人正、反义转化生长因子beta1基因真核表达载体的构建

目的 :构建人转化生长因子beta 1(TGFβ1)正、反义基因真核表达载体。方法 :从人胎脑的cDNA文库中扩增出TGFβ1共 12 84bp长的DNA片段后 ,与T载体直接进行高效连接 ,并测序证实无突变。接着利用TGFβ1引物两端所设计的酶切位点将目的片段定向亚克隆到真核表达载体pcDNA3.1( )。构建的正反义表达重组体 (pcDNA3.1 TGFβ1和pcDNA3.1 antiTGFβ1)经限制性内切酶消化证实其中有目的片段完整插入。结果 :本实验成功构建了人正、反义TGFβ1基因真核表达载体。结论 :人正、反义TGFβ1基因真核表达载体的构建 ,为今后研究腹膜纤维化的防治奠定了实验基础。     

Down-regulation amyloid β-protein 42 production by interfering with transcript of presenilin 1 gene with siRNA

AIM:ToinvestigatethepathogenesisofAβ42yieldingandnewdrugtargetsaswellasthepossibilityofRNAinterference(RNAi)techniquefortreatmentofAlzheimerdisease(AD).METHODS:HumanADpresenilin1(PS1)cDNAsequencewasobtainedfromNCBIwebsite.ThethreesitesofRNAiactionandonemissensecontrolsitewereselectedinPS1cDNAthroughonlinedesignofAmbioncompany.Toconfirmspecificityofthesesites,weconductedaBLASTsearchoftheIMAGEESTlibrary.Thecorrespondingdouble-strandedDNAwasusedtoconstructpSilencer3.1-H1p…     

脊髓源星形胶质细胞GFAP表达抑制真核表达载体构建及最佳短发夹样RNA分子筛选

目的构建并筛选大鼠胶质原纤维酸性蛋白（GFAP）表达抑制短发夹样RNA（shRNA）真核表达载体。方法针对GFAP基因全编码序列设计并合成三对9bp茎环结构、19bp干扰序列特异性shRNA模板，体外定向克隆构建特异性重组质粒真核表达载体；通过体外大鼠脊髓源星形胶质细胞GFAP表达抑制模型，脂质体介导RNA干扰分子转染，实时荧光定量RT—PCR及Wesem blot技术观察RNA干扰后原代星形胶质细胞GFAP表达抑制效果．筛选最佳GFAP表达干扰抑制真核表达载体。结果序列测定证实GFAP—shRNA重组质粒真核表达载体构建成功，三对shRNA模板在mRNA及蛋白表达水平抑制靶基因表达效率分别为81％、63％、56％。结论高效率的GFAP—shRNA真核表达载体在大鼠原代星形胶质细胞GFAP表达抑制模型中能高效抑制GFAP基因表达，为后续多靶点RNA干扰技术在脊髓损伤胶质瘢痕抑制基因治疗中的应用奠定了前期基础。     

S_(180)肿瘤核糖核酸体外对小鼠脾淋巴细胞蛋白质合成的影响

用放射性同位素标记氨基酸参入法研究了肿瘤细胞RNA对小鼠脾淋巴细胞蛋白质合成的影响。结果表明,肿瘤RNA在体外可显著抑制脾淋巴细胞的(~3H)亮氨酸参入,这种抑制效应无肿瘤细胞来源特异性,但有组织来源特异性。其抑制效应的活性组分可能存在于RNase酶解后的Sephadex G-100凝胶过滤第二组分中。     

慢性丙型肝炎患者丙型肝炎病毒核酸与IgM抗体关系的研究

本文检测了１４０例丙型肝炎病毒（ＨＣＶ）抗体（抗－ＨＣＶ）阳性的慢性丙型肝炎（ＣＨＣ）的ＨＣＶ核酸（ＨＣＶＲＮＡ）和ＩｇＭ抗体（抗－ＨＣＶＩｇＭ）两项指标。结果表明，原始诊断为不同临床型的肝炎（ＨＣ）患者，８年随访时，ＨＣＶＲＮＡ和抗－ＨＣＶＩｇＭ阳性率分别为８０．７％和９０．７％（ｕ＝２．３９Ｐ＜０．０５）．在原始诊断不同临床型ＨＣ转慢者中，上述两项指标均未发现统计学上的差别（均为Ｐ＞０．０５）。ＨＣＶＲＮＡ与抗－ＨＣＶＩｇＭ配对比较，符合率为７８．６％。基因分型初步结果表明，河北省固安ＨＣＶ以基因Ⅱ型为主。本研究提示，随访８年的ＣＨＣ患者绝大多数仍有传染性；本文方法检测的抗－ＨＣＶＩｇＭ不能代表早期感染标志，但代表慢性感染活动化或带毒，所以，在不具备检测ＨＣＶＲＮＡ的地方更具有实用价值．     

