常率显著高于开车 8年以下组 (P <0 .0 5 )。　结论 :司机职业可引起男性精液质量异常。     

精子冻融技术及其研究进展

精子冻融是辅助生殖技术的基础,冻融可能造成精子结构损伤和活力功能丧失,因此优化冻融方法十分必要。提高冷冻前精液质量、进行精子优选、添加合适的冷冻保护剂、选择适宜的冷冻复苏方法等均可有效地减少冻融损伤,有利于保存男性生育力。本文对精子冻融的基本原理、方法改良、针对特殊质量精液的冷冻方法及冷冻后精子质量评估进行了综述。     

40岁以上男性精子DNA损伤与其精液常规参数的关系分析

目的分析40岁以上男性精子DNA损伤与其精液常规参数的关系。方法收集本院148例40岁不育男性患者精液以及120例40岁已生育体检男性精液,采用计算机辅助精液分析系统和双尾彗星实验检测常规精液参数及精子DNA损伤情况,分析40岁男性精子DNA损伤与其精液常规参数的关系。结果 40岁组不育男性精液的总活力、前向运动率和顶体反应率显著低于40岁正常组,DNA碎片化指数(DFI)值显著高于40岁正常组,差异具有统计学意义(P0.05);40岁不育男性中,DFI值≤30%组(112例)精子浓度、总活力、正常形态率、前向运动率和顶体反应率均显著高于DFI30%组(36例)(P0.05),其DFI值与精子浓度、总活力、正常形态率、前向运动率和顶体反应率呈显著负相关;40岁不育男性中,精子单链损伤比例高组(108例)精子浓度、总活力、正常形态率、前向运动率和顶体反应率均显著高于双链损伤比例高组(40例)(P0.05),其双链损伤指数/DFI(DSB-DFI/DFI)值与精子浓度、总活力、正常形态率、前向运动率和顶体反应率呈显著负相关,而单链损伤指数/DFI(SSB-DFI/DFI)值与精液各参数无显著相关性。结论 40岁不育男性精液参数异常和DNA损伤情况较同龄已育男性严重,精液DNA碎片化指数和双链损伤类型对40岁以上不育男性生殖情况具有一定评估价值。     

精子DNA损伤与辅助生殖技术

随着辅助生殖技术的广泛开展,精子评估已由传统的精液常规分析向细胞、分子水平深入发展。精子DNA损伤是反映男性生育力的一个新指标。精子DNA损伤的发生机制包括精子染色质包装与分离异常、氧化应激、细胞凋亡异常等。精子染色质结构分析是目前检测精子DNA损伤最常用的方法之一。精子DNA损伤可能与辅助生殖技术治疗结局、复发性自然流产、增加ICSI后代遗传风险相关。采取口服抗氧化药物、取睾丸精子行ICSI、预冻存精子、去除病因以及中医中药等治疗对策可能会降低精子DNA损伤程度,进而提高辅助生殖技术成功率。本文主要就精子DNA损伤的机制与检测方法、DNA损伤与生殖结局以及辅助生殖技术中与DNA损伤相关的治疗对策作一综述。     

精子功能与氧自由基

本文综述了精液内氧自由基的产生系统和拮抗系统 ,以及氧自由基对精子功能和形态的影响。精液中一定水平的氧自由基是精子获能和发生顶体反应所必需的 ,但氧自由基生成量增加 ,超过精子所需的生理剂量 ,则会导致精子受损 ,影响人类受精和家畜繁殖     

