利用比较基因组杂交技术分析横纹肌肉瘤中染色体基因组的变化特征

目的 探讨横纹肌肉瘤(RMS)基因组DNA的变化特征.方法 在一步法RT-PCR检测所有标本PAX3/PAX7-FKHR mRNA融合基因表达情况的基础上,应用比较基因组杂交技术分析25例原发性RMS患者,其中腺泡状横纹肌肉瘤(ARMS)10例,胚胎性横纹肌肉瘤(ERMS)12例,多形性横纹肌肉瘤(PRMS)3例.根据融合基因表达情况、组织学分型、临床分期、组织学分级、性别和年龄进行分组比较.同时收集RMS细胞株A204(腺泡型)、RD(胚胎型)作为对照.结果 25例RMSCGH分析结果显示:(1)RMS中发生DNA拷贝数扩增最常见的部位是2p和12q,其他依次为6p、9q、10q、1p、2q、6q、8q、15q和18q(30%),发生DNA拷贝数丢失最常见的部位是3p、11P和6p(30%).(2)ARMS扩增最常见的染色体臂是12q、2p、6、2q、4q、10q和15q(30%),缺失最常见的染色体臂是3p、6p、1q、5q(30%);ERMS扩增最常见的染色体臂是7p、9q、2p、18q和1 p、8q(30%),缺失最常见的染色体臂是11p.基因组变化在不同的组织学分类中差异无统计学意义(P0.05).(3)伴有融合基因组扩增最常见的染色体臂是12q、2、6、10q、4q和15q(30%),缺失最常见的染色体臂是3 p、6p、5q(30%);不伴有融合基因组扩增最常见的染色体臂是2p、9q、18q(30%),基因组缺失最常见的染色体臂是11p和14q(30%).12q扩增在这两组间的差异有统计学意义(P<0.05),并多见于伴有融合基因的RMS.(4)在临床分期分组中,9q扩增在Ⅱ期和Ⅲ～Ⅳ期间差异有统计学意义(P<0.05),且多见于Ⅱ期患者.结论 (1)2p、12q、6p、9q、10q、lp、2q、6q、8q、15q、18q扩增及3p、11p、6p缺失可能与RMS发病相关;(2)12q扩增可能与融合基因相关;(3)9q扩增可能与RMS发病早期有关.     

RNAi沉默HMGA1基因对肝癌细胞凋亡和细胞周期的影响

目的:筛选HMGA1基因沉默稳定转染肝癌细胞株,检测沉默后肿瘤细胞凋亡、周期和增殖的变化。方法:靶向HMGA1的RNAi载体转染肝癌细胞,G418筛选得到稳定转染细胞株并鉴定;MTT法和FACS检测细胞增殖、凋亡及细胞周期。结果:成功建立基因沉默HMGA1稳定细胞株;HMGA1基因沉默后,细胞凋亡百分率(29.46±3.04%)明显高于质粒对照组和空白对照组(1.96±0.76%和2.04±0.70%,P<0.01);G0-G1期细胞百分率(75.21±2.35%)明显高于质粒对照组和空白对照组(37.98±3.02%和39.23±3.63%,P<0.01),G2-S期(24.79±2.35%)显著低于质粒对照组和空白对照组(62.03±3.01%和60.77±3.63%,P<0.01);MTT检测显示,HMGA1沉默细胞增殖与质粒对照组和空白对照组相比显著降低(P<0.01或0.05)。结论:基因沉默HMGA1可促进肿瘤细胞凋亡,抑制细胞增殖。     

HLAⅠ、Ⅱ类基因与肾综合征出血热重型与轻型的相关性研究

目的 探讨HLAⅠ、Ⅱ类基因多态性与肾综合征出血热（hemorrhagic fever with renal syndmme，HFRS）患者临床特征的相关性。方法 应用PCR-SSO（polymerase chain reaction using sequencespecific oligonucleotides）基因分型技术对中国北方地区56例汉族HFRS患者HLAⅠ、Ⅱ类基因频率分布进行了检测。结果 HFRS重型组HLA-B35携带率为20％，HLA-B62及HLA-DR11携带率皆为15％．与轻型组比较差异皆具有统计学意义（P〈0．05）。结论 HLAⅠ类基因中HLA-B35、HL4-B62及HLAⅡ类基因中HLA-DR11与中国北方地区汉族人群HFRS患者病情严重性密切相关。     

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。