血清抗精子抗体阳性对青春期雄性大鼠睾丸组织及睾丸生殖细胞中Fas/Fas-L凋亡途径的影响

目的:通过人工免疫雄性大鼠获得抗精子抗体(AsAb)介导的血清AsAb阳性的免疫性不育大鼠动物模型,观察AsAb对青春期大鼠睾丸组织及睾丸生殖细胞中Fas/Fas-L凋亡途径的影响。方法:5周龄(青春期)雄性Wistar大鼠30只,其中10只处死,取精子制备精子悬液免疫大鼠,余20只动物随机分为实验组(10只)和对照组(10只),4周后摘取睾丸。光镜观察睾丸组织改变,免疫组化法检测Fas、Fas-L和Caspase-3蛋白的表达。结果:实验组睾丸组织切片呈凋亡样改变,实验组Fas、Fas-L及Caspase-3蛋白的OD值(176.97±4.58,187.52±7.76,157.65±7.38)较对照组(161.87±5.37,150.27±8.65,120.37±6.76)显著增高(P<0.01)。结论:同种精子免疫大鼠,可成功制作AsAb介导的免疫性不育模型;AsAb影响雄性大鼠的生育力,其机制可能与Fas/Fas-L凋亡途径中Fas、Fas-L和Caspase-3蛋白的表达升高有关。     

国外期刊摘译

应用荧光原位杂交分析行ICSI的无精子症患者睾丸和附睾精子的染色体异常RodrigolL ,etal.HumanReprod ,2 0 0 4 ,19(1) :118- 12 3.文献报道不育男性精液中精子染色体发生数目异常的几率较正常人增大。然而 ,却没有有关睾丸和附睾精子细胞遗传学方面的研究报告 ,并且有关这方面的研究结果仍存在争议。本文作者采用荧光原位杂交技术分析了 2 7个标本 ,9个来自梗阻性无精症患者 (13个标本均采自睾丸 ;2个采自附睾 ) ,13个来自非梗阻性无精症患者 (13个标本均采自睾丸 ) ,5个为来自具有生育能力且精子活率正常的男性 (作为对照组 ) ,并对每…     

尿毒症对男性生育能力的影响

目的 探讨尿毒症对男性患者生育能力的影响.方法 检测40例尿毒症患者和40例不育患者以及40例正常生育者的精液,并根据公式:精子密度(×106/ml)×精子活动力×精子正常形态率计算出生育力指数.结果 正常生育者生育力指数为11.10(15.24),不育者的生育力指数为4.18(6.36),而尿毒症患者的生育力指数仅为0.23(0.76).结论 尿毒症患者的生育能力明显下降,造成"继发性"不育、应引起人们的重视.     

股部皮下埋藏睾丸对精子发生影响的实验研究

目的 探讨采用股部埋藏睾丸的方式修复阴囊皮肤缺损对睾丸精子发生的影响。方法 以育龄期贵州小型猪为实验动物，随机将雄性小型猪分为实验组和对照组。实验组动物手术切开阴囊制作股部埋藏睾丸动物模型，对照组手术处理同实验组但睾丸不移位。3个月后实验组、对照组随机各取2只行睾丸活检，观察睾丸的病理变化。然后将实验组、对照组6只分别与雌猪配对喂养3个月，观察有无生育，并在动物模型建立6个月后再次对所有实验组和对照组动物进行睾丸活检。结果 术后3个月和6个月实验组病理检查均发现睾丸曲细精管内各级生精细胞明显减少，未见成熟精子；而对照组睾丸曲细精管内各级生精细胞正常，可见成熟精子。术后3个月后，实验组动物与雌性动物配对喂养3个月无生育；而对照组生育正常。结论 猪股部埋藏睾丸的方法影响睾丸的精子发生，建议临床上最好不用此方法修复阴囊皮肤缺损，尤其是对希望保留生育能力的患者。     