Effects of spinal cord injury on semen parameters

OBJECTIVE/BACKGROUND: Neurogenic reproductive dysfunction in men with spinal cord injury (SCI) is common and the result of a combination of impotence, ejaculatory failure, and abnormal semen characteristics. It is well established that the semen quality of men with SCI is poor and that changes are seen as early as 2 weeks after injury. The distinguishing characters of poor quality are abnormal sperm motility and viability. In the majority of the men with SCI, the sperm count is not abnormal. We elaborate on the effects of the SCI on semen parameters that may contribute to poor motility and poor viability. METHODS: Review. DESIGN: PubMed and MEDLINE databases were searched using the following key words: spinal cord injuries, fertility, sexual dysfunction, and spermatogenesis. All literature was reviewed by the team of authors according to the various stages of sperm development and transport in the male reproductive cycle. FINDINGS: The cause of asthenozoospermia appears to be multifactorial. CONCLUSION: Current literature does not support the preeminence of a single factor relating to neurogenic reproductive dysfunction in men with SCI. After SCI, there is ample evidence of disturbance of sperm production, maturation and storage, and transport due to an abnormal neuroendocrine milieu. Semen quality seems to be primarily affected by changes to the seminal plasma constituents, type of bladder management, and the neurogenic impairment to the ejaculatory function. Further focused and structured studies are required.     

Penile vibratory stimulation and electroejaculation in the treatment of ejaculatory dysfunction

The purpose of this review is to present the current understanding of penile vibratory stimulation (PVS) and electroejaculation (EEJ) procedures and its clinical use in men with ejaculatory dysfunction. Unfortunately, the record of treating such individuals has been quite poor, but within recent years development and refinement of PVS and EEJ in men with spinal cord injury (SCI) has significantly enhanced the prospects for treatment of ejaculatory dysfunction. The majority of spinal cord injured men are not able to produce antegrade ejaculation by masturbation or sexual stimulation. However, approximately 80% of all spinal cord injured men with an intact ejaculatory reflex arc (above T10) can obtain antegrade ejaculation with PVS. Electroejaculation may be successful in obtaining ejaculate from men with all types of SCI, including men who do not have major components of the ejaculatory reflex arc. Because vibratory stimulation is very simple in use, non-invasive, it does not require anaesthesia and is preferred by the patients when compared with EEJ, PVS is recommended to be the first choice of treatment in spinal cord injured men. Furthermore, EEJ has been successfully used to induce ejaculation in men with multiple sclerosis and diabetic neuropathy. Any other conditions which affect the ejaculatory mechanism of the central and/or peripheral nervous system including surgical nerve injury may be treated successfully with EEJ. Finally, for sperm retrieval and sperm cryopreservation before intensive anticancer therapy in pubertal boys, PVS and EEJ have been successfully performed in patients who failed to obtain ejaculation by masturbation. Nearly all data concerning semen characteristics in men with ejaculatory dysfuntion originate from spinal cord injured men. Semen analyses demonstrate low sperm motility rates in the majority of spinal cord injured men. The data give evidence of a decline in spermatogenesis and motility of ejaculated spermatozoa shortly after (few weeks) an acute SCI. Furthermore, it is suggested that some factors in the seminal plasma and/or disordered storage of spermatozoa in the seminal vesicles are mainly responsible for the impaired semen profiles in men with chronic SCI. Home insemination with semen obtained by penile vibratory and introduced intravaginally in order to achieve successful pregnancies may be an option for some spinal cord injured men and their partners. The majority of men will further enhance their fertility potential when using either penile vibratory or EEJ combined with assisted reproduction techniques such as intrauterine insemination or in-vitro fertilization with or without intracytoplasmic sperm injection.     

Characterization of ultrastructural and metabolic abnormalities in semen from men with spinal cord injury

