用细胞原位杂交方法检测NPY mRNA在PC12细胞中的表达

目的：建立ＮＰＹ ｍＲＮＡ物细胞原位杂交检测方法,并以ＮＧＦ为诱导因子研究Ｄｅｘ对大鼠嗜铬细胞瘤ＰＣ１２细胞中ＮＰＹｍＲＮＡ表达的影响。方法：采用细胞原位杂交方法。方法：ＮＧＦ具有诱导ＮＰＹ基因表达的生物学效应且呈剂量效应关系;Ｄｅｘ对ＮＰｘＲＮＡ表达具有双相调节效应,即早期（〈８ｈ）可促进ＮＧＦ的诱导作用,而晚期（〉１６ｈ）则具有抑制作用（Ｐ〈０．０１）,但单独Ｄｅｘ对ＮＰＹｍＲＮＡ表达无显著影     

Localization of parvalbumin mRNA in rat brain by in situ hybridization histochemistry

Summary Parvalbumin mRNA was localized in rat brain by in situ hybridization using a ³⁵S labelled rat parvalbumin cDNA and a synthetic oligodeoxyribonucleotide (corresponding to base sequences 140 to 183 of rat parvalbumin cDNA). Strongest hybridization signals were detected in the Purkinje cells of the cerebellum and in neurones of the reticular nucleus of the thalamus. Signal was also detected in the cerebral cortex, hippocampus, basal ganglia and brain stem in agreement with the distribution of parvalbumin immunoreactivity.     

52 例病毒性肝炎患者肝组织中丙型肝炎病毒 RNA 和 HCAg-NS3 的测定

应用地高辛标记探针原位杂交法和单克隆抗ＨＣＶ－ＮＳ３－ＨＲＰ建立直接酶标免疫组化法分别测定５２例肝炎患者肝组织ＨＣＶＲＮＡ和ＨＣＡｇ－ＮＳ３。结果抗ＨＣＶ阳性组ＨＣＶＲＮＡ检出率５７．１％（１６／２８），ＨＣＡｇ－ＮＳ３检出率５３．６％（１５／２８）；抗ＨＣＶ阴性组其两项检出率均为１２．５％（３／２４）。肝组织中ＨＣＶＲＮＡ阳性物呈蓝紫色细小颗粒存在于肝细胞核或胞浆内，其在肝小叶中的分布可分为３型，即弥漫型、局灶型、散在型。肝组织中ＨＣＡｇ－ＮＳ３阳性物呈棕黄色细小颗粒分布于肝细胞核或胞浆内，以单个或数个阳性细胞散布于肝小叶中。２３例ＨＣＶＲＮＡ或／和ＨＣＡｇ－ＮＳ３阳性病例以肝炎后肝硬化（ＬＣ）病例占多数（１４／２３），其次为慢性重型肝炎（ＣＳＨ）和中度慢性肝炎（ＣＡＨ）。此两种检测方法具有较高符合率（９０．４％，４７／５２），表明病毒核酸及其表达产物均存在于肝细胞内，与ＨＣＶ感染密切相关。这为ＨＣＶ感染诊断提供了直接依据，有利于研究ＨＣＶ感染中病毒复制、慢性化进程、抗病毒治疗监测及重叠感染时病毒相互关系。     

Cell kinetic study of the endometrium by nonisotopic in situ hybridization for histone H3 messenger RNA and immunohistochemistry for Ki‐67 and for estrogen and progesterone receptors

