汉坦病毒感染细胞整合素受体信号转导相关差异表达基因筛选

通过基因芯片技术观察汉坦病毒感染细胞后整合素受体信号转导相关基因的差异表达,以探讨汉坦病毒进入细胞的分子机制。用汉滩病毒76—118株感染原代人胚肺成纤维细胞,同时设立正常对照细胞,于感染后第6h、24h和96h同时抽提感染组与正常对照组细胞总RNA,利用荧光标记dUTP逆转录制备cDNA探针,与人类受体及信号转导类表达谱芯片杂交,并对Cy3、Cy5荧光信号做扫描分析。结果显示有37项基因出现差异表达,其中31项基因在感染后6h出现差异表达,13项下调、18项上调;但在感染后24h和96h分别只有5项和4项基因出现表达差异;而且整合素信号转导通路中的重要基因如一些蛋白激酶相关基因在感染6h表达增强,丝化增强子1在感染6h和96h表达下调,微管相关蛋白1A在感染6h表达上调、感染24h和96h表达下调。这些进一步说明汉坦病毒感染细胞与整合素有关。     

Ultrasonication of insulin-loaded microgel particles produced by internal gelation: Impact on particle's size and insulin bioactivity

Alginate-dextran sulfate (ADS) microgel has been used to protect insulin from gastrointestinal attack and as a carrier to promote insulin permeation through intestinal epithelium. The throughput of ADS submicron particles generation by emulsification/internal gelation is limited by its wide size distribution.     

Site du Paléolithique inférieur de Bogatyri/Sinyaya Balka dans la péninsule de Taman,Kraï de Krasnodar,Russie

The paper aims to present new multidisciplinary results obtained from the study of the Lower Palaeolithic Bogatyri/Sinyaya Balka site (the Taman peninsula), organized after 2010. The particular attention is given to the estimation of the age of the site and to the unique way of adaptation of the Homo erectus to the special environmental conditions of the Taman peninsula.     

Conformation-dependent antibody response to Pseudomonas aeruginosa outer membrane proteins induced by immunization in humans

Outer membrane proteins (OMPs) of pathogenic bacteria have been used as protective antigens in developing bacterial vaccines. In the present study, we compared the antibody responses to a Pseudomonas aeruginosa OMP vaccine elicited in humans and rabbits by immunization. Immunization with the vaccine induced high titers of serum IgG antibody both in rabbits and humans but reactivities of the induced antibodies with the OMPs were different. The rabbit immune sera recognized most of the OMPs in the vaccine both in immunoblot and immunoprecipitation analyses. In contrast, a great variation in band pattern and intensity was observed among the human immune sera in immunoblot analysis, but not in immunoprecipitation analysis. Denaturation of the OMPs did not affect the binding activity of the rabbit immune sera as determined by ELISA, but substantially reduced those of the human immune sera and anti-OMP IgG purified from a pooled normal human plasma. These data suggest that antibody response to P. aeruginosa OMPs elicited by immunization in humans is mainly directed against discontinuous or conformation-dependent epitopes, which should be taken into account in developing vaccines, especially for OMP-derived synthetic peptides.     

Influence of Exon Duplication on Intron and Exon Phase Distribution

Nonrandomness in the intron and exon phase distributions in a sample of 305 human genes has been found and analyzed. It was shown that exon duplications had a significant effect on the exon phase nonrandomness. All of the nonrandomness is probably due to both the processes of exon duplication and shuffling. A quantitative estimation of exon duplications in the human genome and their influence on the intron and exon phase distributions has been analyzed. According to our estimation, the proportion of duplicated exons in the human genome constitutes at least 6% of the total. Generalizing the particular case of exon duplication to the more common event of exon shuffling, we modeled and analyzed the influence of exon shuffling on intron phase distribution. Received: 28 March 1997 / Accepted: 9 July 1997     

The rhetoric of Islamophobia: an analysis of the means of persuasion in Hege Storhaug’s writings on Islam and Muslims

Few actors have had a greater impact on the “framing of Muslims” as a social and political “problem” in Norway since 2001 than Hege Storhaug of the government- and corporate billionaire funded civil society organization Human Rights Service (HRS). Using the methodological tools of the “rhetorical branch” of Critical Discourse Analysis (CDA), and applying the Aristotelian concepts of ethos, logos and pathos, we analyze the bestselling popular title on Islam and Muslims ever published in Norway, namely Storhaug’s self-published 2015 title “Islam – The Eleventh Plague”. We argue that Storhaug’s popular success must be understood in light of her rhetorical appeals to femonationalism, the critique of religion and “Enlightenment” values. We show how she in her writings incites fear of the Muslim “Other” through specific rhetorical devices and a positioning of herself as a defender of the “nation” and the “people” – against national and international “elites”.     