睾丸性不育动物模型的建立

目的:探讨制作睾丸性不育动物模型的方法。方法:①X-射线局部照射:取8～10周龄的BALB/c雄鼠70只,分实验组1、2、3、4、5、6和对照组各10只。实验组1、2、3、4、5、6分别用1000、1200、1400、1600、1800、2000cGy6种不同的剂量照射睾丸局部10min;对照组不做任何处理。做受孕试验。②环磷酰胺腹腔注射:取4～5周龄的雄鼠40只,分实验组1、2、3和对照组各10只。实验组分别以30、5070mg/kg体重剂量的环磷酰胺腹腔注射,每周2次,连续注射5周;对照组腹腔注射生理盐水。做受孕试验。③达菲林腹腔注射:取8～10周龄的雄鼠20只,分实验组和对照组各10只。实验组腹腔注射0.375mg/ml达菲林0.4m,l对照组腹腔注射生理盐水0.4ml。做受孕试验。④通过脱氧核苷酸末端转移酶介导的缺口末端标记法(TUNEL)、苏木精-伊红(HE)染色及免疫组化等病理学检查鉴定。未使雌鼠受孕的雄性小鼠定为睾丸性不育,共30只。结果:①X-射线局部照射:实验组1、2的雄鼠分别于X-射线照射后10、15d使雌鼠受孕;实验组3、4的雄鼠,经观察3个月也未能使雌鼠受孕;实验组5、6的雄鼠,于X-射线照射后第5d和第2d死亡;对照组雄鼠皆于3d内使雌鼠受孕。②环磷酰胺腹腔注射:实验组1雄鼠体重增加7g左右,停药后9～14d恢复生育能力,使雌鼠受孕;实验组2雄鼠体重增加4g左右,停药后观察3个月未使雌鼠受孕;实验组3雄鼠体重不增,呈消瘦状,分别于用药第3、4、5周相继死亡;对照组雄鼠皆于3d内使雌鼠受孕。③达菲林腹腔注射:实验组雄鼠于停药后3周左右使雌鼠受孕;对照组雄鼠皆于3d内使雌鼠受孕。④TUNEL结果:对照组睾丸组织偶见凋亡细胞,占(0.71±0.12)%;睾丸性不育组睾丸组织凋亡细胞数量明显增加,占(10.36±1.48)%,两者比较差异有显著性(P<0.05)。HE染色:对照组睾丸组织正常,生精小管呈饱满椭圆形,各级生精细胞排列有序,内含大量精子细胞;睾丸性不育组生精小管萎缩,细胞排列疏松,未见各级生精细胞,只有支持细胞,各生精小管间腔隙扩大,间质细胞减少。免疫组化染色:CD29、Hsp90α、CD117的阳性表达率对照组分别为(50.3±5.2)%、(41.6±3.5)%、(73.6±3.7)%,而睾丸性不育组明显降低,分别为(1.3±0.2)%、0%、(1.6±0.3)%(P<0.01)。p53阳性表达率对照组为(19.7±0.8)%,而睾丸性不育组明显增高,为(39.4±2.9)%(P<0.01)。结论:X-射线睾丸局部照射、环磷酰胺腹腔注射法均可制作睾丸性不育动物模型。     

病毒感染与男性不育关系的研究进展

近年来研究发现病毒感染与不孕不育存在一定的联系,人类免疫缺陷病毒、乙肝病毒等感染不仅损伤生殖系统的功能、影响生育能力,而且也能够通过性接触传播而导致不育,病毒的遗传物质可通过干扰精卵正常受精或影响胚胎的正常着床导致不孕。在男性不育方面,病毒主要通过损伤睾丸功能和影响精子浓度、活力、形态以及DNA完整性使精子质量降低,影响正常精卵结合,进而导致男性生育能力下降甚至不育。本文就相关病毒感染与男性不育的关系做一综述。     

不同剂量昆明山海棠对雄大鼠生殖系统的形态学研究

雄性SD大鼠60只,随机分成6组,二组对照,其余4组用昆明山海棠稀醇提取物(ATH)灌胃给药,每日分别为2.0g/kg;1.0g/kg;及0.3g/kg(二组),每周六次。用药6周至完全不育后剖杀用药的前三组及一组对照。取睾丸、附睾、心、肝、脾、肺、肾、胃,行常规HE及PAS+苏木素染色,部分鼠做DNA、RNA组化反应。取附睾尾评价精子数据。0.3g/kg组及另一组对照继续以每日0.2g/kg的维持量灌胃16周以保持不育,停药5周后证实其生育力恢复后剖杀,观察指标同前。结果表明:2.0g/kg组曲细精管平均损伤率为55.4%,1.0g/kg组曲细精管损伤明显少于前组(损伤率6.2％),小剂量鼠无论给药6周或22周睾丸和附睾形态与对照无异。结果提示:大鼠睾丸曲细精管的损伤范围和损伤的程度与ATH的剂量大小有关,而与作用时间的长短关系不大。最低抗生育剂量时,对ATH最敏感的部位是变态期精子细胞及附睾精子,以此剂量长期给药对睾丸附睾无明显影响,所致不育可以恢复。     

