bcl—2基因在小鼠睾丸及试验隐睾中表达的研究

目的 研究ｂｃｌ－２基因在正常小鼠睾丈中的定位表达及试验隐睾所导致的改变。方法 以Ｗｅｓｔｅｒｎ－ｂｌｏｔｔｉｎｇ印迹法从蛋白水平检测ｂｃｌ－２基因在正常小鼠睾及及试验隐睾中的表达、变化;地高辛标记ｂｃｌ－２基因ｃＤＮＡ探针,石蜡组织切片原位杂交技术,从ｍＲＮＡ水平检测ｂｃｌ－２基因在小鼠精上皮中的定位及试验隐睾所导致的改变。结果 ｂｃｌ－２蛋白在正常小鼠睾丸中有较丰富的表达,试验隐睾导致其表达明显降低;ｂｃｌ－２基因在生精细胞中有较高水平的ｍＲＮＡ转录,表达细胞主要为各级生精细胞,支持细胞和间质细胞未见表达,隐睾术后仍然保持较高水平的ｍＲＮＡ转录。结论 ｂｃｌ－２蛋白可能在生精细胞主动退化的调节中起着重要作用。     

抗凋亡Bcl－2蛋白在人骨髓造血组织中的定位与分布

通过双际记免疫组织化学技术，探讨了抗细胞凋亡的Ｂｃｌ－２蛋白在１２例反应性骨髓组织中的定位与分布。结果显示，ｂｃｌ－２特异性单克隆抗体与荆豆素ＵＥＡ－１标记的骨髓造血细胞彼此分离、互不重叠。提示人骨髓中Ｂｃｌ－２蛋白阳性造血细胞源自粒系或淋巴系，而不是红系或巨核系。     

UV-C诱导的体外凋亡SMC内Bcl-2蛋白表达的研究

目的 观察经ＵＶ－Ｃ照射后的凋亡ＳＭＣ内Ｂｃｌ－２蛋白表达的情况。方法 应用组织贴块法进行体外ＳＭＣ培养，ＳＰ免疫组织化学技术和图像分析技术检测经ＵＶ－Ｃ照射后的ＳＭＣ内Ｂｃｌ－２蛋白的表达。结果 ＵＶ－Ｃ照射体外ＳＭＣ后，细胞出现典型的凋亡形态学和生化指标的改变；凋亡ＳＭＣ胞浆内Ｂｃｌ－２表达下降和出现胞浆内Ｂｃｌ－２的迁移和再分布入核。结论 ＵＶ－Ｃ照射可引起体外ＳＭＣ凋亡，且Ｂｃｌ－２可能参     

大肠腺瘤及癌中细胞凋亡和bcl—2,bax基因的表达

探讨细胞凋亡及其调控基因ｂｃｌ－２和ｂａｘ与大肠癌发生发展之间的关系。方法：采用Ｆｅｕｌｇｅｎ染色，ＴｄＴ酶介导的生物素化ｄＵＴＰ缺口末端标记技术和免疫组织Ｓ－Ｐ法，分别检测大肠正常粘膜、腺瘤、癌及癌旁非腺瘤性不典型增生中细胞凋亡率和ｂｃｌ－２、ｂａｘ基因的表达。     

肝细胞癌和肝硬变中bcl-2蛋白表达与细胞凋亡的关系

目的：研究肝细胞癌和肝硬变中ｂｃｌ－２蛋白表达与细胞凋亡的关系。方法：应用原位脱氧核糖核酸末端转移酶标记法和Ｓ－Ｐ免疫组化技术检测２８例肝硬变和３５例肝细胞癌组织中凋亡细胞的分布、密度及ｂｃｌ－２蛋白的表达。结果：肝细胞癌组织中凋亡细胞密度显著低于肝硬变，且随其恶性程度的增高呈逐渐降低趋势；凋亡细胞在肝硬变中多分布于假小叶周边区域。ｂｃｌ－２蛋白在肝细胞癌组织中的表达强度明显高于肝硬变，但表达的阳性率差异不明显。结论：ｂｃｌ－２通过其表达产物调控肝硬变和肝细胞癌中的细胞凋亡，在肝细胞癌发生中起重要作用，但ｂｃｌ－２蛋白并非细胞凋亡的唯一调控因素     