BACKGROUND: In men with spinal cord injury (SCI), fertility is impaired because of a combination of ejaculatory dysfunction and poor semen quality. We hypothesized that ultrastructural and metabolic abnormalities of sperm could be an underlying factor in infertility in these men. METHODS: To investigate mechanisms contributing to the abnormal sperm parameters, we analyzed seminal constituents and sperm ultrastructure in ejaculates from 7 men with SCI and compared them with 5 control subjects. Sperm adenosine triphosphate (ATP) content was measured using a firefly luciferase bioluminescent assay kit, and ultrastructure was evaluated by electron microscopy. Seminal oxidative stress (8-iso-prostaglandin F-2alpha [8-iso-PGF2alpha]) and transforming growth factor-beta1 (TGF-beta1) levels were measured by specific enzyme-linked immunoabsorbent assay kits. RESULTS: Semen samples from men with SCI showed azoospermia (3) and severe asthenospermia (2; motility 0-5%). A majority (65%) of sperm from asthenospermic samples showed degenerative changes and significant axonemal defects. Incubation of normal sperm with SCI seminal plasma induced a concentration-dependent decrease in sperm motility (43%) accompanied by a significant drop in intracellular ATP content (33%). Semen samples from men with SCI exhibited levels of 8-iso-PGF2alpha 3.5-fold higher than those from controls and levels of TGF-beta1 that were 10% higher than those from controls. CONCLUSION: Our results suggest that seminal constituents of men with SCI are detrimental to sperm movement and that ultrastructural degenerative changes may contribute to the impaired sperm motility and viability seen in these patients. These preliminary results must be confirmed in larger patient populations and longitudinal studies.     

Seminal vesicle aspiration in spinal cord injured men: insight into poor sperm quality

PURPOSE: In men with spinal cord injury poor quality semen is seen when performing electroejaculation and penile vibratory stimulation. We determined whether sperm stasis within the seminal vesicles is a potential cause of this problem. MATERIALS AND METHODS: Seminal vesicle aspiration was performed immediately before electroejaculation or penile vibratory stimulation in men with aspermia secondary to spinal cord injury. Sperm count and quality of seminal vesicle aspiration and subsequent ejaculation were compared with historical ejaculated counts, ultrasound findings and patient characteristics. RESULTS: Mean total number of right plus left seminal vesicle sperm plus or minus standard deviation was 511 +/- 960 x 10(6). Mean total number of sperm obtained by seminal vesicle plus electroejaculation or penile vibratory stimulation was 918 +/- 1,261 x 10(6). Average motility and viability of the seminal vesicle aspirated sperm were 1.3 and 3.2%, respectively. Average motility of the ejaculated sperm was 26.4% after seminal vesicle aspiration versus 16.3% in previous ejaculation induction procedures performed in the same patients. Seminal vesicle aspirated sperm represented 66% of the total number of sperm obtained during the session and was equal to 49% of the sperm obtained at previous electroejaculation or penile vibratory stimulation sessions. The period of abstinence correlated only with ejaculate count (simple regression p = 0.009). No other clinical characteristics had any effect on sperm count or quality. CONCLUSIONS: Large numbers of poor quality sperm are present within the seminal vesicles of spinal cord injured men and these sperm comprise a large portion of the specimens collected by electroejaculation or penile vibratory stimulation. This phenomenon is independent of the period of abstinence, implicating disordered storage of sperm due to spinal cord injury rather than infrequent ejaculation. The large number of senescent sperm within the seminal vesicles appears to be a primary cause of poor sperm quality in spinal cord injured men.     

Beneficial effects of vitamin E in sperm functions in the rat after spinal cord injury

Male infertility as a result of spinal cord injury (SCI) is associated with abnormal semen qualities including low sperm counts and poor sperm motility and morphology. Clinical studies suggest that reactive oxygen species (ROS)-related events might contribute to abnormal sperm functions after SCI. The current study examined whether impaired sperm functions after SCI can be ameliorated by an antioxidant, vitamin E. Vitamin E feeding of spinal cord transected (SCX) rats during the acute (maintenance) and chronic (restoration) phases of the injury partially preserved sperm viability and mitochondrial potential; similar effects were only seen in spinal cord contused (SCC) rats during the chronic phase. A beneficial effect of vitamin E on sperm motility, however, was only observed in SCX rats during the chronic phase of the injury. These results suggest that ROS-related events might account for some of the effects of cord injury on sperm functions, depending on the extent of injury and time postinjury. Furthermore, we found that sperm heads from SCC and SCX rats were less condensed compared to those from sham control rats. Such effects were attenuated by vitamin E, suggesting that ROS-related events may also contribute to abnormal sperm morphology after SCI. Partial restoration of male accessory gland weights in those rats fed vitamin E further suggests its beneficial effects on the functions of these glands. Conclusion: Vitamin E feeding attenuated some of the effects of spinal cord injury on sperm functions and male accessory glands in the rat. These results support a role of ROS-related events in deterioration of semen quality after cord injury. Further understanding of the underlying mechanisms for effects of vitamin E on sperm functions and male accessory glands will provide scientific rationale for the use of vitamin E or other antioxidant as therapeutic means to preserve sperm functions and semen quality in SCI men.     

