Chronic schizophrenia is associated with over-expression of the interleukin-2 receptor gamma gene

Altered immune response, including low-grade inflammatory processes, is involved in the pathogenesis of schizophrenia, a chronic psychiatric disorder with complex etiology. Distinct gene variants of a number of pro-inflammatory and chemotactic cytokines together with their receptors associate with this disorder. Interleukin-2 receptor gamma (IL-2RG) represents an important signaling component of many interleukin receptors and so far, no data on the functional state of this receptor in schizophrenia have been reported. The aim of this study was to investigate mRNA expression of the IL2RG gene (IL2RG) in schizophrenia patients in comparison with healthy subjects (controls). Total RNA was isolated from peripheral blood of 66 schizophrenia patients and 99 healthy subjects of Armenian population. The mRNA expression was determined by quantitative real-time polymerase chain reaction (RT-PCR) using PSMB2 as housekeeping gene. IL2RG mRNA expression was upregulated in peripheral blood of patients in comparison with controls (patients vs. controls, median [interquartile range]: 2.080 [3.428–1.046] vs. 0.324 [0.856–0.000], p<0.0001). In conclusion, our findings suggest that over-expression of the IL2RG gene may be implicated in altered immune response in schizophrenia and contribute to the pathomechanisms of this disorder.     

脑胶质瘤环磷腺苷效应元件结合蛋白的表达

目的探讨环磷腺苷效应元件结合蛋白(c AMP-response element binding protein,CREB)在不同级别脑胶质瘤组织中的表达及其临床意义。方法采用免疫组化法检测CREB在5例颅脑损伤减压术后脑组织切片,55例不同分化程度脑胶质瘤组织切片中的表达,并采用Western blot、RT-PCR检测CREB在10例颅脑损伤减压术后脑组织标本,30例不同分化程度脑胶质瘤组织中的表达。结果免疫组化法测得CREB的阳性表达率分别为对照组2/5,Ⅰ-Ⅱ级胶质瘤10/15,Ⅲ级胶质瘤11/12,Ⅳ级胶质瘤28/28,免疫组化显示数据总体上有统计学差异(H=28.183,P0.05)。RT-PCR法检测2~(-ΔΔCt)值,对照组:1.00±0.00,Ⅰ-Ⅱ级胶质瘤:1.35±0.07,Ⅲ级胶质瘤:2.88±0.11,Ⅳ级胶质瘤:3.75±0.20。脑胶质瘤组织中CREB的表达均高于对照组脑组织,且各级别癌组织间有统计学意义(F=1.208,P0.05)。Western blot法测得对照组:0.311±0.014,Ⅰ-Ⅱ级胶质瘤:0.469±0.026,Ⅲ级胶质瘤:0.641±0.028,Ⅳ级胶质瘤:0.896±0.024,各组间有统计学差异(F=1.123,P0.05)。结论 CREB在不同分化程度脑胶质瘤组织中均存在过表达现象,与肿瘤的分化程度呈正相关,可能在胶质瘤发生发展过程中起着重要的调节作用。     

蛇毒蛋白基因克隆与高效表达的研究进展

蛇毒是从毒蛇的毒腺中分泌出来的毒液,属于生物毒素。其化学成分很复杂,是一类有酶活性和其他生物活性的蛋白质和多肽的混合物．具有广泛的生物学活性．可作为天然的药用资源。目前通过捕蛇或养蛇提取蛇毒存在局限性．所以采用基因工程生产高纯度的蛇毒蛋白成为当前及今后的研究方向。本文就蛇毒蛋白基因克隆与高效表达的研究进展作一综述。     

过表达 mimecan对人脐静脉内皮细胞增殖和凋亡的作用初探

目的 应用反转录病毒载体技术构建稳定过表达mimecan的人脐静脉内皮细胞株（HUVEC）,并观察过表达mime-can后HUVEC增殖及凋亡的变化.方法 应用反转录病毒载体构建稳定过表达mimecan的HUVEC细胞株,real-time PCR和Western blot法检测mRNA和蛋白水平的表达.应用CCK-8方法检测过表达mimecan后HUVEC增殖的变化,应用Annexin V-PE染色观察过表达mimecan后HUVEC凋亡的变化.结果 构建了稳定过表达mimecan的HUVEC细胞株.过表达mimecan后,HUVEC增殖能力降低,凋亡率增加.结论 过表达mimecan抑制HUVEC的增殖,促进其凋亡.     

