Anti-tuberculosis drug-induced liver injury (ADLI) is a serious complication of tuberculosis treatment, yet its molecular mechanisms remain poorly understood. Disruptor of telomeric silencing 1-like (DOT1L) is a unique histone methyltransferase that catalyzes monomethylation (me1), dimethylation (me2) and trimethylation (me3) of histone H3 at lysine 79 (H3K79), despite lacking a SET domain. Beyond its methyltransferase activity, DOT1L participates in various biological processes-including the cell cycle, DNA damage response, and gene transcription-both in a methylation-dependent and -independent manner. Notably, dysregulation of DOT1L has been closely associated with the pathogenesis of liver diseases. In this study, we established an ADLI mouse model using a combination of isoniazid, rifampicin, and pyrazinamide, and observed significant upregulation of DOT1L expression in the liver tissues of ADLI mice. Intervention with the specific DOT1L inhibitor SGC0946 markedly alleviated liver injury. Mechanistically, we found that DOT1L catalyzes H3K79 hypermethylation at the p53 gene promoter, leading to activation of the p53-Bax/Bcl-2 apoptosis signaling pathway and subsequent apoptosis of hepatocytes. In contrast, SGC0946 treatment reduced H3K79 methylation levels at the p53 promoter and suppressed the p53-Bax/Bcl-2 pathway, thereby attenuating liver injury. Our findings highlight the importance of DOT1L expression homeostasis in liver physiology and reveal a novel epigenetic mechanism by which DOT1L participates in ADLI progression through transcriptional regulation of p53. These results position DOT1L as a potential therapeutic target for the prevention or treatment of ADLI.

BackgroundThis study aimed to explore the distinct characteristics of the gut microbiota in tuberculosis (TB) patients who experienced liver injury following anti-TB treatment compared with those who did not.MethodWe employed a nested case-control study design, recruiting newly diagnosed pulmonary TB patients at Tangshan Infectious Disease Hospital. Participants were categorized into the Antituberculosis Drug-Induced Liver Injury (ADLI) group and the Non-ADLI group based on the occurrence of liver injury after treatment. Both groups received identical anti-TB regimens. Stool samples were collected from patients who developed liver injury within 2–3 weeks of starting treatment, alongside matched controls during the same timeframe. The samples underwent 16S rDNA sequencing, and clinical data and blood samples were also collected for further analysis. At the same time, we constructed mouse models to explore the effects of different anti-tuberculosis drugs on gut microbiota.ResultsFollowing anti-TB treatment, we observed a decrease in microbial diversity and significant structural changes in the gut microbiota of TB patients (P < 0.05). At T1, the Non_ADLI_T1 group presented relatively high levels of Phascolarctobacterium, Anaerofustis and Mailhella. In contrast, the ADLI_ T1 group presented elevated levels of Bacteroides, Veillonella, Clavibacter, Corynebacterium, Anaerococcus, Gardnerella, Peptostreptococcus and Lautropia. At T2, the ADLI_T2 group presented increased levels of Enterococcus, Faecalibacterium, unclassified_f__Burkholderiaceae, Cardiobacterium, Ruminococcus_gnavus_group and Tyzzerella_4 than did the Non_ADLI_T2 group. Additionally, the ADLI_T2 group presented decreased levels of Prevotella_9, Akkermansia, Erysipelotrichaceae_UCG-003, Rubrobacter and norank_f__Desulfovibrionaceae than did the Non_ADLI_T2 group. In animal experiments, similar changes to those in the human population were observed in the mouse model compared to the control group. Any single anti-tuberculosis drug or two-drug combination or three-drug combination can cause dysbiosis of the mouse gut microbiota. The signature genera between groups are different and related to the type of anti-tuberculosis drug.ConclusionAnti-tuberculosis treatment induces dysbiosis in the gut microbiota of TB patients. Notably, there are significant differences in microbiota characteristics between TB patients with and without liver injury at both onset and during treatment. There are some differences in the characteristics of bacterial flora in liver injury caused by different drugs.

