重组人白细胞介素10的融合表达及鉴定

目的 研究重组人白细胞介素10(rhIL—10)载体在大肠杆菌B121(DE3)pLyse细胞中的表达，为进一步研究IL-l0在动脉粥样硬化中的作用机制奠定基础。方法 用构建成功的IL-l0—PCRT7/NT-TOPO质粒转化大肠杆菌BL21(DE3)pLyse细胞，并通过SDS-PAGE鉴定融合表达蛋白。结果 PCRT7/NT—TOPO质粒载体成功载入rhIL-l0基因；在异丙基硫代—β-D-半乳糖苷(IPTG)诱导下表达的蛋白质主要以包涵体形式存在。结论 在IPTG诱导下，重组的IL-l0—PCRT7/NT—TOPO质粒载体在大肠杆菌BL2l(DE3)pLyse细胞内成功表达。     

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。     

刺玫果对老龄小鼠衰老指标SOD、MDA的影响作用

目的 :研究刺玫果提取物 (喷雾干燥粉 )对衰老小鼠脑、肝组织中超氧化物歧化酶 (SOD)和丙二醛(MDA)含量的影响。方法 :SOD测定为黄嘌呤氧化酶法 ,MDA测定为硫代巴比妥酸法。结果 :刺玫果提取物可显著增强老龄小鼠脑和肝SOD活性 ,降低MDA含量。结论 :刺玫果提取物具有一定的抗衰老作用     

中国人混合型高脂血症患者脂蛋白脂酶常见基因变异的筛查

目的 研究中国人混合型高脂血症（CHL）患者脂蛋白基因变异情况。方法 应用多聚酶链反应（PCR）对广东地区156例混合型高脂血症患者脂蛋白脂酶基因的多态性进行了研究,检测的LPL基因突变位点有：-93T→Asn及Asn291→Swer突变。结果 156例混合型高脂血症患者均未检出上述突变基因。结论 在欧洲白种人存在的与混合型高脂血症相关联的LPL常见的基因变民可能与中国人混合型高脂血症无关。     

Genotype-phenotype correlations in RHOBTB2-associated neurodevelopmental disorders

PurposeMissense variants clustering in the BTB domain region of RHOBTB2 cause a developmental and epileptic encephalopathy with early-onset seizures and severe intellectual disability.MethodsBy international collaboration, we assembled individuals with pathogenic RHOBTB2 variants and a variable spectrum of neurodevelopmental disorders. By western blotting, we investigated the consequences of missense variants in vitro.ResultsIn accordance with previous observations, de novo heterozygous missense variants in the BTB domain region led to a severe developmental and epileptic encephalopathy in 16 individuals. Now, we also identified de novo missense variants in the GTPase domain in 6 individuals with apparently more variable neurodevelopmental phenotypes with or without epilepsy. In contrast to variants in the BTB domain region, variants in the GTPase domain do not impair proteasomal degradation of RHOBTB2 in vitro, indicating different functional consequences. Furthermore, we observed biallelic splice-site and truncating variants in 9 families with variable neurodevelopmental phenotypes, indicating that complete loss of RHOBTB2 is pathogenic as well.ConclusionBy identifying genotype-phenotype correlations regarding location and consequences of de novo missense variants in RHOBTB2 and by identifying biallelic truncating variants, we further delineate and expand the molecular and clinical spectrum of RHOBTB2-related phenotypes, including both autosomal dominant and recessive neurodevelopmental disorders.     

白藜芦醇通过Fas依赖途径诱导K562细胞凋亡

目的探讨白藜芦醇对K562细胞的凋亡诱导效应和可能的作用机制。方法应用噻唑蓝(MTT)比色法、WrightGiemsa染色、DNA琼脂糖凝胶电泳和细胞周期分析法检测K562细胞凋亡；采用流式细胞术(FCM)测定细胞Fas蛋白表达水平；采用RT-PCR法检测Caspase-3 mRNA表达水平，采用比色法检测Caspase-3活性。结果白藜芦醇可抑制K562细胞增殖，并呈现典型的凋亡形态改变；DNA电泳可见明显的梯状条带；细胞周期分析显示S期细胞数明显增多，出现S／G2期阻滞。40μmol／L白藜芦醇作用48h到72h后，Fas阳性率由4．1％上升到45．6％；Caspase-3 mRNA表达水平明显升高，活性明显增强。结论白藜芦醇通过Fas依赖性Caspase-3激活途径诱导K562细胞凋亡。     

碱性成纤维细胞生长因子对卵巢癌的影响

目的观察Ras-Raf-ERK 1/2途径介导的碱性成纤维细胞生长因子(bFGF)对卵巢癌细胞系CAOV3细胞增殖和凋亡影响,探讨bFGF及其信号转导途径与卵巢癌发生发展关系。方法以bFGF和促细胞分裂剂激活性蛋白激酶1(MEK1)抑制剂PD98059处理CAOV3细胞,用四甲基偶氮噻唑蓝(MTT)法检测细胞增殖情况,流式细胞术检测细胞凋亡情况,蛋白印迹(Western blot)检测细胞外信号调节蛋白激酶1/2(ERK1/2)的活性。结果bFGF处理后细胞增殖比明显增加,且与bFGF水平呈剂量依赖关系,bFGF浓度为75 ng/ml时细胞增殖比最高为140%;bFGF使无血清诱导的凋亡细胞比例下降[(33.20±5.32)%~(2.38±3.36)%];bFGF诱导ERK1/2活性增高。PD98059可抑制bFGF的这些作用。结论bFGF通过Ras-Raf-ERK 1/2途径介导,促进卵巢癌CAOV3细胞增殖,抵抗无血清诱导凋亡。在卵巢癌发生发展过程中,bFGF信号传递发挥了重要作用。     

C-reactive protein does not impair insulin suppression of glucose release in primary hepatocytes

Recent studies have suggested that CRP may interfere with insulin signaling in skeletal muscle and endothelial cells. The aim of this study was to determine whether highly purified CRP increased the rate of glucose appearance in primary hepatocytes in the absence or presence of insulin. Primary rat hepatocytes were provided glucose-free media containing 10 mM lactate, 1 mM pyruvate, 0, 1 or 10 nM insulin, and 0 or 10 μg/ml of purified CRP for 6h. Purified CRP did not increase glucose release in the absence of insulin and did not reduce the ability of insulin to suppress glucose release.