经皮睾丸微穿刺活检后冷冻精子的卵胞质内单精子注射

目的:对经皮睾丸微穿刺活检后冷冻保存精子卵胞质内单精子注射(ICSI)治疗非梗阻性无精子症所致不育进行临床总结,并对其影响治疗结果的因素进行探讨。方法:对62例非梗阻性无精子症患者进行经皮睾丸微穿刺活检,发现活动精子者(35例)对睾丸活检组织进行冷冻保存;女方促排卵常规使用促性腺激素释放激动剂(GnRHa)/卵泡刺激素(FSH)/人绒毛膜促性腺激素(hCG)方案,B超监测卵泡发育情况并引导经阴道取卵,冷冻的睾丸组织解冻后行ICSI,良好胚胎进行移植。结果:取卵周期为35个,冷冻的睾丸精子解冻后行ICSI,35个周期进行常规胚胎移植。13例临床妊娠。启动周期、取卵周期与移植周期临床妊娠率均为37.14%(13/35)。结论:经皮睾丸微穿刺活检后ICSI是治疗非梗阻性无精子症所致不育的最主要和有效的方法;活检后对有活动精子的睾丸组织进行冷冻保存不影响治疗结果,可以减少患者睾丸活检的次数,减轻患者痛苦。     

抗精子抗体对小鼠睾丸超微结构的影响

目的研究免疫性不育雄性小鼠的睾丸超微结构的特点,探讨抗精子抗体（AsAb）对睾丸生精微环境的影响。方法用同种精子及福氏佐剂免疫昆明种雄性小鼠。ELISA法检测AsAb,以此验证获得免疫性不育动物模型,同时设立对照组;解剖小鼠取其精子观察精子质量差别,透射电镜观察各组睾丸的超微结构。结果人工免疫后的雄鼠血清AsAb均为阳性。正常对照组均为阴性;模型组小鼠精子质量与对照组比较显著下降,生精上皮超微结构受损严重。结论用同种精子及福氏佐剂免疫动物,可成功制作抗精子抗体介导的免疫性不育动物模型;AsAb可通过攻击睾丸生精微环境影响雄性小鼠的生育力。     

睾丸精子辅助生育30例报告

本生殖中心自 1 997年至今 ,用睾丸精子卵浆内单精子注射 (intracytoplasmic sperm injec-tion,ICSI)为 30对无精子症不育夫妇行辅助生育治疗 32个周期 ,报道如下。一、资料与方法30例无精子症不育、染色体核型正常的患者平均年龄 35 (2 2～ 45 )岁 ,治疗前均检查血卵泡刺激素 (FSH)、黄体生成素 (L H)、睾酮 (T)、泌乳素 (PRL )水平及睾丸活检了解睾丸功能及生精状况。首先对女方促超排卵治疗 [1] ,女方平均年龄 31 (2 1～ 39)岁 ,当卵泡发育成熟时 ,以阴道B超介导下取卵 ,卵子的处理及 ICSI方法见文献 [2 ]。获得卵子后即行睾丸精…     

输精管结扎术后附睾瘀积综合征发病机制、诊断与治疗探讨

为了阐明输精管结扎术后附睾瘀积综合征的发病机制，探讨诊断与治疗方法，收集１０例病人，行血清及精浆抗精子抗体测定及Ｂ超、Ｃ超、ＭＲＩ等项硷查。手术采用附睾一近睾端输精管。结扎结节整块切除术。术中取附睾液行细菌培养及睾丸活检，标本行光镜和电镜检查。术后随访，９例治愈，１例无效。瘀积附睾的组织学检查表明是附睾对外漏精子的无菌性炎症反应。睾丸活检，生精过程正常２侧，其同侧结扎断端有精子肉芽肿形成，提示输精管结扎断端精子肉芽肿的形成对睾丸有保护作用。     