Aortal collagen polymorphism in monkey and man

Aortal collagen typing in monkey and man showed the presence of types I, HI and V in human aorta and types I and III in monkey aorta. Type III collagen was found to be a predominate type in both species. The molecular weight of type III collagen was similar in these species while type I collagen was different. Both monkey and human collagen types I and III were found to be immunogenic. Type I collagen was significantly increased while type III was decreased in human atherosclerotic plaque. Collagen typing in fatty streak remained unaltered.     

内皮细胞特异性启动子pvWF和pVE的克隆及表达

目的:克隆并验证内皮细胞(Endothelial Cells,ECs)特异性启动子,为转染人胚胎干细胞(h ESC)后实时监测ECs的定向分化情况以及利用干细胞实施血友病A的基因治疗研究提供基础。方法:通过酶消化法原代分离人脐静脉内皮细胞(HUVECs),结合RT-PCR和免疫荧光验证分离后的HUVECs表达内皮细胞特异性标志基因血管性血友病因子(v WF)和血管内皮钙粘素(VE-cadherin/CDH5)。抽提HUVECs的g DNA,通过PCR扩增内皮细胞特异性表达基因v WF和VE-cadherin转录起始位点上游不同大小的启动子片段,将其取代报告基因载体p EGFP-N1中的广谱启动子CMV,构建4个质粒,即pv WF-1、pv WF-2、p VE-1、p VE-2,分别转染HUVECs和h ESCs,48 h后观察并比较各启动子片段启动绿色荧光蛋白GFP表达情况,筛选最具特异性及转录活性的启动子片段。结果:通过酶消化法,本研究成功分离出具有典型上皮样细胞的HUVECs。RT-PCR和免疫荧光结果表明HUVECs特异性表达v WF和VE-cadherin。酶切及测序证实所构建的4个含ECs特异性启动子片段的质粒与理论序列相符,通过核转染至HUVECs及h ESCs后,48 h后观察到所克隆的VE-cadherin 2105bp启动子片段具有内皮细胞表达的特异性和较强的转录活性。结论:本研究成功筛选出具有内皮细胞表达特异性及较强转录活性的启动子片段。     

A Mass Spectrometric Study for Comparative Analysis and Evaluation of Metabolite Recovery from Plasma by Various Solvent Systems

A solvent system that extracts a maximum number of metabolites belonging to diverse chemical classes from complex biofluids, such as plasma, may offer useful inputs to understand the metabolic and physiological state of an individual. The present study compared seven solvent systems for extraction of metabolites from plasma. The extracts were analyzed by mass spectrometry (MS) and MS/MS (MS2) using a quadrupole time-of-flight liquid chromatography/MS system in positive and negative modes of ionization. Metabolites with molecular mass below 400 were identified using Human Metabolome Database MS2 and MS search interfaces. The acetone/isopropanol (2:1) system yielded promising results in positive ionization mode, as the maximum number of MS and MS2 features was detected in the extract. It was found to be superior in extraction of various classes of metabolites, especially organic acids, nucleosides and nucleoside derivatives, and heterocyclic molecules. Glycerophosphocholines in the mass range of 400–700 were found to be efficiently extracted by the methanol/chloroform/water (8:1:1) system. In negative mode as well, the maximum number of MS2 features was detected in methanol/chloroform/water and acetone/isopropanol extracts. The fingerprints of molecular features obtained in the negative and positive modes differed from each other to a significant extent.     

Evidence for Feedback Regulation of Glutamate Decarboxylase by γ-Aminobutyric Acid

Abstract: The concentration of γ-aminobutyric acid (GABA) in the human ovary and the capacity of a membrane preparation from the same organ to bind [³H]GABA specifically were examined. The GABA concentration in the ovary was found to be 214 ± 66 nmol/g frozen tissue (mean ± SEM of six independent determinations). Moreover, a single population of high-affinity GABA binding sites has been identified in the ovarian membranes. The apparent dissociation constant ( K _d) and maximum binding capacity ( B _max) were 38.3 n M and 676 fmol/mg protein, respectively. The specific binding of [³H]GABA was displaced by muscimol, unlabelled GABA, or (+)bicuculline, but was unaffected by (±)baclofen and picrotoxin. The present results show that GABA and an extremely high density of GABA_A receptor binding sites are present in the human ovary, indicating a physiological significance of this amino acid in the female reproductive system.