早期人胎儿睾丸组织异种异体生长发育的研究

目的 采用免疫缺陷小鼠为受体的睾丸组织移植技术,探索小于3月龄的人类胎儿未成熟睾丸组织在异种异位生长发育的可行性,为睾丸移植技术用于人类生精机制研究提供更为易行的供体来源。 方法 将9例10～13周流产胎儿的睾丸组织分别移植到6只雄性去势免疫缺陷小鼠背部皮下,根据移植物的生长状态以及受体的健康状况于不同时间取出移植物。通过对睾丸组织移植前后的组织学观察,对人类未成熟睾丸组织在异种异位的生长发育情况进行分析。 结果 3个月左右的胎儿睾丸约为5～7mg,移植后取出的最大移植物重量为84.1mg。9例未成熟睾丸组织移植后观察到有6例在受体中继续生长发育,其中的生精小管直径由移植时的(44.26±3.14)μm发育成为(77.69±7.47)μm。小管中无规则分布的原始生殖细胞和原始支持细胞开始向基底膜部位迁移,其中部分原始生殖细胞已定位于基底膜处,并具有精原细胞的特征;支持细胞也由幼稚状态发育为具有丰富胞质的成熟型细胞,有序排列在精原细胞周围,形成壁龛样结构。 结论 将较易获得的早期流产胎儿睾丸组织移植到免疫缺陷小鼠体内后,睾丸组织可继续生长发育,因此可作为人类睾丸组织生长发育研究的供体组织。     

细辛对D-半乳糖所致衰老小鼠睾丸的形态学研究

本文研究了细辛对D－半乳糖所致衰老小鼠的抗衰老作用，应用光镜电镜技术测定小鼠睾丸的重量、生精小管直径、生精上皮细胞数、随龄变化的间质细胞数，同时观察了细辛对上述指标的影响。结果 1．随增龄、睾丸重量减轻，生精小管直径缩小。衰老时租精细胞缺如，仅有支持细胞或散在分布，间质细胞随龄递减。2．细辛可以使小鼠的曲细精管增粗，生精过程活跃，生精细胞增多，间质细胞增多。结论 细辛有一定抗衰老作用。     

睾丸动脉血流障碍对小鼠睾丸生精上皮和血-睾屏障的影响

目的:探讨睾丸动脉血流障碍对小鼠睾丸生精上皮和血-睾屏障的影响.方法:成熟雄性BALB/c小鼠,随机分为对照组和6个实验组,对照组仅暴露睾丸动脉而不进行结扎,实验组显微镜下行单侧睾丸动脉结扎, H-E染色观察睾丸组织的病理改变,电镜观察血-睾屏障的变化.结果:对照组睾丸生精小管无明显病理改变;1h组睾丸生精小管上皮细胞排列不均匀,细胞界限清楚,线粒体轻微肿胀,核周隙轻微增宽,睾丸生精小管的基膜完整;2h和3h组生精小管生精细胞层数减少,线粒体轻微空泡改变,线粒体嵴扩张,支持细胞内质网扩张明显,有局部液化灶,生精小管基膜轻微波纹状;睾丸间质可见炎性细胞浸润,多为分叶的中性粒细胞;4h和5h组睾丸间质血管中充血明显,血管内壁有大量的炎性细胞贴壁现象,血管壁增厚呈玻璃样变,精原细胞有核碎裂现象,细胞界限不清,基膜间隙增宽,基膜皱折,精子细胞顶体位置未见顶体,精子细胞局部有空泡,界限不清,细胞膜不完整;6h组睾丸生精小管内主要为精原细胞和初级精母细胞,精原细胞与基膜脱离,基膜有"分层"现象,支持细胞、精母细胞肿胀明显,细胞膜不完整,界限不清,生精小管内可见多核巨细胞.结论:单侧睾丸动脉结扎/再通可引起睾丸生精细胞的缺血性损伤和坏死,生精上皮细胞和血-睾屏障的病理变化随缺血时间的延长而加重.     