角化棘皮细胞凋亡和bcl—2,bax表达及其相互关系

目的：探讨角化棘皮瘤（ｋｅｒａｔｏａｃａｎｔｈｏｍａ，ＫＡ）在细胞凋亡方面的生物学特点。方法：用末端脱氧核苷酰转移酶（ＴｄＴ）介导的ｄ－ＵＴＰ生物素缺口末端标记技术（ＴＵＮＥＬ），原位检测２０例角化棘皮瘤中凋亡细胞；应用免疫组织化学技术检测皮损部位ｂａｘ和ｂｃｌ－２基因产物的表达．结果：ＫＡ中８０％（１６／２０）瘤中心出现凋亡细胞。３０％（６／２０）肿瘤边缘ｂｃｌ－２呈弱阳性染色。全部病例ｂａｘ染     

生发中心淋巴细胞凋亡与小结树突细胞关系的研究

研究人淋巴结和扁桃体生发中心内的凋亡细胞与小结树细胞之间的关系。方法用ＴＵＮＥＬ和免疫化学方法分别检测凋亡细胞,ＦＤＣ和Ｂｃｌ－２蛋白的表达。结果在生发中心有散在的凋亡细胞且多被巨噬细胞所吴噬;     

胶质瘤bcd—2基因表达水平与其细胞增殖和凋亡关系的研究

目的 探讨胶质瘤细胞ｂｃｌ－２基因表达水平与肿瘤恶性程度、细胞增殖活性及凋亡程度的关系。方法 以６９例不同级别的人胶质瘤组织为研究对象,用原位杂交及免疫组化染色ＡＢＣ法分别检测ｂｃｌ－２ｍＲＮＡ、ｂｃｌ－２蛋白和增殖细胞核抗原（细胞增殖活性标记物）的表达,并用３’末标记法做原位细胞凋亡检测。结果 ６４例（９２．８％）表达ｂｃｌ－２ｍＲＮＡ,６０例（８７．０％）表达ｂｃｌ－２蛋白,两者的表达水平呈正     

Bcl-2表达与小鼠腹腔巨噬细胞凋亡的关系

利用免疫荧光技术和激光扫描共聚焦显微镜（共焦镜）技术检测了地塞米松介导小鼠腹腔巨噬细胞凋亡时调亡抑制基因Ｂｃｌ－２表达的时空变化。结果显示,免疫荧光技术和共焦镜技术可对细胞内的蛋白表达作定量和定位检测,凋巨噬细胞内Ｂｃｌ－２逐渐减少,核中Ｂｃｌ－２相对量逐渐减少,胞浆中Ｂｃｌ－２相对量逐渐增多,Ｂｃｌ－２表达和地塞米松介导的巨噬细胞凋亡呈极显著负相关。结果表明,地塞米松介导巨噬细胞凋亡时,凋亡抑制     

胶质瘤p53、bcl-2、MDM2表达与细胞增殖和凋亡的关系

目的：研究胶质瘤中ｐ５３ 、ｂｃｌ２ 、ＭＤＭ２ 表达与细胞增殖和凋亡的关系。方法：对４８ 例胶质瘤用免疫组化法检测ｐ５３、ｂｃｌ２、ＭＤＭ２ 蛋白表达;以Ｋｉ６７ 标记指数和ＡｇＮＯＲ 计数检测其增殖活性;用ＴＵＮＥＬ 法检测细胞凋亡。结果：ｐ５３、ｂｃｌ２、ＭＤＭ２ 蛋白表达阳性率分别为４７－９ ％ 、３５－８ ％ 及１２－５ ％ ,细胞增殖活性随肿瘤ｐ５３ 、ｂｃｌ２、ＭＤＭ２ 表达水平增高而增高,细胞凋亡则相反。结论：ｐ５３、ｂｃｌ２、ＭＤＭ２ 蛋白过表达与细胞增殖失控、凋亡抑制关系密切,在胶质瘤恶性进展中起重要作用。     

镉对大鼠前列腺损伤及锌保护的超微结构与硫胺素焦磷酸酶细胞化学的研究

目的　研究镉对大鼠前列腺腹侧叶的超微结构影响及锌保护作用。　方法　小剂量氯化镉 (2 m g/ kg体重 )及锌腹腔内注射后用透射电镜观察及硫胺素焦磷酸酶 (TPPase)电镜细胞化学定量分析。　结果　镉注射后 4h,前列腺上皮主细胞出现轻微的超微结构改变 ,3～ 7d呈明显退变 ,15 d退变减轻 ,出现粗面内质网 (RER)形成的同心圆性小体 ,30～ 6 0 d基本恢复正常。镉注射后 4h,TPPase反应产物较正常组减少 ,3d和 15 d进一步减少。锌保护组较相应的单纯镉组的超微结构损伤轻 ,且 TPPase反应产物也较多。　结论　镉对大鼠前列腺腹侧叶上皮主细胞早期直接损伤较轻 ,所致 TPPase反应产物减少明显。锌对镉所致主细胞超微结构损伤及 TPPase活性降低均有保护作用     