ABSTRACT This study aims to elucidate additional mutation loci associated with fluoroquinolone (FQ) resistance and evaluate the discriminatory capacity of mutation loci and allele mutation frequencies in identifying FQ-resistant Mycobacterium tuberculosis (MTB) isolates. A random selection of isolates was extracted from an ongoing collection. Drug resistance was determined using the resazurin microtiter assay (REMA) as the gold standard. Mutation loci and the burden of mutations in the quinolone resistance-determining region (QRDR) were elucidated through whole-genome sequencing (WGS). Novel amino acid mutations, namely, G520D and G520T, were identified in the gyrB and associated with FQ resistance. In the context of distinguishing FQ-resistant isolates, the AUC for the QRDR mutation frequency burden (0.969) surpassed that of the mutation locus (0.929), and this difference was statistically significant ( P = 0.03). Furthermore, using the resistance mutation locus as a reference, setting the QRDR mutation frequency burden threshold at 1.31% resulted in a 3.60% increase in the accuracy of classifying FQ-resistant isolates (NRI = 3.60%, P < 0.001). The QRDR mutation frequency burden appears to offer superior diagnostic efficacy in discriminating FQ-resistant isolates compared to qualitative detection of mutant loci. IMPORTANCE Fluoroquinolone (FQ) drugs are recommended as second-line drugs for the treatment of multidrug-resistant tuberculosis. With the massive use of FQ drugs in the clinical treatment of tuberculosis (TB), there is an increasing rate of drug resistance to FQ drugs. In this study, we identified and demonstrated novel amino acid mutations associated with FQ resistance in Mycobacterium tuberculosis (MTB), and we quantified the mutation sites and identified the quinolone resistance-determining region (QRDR) mutation frequency burden as a novel diagnostic method for FQ resistance. We hope that the results of this study will provide data support and a theoretical basis for the rapid diagnosis of FQ-resistant MTB.

The escalating frequency of environmental pollution incidents has raised significant concerns regarding the potential health impacts of pollutant fluctuations. Consequently, a comprehensive study on the role of pollutants in the prevalence of viral hepatitis is indispensable for the advancement of innovative prevention strategies. Monthly incidence rates of viral hepatitis from 2005 to 2020 were sourced from the Chinese Center for Disease Control and Prevention Infectious Disease Surveillance Information System. Pollution data spanning 2014-2020 were obtained from the National Oceanic and Atmospheric Administration (NOAA), encompassing pollutants such as CO, NO2, and O3. Time series analysis models, including seasonal auto-regressive integrated moving average (SARIMA), Holt-Winters model, and Generalized Additive Model (GAM), were employed to explore prediction and synergistic effects related to viral hepatitis. Spearman correlation analysis was utilized to identify pollutants suitable for inclusion in these models. Concurrently, machine learning (ML) algorithms were leveraged to refine the prediction of environmental pollutant levels. Finally, a weighted quantile sum (WQS) regression framework was developed to evaluate the singular and combined impacts of pollutants on viral hepatitis cases across different demographics, age groups, and environmental strata. The incidence of viral hepatitis in Beijing exhibited a declining trend, primarily characterized by HBV and HCV types. In predicting hepatitis prevalence trends, the Holt-Winters additive seasonal model outperformed the SARIMA multiplicative model ((1,1,0) (2,1,0) [12]). In the prediction of environmental pollutants, the SVM model demonstrated superior performance over the GPR model, particularly with Polynomial and Besseldot kernel functions. The combined pollutant risk effect on viral hepatitis was quantified as beta WQS (95% CI) = 0.066 (0.018, 0.114). Among different groups, PM2.5 emerged as the most sensitive risk factor, notably impacting patients with HCV and HEV, as well as individuals aged 35-64. CO predominantly affected HAV patients, showing a risk effect of beta WQS (95% CI) = - 0.0355 (- 0.0695, - 0.0016). Lower levels of PM2.5 and PM10 were associated with heightened risk of viral hepatitis incidence with a lag of five months, whereas elevated levels of PM2.5 (100-120 mu g/m3) and CO correlated with increased hepatitis incidence risk with a lag of six months. The Holt-Winters model outperformed the SARIMA model in predicting the incidence of viral hepatitis. Among machine learning algorithms, SVM and GPR models demonstrated superior performance for analyzing pollutant data. Patients infected with HAV and HEV were primarily influenced by PM10 and CO, whereas SO2 and PM2.5 significantly impacted others. Individuals aged 35-64 years appeared particularly susceptible to these pollutants. Mixed pollutant exposures were found to affect the development of viral hepatitis with a notable lag of 5-6 months. These findings underscore the importance of long-term monitoring of pollutants in relation to viral hepatitis incidence.