We investigated the cell kinetics of the endometrium in hysterectomy specimens taken for leiomyoma from 22 women with regular ovulatory menstrual cycles. Formalin‐fixed, paraffin‐embedded tissue sections were examined for proliferating activity using histone H3 messenger RNA in situ hybridization (H3 mRNA‐ISH) and immunostaining for the Ki‐67 antigen. The relationship of the proliferative activity of endometrial cells to the immunohistochemical expression of the estrogen receptor (ER) and the progesterone receptor (PR) was also examined. During the menstrual cycle, H3 mRNA expression was observed in both the epithelial cells and the stromal cells of the endometrium. In the functional layer, the labeling indices for H3 mRNA (H3 mRNA‐LIs) in the epithelial cells peaked in the late proliferative phase, decreased sharply in the early secretory phase, and remained unchanged thereafter. On the other hand, H3 mRNA‐LIs of stromal cells displayed two peaks: one in the midproliferative phase and the other in the late secretory phase, the former peak being the greater. In the basal layer, epithelial cells and stromal cells showed low H3 mRNA‐LIs and no significant variation throughout the menstrual cycle. The H3 mRNA‐LIs correlated well with the Ki‐67‐LIs and were lower than the corresponding Ki‐67‐LIs. The regression coefficient (H3 mRNA‐LIs against the Ki‐67‐LIs) was 0.33 for epithelial cells and 0.49 for stromal cells, suggesting that the cell cycle time was longer for epithelial cells than for stromal cells. The proliferative activity of endometrial cells showed close relationships with the expressions of ER and PR in the endometrium. When used in combination with other proliferative markers in paraffin‐embedded tissue sections, H3 mRNA‐ISH could open broader perspectives on the cell kinetics of the endometrium. Anat Rec 266:234–240, 2002. © 2002 Wiley‐Liss, Inc.     

锰对大鼠生精细胞Caspase-3 mRNA调控及支持细胞波形蛋白表达的影响

目的 探讨锰对大鼠生精细胞Caspase-3 mRNA调控及支持细胞波形蛋白（vimentin）表达的影响。 方法 将正常雄性SD大鼠48只,随机分为1个对照组和2个染锰组,给予各组大鼠腹腔注射生理盐水和15mg/kg、30mg/kg氯化锰。分别染锰4周和6周,随机处死各组大鼠8只。应用末端脱氧核糖核酸转移酶介导的dUTP缺口末端标记技术（TUNEL） 、原位杂交法和免疫组织化学链酶亲和素生物素过氧化物酶复合物（SABC）法进行检测研究。 结果 1. 与空白对照组比较,15mg/kg、30mg/kg染锰组生精细胞凋亡指数（AI）均升高（ P<0.05, P<0.01）,Caspase-3 mRNA阳性细胞率均显著升高（ P<0.01）,支持细胞波形蛋白（vimentin）阳性细胞率均显著降低（ P<0.01）。2. 染锰剂量相同,染锰6周组与染锰4周组比较,生精细胞AI与Caspase-3 mRNA阳性细胞率均显著升高（ P<0.01）,支持细胞vimentin阳性细胞率均显著降低（ P<0.01）。3. 染锰时间相同,30mg/kg MnCl2组与15mg/kg MnCl2组比较,生精细胞AI与Caspase-3 mRNA阳性细胞率均显著升高（ P<0.01）,支持细胞vimentin阳性细胞率均显著降低（ P<0.01）。4. 各组大鼠生精细胞AI和Caspase-3 mRNA阳性细胞率呈正相关（ r =0.842, P<0.01）,与支持细胞vimentin阳性细胞率呈负相关（ r =-0.859, P<0.01）,各组大鼠生精细胞Caspase-3 mRNA阳性细胞率和支持细胞vimentin阳性细胞率呈负相关（ r =-0.975, P<0.01）。 结论 染锰大鼠生精细胞Caspase-3 mRNA表达增加导致生精细胞凋亡增加;支持细胞vimentin表达下降;这可能是锰生殖毒性的重要分子机制之一。     

The mRNA expression of AMPA type glutamate receptors in the primary motor cortex of patients with amyotrophic lateral sclerosis: an in situ hybridization study

The pathogenetic mechanisms leading to progressive neurodegeneration in amyotrophic lateral sclerosis (ALS) have not been fully elucidated. One possible factor responsible for the selective motor neuron loss in the motor cortex, brain stem and spinal cord is glutamate-induced excitotoxicity particularly mediated via alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) type glutamate receptors. Data about the expression pattern of AMPA receptors in the primary motor cortex are lacking so far. The pharmacological and physiological properties of AMPA receptors are defined by the heteromeric composition of the four different receptor subunits. Different expression patterns of these subunits at motor neurons may provide a molecular basis for increased vulnerability to excitotoxic damage. Using in situ hybridization histochemistry we did not detect any significant differences in the distribution of AMPA receptor mRNA in the motor cortex of ALS patients compared to controls.