目的 探究长链非编码RNA SNHG12在乙肝病毒X蛋白(HBx)诱导肝癌发生过程中的作用。方法 把课题组构建的肝前体细胞14-19、EGFP-14-19、HBx-EGFP-14-19通过小鼠肝门静脉注射到体内;采用 qRT-PCR、Western blot 方法检测30 d,90 d,180 d,360 d小鼠肝脏组织及细胞模型中HBx、SNHG12以及下游调节基因的mRNA和蛋白表达情况;使用si-SNHG12干扰其表达,并通过CCK-8、划痕实验、transwell实验、流式细胞术观察其对体外表型影响;检测SNHG12及下游的 mRNA 和蛋白表达;HE染色观察小鼠肝组织切片。结果 qRT-PCR结果表明SNHG12、Notch1、Hes1在HBx-EGFP-14-19细胞及各时间段的小鼠肝组织中均上调(F=48.808,P<0.000 1;F=13.322,P<0.000 1);Western blot结果显示在HBx过表达细胞及动物模型中,受HBx诱导SNHG12表达升高后Notch1信号通路被激活,促凋亡因子Bax下调,抗凋亡因子Bcl-2上调,细胞周期因子CDK2和Cyclin E上调;抑制SNHG12表达后,qRT-PCR、Western blot实验结果显示SNHG12(t=22.746,P<0.000 1)及其上述基因表达均扭转,CCK-8实验显示细胞增殖受到明显抑制,流式细胞术检测显示细胞凋亡增多,划痕及transwell实验表明细胞迁移及侵袭减弱。结论 HBx通过上调SNHG12诱导Notch1表达,导致细胞增殖、周期和凋亡异常,从而促进肝癌的发生。

Objective In order to explore the role of long non-coding RNA SNHG12 in hepatitis B virus X protein (HBx) induced hepatocarcinogenesis. Methods The liver precursor cells 14-19, EGFP-14-19 and HBx-EGFP-14-19 constructed by the research group was injected into the body through the hepatic portal vein of KM mice; qRT-PCR and Western blot were used to detect the mRNA and protein expression of HBx, SNHG12 and downstream regulatory genes in liver tissues of 30 d, 90 d, 180 d and 360 d mice and cell models. Si-SNHG12 was used to interfere with SNHG12 expression in vitro, the mRNA and protein expression of SNHG12 and downstream genes were detected, and its effect on phenotype in vitro was observed by CCK-8, flow cytometry, scratch test and transwell test. HE staining was used to observe the liver sections of mice. Results qRT-PCR showed that SNHG12, Notch1 and Hes1 were up-regulated in HBx overexpression cells and mouse liver tissue at each time point (F=48.808,P<0.000 1;F=13.322,P<0.000 1); the results of Western blot showed that in HBx over-expressing cells and animal models, the expression of SNHG12 was increased induced by HBx, resulting in the activation of Notch1 signal pathway, the down regulation of pro-apoptotic factor Bax, anti-apoptotic factor Bcl-2, and the up-regulation of cell cycle factors CDK2, cyclin E. After inhibiting the expression of SNHG12, the results of qRT-PCR and Western blot showed that SNHG12 (t=22.746,P<0.000 1) and the above genes were inhibited; CCK-8 experiment showed that cell proliferation was significantly inhibited, flow cytometry showed that cell apoptosis increased, scratch experiment and transwell experiment showed that cell migration and invasion decreased. Conclusions HBx induced Notch1 expression by up regulating SNHG12, resulting in abnormal cell proliferation, cycle and apoptosis, so as to promote the occurrence of liver cancer.

