家兔输精管结扎后自身免疫反应与血脂变化的观察

为观察家兔输精管结扎后对精子抗原的自身免疫反应与血脂变化的关系,将输精管结扎后7、9个月日本大耳白雄兔20只,随机分为“结扎·胆固醇组”和“结扎·普食组”,另选月龄相近的同种雄兔20只,分为“对照·胆固醇组”和“对照·普食组”。结果表明,在普食条件下,输精管结扎不引起血脂升高,在高脂饲料条件下,输精管结扎组血脂升高较对照组者为著;实验性高脂血症对结扎兔的体液免疫反应没有抑制作用,面对细胞免疫有较弱的抑制作用。     

输精管结扎对实验性动脉粥样硬化的影响

输精管结扎术后7和9个月家兔各10只,随机分为输精管结扎基础饲料(V-S)组和输精管结扎胆固醇(V-Ch)组,同种雄兔20只随机分为对照基础饲料(C-S)组和对照胆固醇(C-Ch)组。实验结果表明,V-S组血脂、脂质过氧化物含量与C-S组比较无差异,主动脉和冠动脉均无脂质斑块形成。在持续高脂负荷后,V-Ch组总脂、β-脂蛋白水平显著地高于C-Ch组,但是主动脉、冠动脉的病变面积和程度则无差异。这可能与V-Ch组血清脂质过氧化物含量与C-Ch组比较无增高有关,也可能与V-Ch组虽有抗精子抗体产生,但无循环免疫复合物形成有关。     

输精管吻合不育家兔皮质质醇与IL—1、TNF—α变化

应用显微外科技术以来 ,输精管吻合解剖学成功率可达95 %以上 ,但生育力恢复却只有 5 0 %～ 60 %。吻合不育的内分泌变化机制值得研究。本文研究了输精管吻合不育家兔血清皮质醇变化 ,并检测了血清 IL- 1活性和 TNF- α含量。1　材料与方法1.1　动物模型　日本大耳白雄兔 3 1只 ,体重 2 .5～ 3 .0 kg,随机分为 :1输精管吻合组 14只雄兔于输精管结扎 12月后 ,行显微外科吻合术。 3月后与成年雌兔交配 ,观察 2月后根据配偶妊娠与否分为吻合育组 ( VFG)和吻合不育组( VIG) ,各 7只。 2输精管结扎组 ( VG) 8只雄兔只接受结扎手术和假吻合…     

兔输精管结扎术三个月对精子发生的影响

关于输精管结扎对精子发生的影响 ,以往的研究报告不尽一致。本文拟采用体视学方法来定量研究兔输精管结扎术三个月对精子发生的影响。 6只青春期兔作对照 ,8只青春期兔作双侧输精管结扎术 ,三个月后取出一侧睾丸及附睾作形态定量分析。运用 2 5 μm厚甲基丙烯酸树脂切片及无偏、有效的体视学新工具——光学体视框来测量生精细胞 (核 )数。结扎组较正常组睾丸平均体积明显缩小 ;各级生精细胞数量也明显减少 ;支持细胞数无改变 ;但结扎组中有 2例的睾丸体积和生精细胞数接近正常。输精管结扎术后三个月 ,大白兔精子发生受到抑制 ,但有明显个…     

家兔附睾管尾段吸收功能的研究

家兔输精管低位结扎后附睾管尾段结构出现明显变化，外形和光电镜观察显示其管径增大，管腔扩张，腔内精子密集。附睾的平均重量增加。主细胞顶部胞膜向胞质内凹陷增多，并形成施工吞饮小泡，核上区出现许多与吸收作用有关的小泡、大泡、多泡体、溶酶体和线粒体等超微结构。出现上述变化的时间是术后的第３个月。输精管中位结扎组的变化情况基本相似于低位结扎组。输精高位结扎后因不能排出的睾网液可在附睾的近段被大部吸收，少量的     

输精管结扎与实验性高血脂对听觉脑干电位及血清过氧化脂质的影响

输精管结扎12月以上日本大耳白兔16只,随机分为输精管结扎基础饲料(V-S)组和输精管结扎胆固醇(V-Ch)组。体重相近的同种雄兔16只,分为对照基础饲料(C-S)和对照胆固醇(C-Ch)组。听觉脑干电位检测结果表明:气导ABR阈值,输精管结扎组(V-S.V-Ch)与各门对照组比较无差异(P<0.05),而高胆固醇饲料组(C-Ch.V-Ch)则显著地高于对照组(P<0.01),并且与TC水平正相关(P<0.01);各组Ⅰ、Ⅲ、Ⅴ波潜伏期无延长,波间期各组比较也无显著差异,(P>0.05)。血清过氧化脂质含量与TC水平正相关(P<0.01)而与血清抗精子抗体滴度无相关(P>0.05)。本实验证明,实验性高血脂使气导ABR阈值及血清过氧化脂质增高,而输精管结扎对其无影响。     

