新生期注射谷氨酸单钠对大鼠其脑钙结合素表达的影响

为探讨谷氨酸单钠（monosodium glutamate,MSG）对脑钙结合素（calbindin-28kD,CB）表达的影响，选用新生SD大鼠29只，分为实验组和对照组，实验组设3g/kg和1g/kg2个剂量组，生后第3－9天经腹腔注射MSG，对照组设实验对照（注射生理盐水12mL/kg）和空白对照，满10周龄时4％甲醛灌注固定，取脑做冰冻冠状切片进行钙结合素的免疫组化反应。计算各组大鼠丘脑、下丘脑、海马及大脑基底核算脑区钙结合素阳性神经元数量。发现钙结合素能神经元分布数量具有脑区差异，且新生期注射谷氨酸单钠可使成年后某些核团的钙结合素阳性神经元增多，表明钙纱能神经元可抗谷氨酸神经毒性。     

人心房成纤维细胞体外组织块贴壁培养与鉴定

从建立体外循环将进行心脏手术的风湿性心脏病患者体内无菌下取心脏停跳前右心耳组织，采用组织块贴壁法进行心房成纤维细胞的分离培养，通过光学显微镜观察培养后的细胞形态，并通过免疫细胞化学的方法鉴定所分离培养的细胞。贴壁4—6d后可见长梭形贴壁生长的细胞游离出组织块，细胞生长迅速，2—3d后即呈融合状态，细胞排列紧密，有的交叉重叠生长，胞体较大，细胞浆透明，细胞核较大，呈椭圆形，无自发性搏动。免疫细胞化学检测显示所培养细胞波形蛋白呈阳性反应。通过组织块贴壁法顺利分离人心房成纤维细胞。     

蛋白激酶C单克隆抗体的制备

应用杂交瘤技术制备了蛋白激酶C(PKC)的单克隆抗体,用蛋白A-Sepharose-CL4B协同沉淀复合物分析单抗识别的蛋白,分子质量与PKC相同。采用该抗体对正常和转化的C_3H_(10) T_(1/2)细胞进行免疫荧光观察,发现它们的PKC含量明显不同。但荧光分布都主要集中在细胞质和细胞膜部分。     

Developmental study on the immunocytochemical localization of NOV protein-containing neurons in the rat brain

The localization and development of nephroblastoma overexpressed gene (nov) protein-immunoreactive neurons in the brain of E8-P300 rats have been studied using immunocytochemistry and image analysis. Results are as follows: No NOV protein-immunoreactive cells were detected in the rat brain during prenatal development. A few of positive cells were detected at the early postnatal stage. However, the number and the immunoreactivity of these cells increased gradually at later stages. NOV-immunoreactive cells were widely distributed in the rat brain during P30–P60. The number of immunoreactive cells and their intensity also peaked within this stage. The number and staining intensity of NOV-positive cells decreased gradually with age. The positive cells were mainly located in cingulate cortex, striatum, hippocampus, hypothalamus, cerebellum and brain stem. The present results indicate thatnov may play an important role in the development and differentiation of brain as well as maintaining the function of brain.     

秀丽小杆线虫Caenorhabditis elegans pkc-2基因cDNA的克隆和表达

蛋白激酶C在秀丽小杆线虫中具有调节肌细胞渐进性萎缩的功能.为了揭示它的调节机制, 本研究克隆了秀丽小杆线虫中蛋白激酶C pkc-2基因的cDNA pkc-2-c,构建了含该pkc-2基因cDNA亚型的重组质粒pPD 118.20-pKG 63;揭示了该cDNA在秀丽小杆线虫体壁肌细胞中的定位.     

Synthesis dynamic and developmental profile of prothoracicotropic hormone between diapause- and nondiapause-destined individuals in Helicoverpa arrnigera

Biosynthesis and secretion of prothoracicotropic hormone （PTTH） of diapause- and nondiapausedestined individuals in Helicoverpa armigera were studied using whole-mount immunocytochemistry and enzyme-linked immunosorbent assay （ELISA）. The immunocytochemistry revealed that PTTH is expressed in two pairs of lateral neurosecretory cells of the brain. The presence of immunoreactivity has not significant difference between the brains of the diapause- and nondiapause-destined 6th instar larvae. However, the obvious differences of expressional pattern from day 4 pupae were observed between the two types. PTTH titers in hemolymph from the 6th instar larvae to pharate adults were measured by the ELISA. Although there were similar titer changes between the two types of individuals at the larval stage, a significant difference from developmental expression was detected at the pupal stage, suggesting that the expression and secretion of PTTH does play a crucial role in regulation of pupal diapause of H. armigera.     

杆状病毒表达的hZP3 B细胞表位嵌合肽的免疫活性研究

为研究杆状病毒系统表达的人透明带ZP3的B细胞表位嵌合肽(BCP)的免疫原性及其抗血清与人透明带的免疫交叉反应，重组z3bcp病毒用于感染sf9细胞以表达BCP。Tricine—SDS—PAGE用于分离感染液中的小相对分子质量表达产物，表达产物用于免疫小鼠制备抗血清，免疫细胞化学试验用于检验抗血清与人卵泡透明带的交叉反应性。结果显示，矽细胞被z3bcp病毒感染后，培养液中分离出一条相对分子质量约为6000的特异带，而正常矽细胞中没有等同的带。免疫细胞化学试验结果显示，该特异肽的抗血清能与人透明带特异结合，表明人透明带BCP嵌合肽在杆状病毒系统中成功表达，表达产物具有抗原性，抗血清能与人透明带特异结合。     

茶多酚对肿瘤细胞多药耐药性逆转作用的研究

用免疫组化法和流式细胞仪对肿瘤细胞系MCF－7和MCF－7／Adr的P－糖蛋白表达水平进行定性定量研究。用噻唑蓝比色法（MTT）研究茶多酚的细胞毒性及其对耐药性的逆转作用,并与Pgp抑制剂－奎尼定进行了比较。免疫组化法检测P－糖蛋白表达水平,MCF－7／Adr呈强阳性,而CF－7呈阴性;流式细胞仪定量检测结果MCF－7／Adr细胞系细胞阳性率为15％,MCF－7细胞系细胞阳性率为1.8%。MCF－7／Adr细胞系存在Pgp的过度表达。噻唑蓝比色法（MTT）检测结果茶多酚、奎尼定的加入对阿霉素对MCF－7的细胞毒性几乎没有影响。而茶多酚、奎尼定的加入明显增加了MCF－U／Adr对阿霉素的敏感性。免疫组化与流式细胞技术结合,可用于肿瘤细胞系P－糖蛋白表达水平的定性定量检测。茶多酚不仅具有一定的抗癌活性,还与奎尼定一样具有多药耐药性逆转作用。     

SOME CHARACTERISTICS OF RAT NIGROSTRIATAL DOPAMINERGIC NEUROCONNECTION DURING DEVELOPMENT——A COMBINED IMMUNOCYTOCHEMICAL AND ELECTRON-MICROSCOPIC STUDY

The characteristics of the nigrostriatal dopaminergic neuroconnections in developing rats are studied by combined immunocytochemical and electron-microscopic techniques, with an antibody to tyrosine hydroxylase (TH). From the embryonic day 21 on, some of the THpositive nerve fibers are densely packed in the striatum to form a patch-like dopamine island. The percentage of the TH-positive nerve terminals among the labeled profiles is much higher inside than outside the dopamine island (P<0.01). On the other hand, the TH-positive terminals mainly form symmetrical axon-dendritic synapses, while most of the TH-negative terminals form asymmetrical axon-spinous synapses. The functional significance of the characterized dopaminergic connection is discussed.