目的 探讨LncRNA NEAT1 作为ceRNA解除miR-205-5p对E2F1 的靶向作用.方法 培养MH-S肺巨噬细胞,转染siCtrl、silncRNA NEAT1、silncRNA NEAT1+miR-205-5p抑制剂后,qRT-PCR检测E2F1 的mRNA转录水平,WB检测E2F1 的蛋白水平;并用RIP和RNA pull-down检测lncRNA NEAT1 和miR-205-5p的结合情况.建立矽肺和对照小鼠模型,并尾静脉注射小鼠模型(siCtrl、silncRNA NEAT1、silncRNA NEAT1+miR-205-5p抑制剂),qRT-PCR检测E2F1 的mRNA转录水平,WB 检测 E2F1 的蛋白水平.结果 与 siCtrl 组(1.15±0.10)比较,转染 silncRNA NEAT1 后E2F1 表达(0.38±0.08)下调;与 silncRNA NEAT1 组比较,转染 silncRNA NEAT1+miR-205-5p 抑制剂后E2F1 表达(0.58±0.10)上调,差异有统计学意义(P<0.01).抗AGO2 抗体可以将lncRNA NEAT1 沉淀下来,表明lncRNA NEAT1 可以与 AGO2 形成复合物,竞争性结合 miR-205-5p.与 NEAT1-Mut 组(1.04±0.05)、NEAT1-NC 组(0.99±0.04)比较,NEAT1 处理后miR-205-5p(3.90±0.10)显著增加,差异有统计学意义(P<0.01).结论 LncRNA NEAT1 可作为ceRNA,解除miR-205-5p对E2F1 的靶向作用.

目的 探讨尘肺患者炎症因子改变及与颈动脉硬化的关系.方法 选取2015年9月—2021年9月开滦总医院林西医院诊治的尘肺患者119例为观察组,其中Ⅰ期患者73例,Ⅱ期患者29例,Ⅲ期患者17例,同期健康检查者51例为对照组,均进行炎症因子[白细胞介素-6(Interleukin-6,IL-6)、C反应蛋白(C-reactive protein,CRP)、同型半胱氨酸(Homocysteine,Hcy)、胱抑制素C(Cystisin C,Cys C)]检测和颈部超声检查[内膜中层厚度(Intima-media thickness,IMT)].分析尘肺患者炎症因子的改变情况,分析炎症因子与颈动脉硬化的关系.结果 尘肺患者Ⅰ、Ⅱ、Ⅲ期的炎症因子(IL-6、CRP、Hcy、Cys C)水平和 IMT 分别为[(16.29±3.17)ng/L、(7.91±1.26)mg/L、(13.84±3.97)μmol/L、(1.79±0.43)mg/L、(1.05±0.24)mm]、[(27.62±5.42)ng/L、(15.68±3.09)mg/L、(25.93±6.01)μmol/L、(3.52±0.84)mg/L、(1.28±0.19)mm]、[(48.07±8.35)ng/L、(38.64±6.37)mg/L、(46.78±7.25)μmol/L、(5.80±1.02)mg/L、(1.67±0.32)mm],均高于对照组[(5.98±1.04)ng/L、(2.23±0.85)mg/L、(5.61±1.12)μmol/L、(0.81±0.16)mg/L、(0.86±0.13)mm],差异均有统计学意义(均P＜0.01),尘肺Ⅱ期和Ⅲ期患者的炎症因子(IL-6、CRP、Hcy、Cys C)水平和IMT均高于尘肺Ⅰ期患者(均P＜0.05),尘肺Ⅲ期患者的炎症因子(IL-6、CRP、Hcy、Cys C)水平和IMT均高于尘肺Ⅱ期患者(均P＜0.05).IMT增厚和斑块的尘肺患者炎症因子(IL-6、CRP、Hcy、Cys C)水平均高于IMT正常的尘肺患者(均P＜0.05),IMT斑块的尘肺患者炎症因子(IL-6、CRP、Hcy、Cys C)水平均高于IMT增厚的尘肺患者(均P＜0.05).IMT出现增厚和斑块时,炎症因子(IL-6、CRP、Hcy、Cys C)与IMT均呈正相关(均P＜0.05).结论 尘肺患者的炎症因子水平较高,可能较易发生颈动脉硬化.