输精管吻合术后不育原因的实验研究精子密度与睾丸功能

14只成年日本大耳白兔,双侧输精管结扎12月后,以显微外科技术行双侧输精管吻合术,3月后与雌兔配对交配,观察2个月,根据妊娠与否分为输精管吻合育组(VFG)和输精管吻合不育组(VIG),各7只。另设输精管结扎组(VG)和假手术组(SOG)作为对照。结果表明,(1)VFG的精子密度与SOG比较虽为低值(P<0.01),但显著地高于VIG(P<0.01)。(2)精子密度与睾丸ACE活力、Na~+,K~+-ATPasc活力、Mg~(++)-ATPasc活力、睾丸cAMP含量均呈显著的正相关。(3)精子密度、cAMP含量与ABP呈明显的负相关。(4)VFG血清睾酮含量与VIG比较有显著差异(P<0.05)。VFG、VIG中血清睾酮水平与精子密度呈明显的正相关(r=0.60、P<0.05)。     

家兔高位输精管结扎后其近端结构变化的观察

家兔高位输精管结扎后其近端结构出现明显变化。光镜和电镜观察显示，管的近端管径增粗，管腔扩张，腔内充满精子。柱状上皮细胞顶部胞膜内陷增多，胞质内含有较丰富的有衣小泡、多泡体、溶酶体和线粒体等。出现变化的时间是术后的第３个月。中位输精管结扎后其近端结构的变化基本相似于高位结扎。低位输精管结扎后无变化。本研究结果提示，家兔输精管不单纯是输送精子的管道，还有较强的摄取某些物质的作用。     

超声波对家犬实验性附睾淤积症治疗的电镜观察

用超声波处理家犬附睾淤积区8 min(功率0.75 W/cm~2),每日1次,连续7 d。结果发现:由输精管结扎所引起的附睾超微结构的异常改变或明显改善,或完全复原;同时造成附睾主细胞新的损伤。本实验通过输精管结扎组与对照组、超声波治疗组与输精管结扎组的比较,既为超声波对输精管结扎并发附睾淤积症疗程治疗近期效果提供了形态学依据,也提示进一步探索该物理因子作用最适宜的“量”的必要性。本文还对超声波治疗输精管结扎并发附睾淤积症的机理进行了讨论。     

具有杀精子功能的生物材料甲基丙烯酸乙醋—甲基丙烯酸—甲基丙烯酸羟乙酯与动物实验

目前男性避孕采用的结扎法和粘堵法都是用堵塞输精管的办法来实现的,容易引起附睾郁积等副作用,而且这两种方法都是绝育。 本文介绍一种即亲水又疏水,其水肿胀度可任意控制的三元共聚物甲基丙烯酸乙酯—甲基丙烯酸—甲基丙烯酸羟乙酯(HFMC—1)的合成及其动物实验结果。共聚物溶液注入输精管内能很快地聚集一起,附着在输精管内,能长时间提供足以杀死精子的酸性环境,使通过的精子活力降低,从而达到避孕的目的。所以使用该共聚物进行男性避孕,具有不堵塞和生育可复的特点。动物实验结果表明:HFMC—1具有杀精子的作用。注射HFMC—1后的雄兔,其精子存活率由85％左右下降至10％左右,十个月以后,精子存活率仍能控制在40％左右。     

Antisperm antibody titres, immune complex deposition and immunocompetence in long-term vasectomized mice

Experiments were performed to determine sperm antigen-specific and non-specific immunological changes in long-term vasectomized BDF₁ mice. Circulating antisperm antibodies, detected by immunofluorescence assay, were observed as early as 1 month post-vasectomy; antisperm titres increased with time and were highest in animals that had been vasectomized for over 2 years. Several aged sham-vasectomized mice also had significant antisperm antibody titres, but the development of antisperm antibodies was significantly different from that of the vasectomized group. Tests of general immunocompetence, performed on vasectomized and sham-vasectomized mice at various intervals up to 2·5 years post-surgery, revealed a decrease in mitogenic responsiveness over time in both groups, but no difference between age-matched groups in lymphocyte responses to mitogenic or allogeneic stimulation in vitro, or to in vivo challenge with picryl chloride (delayed hypersensitivity response) or immunization with foreign antigen (humoral response). Most vasectomized mice developed epididymitis and epididymal sperm granulomas by 9 months post-surgery. Patchy regions of hypospermatogenesis were observed in some testes as early as 3 months post-vasectomy, and spermatogenesis was markedly impaired in all long-term vasectomized mice examined. Orchitis lesions characterized by immune complex deposition and lymphocytic infiltration were found in testes from three out of 14 long-term vasectomized mice, as compared to none of the sham-operated group. Studies of vasectomy-associated immune complex deposition in the kidney were inconclusive because aged animals from both vasectomized and sham-vasectomized groups had similar patterns of immunoglobulin and complement deposition in the glomerular basement membranes.     