Astrocyte RNA in relation to neuronal RNA depletion in Alzheimer's disease

Summary A new double-staining procedure, in which the techniques of immunocytochemistry of glial fibrillary acidic protein (GFAP) and quantitative microdensitometry of azure B-RNA were combined, was used to study nucleic acid alterations in fibrous astrocytes in Alzheimer's disease (AD). RNA contents of GFAP-positive cells of the hippocampal endplate (Rose's H₃-H₅ fields) and the dentate gyrus molecular layer were determined in ten autopsy-proven AD patients (ages 51–88) and ten age-matched, non-demented control. In addition, RNA contents of pyramidal neurons of the endplate were examined. While there were no differences in RNA contents of astrocytes of either region between AD patients and controls, neuronal RNA was markedly depleted. These data suggest that astrocytes maintain protein synthetic capabilities in AD and that RNA loss is limited to the neuronal compartment.Supported by Grants 1P01-AG05119 and 1P50-AG05144 from the National Institutes of Health and by a Small Research Project Award from the University of Kentucky Medical Center     

丙型肝炎患者外周血单核细胞中丙型肝炎病毒复制的研究

９例临床诊断为丙型肝炎患者，研究其外周血单核细胞中ＨＣＶＲＮＡ的存在及复制。９例患者血清标本抗－ＨＣＶ及ＨＣＶＲＮＡ均为阳性，采用高敏感的逆转录一套式ＰＣＲ法测定其外周血单核细胞中ＨＣＶ正、负链ＲＮＡ，结果９例患者外周血单核细胞中７例ＨＣＶ正链ＲＮＡ阳性，３例ＨＣＶ负链ＲＮＡ阳性，证实部分丙肝患者外周血单核细胞中存在ＨＣＶ的复制，表明肝细胞并非为ＨＣＶ感染与复制的唯一场所。     

Expression of IGF-I and IGF-II mRNA in breast tissue

Summary Expression of IGF-I and IGF-II was studied in human breast cancer tissues by in situ hybridization. IGF-I mRNA was detected only in stromal cells adjacent to normal breast epithelial cells. Stromal cells associated with the tumor cells did not contain IGF-I, nor did malignant or benign breast epithelial cells. In contrast, IGF-II mRNA was found in both the malignant epithelial cells and their adjacent stromal cells. These data imply that stromal cells associated with breast epithelium may switch expression from IGF-I to IGF-II during breast cancer evolution. This appearance of IGF-II expression may identify cancer-associated stromal cells that have a fetal phenotype.     

Robust fixed‐order H2 controller for micro air vehicle—design and validation

Design and real‐time validation of a robust fixed‐order H₂ optimal controller for micro aerial vehicle, named Sarika‐2, is presented. Strengthened discrete optimal projection equations, which approximate the first‐order necessary optimality condition, are used for the controller design. Effect of low‐frequency gust disturbance and high‐frequency sensor noise is alleviated through the output sensitivity and control sensitivity minimization. The novelty of this paper is that a single robust H₂ controller, which is designed at the central operating point, ensures simultaneous stabilization of the radio‐controlled aircraft over the entire cruise speed range of 16–26 m/s. The controller is implemented on a digital‐signal‐processor‐based flight computer, and subsequently, it is validated through the real‐time hardware‐in‐loop‐simulation. The responses obtained from the hardware‐in‐loop‐simulation compares well with those obtained from the off‐line simulation. Copyright © 2006 John Wiley & Sons, Ltd.     

2型糖尿病(消渴病)微观辨证的临床研究

目的　为糖尿病的临床辨证、治疗用药提供客观依据和帮助。方法　观察 2型糖尿病 (消渴病 ,diabetesmellitus)不同证型患者各 30例 ,正常对照组 30例 ,检测血清一氧化氮 (NO)、血浆α -颗粒膜蛋白 (GMP 14 0 )、血浆D -二聚体 (D D)、血清白细胞介素 - 6 (IL 6 )这些微观指标的变化 ,探讨其与中医辨证分型的关系。结果　①血清NO值在各组中的变化是阴虚热盛组 >正常对照组>气阴两虚组 >气滞血瘀组 ,每两组间具有显著性差异 (P <0 0 5 ) ,其中气滞血瘀组与其它 3组具有非常显著性差异 (P <0 0 1)。②糖尿病各组患者血浆GMP 14 0、D D值均较正常对照组有非常显著性增高 (P <0 0 1) ,在糖尿病组各证型中 ,呈阴虚热盛组 <气阴两虚组 <气滞血瘀组变化趋势 ,且血浆GMP 14 0和D D的变化呈正相关 (r=0 6 6 )。③血清IL 6在糖尿病组较正常对照组升高 ,其变化呈阴虚热盛组 <气阴两虚组 <气滞血瘀组趋势 ,每两组间具有非常显著性差异 (P <0 0 1)。结论　糖尿病发病以正气不足为内在依据。随着病程的进展 ,脾肾亏虚及血瘀加重是必然趋势。治疗应重视补脾肾和活血化瘀