Manganese dioxide nanoparticles (MnO 2 -NPs) have a wide range of applications in biomedicine. Given this widespread usage, it is worth noting that MnO 2 -NPs are definitely toxic, especially to the brain. However, the damage caused by MnO 2 -NPs to the choroid plexus (CP) and to the brain after crossing CP epithelial cells has not been elucidated. Therefore, this study aims to investigate these effects and elucidate potential underlying mechanisms through transcriptomics analysis. To achieve this objective, eighteen SD rats were randomly divided into three groups: the control group (control), low-dose exposure group (low-dose) and high-dose exposure group (high-dose). Animals in the two treated groups were administered with two concentrations of MnO 2 -NPs (200 mg kg −1 BW and 400 mg kg −1 BW) using a noninvasive intratracheal injection method once a week for three months. Finally, the neural behavior of all the animals was tested using a hot plate tester, open-field test and Y-type electric maze. The morphological characteristics of the CP and hippocampus were observed by H&E stain, and the transcriptome of CP tissues was analysed by transcriptome sequencing. The representative differentially expressed genes were quantified by qRT-PCR. We found that treatment with MnO 2 -NPs could induce learning capacity and memory faculty decline and destroy the structure of hippocampal and CP cells in rats. High doses of MnO 2 -NPs had a more obvious destructive capacity. For transcriptomic analysis, we found that there were significant differences in the numbers and types of differential genes in CP between the low- and high-dose groups compared to the control. Through GO terms and KEGG analysis, high-dose MnO 2 -NPs significantly affected the expression of transporters, ion channel proteins, and ribosomal proteins. There were 17 common differentially expressed genes. Most of them were transporter and binding genes on the cell membrane, and some of them had kinase activity. Three genes, Brinp, Synpr and Crmp1, were selected for qRT-PCR to confirm their expression differences among the three groups. In conclusion, high-dose MnO 2 -NPs exposure induced abnormal neurobehaviour, impaired memory function, destroyed the structure of the CP and changed its transcriptome in rats. The most significant DEGs in the CP were within the transport system.

目的 采用16S rDNA测序技术分析利福平(RFP)致大鼠抗结核药物性肝损伤(ADLI)中肠道菌群的变化.方法 将24只雄性SD大鼠随机均分为对照组(D0组)、RFP灌胃10 d组(R10组)、RFP灌胃28 d组(R28组).用随机数字表法每组选取4只大鼠的粪便标本进行16S rDNA测序.结果 与D0组比较,R10组和R28组 α 多样性和 β 多样性均发生改变(P<0.05),门水平上拟杆菌门(Bacteroidetes)丰度增加,厚壁菌门(Firmicutes)丰度减少(P<0.05);属水平上unclassified_f_Prevotellaceae、特氏菌属(Blautia)、Prevotel-laceae_NK3B31_group、Erysipelotrichaceae_uCG-003、Fournierel-la丰度增加,乳杆菌属(Lactobacillus)、Romboutsia、Rumino-coccaceae_uCG-014丰度减少(P<0.05).结论 RFP致大鼠ADLI过程中,肠道菌群数量减少,结构和组成均发生改变,有益菌减少,致病菌增多,但其菌群多样性未见改变.