Oxidative stress‐mediated alterations on sperm parameters in male Wistar rats exposed to 3G mobile phone radiation

In recent years, there has been significant increase in mobile phone users. With this, health concerns associated with the exposure to electromagnetic radiation are also increasing. Continuous exposure to electromagnetic (EM) radiation generated from mobile phone is one of the probable reasons behind increasing male infertility. EM radiations induce oxidative stress that leads to numerous changes in reproductive parameters. With this hypothesis, we studied the effect of 3G mobile phone radiations on the reproductive system of male Wistar rats. Adult rats were divided into two groups: control and radio frequency‐exposed. The animals were exposed to 3G mobile phone radiation for 45 days (2 hr/day) in specially designed exposure setup under standard conditions. Various biochemical and physiological parameters such as sperm count, sperm morphology, mitochondrial activity, lipid peroxidation, reactive oxygen species level and histopathological analysis were studied. Histopathological examination revealed a reduction in spermatogenic cells and alterations in sperm membrane. Significant increase in ROS and lipid peroxidation level with simultaneously decrease in sperm count, alterations in sperm tail morphology were observed in the exposed group. In conclusion, exposure to mobile phone radiations induces oxidative stress in male Wistar rats which may lead to alteration in sperm parameters and affects their fertility.     

Gonadotropin therapy in males with hypogonadotropic hypogonadism: Factors affecting induction of spermatogenesis after gonadotropin replacement

Sixteen patients with hypogonadotropic hypogonadism received gonadotropin replacement therapy. Two patients treated with HCG alone showed induction of spermatogenesis 2 and 12 months after the start of treatment. Three subjects receiving combination therapy showed sperm appearance 6–28 months after treatment. The patients showing sperm appearance, whose testicular volume was ≥4 ml, showed a higher sperm count and impregnated their partners, although no relationship was found between pretreatment testicular volume and sperm appearance. The response to HCG test correlated with sperm appearance after gonadotropin therapy. Sperm appearance was not observed in any subject except for one who showed no response to luteinizing hormone-releasing hormone (LH-RH) test and none of the patients without response of FSH to LH-RH demonstrated any induction of spermatogenesis. In conclusion, the responses to LH-RH test and possibly to HCG test could predict the induction of spermatogenesis after gonadotropin replacement therapy, and a large testicular volume is associated with post-treatment fertility.     

Azoospermia and maturation arrest: malfunction of valves in erect poster of humans leads to hypoxia in sperm production site

Maturation arrest (MA) of spermatogenesis is diagnosed on histology as interruption of spermatogenesis before the final stage without impairment of Sertoli or Leydig cells. It is considered a condition of irreversible or absolute infertility. Varicocele, which represents impairment in the testicular venous drainage system, has been shown to be a bilateral disease. Malfunction of the valves increase the hydrostatic pressure in the testicular venous system that exceeds the pressure in the arterial system leading to hypoxia in the testicular microcirculation and in the seminiferous tubules, the sperm production site. Sperm production deteriorates, and ultimately progresses to azoospermia. Our prediction was that MA, if genetic factors are excluded, is the final stage of long standing hypoxia. This would indicate that MA is not always an independent disease entity, but may represent progressive process of deterioration of the testicular parenchyma beyond azoospermia. By histology and electron microscopy, our prediction confirmed, at least partially, that MA is associated with degenerative ischaemic changes in the seminiferous tubules. Adequate treatment of bilateral varicocele by microsurgery or super-selective sclerotherapy of the internal spermatic veins including associated network of venous bypasses, vertically oriented, may resume the flow of oxygenated blood. If irreversible damages did not occur and ischaemia is not too long standing, limited sperm production may be restored, at least partially.     

