Objective·To detect the differences in m6A methylation modification and gene expression of liver tissue mRNA in high-fat diet-induced mouse models of non-alcoholic fatty liver disease (NAFLD) using microarray technology.Methods·The NAFLD models were established in 6-8 weeks old male C57BL/6J mice fed with high-fat chow for 16 weeks (high-fat group, n=10). The basal group (n=10) was given 10% fat diet. Hematoxylin-eosin (H-E) staining was used to assess the histopathological changes in liver tissue and to determine the success of the NAFLD models. The changes of mRNA m6A methylation and expression levels in the liver tissues of the two groups were detected by using methylated RNA immunoprecipitation (MeRIP) and microarray expression profiling.Results·The livers of the mice in the basal group were bright red with few fat deposits, while the livers of the mice in the high-fat group were yellowish with diffuse infiltration and fusion of lipid droplets in the hepatocytes by H-E staining, suggesting that the high-fat diet-induced NAFLD models were successfully constructed. The results of the MeRIP-microarray showed that the m6A methylation levels of 320 genes in the livers of mice in the high-fat group were significantly altered compared with those in the basal group (P<0.05 and fold change>1.5), of which 108 genes were up-regulated and 212 genes were down-regulated. Genes with significant differences in m6A methylation levels between the two groups were intersected with those with differentially expressed mRNAs, and 163 genes were found to have significant differences in both m6A methylation level and mRNA expression level.Conclusion·The change in m6A modification of liver tissue mRNA in the high-fat diet-induced mouse models of NAFLD is significant and the change is associated with the gene expression of mRNA.

目的:探讨两种不同的高脂饲料在C57/BL6J小鼠肥胖和胰岛素抵抗(IR)模型制备过程中的效果差别,以期为更好的制备C57/BL6J小鼠肥胖和IR模型提供理论指导.方法:以雄性C57/BL6小鼠为实验材料,随机分为三组,分别为基础组,喂食国内A公司啮齿动物基础饲料;模型1组,喂食国内A公司生产的啮齿动物高脂饲料(HFD-A组);模型2组,喂食美国ResearchDiets生产的啮齿动物高脂饲料(HFD-R组).至饲养12周比较组间体重、血脂指标,腹腔注射葡萄糖耐量(IPGTT)和胰岛素耐量(IPITT)等指标的差异.结果:随着饲养时间的延长三组动物体重均逐渐上升,至饲养12周基础组平均体重为(31.78±1.36)g,HFD-A组为(41.56±1.67)g,HFD-R组为(46.38±1.98)g,差异显著(P<0.05);至饲养12周时,HFD-A组和HFD-R组肥胖模型成功率均为100％(12/12),无统计学差异(P>0.05);与基础组比较,HFD-R组葡萄糖(Glu),甘油三酯(TG),胆固醇(CHOL),高密度脂蛋白(HDL),低密度脂蛋白(LDL)和游离脂肪酸(NEFA)等多项指标显著改变,差异有统计学意义(P<0.05),而HFD-A组仅Glu和LDL升高显著(P<0.05).与基础组比较,HFD-R组的IPGTT和IPITT检测结果均显著升高(P<0.05),而HFD-A组虽有升高趋势但是差异无统计学意义(P>0.05).结论:两种高脂饲料均能够在饲养12周时成功制备C57/BL6J小鼠肥胖模型,但是只有HFD-R组达到了IR模型的标准.

目的:探讨肝硬化患者血常规指标变化的临床诊断价值.方法:随机抽取2019年1月～6月我院100例肝硬化患者作为研究对象,行血常规检测,并与100例健康查体者(对照组)进行RBC、 WBC、 PLT、 Hb、红细胞比容(HCT)、平均红细胞体积(MCV)、血小板分布宽度(PDW)比较.结果:检测结果与对照组比较,肝硬化患者RBC、 WBC、 PLT、 Hb、 HCT明显降低,差异有统计学意义(P＜0.01);肝硬化患者MCV、 PDW显著增高,差异有统计学意义(P＜0.05).结论:血常规指标可反映肝硬化的肝损害程度,及时进行血常规检测对肝硬化的早期诊断有重要临床价值,对肝硬化患者病情变化、临床治疗及预后的判断有一定临床意义.

目的:初步探讨肝癌细胞株、 正常肝细胞株中DEC1mRNA的表达水平和DEC1蛋白的表达水平和DEC1蛋白亚细胞定位.为进一步发现DEC1与原发性肝癌发生发展的关系提供依据.方法:选取人肝癌细胞株SMMC-7721,HepG-2,Bel-7402,Hep-3B细胞和人正常肝细胞株HL-7702.用RT-PCR、 流式细胞术和免疫细胞化学方法检测DEC1mRNA及蛋白的表达水平.结果:其中HL-7702细胞中的DEC1 mRNA表达水平高于其他各肝癌细胞株,但差异没有统计学意义(P>0.05).各细胞株中DEC1蛋白在细胞核与细胞浆均表达阳性.但以HL-7702核表达较强.流式细胞术结果显示DEC1在肝癌细胞株Bel-7402中的标记率为56％.结论:DEC1在各株肝细胞中转录水平表达的差异并不明显.各细胞株中DEC1蛋白在细胞核与细胞浆均表达阳性.

目的:DEC1属于bHLH类转录因子家族成员,其在肝细胞中广泛表达.DEC1通过调节肝脏内的新陈代谢、 能量平衡、 生理功能等作用参与肝脏疾病的发生和发展过程.

中暑是指在高温、 高湿的环境中, 体温调节中枢功能障碍、 汗腺功能衰竭、 水电解质紊乱等为特征的一种疾病[1]. 根据其发病机制与临床表现等可将中暑分为三个不同的类型, 即热痉挛、 热衰竭、 热射病,患者可能有一种或多种类型的临床表现, 其中热射病为一种致命性疾病, 具有极高的致死率[2].

目的 信号转导和转录激活蛋白5 (signal transduction and transcriptional activation protein,STAT 5),包括STAT 5a和STAT 5b,被认为在肿瘤的多种生物学特性中具有重要作用.本研究旨在检测胃癌组织STAT 5b的表达,分析STAT 5b在胃癌细胞对吉非替尼化疗敏感性中的作用及可能机制.方法 选取2013-03-01-2013-12-31在山东大学附属千佛山医院住院的69例原发性胃癌患者作为研究对象.免疫组化法检测69例胃癌患者癌和癌旁组织中STAT5b的表达,体外培养胃癌MGC-803和MKN-45细胞,分别暴露在吉非替尼和(或)STAT 5b siRNA中.结果 胃癌组织STAT 5b阳性表达率为71.0％(49/69),胃癌旁组织中阳性表达率为39.1％(27/69),差异有统计学意义,P=0.001.4 mmol/L吉非替尼作用24h后MGC-803和MKN-45的存活率分别为(54.01±6.72)％和(56.13±5.02)％,明显降低且呈现时间和浓度依赖性;细胞凋亡率分别为(5.1±1.1)％和(5.9±1.3)％,与对照组(2.1±1.2)％和(1.6±1.1)％相比显著升高,P＜0.05;STAT 5b的蛋白表达水平显著下调.STAT 5b siRNA协同吉非替尼作用MGC-803和MKN-45后的细胞凋亡率分别为(23.4±3.2)％和(19.7±3.16)％,显著低于STAT 5b siRNA和吉非替尼单独作用组.结论 与癌旁组织比较,STAT 5b在胃癌组织中的表达显著升高,干扰STAT 5b的表达能够增强胃癌细胞对吉非替尼的化疗敏感性,其机制与促进细胞凋亡有关.