增塑剂邻苯二甲酸二丁酯致小鼠睾丸组织的病理学改变

背景：目前国内外研究邻苯二甲酸二丁酯对生殖损害研究对象多为大鼠,且不同时间点下小鼠病理组织学未见,以小鼠为移植对象没有明确移植时间。 目的：探讨邻苯二甲酸二丁酯致小鼠睾丸组织病理学改变,并找出其改变最大的时间点,为移植做准备。 方法：妊娠balb/c小鼠20只随机分成3组,分别为正常对照组6只,玉米油对照组6只,DBP组8只。自妊娠12~21 d,分别经口给予邻苯二甲酸二丁酯和玉米油,分别在小鼠出生后4~8周每间隔1周观察仔代雄小鼠睾丸的组织病理学改变,找到变化最大的时间点。 结果与结论：邻苯二甲酸二丁酯组染毒小鼠睾丸出现明显的生理、病理和电镜下的改变。邻苯二甲酸二丁酯可引起雄性仔鼠性分化异常,睾丸生精上皮损害和生精过程障碍,从而对雄性仔鼠生育力产生不利影响,在5,6周小鼠睾丸组织损害最大,可以作为移植变化最大时间进行选择。     

蒺藜提取物对小鼠损伤睾丸组织结构的影响

目的:观察蒺藜提取物对小鼠损伤睾丸光镜结构和超微结构的影响,探讨蒺藜对小鼠损伤睾丸的保护修复作用.方法: 健康雄性小鼠随机分为对照组、模型组和给药组,再分热处理后6d和14d两个时间组;建立睾丸热损伤模型,给予治疗剂量的蒺藜提取物;光镜标本和透射电镜标本制备,镜下观察.结果: 模型组与对照组相比,生精小管细胞减少,排列紊乱,可见损伤坏死细胞;给药组与模型组相比,生精小管细胞明显增多,排列较整齐,细胞结构基本正常.结论: 蒺藜对小鼠损伤睾丸具有保护修复作用.     

生后发育过程中睾丸神经生长因子基因的表达

为了通过观察小鼠生后发育过程中睾丸组织神经生长因子 ( NGF)基因表达的变化藉以探讨 NGF在睾丸发育、精子发生过程中的生物学作用 ,本研究采用 RT-PCR方法 ,半定量分析生后 1d、1周、2周、4周及 8周小鼠睾丸组织中 NGF m RNA的表达变化 ;以地高辛标记的βNGF c DNA为探针 ,采用原位杂交法观察 NGF m RNA在幼年及成年小鼠睾丸组织中的分布。 RT-PCR结果显示 ,小鼠生后 1d、1周、2周、4周、8周睾丸组织中均有 NGF m RNA的表达 ,以生后 1周时的表达量为最高。原位杂交结果表明 ,幼年鼠 NGF m RNA杂交信号位于睾丸的间质之中 ,而成年鼠 NGF m RNA杂交信号则主要位于睾丸生精小管管壁和管腔中。结果提示 :NGF m RNA在小鼠出生时即有表达 ,其表达量及在睾丸组织中的分布 ,随小鼠的发育而有变化     

β_1整合素在生后不同年龄阶段小鼠睾丸生精细胞分布的研究

目的　观察 β1 整合素在出生后不同年龄阶段小鼠睾丸生精细胞中的免疫组化定位。方法　用免疫组织化学观察 β1 整合素在生后 1d、4d、7d、1 1d、2周、3周、4周、5周、6周和成年昆明小鼠睾丸生精细胞中的分布。结果　出生后 1～ 1 0d的小鼠睾丸生精小管上皮细胞未见 β1 整合素的表达 ;出生后 1 1d到成年小鼠的不同年龄阶段均可检测到 β1 整合素在睾丸生精小管上皮细胞中的表达。在生后 1 1d到 4周小鼠 ,β1整合素主要表达于增殖能力旺盛的精原细胞的胞膜 ,5周后 β1整合素主要表达于精原细胞和拉长期精子细胞头部细胞膜。结论　β1 整合素是生后 1 1d～ 4周小鼠精原细胞的表面标志     

杜仲对D-半乳糖所致衰老小鼠睾丸的形态学研究

目的 本实验以D－半乳糖所致衰老小鼠为研究对象，研究了杜仲的抗衰老作用。方法 应用光镜电镜技术观察小鼠睾丸的重量、生精小管直径、生精上皮细胞数、间质细胞数的随龄变化，同时观察了杜仲对上述指标的影响。结果 1、随增龄，睾丸重量减轻，生精小管直径缩小。衰老时生精细胞缺如，仅剩支持细胞或见散在分布，间质细胞随龄递减。2、杜仲具有一定的抗衰老作用，使生精过程活跃，生精细胞增多，间质细胞增多不明显，生精小管直径改变不显著。结论 杜仲具有一定的抗衰老作用。     