Semen characteristics and inflammatory mediators in infertile men with different clinical diagnoses

This study was aimed at investigating whether semen characteristics in different clinical diagnoses of infertility are associated with PMN elastase, IL-6, IL-8, IL-1beta and TNFalpha levels detected in seminal plasma. Sixty-eight patients were divided into groups according to their clinical diagnosis: idiopathic infertility (group I), varicocele with infections (group II), varicocele (group III), infections (group IV), controls (group V). Physical examination and scrotal Eco-color Doppler was used to detect the varicocele. Patients with positive bacteriological semen analysis were considered as having an infection of the male reproductive tract. Samples were examined by light microscopy and transmission electron microscopy (TEM). TEM data were quantified with a mathematical formula furnishing a fertility index and the percentage of sperm apoptosis, immaturity and necrosis. PMN elastase/alpha1-PI complex levels were determined by ELISA and IL-6, IL-8, IL-1beta, TNFalpha by Bio-Plex Cytokine assay. Sperm concentration (I-II: p < 0.005; III-IV: p < 0.0001), motility (I-IV: p < 0.0001) and the fertility index (I: p < 0.005; II-IV: p < 0.0001) were significantly lower in the groups vs. controls, whereas sperm pathologies, except for apoptosis, were significantly higher in group I and apoptosis and necrosis were higher in group III. An increase in immaturity (p < 0.005) with a decrease in necrosis (p < 0.005) were observed in group III vs. group IV. Significantly higher levels of inflammatory mediators were detected in groups III and IV vs. controls. Despite a broad relationship among different inflammatory mediators, no correlation was found among them and the semen parameters, including indices from TEM analysis. In conclusion, patients with idiopathic infertility showed altered semen quality and normal levels of inflammatory mediators. Genitourinary infection and varicocele induced an inflammatory effect which could play a detrimental role in spermatogenesis, revealed by a decrease in sperm motility and the fertility index, concomitant with an increase in immaturity mainly in varicocele and necrosis in infection.     

Evaluation of spermatogenesis and fertility in F1 male rats after in utero and neonatal exposure to extremely low frequency electromagnetic fields

Aim: To determine whether in utero and neonatal exposure to a 60Hz extremely low frequency electromagnetic field (EMF) results in spermatotoxicity and reproductive dysfunction in the F1 offspring of rats. Methods: Age-matched,pregnant Sprague-Dawley rats were exposed continuously (21h/day) to a 60 Hz EMF at field strengths of 0 (sham control), 5, 83.3 or 500 μT from day 6 of gestation through to day 21 of lactation. The experimentally generated magnetic field was monitored continuously (uninterrupted monitoring over the period of the study) throughout the study. Results: No exposure-related changes were found in exposed or sham-exposed animals with respect to the anogenital distance, preputial separation, testis weight, testicular histology, sperm count, daily sperm production,sperm motility, sperm morphology and reproductive capacity of F1 offspring. Conclusion: Exposure of Sprague-Dawley rats to a 60Hz EMF at field strengths of up to 500μT from day 6 of gestation to day 21 of lactation did not produce any detectable alterations in offspring spermatogenesis and fertility.     

Probability of sperm detection in nonobstructive azoospermic men undergoing testicular sperm extraction procedures unrelated to clinical parameters

The study was conducted to evaluate the significance of preoperative clinical parameters for detection of mature testicular sperm cells in nonobstructive azoospermic men. Sixty-five consecutive men with nonobstructive azoospermia underwent testicular sperm extraction procedures. Testicular samples were analyzed histologically with patterns classified as mature spermatogenesis (normal or partial), arrest of spermatogenesis, and Sertoli cell only. Testicular sperm cells were isolated for use in an IVF/ICSI program. Histologic patterns and detection rate of sperm cells were correlated to clinical characteristics. Mature sperm cells were found in all levels of serum FSH. The men were divided into 3 groups based on their clinical characteristics (serum FSH level and testicular size). The distribution of the different testicular histologic patterns, as well as detection rate of sperm cells, was similar in all groups. No correlation was found between serum levels of FSH, LH, prolactin, or testosterone and sperm presence. None of these parameters, nor the testicular size and consistency, can serve as predictive variables of the histological pattern or the presence of mature sperm cells in the testicular biopsies in cases of nonobstructive azoospermia. Until an effective predictive tool is available, a trial of sperm retrieval is recommended for all azoospermic men independent of their clinical characteristics.     

Does brachytherapy of the prostate affect sperm quality and/or fertility in younger men?