输精管结扎后家兔的白细胞粘附抑制试验和免疫器官的形态学观察

日本大耳白成熟雄兔40只,对照组17只,实验组23只,行双侧输精管结扎术。LAIT检测结果:实验组23只中,阳性者10只,其阳性率为43.5%,对照组17只全部阴性,两组比较,P值小于0.01,差异显著;胸腺、脾脏、淋巴结的形态学观察,发现实验组淋巴组织增生,巨噬细胞吞噬现象活跃,这与LAIT的结果是相一致的。     

Detection of anti-sperm antibodies in serum, seminal plasma and cervical mucus by the immunobead test

Anti-sperm antibodies in serum and seminal plasma were detected by means of an indirect immunobead test (IBT). Immunobeads with separate specificites for each immunoglobulin class (IBT-IgG, IBT-IgM, and IBT-IgA) were used. Semen parameters were controlled in all sperm donors and Biggers-Whitten-Whittingham (BWW) medium supplemented with human serum albumin (HSA) was used to increase sperm motility. This technique was tested with high titre anti-human sperm sera induced in rabbits. Sperm tails showed a good response by IBT. We included in this study 178 men and 35 women evaluated for infertility and the sera were also tested by the Tray Agglutination Test (TAT). Although the presence of semen markers such as agglutination or trembling of spermatozoa is meaningful even by itself, the percentage of anti-sperm antibodies was increased in the patients with markers, both using IBT (21.4%) and using TAT (35.7%). At high titres of specific immunoglobulins (rabbit antisera and vasectomized men), the correlation between IBT and TAT techniques was better than in sera with very low titres, in which more positive TAT's were detected.     

Autoimmunity to sperm antigens in vasectomized men.

Sera from vasectomized men were tested for the presence of antibodies directed against sperm antigens. A high percentage (about 55%) of the vasectomized men developed agglutinating antibodies. A lower percentage (22%) also developed low titres of antibodies to human protamine, as detected in the indirect IFT on swollen sperm heads and 22% developed cytotoxic antibodies. A correlation was found between the presence of anti-protamine antibodies and the presence of agglutinating and of cytotoxic antibodies. This correlation, and also the fact that they developed after vasectomy, indicates that the formation of antibodies against human protamine is a result of an autoimmune response to spermatozoa. The indirect IFT was also performed on 'normal' unswollen spermatozoa. All the sera were positive at least on one of the sperm antigens located in the acrosome, equator, or post-nuclear region, but no increase nor decrease in titre was found after vasectomy.     

The predominance of IgG1 and IgG3 subclass antisperm antibodies in infertile patients with serum antisperm antibodies.

Mouse monoclonal antibodies (MAb) specific for each of the four human IgG subclasses and immunofluorescence flow cytometry were used to evaluate the subclass of the IgG antibody response to sperm in serum samples from 13 men and 6 women with a high titer (greater than 1:15,625) of IgG antisperm antibodies (ASA] determined by an indirect immunobead test. Five sera without ASA were also studied as a control. All 19 (100%) of the ASA-positive sera contained immunoglobulin (Ig)G ASA of the IgG1 and IgG3 subclasses. A 1:1 correlation was observed between the presence of IgG1 and IgG3 ASA. IgG2 was essentially undetectable, while IgG4 reactivity, although less intense than IgG1 and IgG3, was more prominent in the sera from the five vasectomized men. The ability of the IgG1 and IgG3 ASA-positive sera to deposit complement (C) on sperm was demonstrated by the concomitant binding to antibody-laden sperm of polyclonal antibodies to the membrane attack complex (C5b-9) of C. Both C-fixing and non-C-fixing ASA-positive sera were found to possess IgG1 and IgG3 antisperm antibodies. The predominance of IgG1 and IgG3 subclasses suggested a T-cell dependent immune response to sperm antigens.     