目的 研究泛素特异性蛋白酶7(USP7)在肝细胞中对凋亡因子的调控作用.方法 将HepG2.2.15细胞随机分为3组:对照组,转染组和阴性对照组.对照组为常规培养的细胞;转染组转染si-USP7,阴性对照组使用10％siRNA-mate转染试剂进行处理,各组均处理72 h.实时荧光定量聚合酶链反应(Q-PCR)检测USP7、P53、B淋巴细胞瘤-2相关X蛋白(Bax)的mRNA表达水平;蛋白质印迹法检测U S P7、P53蛋白的表达情况;酶联免疫法检测谷丙转氨酶(GPT)、谷草转氨酶(GOT)、乙型肝炎病毒(HBV)DNA、乙型肝炎表面抗原(HBsAg)和乙型肝炎核心e抗原(HBeAg)的表达量.结果 对照组、转染组、阴性对照组USP7 mRNA表达水平分别为1.74±0.01,1.00±0.06和1.73±0.01;P53 mRNA表达水平分别为1.94±0.01,0.81±0.03和1.92±0.01;Bax mRNA表达水平分别为6.28±0.04,1.81±0.16,6.26±0.04;蛋白结果 的趋势与基因一致.以上3组的HBV-DNA表达量分别为(1032.68±2.96),(541.92±6.78)和(1035.44±1.86)pg·mL-1;HBsAg表达量分别为(905.19±1.41),(246.23±7.04)和(902.56±4.74)U·mL-1;HBeAg表达量分别为(45.28±0.06),(10.09±0.68)和(45.82±0.36)U·mL-1.以上指标,转染组与对照组比,差异均有统计学意义(均P<0.05).结论 USP7的下调能减少P53的表达,使Bax表达下降,并降低乙型肝炎病毒的复制和细胞凋亡.

We aimed to elucidate the differences in genomic methylation patterns between ADLI and non-ADLI patients to identify DNA methylation-based biomarkers. Genome-wide DNA methylation patterns were obtained using Infinium MethylationEPIC (EPIC) BeadChip array to analyze 14 peripheral blood samples (7 ADLI cases, 7 non-ADLI controls). Changes in the mRNA and DNA methylation in the target genes of another 120 peripheral blood samples (60 ADLI cases, 60 non-ADLI controls) were analyzed by real-time polymerase chain reaction and pyrosequencing, respectively. A total of 308 hypermethylated CpG sites and 498 hypomethylated CpG sites were identified. Significantly, hypermethylated CpG sites cg06961147 and cg24666046 in TANC1 associated with ADLI was identified by genome-wide DNA methylation profiling. The mRNA expression of TANC1 was lower in the cases compared to the controls. Pyrosequencing validated these two differentially methylated loci, which was consistent with the results from the EPIC BeadChip array. Receiver operating characteristic analysis indicated that the area under the curve of TANC1 (cg06961147, cg24666046, and their combinations) was 0.812, 0.842, and 0.857, respectively. These results indicate that patients with ADLI have different genomic methylation patterns than patients without ADLI. The hypermethylated differentially methylated site cg06961147 combined with cg24666046 in TANC1 provides evidence for the diagnosis of ADLI.

目的 旨在通过Meta分析研究临床实验室检测的生物标志物,包括血清C反应蛋白(CRP)、血清淀粉样蛋白(SAA)、乳酸脱氢酶(LDH)和D-二聚体(DD)与新型冠状病毒肺炎(COVID-19)不良预后之间的关联.方法 在PubMed(Medline)、Web of Science和Cochrane中检索截止到2021年3月2日关于COVID-19的研究.研究纳入报告COVID-19患者的实验室检测项目和结果的原创文章,包括死亡、急性呼吸窘迫综合征(ARDS)、重症监护病房(ICU)的护理需求和严重COVID-19.确定不同疾病严重程度生物标志物水平的均数和标准差,通过随机效应模型进行Meta分析.结果 共32项研究中汇总了7739名COVID-19患者,CRP与COVID-19的不良预后显著相关[SMD=0.98,95％CI=(0.85,1.11),P<0.001],血清淀粉样蛋白(SAA)升高与COVID-19的预后不良结局增加相关[SMD=1.06,95％CI=(0.39,1.72),P=0.002],LDH升高与预后不良结局相关[SMD=1.18,95％CI=(1.00,1.36),P<0.001],预后不良结局患者的DD水平较高[SMD=0.91,95％CI=(0.79,1.02),P<0.001].结论 Meta分析表明,血清CRP、SAA、LDH和DD升高与COVID-19患者的预后不良有关.