Regional variations in endoplasmic reticulum in the vas deferens of normal and vasectomized rats

The fine structure of the epithelium of different parts of the rat vas deferens was studied in normal rats and at intervals of up to nine months after vasectomy. The cytology of the columnar epithelial cells showed regional variations particularly in the type and extent of endoplasmic reticulum. Cells of the proximal part of the vas deferens were characterized by basal and perinuclear granular endoplasmic reticulum, a large Golgi apparatus, and numerous apical microvilli, vesicles, and vacuoles. In contrast, the columnar cells of the distal portion of the vas deferens contained large amounts of smooth endoplasmic reticulum, much of which assumed the form of large whorls of smooth membranes. In the distal segment the number of microvilli, vesicles, and vacuoles was less but mitochondria were more numerous than in the proximal part. Cells in the middle portion of the vas deferens showed some features of both proximal and distal parts. These structural characteristics, along with previous experiments, suggest that in the proximal portion of the vas deferens absorption of material from the lumen is an important function, while the cells of the distal part may synthesize steroids. Thus, these regional differences in ultrastructure probably reflect regional differences in function along the length of the vas deferens. After vasectomy the regional specializations were retained and did not appear to be altered. The possibility that this structural and functional sequence is significant for normal fertility and its relation to attempts to reconnect the vas deferens after vasectomy are considered.     

Permeability of the diaphragmatic mesothelium: The ultrastructural basis for “stomata”

The ultrastructure of the hamster efferent ducts and epididymis was studied and the results were correlated with previously published data on the composition of luminal fluid obtained by micropuncture. Samples of the efferent ducts and parts of the epididymis designated initial segment, caput, corpus, proximal cauda, distal cauda, and “epididymal vas” were prepared. The efferent ducts contained principal cells characterized by a profusion of apical vesicles and numerous very large vacuoles that were distributed throughout the cytoplasm. Ciliated cells had few vesicles and vacuoles. Occasional cells contained many particles resembling glycogen. In the epididymis, the following trends were observed. The height of the epithelium and the size of the principal cells declined from initial segment to distal cauda. Apical vesicles and vacuoles with a light content were extremely numerous in principal cells of the initial segment and decreased progressively in the more distal regions. In the initial segment, basal and perinuclear rough endoplasmic reticulum was abundant and was distended with a material that resembled newly synthesized protein. Further distally in the epididymis cisternae of the rough endoplasmic reticulum were narrow and contained little intracisternal material. Light cells containing many vesicles, vacuoles, and lysosome-like structures were very prominent in the caudal segments. The epithelium of the epididymal vas had features intermediate between cauda epididymidis and ductus deferens. The cytoplasmic droplet in luminal sperm began to migrate caudally between the caput and corpus epididymidis and reached the posterior extremity of the middle piece in the distal cauda. Some degenerating sperm were observed in the lumen of the distal segments of the epididymis. The abundance of cytoplasmic vesicles and vacuoles in principal cells of the efferent ducts and initial segment of the epididymis correlated with the site of greatest fluid absorption as determined by micropuncture studies, suggesting that these structures are involved in absorption of fluid from the lumen. Between the caput and distal cauda epididymal segments, where absorption of sodium and potassium but not of fluid occurred, there were few vesicles and vacuoles in principal cells, but the “light” cells were large and numerous and contained many vacuoles. The principal cells of the initial segment were best equipped with rough endoplasmic reticulum to synthesize a protein.     

Light microscopical and ultrastructural studies on the vas deferens of the lizard Mabuya carinata

Adult male lizards (Mabuya carinata) were studied during breeding and non breeding seasons to determine the regional and seasonal differences if any in the vas deferens and to compare ultrastructural features of luminal epithelial cells with those of endotherms. The vas deferens of the lizard is a convoluted tube extending from the epididymis to the hemipenis passing over the kidney. Based on morphometric data of luminal diameter and epithelial cell height three distinct regions viz; proximal, middle and distal regions were identified in the vas deferens. The epithelium is surrounded by a thin layer of lamina propria, many layers of circular smooth muscle fibers and an outer layer of visceral pleuro peritoneum. Based on cell and nuclear morphology and ultrastructure, five different cell types viz; principal cell, basal cell, mitochondria rich cell, halo cell and narrow cell were identified in the epithelium during both breeding and non breeding season. Principal cells and basal cells were more abundant in both seasons. The types of luminal epithelial cells of vas deferens of M. carinata and their ultrastructural features are similar to those of mammals. Further, vas deferens of M. carinata differs from mammals in having only circular smooth muscles in contrast to circular and longitudinal muscles of mammalian vas deferens. To the best of our knowledge this is the first report describing cell types of vas deferens, their ultrastructure and ultrastructural seasonal variations in reptiles.     