Study of sperm antibodies in the sera of vasectomized males

Agglutination and immobilization tests were performed to detect the presence of sperm antibodies in the sera of 100 vasectomized men. Overall, 52% of subjects had sperm antibodies. 50% showed agglutinating antibodies, while immobilizing antibodies were present in 40%. Of the 52 positive cases, 23 showed agglutinating antibody alone in their sera. 73 had both agglutinating and immobilizing antibodies, and 3.8% had sperm immobilizing activity alone. The age incidence of the presence of sperm antibodies was as follows: under 35 years, 66%; 36-40 years, 50%; 41-45 years, 50%; and 46 years and above, 33.3%. The highest percentage of presence of sperm antibodies was recorded in men who had been vasectomized less than 1 year previously (75%). Significantly high titers of sperm agglutinating activity were found in 20% of the positive cases, while high titers of sperm immobilizing activity were noted in 40% of positive cases. It has been suggested that the development of sperm antibodies after vasectomy may prevent subsequent successful restoration of fertility by reanastomosis.     

Effects on the efferent ducts in Macaca mulatta.

Immunopathologic findings in efferent ducts of 36 rhesus macaques vasectomized as many as 12 years earlier and of 11 age-matched control animals were compared. Electron-microscopic observation of these ducts revealed changes after vasectomy. The epithelium shortened from a prevasectomy height of 25 mu to 14 mu as the ducts stretched after vasectomy. The number of sperm and macrophages in the lumen increased. The basement membrane was 300-700 A wide in nonvasectomized animals but several times that in animals vasectomized 6 or more years before; the mean width significantly increased with time after vasectomy. Numerous electron-dense immune complexes were found within the thickened lamina in 33% of vasectomized animals and in none of the controls. The mean size of the electron-dense areas varied from 0.01 sq mu in a monkey vasectomized 3 years earlier to 0.18 sq mu in an animal vasectomized 7 years earlier; the mean area significantly increased with time after vasectomy. Frozen sections of testis and epididymis were evaluated through the use of fluorescein-conjugated antibodies. Of the nonvasectomized controls, 18% showed immune deposits. Of the vasectomized animals, 53% revealed C3 deposition in the basement membrane surrounding the efferent ducts. The presence of electron-dense deposits plus the finding of putative immune complexes as revealed by immunofluorescence suggested that vasectomy enhances leakage of sperm antigens, particularly in the region of the efferent duct.     

An Indirect Enzyme-Linked Immunosorbent Assay (ELISA) for the Detection and Quantitation of Antisperm Antibodies

ABSTRACT: Several recent comparative investigations using various assays to detect and quantitate levels of antibody to human spermatozoa have produced widely varying results. In an attempt to reduce test variability, an indirect enzyme-linked immunosorbent assay (ELISA) was devised to measure antisperm antibodies. A standardized protocol was adapted employing sperm adsorption to polystyrene microtiter plates, at a density of 10⁵ sperm per well, serum and enzyme-conjugate incubation conditions at 37°C for 60 min, and three ten-minute washes between incubations using phosphate-buffered saline containing Tween-20. Using antihuman sperm antisera generated in rabbits, the ELISA was shown to yield significantly detectable antibody at dilutions of 1/16,384. The ELISA demonstrated approximately 89% reproducibility (ie, 100% minus the coefficient of variation) for an “intraassay” study wherein 300 determinations were performed on the same day on sperm from ten donors. However, when sperm from one donor were used in 30 determinations during ten assays over a six-month period, “interassay” reproducibility was approximately 51%. The ELISA was compared with macroagglutination, microagglutination, and immobilization tests, using rabbit antisperm serum and human sera from vasectomized males. Results of this study indicated that the ELISA was more sensitive, less subjective, and easier to perform than these other commonly used techniques.     

Studies on the immunogenicity of protamines in humans and experimental animals by means of a micro-complement fixation test.

A complement fixation study with human, monkey and rabbit sera, using purified sperm nuclear basic proteins as antigens, led to the following conclusions. (1) Protamines, the sperm-specific basic nuclear proteins, may be immunogenic in mammalians. (2) Antibodies detected in the indirect immunofluorescence test on human swollen sperm heads in sera from infertile and vasectomized men, are directed primarily against human protamines. (3) The results obtained suggested that differences in the immunization site and/or in the configuration of the immunizing protamine, may lead to the formation of antibodies directed against different antigenic determinants. Autoimmunity to protamines, following vasectomy or in infertile men, is accompanied by the formation of antibodies cross-reacting with common antigenic determinants present in protamines of other species. Induction of immunity to protamines by means of immunization with protamines-RNA complexes (in rabbits), or protamine-insulin complexes (in humans), leads to the formation of antibodies reacting more specifically with the immunizing protamine, showing only slight cross-reaction with other protamines. (4) The histone-like fraction present in mature human spermatozoa is composed mainly of histone fraction H2B.