目的 通过收集乳腺癌患者化疗期间的各项临床数据,发现乳腺癌患者化疗时导致肝功能异常的影响因素,并建立风险预测模型.方法 对入院的乳腺癌患者进行流行病学问卷调查、临床生化检测和血样标本的收集,并分析乳腺癌患者化疗期间的年龄、肿瘤分型、肿瘤TNM分期、化疗方案等数据,使用决策树构建乳腺癌化疗药物性肝损伤风险预测模型.采用EpiData 3.0建立数据库,R软件进行统计分析.结果 共收治675例乳腺癌患者,均为女性.按照研究对象的纳入与排除标准,发现人口学信息、临床生化指标以及血样标本均收集完整的乳腺癌患者640例.经化疗后,106例乳腺癌患者发生肝损伤,占比16.56％.60～ 80岁年龄组发生药物性肝损伤的患者比例(22.82％)高于其他年龄组,且肝损伤的发生率随着年龄的增加呈上升趋势(P＜0.05).已绝经的乳腺癌患者及有长期服药史的乳腺癌患者发生肝损伤的比例分别为21.17％和17.32％,高于未绝经和无长期服药史的肝损伤患者,差异有统计学意义(P＜0.05).肿瘤病理学分型、肿瘤TNM分期及化疗方案均与乳腺癌患者药物性肝损伤发生率相关,其中化疗方案中联合用药组的肝损伤发生率为20.67％,明显高于其他药物组别,差异有统计学意义(P＜0.05).乳腺癌化疗药物性肝损伤风险的决策树模型筛选出3个预测肝损伤风险的观测指标为年龄、肿瘤TNM分期和联合用药,这三个指标对乳腺癌化疗药物性肝损伤风险预测的正确率为92.19％,其中灵敏度和特异度分别为86.36％和93.40％.结论 年龄、是否绝经、长期服药史、肿瘤病理学分型、肿瘤TNM分期及化疗方案是乳腺癌患者化疗期间发生肝损伤的影响因素.结合以上影响因素建立了乳腺癌化疗药物性肝损伤风险预测模型,从而实现患者个体化治疗,保障乳腺癌患者化疗顺利进行.

Understanding factors associated with disease severity and mortality from coronavirus disease (COVID-19) was critical for effective risk stratification. We aimed to investigate the association between biomarkers of clinical laboratory tests, including serum C-reactive protein (CRP), serum amyloid protein (SAA), lactate dehydrogenase (LDH), and D-dimer (DD) and poor prognosis of COVID-19. We have searched many studies on COVID-19 on PubMed (Medline), Web of Science and Cochrane until 1 March 2021. The interest of this study was original articles reporting on laboratory testing projects and outcome of patients with COVID-19 that comprises mortality, acute respiratory distress syndrome (ARDS), need for care in an intensive care unit (ICU), and severe COVID-19. After synthesizing all data, we performed meta-analysis of random effects, and determined mean difference (MD) and standard mean difference at the biomarker level for different disease severity. A total of 7,739 patients with COVID-19 were pooled from 32 studies. CRP was significantly associated with poor prognosis of COVID-19 (SMD = 0.98, 95% CI = (0.85, 1.11), p < .001). Elevated SAA was associated with an increased composite poor outcome in COVID-19 (SMD = 1.06, 95% CI = (0.39, 1.72), p = .002). An elevated LDH was associated with a composite poor outcome (SMD = 1.18, 95% CI = (1.00, 1.36), p < .001). Patients with a composite poor outcome had a higher DD level (SMD = 0.91, 95% CI = (0.79, 1.02), p < .001). This meta-analysis showed that elevated serum CRP, SAA, LDH, and DD were associated with a poor outcome in COVID-19.