OBJECTIVE: Sperm banking prior to surgical procedures which may affect fertility, such as retroperitoneal lymph node dissection, has been well documented. However, such procedures are usually performed in young men. With older men marrying later in life, or remarrying, we wanted to investigate the effects of radiation on prostate cancer patients who wanted to have children afterwards. MATERIAL AND METHODS: We encountered several patients with prostate cancer who decided to undergo brachytherapy and were planning to have more children. We performed a search using PubMed and Ovid for the period 1966-2001 using the key words "fertility", "sperm banking", "radiation effects", "prostate cancer" and "brachytherapy". RESULTS: Of the four young patients we encountered who underwent brachytherapy, we found no significant change in semen parameters post-therapy, and three of them were able to father a child subsequently without any deleterious side-effects. It has been demonstrated in several reports that external-beam radiation therapy is associated with decreased spermatogenesis due to Leydig cell dysfunction and decreased serum testosterone, as well as having a direct effect on spermatogonia. However, there is a scarcity of literature discussing the effects of prostate brachytherapy on spermatogenesis as the patients involved are usually older and usually do not desire to father any more children. As I has a half-life of 60 days, we used an exposure of 10 mR/h at the symphysis pubis and used integration to find the total dose exposed to the testis as follows: Limits 14 400 to 0, S 10e (-In2/1440.Tdt) where T = 14 400 and 20.75 R = 20.75 cGy. Therefore, the total dose was 20.75 cGy x 0.91 = 18.88 cGy. This value is considered too low to have any significant effect on testicular tissues. CONCLUSIONS: We speculate that the effects of prostate brachytherapy on spermatogenesis in prostate cancer patients are minimal. However, due to the half-life of I, we recommend that these patients should wait for at least 3-4 months before trying to conceive. Furthermore, younger men with prostate cancer may want to consider sperm banking prior to brachytherapy if they want to have children in the future.     

Long-term effect of vasectomy on spermatogenesis in men: a morphometric study

Spermatogenic damage may occur after vasectomy, and the damage is pressure mediated, occurring when the occluded reproductive tract is unable to accommodate additional sperm produced by the testis. This study aimed to determine the long-term effect of vasectomy on spermatogenesis in humans and clarify how the balance between sperm production in the testis and sperm storage in or removal from the tract might be maintained. During inguinal hernia repair, an open biopsy was performed to obtain testicular tissue blocks from 51 Chinese men (aged ≥50 years), of whom 25 (control group) had not undergone vasectomy and 26 (vasectomized group) had undergone bilateral vasectomy 22–42 years before. Methacrylate resin-embedded testicular sections were made, and morphometric studies were performed using light microscopy. In addition, sizes of the testis and epididymis were estimated with ultrasonography. The testicular tissue blocks obtained from one control and seven vasectomized men consisted almost completely of connective tissue. In the other 43 men, significant differences were not found between the two groups in the testicular or epididymal size, qualitative histology or quantitative parameters including the mean diameter or volume fraction of the seminiferous tubules. In conclusion, sperm production and sperm storage/removal reached a static equilibrium after vasectomy, likely due to spermatogenic degeneration or less sperm production as a result of aging or due to vasectomy-induced testicular (interstitial) fibrosis. Thus, complications that might occur in association with overproduction of sperm and distension of the tract would disappear or be relieved with time.     

Sperm ultrastructure and meiotic segregation in a group of patients with chronic hepatitis B and C

Little is known about the effect of chronic hepatitis B and hepatitis C on sperm quality. In this study we analysed sperm quality from selected patients with chronic hepatitis B virus (HBV) or hepatitis C virus (HCV) infections. Semen samples were examined by light and transmission electron microscopy (TEM). TEM data were elaborated with a mathematical formula able to indicate a fertility index and the presence of the three main sperm pathologies: apoptosis, immaturity and necrosis. Meiotic chromosome segregation was investigated by fluorescence in situ hybridisation carried out on sperm nuclei, using probes for chromosomes 18, X and Y. Despite normal sperm concentration, we observed reduced motility. TEM analysis highlighted that 35.7% of patients showed generally good semen quality. However, significantly higher values of apoptosis and necrosis, compared with controls, were observed, demonstrating spermatogenetic alterations. Regarding meiotic segregation, we found an incidence of disomies similar to that observed in control samples, whereas diploidy resulted higher in HCV patients, without reaching statistical significance. In conclusion, sperm quality in the studied group was not impaired, however, apoptosis and necrosis resulted out of normal range and the fertility index was significantly lower in HCV and HBV infected patients versus controls.