Induction of APOBEC3G in human hepatoma cells by interferon-α stimulation

To clarify the role of APOBEC3G (A3G) in cellular defense against hepatitis B virus ( HBV), the expression of A3G in normal human liver and the regulation of the A3G expression in hep-atoma cell line (HuH-7) were investigated. Expression level of APOBEC3s mRNA in human liver was determined by RT-PCR. HuH-7 and HepG2 cells were treated with various concentrations of IFN-α(0 U/ml, 100 U/ml, 500 U/ml, 1000 U/ml)for 12 h. The mRNA levels were measured by a quantitative RT-PCR, the results were normalized relative to the specimens without IFN-αstimulation. Total protein of HuH-7 cells treated with various concentrations of IFN-αfor 48 h was subjected to Western blot analysis. For reporter gene assay, HuH-7 cells were transfected with the reporter plasmids containing IRF-E sites and its mutants with different lengths. Then the cells were treated with or without 1200 U/ml IFN-a for additional 12 h (1000 U/ml) after 24 h of transfection, and the cell lysate was prepared and assayed for luciferase activity. It was found that normal human liver expressed the mRNA of A3G. A3G mRNA expression in HuH-7 and HepG2 cells were up-regulated by IFN-αstimulation in a dose-depen-dent manner. Western blot analysis indicated that A3G protein expression was also enhanced by IFN-αstimulation. Sequence analysis showed the existence of putative sites of IFN regulatory factor element (IRF-E) in 5' region of A3G gene upstream the initiation codon. IFN-αstimulation results in 6- to 8-fold increase in luciferase activity in cells transfected with the plasmid containing IRF-E sites of the 5' upstream sequences, whereas luciferase activity did not change in cells transfected with the plasmid containing mutant IRF-E sites or without IRF-E sites. As a conclusion, A3G are expressed in normal human liver. A3G expression was up-regulated by IFN-αstimulation in hepatoma cells and could be involved in host defense mechanisms against HBV. ERF-E site in 5' region of AP0BEC3G gene upstream the initiation codon plays an important role in this process.     

《中华病理学杂志》五十年发展简史

《中华病理学杂志》自1955年4月28日创刊已50周年。50年来，在党和有关部门的领导下，在读者和作者、全国病理学工作者和海外学者的爱护和大力支持下，经几代编审人员的敬业工作，本刊一直在成长，今日已成为病理学工作者的良朋益友，在传播医学科学信息、反映我国病理学科研成果和国际动向、引导和促进我国病理学发展和国际交流等方面都做出了应有的贡献。     

靶向抑制晶状体上皮细胞增殖的载药毫微球的研究

采用碳二亚胺法使抗人晶状体上皮细胞单克隆抗体与聚乳酸载 5 -氟尿嘧啶毫微球偶联 ,制备出抗人晶状体免疫毫微球。以牛血清白蛋白作乳化剂采用复乳法制备聚乳酸载 5 -氟尿嘧啶毫微球 ,使毫微球表面吸附牛血清白蛋白 ,再将抗人晶状体上皮细胞单克隆抗体与聚乳酸载 5 -氟尿嘧啶毫微球偶联。采用扫描电镜和透射电镜观察微球形貌和结构 ,用动态光散射粒径分析仪测载药毫微球粒径。ELISA法检测偶联后的抗体活性 ,MTT法检测免疫毫微球对晶状体上皮细胞的抑制作用 ,间接免疫荧光法检测该免疫毫微球与晶状体上皮细胞的特异性结合能力和细胞内化过程。结果显示载药毫微球表面光滑 ,平均粒径为 191.0± 0 .2 0 2nm ,其载药率为 8.2 % ,药物利用率为2 4 .6 %。免疫载药毫微球中的单克隆抗体HILE6保留达到原免疫活性 84 % ,2 4h对兔晶状体上皮细胞抑制率可达6 8.4 2 %。该免疫载药毫微球能与晶状体上皮细胞特异性结合 ,30min后可吸附到晶状体上皮细胞上 ,在 4h内进入细胞质最后进入细胞核。该实验为特异性抑制晶状体上皮细胞增殖 ,预防白内障术后并发症 -后囊混浊提供了重要的科学依据。     

家兔实验性动脉粥样硬化血管重塑的形态定量分析

目的 :探讨AS血管重塑的几何形态变化规律。方法 :将 40只新西兰雄性家兔随机等分为正常对照组 (C组 )和动脉粥样硬化模型组 (AS组 ) ,分别于实验的第 4、8、12周末取各组家兔的胸主动脉进行定性观察及定量分析。结果 :随时间延长 ,AS组斑块面积逐渐增大 ,斑块检出率逐渐增加。管腔面积在早期斑块形成时并无改变 ,晚期则明显缩小。斑块面积及管腔面积均分别与内弹性膜包围面积 (IELA)、外弹性膜包围面积 (EELA)呈正相关关系 (P <0 .0 5 ) ,IELA和EELA间也呈显著正相关关系 (P <0 .0 1) ,但斑块面积与管腔面积间并无直线相关关系 (P >0 .0 5 )。结论 :家兔AS病变随时间延长逐渐加重 ,且在斑块形成时伴内弹性膜和外弹性膜的同时扩张 ,管腔狭窄与否主要与IELA和EELA相关。IELA和EELA可作为判断管腔狭窄及评价血管重塑的指标。     

粉防己碱预处理对心肌缺血/再灌注损伤大鼠肿瘤坏死因子-α表达及核因子活化的影响

目的探讨粉防己碱预处理对心肌缺血/再灌注过程中大鼠肿瘤坏死因子-α表达及与核因子活化的影响以及相关关系。方法60只SD大鼠随机分为3组:缺血/再灌注损伤组、假手术组、粉防己碱治疗组,缺血/再灌注损伤组结扎大鼠左前降支造成心肌缺血30min后再灌注24h后处死大鼠;假手术组只穿针不结扎,余步骤同缺血/再灌注损伤组;粉防己碱治疗组在缺血前30min腹腔注射粉防己碱,余步骤同缺血/再灌注损伤组。24h后取心肌标本,用酶联免疫吸附法检测心肌组织中肿瘤坏死因子-α表达水平,凝胶电泳迁移率变化分析检测核因子的活性。结果粉防己碱组与缺血再灌注组相比肿瘤坏死因子-α表达水平明显减低(208.40±25.12 VS 306.65±17.78,P<0.01)而与假手术组相比表达明显增强(208.40±25.12 VS 61.45±9.20,P<0.01)。粉防己碱组核因子的活性明显低于缺血再灌注组(29.58±1.56 VS 40.33±4.39,P<0.01)而与假手术组无显著性差异(29.58±1.56 VS 30.09±3.46,P>0.05)。结论粉防己碱预处理可以使心肌缺血/再灌注损伤过程中肿瘤坏死因子-α生成明显减少,核因子的活化受到有效抑制,从而起到减轻心肌缺血/再灌注损伤的作用。     

不同透明质酸的浓度对明胶-透明质酸薄膜性能的影响

为了考察透明质酸的浓度在明胶-透明质酸共混体系中对薄膜的理化性能及生物相容性的影响,本研究将不同浓度的透明质酸溶液与明胶溶液共混,制备了不同透明质酸浓度的明胶一透明质酸薄膜.实验结果表明,透明质酸的加入明显提高了薄膜的亲水性和柔韧性,并延缓了薄膜的体外降解时间.当透明质酸的浓度在0.01%～0.1%之间时,成纤维细胞在薄膜上生长良好.当透明质酸的浓度大于0.1%时,对成纤维细胞的生长增殖有明显的抑制作用.