人重组Sigma-1受体的高表达及大量纯化

目的 研究人Sigma-1受体高表达及大量纯化的方法 .方法 Sigma-1受体与麦芽糖结合蛋白(MBP)融合在大肠杆菌中表达,TritonX-100抽提,蛋白酶酶切,直链淀粉、Ni2 离子亲合层析纯化,通过SDS-PAGE电泳、125I-碘化可卡因光亲合标记、[H3]-( )-镇痛新结合活性检测.结果 表达纯化的Sigma-1受体相对分子质量约为26kDa的多肽,从表达到最后纯化倍数约为700,能够结合Sigma-1受体特异的光和标志物125I-碘化可卡因,也能结合配体[H3]-( )-镇痛新.结论 实现了均一、有活性的Sigma-1受体大量表达和纯化.     

PPO在MAPK通路中一个新癌基因

目的：在1p36区寻找新的癌基因。用于癌症的诊断和治疗。方法：通过克隆筛选与癌症相关的基因。定量PCR检测谊基因在各种肿瘤组织和细胞株中的表达量。利用基因重组的方法构建该基因的表达质粒，并研究该基因在细胞水平和活体水平对细胞增殖度磷酸化功能的影响。结果：在人类第1号染色体上发现了一个新的癌基因PPO。人类多种肿瘤组织中，谊基因均呈高表达，从细胞水平到活体水平均证实，该基因与增殖和磷酸化有关。结论：我们在1p36区克隆出了一个新的基因，谊基因与细胞增殖及磷酸化功能有关。并命名为PPO（Proliferation and Phosphorylation Oncogene）。PPO能使MAPK通路中ERK2和MEK1／2磷酸化．推测PPO可能是在MAPK通路中一个新的癌基因。     

5-HT3 receptor over-expression decreases ethanol self administration in transgenic mice

 The 5-HT₃ receptor is thought to play a role in the reward pathway and the phenomena of drug abuse by modulating dopamine release in the mesolimbic pathway. Studies involving this receptor have been hampered due to the low level of 5-HT₃ receptors in the CNS. A 5-HT₃ receptor over-expressing mouse was produced to study the role of this receptor in the rewarding properties of drugs of abuse. Over-expression was restricted to the forebrain by controlling gene expression with the Ca²⁺ calmodulin (CAM) kinase IIα promoter. No over-expression was detected in other body organs nor the cerebellum, as measured by ligand binding and Northern analysis. 5-HT₃ receptor over-expressing mice drank less alcohol than non-transgenic mice in a two-bottle free choice test. Over-expression of the 5-HT₃ receptor in these mice resulted in a decrease in ethanol consumption. These mice should prove useful in testing hypothesis regarding a common reward pathway for drugs of abuse and the role 5-HT₃ receptors play in this pathway. Received: 17 June 1998 / Final version: 14 August 1998     

白念珠菌ERG11基因突变及高表达对耐药的影响

目的 探讨白念珠菌ERG11基因突变、高表达及二者的相互作用在氟康唑耐药中的作用.方法 采用PCR法扩增实验菌株的ERG11基因,并进行测序及生物信息学分析,筛查突变位点;抽提实验菌株总RNA,并逆转录合成cDNA,采用实时荧光定量PCR（FQ-RT-PCR）法检测ERG11基因的表达水平.结果 24株临床菌株中,共检测出ERG 11基因存在24个同义突变位点和5个错义突变位点,其中,E174A、T123I、V234F为新发位点;白念珠菌ERG11基因在耐药组中的表达量高于敏感组,差异具有统计学意义未发生错义突变的白念珠菌耐药株ERG11基因的表达量高于发生错义突变的,差异具有统计学意义.结论 白念珠菌ERG 11基因的5个错义突变A114V、E174A、T123I、Y132H、V234F可能与氟康唑耐药形成有关;ERG11基因的高表达可能与氟康唑耐药有关;ERG11基因的错义突变可能对该基因的高表达存在抑制作用.     

抗氯霉素单链抗体的高效生产

[目的]建立抗氯霉素单链抗体(scFvCAP)的大肠杆菌高效生产体系。[方法]采用重叠延伸PCR法组装scFvCAP,并通过优化5’端序列实现高效表达,采用β-环糊精人工伴侣系统实现SDS变性蛋白的复性。[结果]成功组装scFvCAP基因(Genebank ID:GU258048)。在强启动子T7驱动下,scFvCAP基因独立表达时未见显著表达,而通过同义突变所得突变基因scFvCAP mut(Genebank ID:GU258048)即可高效表达。高效表达所形成的包涵体蛋白可溶于1%SDS-8mol/L尿素溶液,经β-环糊精人工伴侣系统复性后所获scFvCAP具有与母本单克隆抗体相似的亲和力。[结论]成功建立了基于大肠杆菌基因工程系统的scFvCAP高效生产体系。