Inflammatory cytokine concentrations are elevated in seminal plasma of men with spinal cord injuries

The semen of most men with spinal cord injury (SCI) contains sperm with abnormally low motility. Studies suggest that the seminal plasma is the source of this condition. The seminal plasma of men with SCI contains an abnormally high number of white blood cells (WBC), specifically, activated T cells. It is known that activated T cells secrete cytokines and elevated concentrations of cytokines can be harmful to sperm. It is not known if the seminal plasma of men with SCI contains elevated concentrations of cytokines. The purpose of this study was to determine if the seminal plasma of men with SCI contained elevated concentrations of cytokines. Using the method of enzyme-linked immunosorbant assay (ELISA), ten cytokines were measured in the seminal plasma of men with SCI as well as healthy non-SCI control subjects. The cytokines of interest were grouped according to Th1 effector functions: interleukin 1 beta, interleukin 2, interleukin 12, tumor necrosis factor alpha, tumor necrosis factor beta, interferon gamma (IL1 beta, IL2, IL12, TNF alpha, TNF beta, INF gamma, respectively) and Th2 effector functions: interleukin 4, interleukin 6, interleukin 10, transforming growth factor beta 1 (IL4, IL6, IL10, TGF beta 1, respectively). The results showed a predominance of Th1 versus Th2 cytokine production in the seminal plasma of men with SCI compared with that of control subjects. This finding suggests an immunologic basis for infertility as a possible avenue of investigation in these men.     

Neutralization of cytokine activity at the receptor level improves sperm motility in men with spinal cord injuries

Men with spinal cord injury (SCI) have a unique semen profile characterized by normal sperm concentrations but abnormally low sperm motility. Previous studies showed that elevated concentrations of cytokines in the seminal plasma of these men contribute to this condition. For example, when elevated concentrations of interlekin-1beta (IL-1beta), IL-6, and tumor necrosis factor-alpha (TNF-alpha) were immunoneutralized in the semen of men with SCI, sperm motility improved. The present study investigated if these cytokines act on sperm cell receptors to inhibit sperm motility. Semen was collected from men with SCI and from healthy non-SCI men. Sperm were separated from the seminal plasma by centrifugation. Eight identical aliquots of 5,000 sperm suspended in 50 microL of seminal plasma were prepared for each subject. Agents were added to the aliquots in order to neutralize IL-1beta, IL-6, and TNF-alpha at the receptor level. In SCI subjects, sperm motility improved in each treatment group compared with the untreated group, but statistical significance was reached only when neutralizing agents to all 3 cytokines were added. Improvement was less pronounced in subjects with close to normal semen cytokine concentrations or close to normal pretreatment sperm motility. In control subjects, IL-1beta, IL-6, and TNF-alpha were within normal values, and addition of receptor blockers to semen had no effect on sperm motility. These data support the hypothesis that cytokines act at the level of the sperm receptor to inhibit sperm motility. These data further support the notion that inactivating semen cytokines leads to improved sperm motility in SCI men. Our goal is to develop this finding into a treatment for low sperm motility in men with SCI.     