细辛和杜仲合剂对D-半乳糖所致衰老小鼠睾丸的形态学研究

目的 本实验研究了细辛与杜仲合剂对D－半乳糖所致衰老小鼠的抗衰老作用。方法 应用光镜电镜技术测定小鼠睾丸的重量、生精小管直径、生精上皮细胞数、间质细胞数的随龄变化，同时观察了细辛、杜仲合剂对上述指标的影响。结果 1．随增龄，睾丸重量减轻，生精小管直径缩小。衰老时生精细胞缺如，仅剩支持细胞或见散在分布，间质细胞随龄递减。2．细辛与杜仲合剂具有一定的抗衰老作用，可以使小鼠的生精小管增粗，生精过程活跃，生精细胞增多，间质细胞增多。结论 细辛与杜仲合剂具有一定的抗衰老作用。     

不同阶段生精小管组织学结构研究

目的探讨不同阶段人体睾丸生精小管面积、生精小管管腔面积变化和生精上皮在不同阶段的组织学特点,及其与生殖的关系。方法:应用常规组织制片技术和图像分析技术。结果①生精小管平均面积变化,从胚胎睾丸形成到青春期前,随睾丸逐渐发育增大,睾丸间质增多,但生精小管面积无明显增大;自青春期生精小管面积迅速增大,25岁左右达到峰值,45岁左右生精小管平均面积缓慢减少,55岁以后显著减少。②生精小管管腔面积变化,从胚胎睾丸形成到青春期前,生精小管几乎无管腔;青春期管腔开始出现并迅速增大,20岁左右达到峰值;于45岁左右管腔面积缓慢减少,55岁以后显著减少。③生精小管的组织学结构变化,从胚胎睾丸形成到青春期前,生精小管上皮由精原细胞和支持细胞组成,但随睾丸发育增大,睾丸间质增多,生精小管上皮和基膜间渐出现明显的间隙;青春期开始,生精小管上皮发育,生精细胞层数增加,管壁各级生精细胞典型,腔面可见精子;55岁后睾丸纤维化明显,生精小管皱缩,随年龄增长,生精上皮细胞数量渐减少,排列紊乱,基膜增厚。结论①生精细胞增殖旺盛是生精小管平均面积迅速增大的原因之一;生精细胞增殖旺盛的阶段是20～30岁,最佳时期是25岁左右。②生精小管管腔的出现与生精细胞的凋亡、基膜扩大的速率远远大于生精细胞的增殖水平有关,而生精小管管腔的出现有利于精子的生成与运输。③衰老睾丸生殖功能的下降与其组织结构的纤维化及生精小管基膜厚度增加等因素有关。     

饮酒对小鼠睾丸生精小管一氧化氮合酶、增殖细胞核抗原表达及细胞凋亡影响的研究

目的　探讨酒精对小鼠睾丸的组织结构、内皮型一氧化氮合酶 (eNOS)、增殖细胞核抗原 (PCNA)及细胞凋亡的影响。　方法　用 5 %、10 %及 15 % 3种不同浓度的酒精作用于 2 2d龄小鼠 ,取睾丸做石蜡切片、HE染色 ;用免疫组织化学方法检测睾丸eNOS、细胞增殖的变化 ;TUNEL法检测细胞凋亡的变化 ,并进行统计学分析。　结果　随着酒精浓度的增大 ,睾丸组织结构发生明显改变 ,生精小管的直径逐渐减小 ,eNOS阳性细胞面积密度逐渐增大 ,单位面积内PCNA阳性细胞和凋亡细胞数目增加 ,高浓度酒精组与其他组差异极显著 (P <0 0 1)。　结论　酒精可使生精小管的直径变小 ,eNOS及PCNA表达增强 ,凋亡细胞增加并随酒精浓度的增大而变化加重。这可能是过量饮酒导致生精细胞减少 ,生殖能力降低的重要因素。     

Intrathymic Tolerance Induction: Determination of Tolerance to Class II Major Histocompatibility Complex Antigens in Maturing T Lymphocytes by a Bone Marrow-Derived Non-Lymphoid Thymus Cell