镉对大鼠附睾的损伤及锌和维生素E的保护作用

用透射电镜结合脂质过氧化物（ＬＰＯ）含量分析法，观察小剂量氯化镉（２ｍｇ／ｋｇ体重）腹腔内注射后１０ｍｉｎ至６０ｄ大鼠附睾头部的损伤改变及锌和维生素Ｅ（ＶＥ）的保护作用。结果表明：注射镉后３０ｍｉｎ，主细胞已出现超微结构改变；３￣７ｄ时主细胞和管周组织及毛细血管退变加重；１５￣３０ｄ时退变有所减轻；６０ｄ时主细胞有恢复趋势而管周组织增厚。镉注射后附睾管周组织内碱性磷酸酶（ＡＬＰａｓｅ）反应产物明显     

Contribution of amniotic membrane to the healing of iatrogenic vas deferens injury

Background/aim Iatrogenic vas deferens injury is one of the most serious complications of operations in the inguinal region. Vasovasostomy is performed as treatment. However, stenosis is common after vasovasostomy. Oligospermia or azoospermia may develop and result in infertility. This study aimed to investigate the effect of amniotic membrane on healing in vas deferens injuries.Materials and methods Four groups consisting of 10 rats each were formed. No procedure was performed in Group-I. In Group-II, the left vas deferens was transected and left to spontaneous healing. In Group-III, the left vas deferens was transected, and end-to-end anastomosis was performed. In Group-IV, the left vas deferens was transected, end-to-end anastomosis was performed, and it was closed with a wrapping of amniotic membrane on the anastomosis line. Rats were sacrificed after 60 days, and each left vas deferens was evaluated. Lumen patency was checked by passing methylene blue through the vas deferens. Subsequently, the vas deferens was evaluated both macroscopically and histopathologically. Data were evaluated using SPSS version 21.0. p < 0.05 was considered statistically significant for all variables.Results The anastomosis lines in Group-IV healed better than those in Group-III, and less stenosis was observed. There were differences between the groups in terms of luminal patency (p = 0.009), adhesions to surrounding tissues (p = 0.02) and separation of the ends of the vas deferens (p = 0.03).ConclusionWe observed improvement on luminal patency and histology of rat vas deferens injury after surrounding human amniotic membrane on the transected and repaired surface. Further studies are needed to apply this promising result on human beings.     

Sympathetic denervation of the smooth muscle of the vas deferens

1. The post-ganglionic nerve fibres to the vas deferens of the guinea-pig and rat were interrupted in vivo by stripping one vas deferens of its serous coat; the other vas deferens was left intact as a control.2. Four to eight days later the stripped vas deferens did not contract in response to electrical transmural stimulation in vitro at 0.1 msec pulse duration. Pulses of 1.0 msec duration produced small contractions which were not abolished by local anaesthetic or adrenergic neurone-blocking drugs.3. Log dose-response curves to noradrenaline were, for stripped vasa deferentia, to the left of those for control vasa. The increase in sensitivity to noradrenaline at 8 days was about sixteenfold for rat vasa and about tenfold for guinea-pig vasa. Tyramine did not contract stripped vasa from guinea-pigs or rats.4. The noradrenaline and adrenaline content of guinea-pig and rat vasa was greatly reduced or abolished 8 days after the stripping operation.5. Fluorescent nerve terminals were usually absent when transverse sections of stripped vasa were examined by fluorescence microscopy after treatment by the formaldehyde condensation method for demonstrating catecholamines.6. It is concluded that post-ganglionic sympathetic denervation is achieved by stripping the vas deferens in vivo of its serous coat and mesenteric attachments.     

Morphology of the vas deferens in an ethanol-drinking strain of rats (UChA and UChB)

Chronic alcoholism alters reproduction and therefore may be responsible for alterations of vas deferens, which are the subject of this analysis in UCh ethanol-drinking rats. The proximal and distal segments of the vas deferens of 20 animals were submitted to macroscopic, light microscopy, electron microscopy and morphometric analysis. The UCh rats showed atrophy of the epithelium of the vas deferens and alterations of the hypothalamus-pituitary axis. Ethanol induces changes in the epithelium of the vas deferens and hypothalamus-pituitary axis of UCh rats.