High seminal platelet-activating factor acetylhydrolase activity in men with spinal cord injury

Spinal cord injury (SCI) causes male infertility, with low sperm motility the major long-term cause. It has been suggested in previous studies that some seminal components may be responsible for the pathological asthenozoospermia. It is hypothesized that platelet-activating factor (PAF) acetylhydrolase (PAFah), which originates in the epididymis and other accessory sexual glands, may be a causative factor. This enzyme catalyzes PAF to acetate and biologically inactive lyso-PAF. PAF is well recognized to be an important phospholipid mediator that stimulates sperm motility and enhances sperm capacitation and fertilization. The present study was designed to analyze differences in PAFah activity in semen of men with SCI and age-matched healthy men. PAFah assay reagent kits were used to measure enzymatic activity by monitoring the production rates of 4-nitrophenol on a spectrophotometer during a given interval. The results showed that subjects with SCI had a higher concentration of PAFah than men in the control group (P < .001). A statistically significant negative correlation was found between enzymatic activity and sperm motility (r(2) = 0.8449; P < .001). Further studies will determine whether seminal vesicle dysfunction in men with SCI leads to abnormal PAFah activity, resulting in low sperm motility.     

Sperm motility in men with spinal cord injuries is enhanced by inactivating cytokines in the seminal plasma

The goal of this study was to determine whether inactivating specific cytokines in seminal plasma improves sperm motility in men affected by spinal cord injury (SCI). For this purpose, we used monoclonal antibodies to interleukin 6 (IL6), interleukin 1 beta (IL1-beta), and tumor necrosis factor alpha (TNF-alpha), all 3 cytokines having been previously detected at high concentrations in the seminal plasma of patients with SCI. In a group of 17 SCI men with low sperm motility (mean +/- SE, 20.1% +/- 3.1%), treatment with the 3 monoclonal antibodies at the median neutralization dose concentrations for 1.0 to 1.5 hours improved sperm motility in all cases. Effectiveness was higher in those specimens with a pretreatment sperm motility between 11% and 30% (from 19.3% +/- 1.4% to 41.9% +/- 4.9%, P < .0002), suggesting that pretreatment sperm motility might represent an indicator of cell damage and, therefore, a factor that influences monoclonal antibody effectiveness. To the best of our knowledge, these results represent the first rational treatment for improving low sperm motility in these severely affected patients.     

Application of 2 vibrators salvages ejaculatory failures to 1 vibrator during penile vibratory stimulation in men with spinal cord injuries

PURPOSE: Penile vibratory stimulation is the treatment of first choice for anejaculation in men with spinal cord injury. Nonresponders to penile vibratory stimulation are usually referred for electroejaculation or surgical sperm retrieval. Compared to penile vibratory stimulation these methods are invasive and usually yield lower total motile sperm, potentially limiting options for assisted reproductive technologies. To avoid these less than ideal options a simple method to salvage penile vibratory stimulation failures would be of benefit to spinal cord injured patients. We investigated the recovery rate when 2 vibrators were used to salvage ejaculatory failures to 1 vibrator in men with spinal cord injury. MATERIALS AND METHODS: A retrospective chart review was performed in 297 spinal cord injured men who underwent a total of 965 trials of penile vibratory stimulation at our center between 1991 and 2006. Only trials with high amplitude vibrators were examined. All men underwent 2 or more penile vibratory stimulation trials using 1 vibrator applied to the dorsum or frenulum of the glans penis. Men failing to ejaculate with 1 vibrator received 1 or more trials in which the glans penis was then sandwiched between 2 vibrators. RESULTS: Of all men 49% and 57% of those whose level of injury was T10 or above responded to penile vibratory stimulation with 1 vibrator. Of failures with 1 vibrator 22% responded to penile vibratory stimulation with 2 vibrators. CONCLUSIONS: Application of 2 vibrators salvaged ejaculatory failures to 1 vibrator during penile vibratory stimulation procedures in men with spinal cord injury. This simple penile vibratory stimulation sandwich method is recommended before referring patients for electroejaculation or surgical sperm retrieval.