目的 探讨替诺福韦酯单药治疗在慢性乙型肝炎(CHB)后肝硬化失代偿期(DCC)治疗中的长期应用价值。方法 随机将84例CHB后DCC患者分为对照组及观察组,每组42例。对照组接受拉米夫定联合阿德福韦酯治疗,观察组接受替诺福韦酯治疗。对比两组12个月内治疗时间内的死亡率及肝癌发生率,并分析两组肝功能、肝硬化指标及Child-Pugh评分变化趋势,同时对比两组治疗过程中HBeAg转阴率、HBV-DNA转阴率及失代偿好转率。此外,对比两组治疗不良反应的发生率。结果 在12个月的治疗时间内,两组死亡率及肝癌发生率比较,差异无统计学意义(P>0.05)。而两组治疗过程中ALT、AST、HA、LN、PCⅢ及Child-Pugh评分呈降低趋势,ALB呈升高趋势(P<0.05);治疗6个月及12个月时,治疗组ALT、AST、HA、LN、PCⅢ及Child-Pugh评分低于对照组,ALB高于对照组(P<0.05)。而两组12个月治疗完成后,HBeAg转阴率比较差异无统计学意义,但观察组HBV-DNA转阴率高于对照组(P<0.05)。此外,两组治疗不良反应发生率比较差异无统计学意义(P>0.05)。结论 在CHB后DCC的治疗中,替诺福韦酯单药治疗方案具有良好的长期治疗效果。

Objective To evaluate the long-term value of tenofovir disoproxil monotherapy in the decompensated cirrhosis(DCC) after chronic hepatitis B(CHB). Methods Eighty-four patients with DCC after CHB were randomly divided into control group and observation group, 42 cases in each group. The control group received lamivudine combined with adefovir dipivoxil, and the observation group received tenofovir disoproxil. Mortality and incidence of liver cancer within 12 months of treatment between the two groups were compared, and the change trend of liver function, liver fibrosis index and child-pugh score in the two groups were analyzed. At the same time,we compared the conversion rate of HBeAg, HBV-DNA and decompensated positive rate between the two groups. In addition, the incidence of adverse reactions were compared between the two groups. Results Within 12 months of treatment, there were no statistically significant differences in mortality and liver cancer incidence between the two groups(P>0.05). And during the treatment, the ALT, AST, HA, LN, PC Ⅲ and Child-Pugh score showed a decrease trend, ALB showed a increase trend(P<0.05). After 6-month and 12-month treatment, ALT, AST, HA, LN, PC Ⅲ Child-Pugh score of treatment group were lower than that of control group, ALB was higher than that of control group(P<0.05). After 12 months of treatment, the negative conversion rate of HBV-DNA in the observation group was higher than that of control group(P<0.05). In addition, there was no statistically difference in the incidence of adverse reactions between the two groups(P>0.05). Conclusion Tenofovir disoproxil monotherapy has a good long-term therapeutic effect in the treatment of DCC after CHB.

目的 探讨拉米夫定治疗对乙肝活动的肺结核病人抗结核治疗中的临床价值。方法 通过回顾性分析159例初治肺结核乙肝表面抗原(HBsAg)阳性、HBV-DNA定量阳性病人,所有病例在抗结核前查肝功能均正常,分为两组,治疗组:在抗结核及护肝治疗过程中,加用拉米夫定抗乙肝病毒治疗;对照组:没用任何抗乙肝病毒药物;分别在抗结核治疗前、治疗2周、4周及8周复查肝功能,对比两组间肝损发生率,及肝损发生时间及严重程度。抗结核治疗4周后复查HBV-DNA定量,对比两组间HBV-DNA定量下降例数。结果 治疗组的肝损发生率仅20.5%,对照组病人抗结核治疗后肝损的发生率为53.1%,两者间差异有统计学意义。治疗组肝损出现时间多为大于4周,而且多数是轻度肝损。治疗组出现肝损中断抗结核治疗的病例数低于对照组。抗乙肝病毒治疗后复查HBV-DNA定量降低例数高于对照组。结论 拉米夫定抗乙肝病毒治疗,能抑制乙肝病毒复制,降低乙肝合并肺结核病人的肝损发生率并减轻肝损严重程度,提高病人对抗结核药物的耐受性。