Two new experimental approaches were established to analyse the influence of the thymus on tolerance induction to major histocompatibility complex (MHC) antigens: The aim of the first experiment was to perform successful transplantation of adult allogeneic thymus tissue into nude mice, an attempt that has been unsuccessful in the past. Tolerance for the MHC genotype of a prospective thymus graft recipient (A) was induced in mice of strain B by injection of (A X B) splenocytes during the neonatal period. Adult thymic tissue obtained from these allogeneic donors (B) were grafted into the nude mice of strain A. The allogeneic thymus was accepted by the nude mice and immunoreconstitution was achieved. Subsequently the recipients developed tolerance to the MHC antigens of the allogeneic thymus donor as proved by mixed lymphocyte cultures and the acceptance of skin grafts. The second experiment was designed to determine which Ia-positive thymic compartment participates in conferring tolerance to MHC antigens in maturing T lymphocytes. Chimaeric thymus grafts were created by transplantation of neonatal thymus (A) into allogeneic nude mice (B) for a period of 8 weeks. The graft was populated with host bone marrow-derived Ia antigen-positive cells. The chimaeric thymuses consisting of type A epithelium but populated with both type A and B lymphocytes and non-lymphoid cells (i.e. Ia-positive macrophages and dendritic cells), were newly transplanted into nude mice of strain A. The engraftment lead to immunological reconstitution and the nude mice acquired tolerance to the MHC antigens expressed by the allogeneic Ia-positive cells populating the chimaeric graft. Irradiation of the chimaeric thymus prior to transplantation allowed transplantation of chimaeric thymus devoid of living thymocytes but still populated with functionally intact Ia-positive non-lymphoid cells. Transplantation of irradiated chimaeric thymuses resulted in immunoreconstitution and induced exactly the same allotolerance pattern as described above. The results demonstrate that not thymus epithelial cells but a bone-marrow-derived non-lymphoid thymus cell, most likely the Ia-antigen-positive thymic macrophage of dendritic cell, is responsible for the induction of tolerance to MHC antigens in developing T lymphocytes.     

环磷酰胺对大鼠睾丸和附睾的影响

目的探讨抗肿瘤药物所致大鼠睾丸和附睾损害,为寻找男性不育症中药治疗的研究提供理论依据。方法选用16只15周龄SD大鼠随机分为对照组和实验组,每组8只;实验组腹腔注射环磷酰胺20mg·kg-1·d-1,连续5d,用药2个月后,应用HE染色法研究大鼠睾丸、附睾远期组织学变化,用原位缺口末端标记法(TUNEL方法)检测生精细胞凋亡。结果实验组大鼠体重、睾丸和附睾重量均显著减轻(P<0.01),睾丸生精小管直径缩小、间距增宽、生精上皮变薄、生精细胞层次和数量减少、生精小管腔多未见精子形成,实验组睾丸生精小管直径、面积、生精上皮细胞数、均显著低于对照组(P<0.01);实验组生精细胞凋亡增多,与对照组比较,生精细胞显著凋亡(P<0.01);附睾管管腔内腔内精子稀少,含有大量脱落细胞,管壁变薄。结论环磷酰胺对大鼠睾丸、附睾远期损害明显,促进生精细胞凋亡。     

Neonatal exposure to diethylstilbestrol causes granulomatous orchitis via epididymal inflammation

Diethylstilbestrol (DES), an endocrine-disrupting chemical, is an infamous artificial estrogenic compound. Although neonatal exposure to DES has been shown to result in inflammation of the male reproductive system, it has not, to our knowledge, been reported to induce testicular inflammation. Here we report that neonatal exposure to DES caused granulomatous orchitis with spermatogenic disturbance in 4 of 17 ICR male mice at 12 weeks of age. In the animals with spermatogenic disturbance, we observed either seminiferous tubules containing only cells with Sertoli cell features (likely Sertoli cell syndrome), or tubule cells in maturation arrest that contained only spermatogonia and/or spermatocytes. Following neonatal DES exposure, 5-week-old mice exhibited inflammation in cauda epididymis; by 8 weeks, the inflammation had spread to all segments of epididymis but not the testis; by 12 weeks, inflammation of the epididymis was observed in all mice. These data indicated that cauda epididymis has increased sensitivity to neonatal DES exposure compared to other segments of epididymis and testis. The data also implied that neonatal DES exposure-induced inflammation in cauda epididymis extended gradually to the testis via corpus and caput during development.     

Histologic changes in the mouse testis after treatment with gossypol tetra-acetic acid

The effect of oral administrations (20 or 40 mg/kg body weight/day, for 21 days) of gossypol tetraacetic acid on the testis of the Parkes strain mouse was investigated. Gossypol treatment did not affect the body weight or testicular weight, but caused a significant depression in the weight of the seminal vesicle. Histologically, the testes in mice treated with gossypol possessed regressed seminiferous tubules showing the exfoliation of germ cells, the occurrence of giant cells, a disorganization of the germinal epithelium, the degeneration of germinal elements, intraepithelial vacuolation and dislocation of the Sertoli cells into the luminal portion. However, the effect of gossypol was not uniform, and normal features were also observed in the majority of the tubules in the testes of the gossypol-treated mice. When quantitatively analysed, the frequency of regressed seminiferous tubules was significantly higher in the testes in the treated mice than the controls. The results suggest that the gossypol treatment induces non-uniform regressive changes in the seminiferous tubules in the mouse testis.