目的 在预测miR-217对泛素特异性蛋白酶7(USP7)靶向作用的基础上探讨USP7在HepG2.2.15细胞中的促炎机制.方法 将HepG2.2.15细胞分为对照组、转染组(siRNA-USP7)和阴性对照组(siRNA-NC).检测细胞上清液中谷丙转氨酶(GPT)及谷草转氨酶(GOT)的含量;用酶联免疫吸附(ELISA)法测定细胞上清液中乙型病毒性肝炎表面抗原(HBsAg)和乙型病毒性肝炎e抗原(HBeAg)的含量;用实时荧光定量逆转录聚合酶链反应(RT-qPCR)测定USP7、核因子κB p65(NF-κB p65)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)mRNA含量;用蛋白质印迹(Western blot)法和ELISA法检测各指标蛋白含量;用免疫共沉淀法验证USP7与NF-κB p65的相互作用.结果 对照组、转染组和阴性对照组的GPT含量分别为(13.45 ±0.07),(6.85 ±0.21)和(13.75±0.35)U·L-1;GOT含量分别为(15.10 ±0.57),(7.55 ±0.07)和(15.70±0.42)U·L-1;HBsAg含量分别为(3766.00±62.39),(2093.50±51.47)和(3727.00±55.73) pg·mL-1;HBeAg含量分别为(49.64±0.65),(27.49 ±0.60)和(49.59±1.13)pg·mL-1;USP7蛋白表达量分别为0.97 ±0.03,0.49±0.01和0.96 ±0.04;NF-κB p65蛋白表达量分别为0.73±0.03,0.46±0.02和0.76±0.01;IL-6蛋白表达量分别为(83.00 ±4.24),(43.00±4.24)和(82.50±3.54)pg·mL-1;TNF-α蛋白表达量分别为(95.00±2.83),(57.00±1.41)和(92.50±4.95) pg·mL-1,转染组与对照组相比,差异均有统计学意义(均P＜ 0.05).免疫共沉淀显示USP7与NF-κB p65能够有效形成复合物.结论 在乙型病毒性肝炎中,USP7呈现高表达,抑制USP7可减轻乙型病毒性肝炎细胞中NF-κB p65、IL-6以及TNF-α的表达,缓解肝细胞炎性损伤.

目的 采用16S rDNA测序技术分析异烟肼(INH)致大鼠抗结核药物性肝损伤(ADLI)时肠道菌群的变化情况.方法 将24只雄性SD大鼠随机均分为对照组(D0组)、INH灌胃10 d组(H10组)、INH灌胃28 d组(H28组),分别于连续灌胃0、10和28 d后收集大鼠新鲜粪便进行16S rD-NA测序,分析肠道菌群的结构和组成,并取肝组织进行HE染色,观察大鼠肝组织病理学变化.结果 与D0组比较,H10组和H28组α多样性未发生改变(均P＞0.05),β多样性发生变化(均P ＜0.05),门水平上Verrucomicrobia丰度减少而Tenericutes丰度增加(均P＜0.05),属水平上Lactoba-cillus、Romboutsia和Akkermansia丰度减少而Ruminococcace-ae_UCG-005、Dubosiella、norank_f__norank_o__Mollicutes_RF39、unclassified_f__Ruminococcaceae、Roseburia和Blautia丰度增加(均P＜0.05).结论 INH致大鼠ADLI时肠道菌群结构和物种组成均发生变化.

Background Anti-tuberculosis drug-induced liver injury (ADLI) limits the treatment of tuberculosis. The mechanisms underlying ADLI are unclear and there are no effective preventative measures to avoid this complication. Methods In this stuy, the protein contents of EZH2, Nrf2, NQO1 and HO-1 were detected by ELISA kit, while those of EZH2 and Nrf2 were determined by Western blot. The Chip experiment was used to detect the level of H3K27me3 in the Nrf2 promoter region.The liver were analyzed histopathologically in vivo using hematoxylin and eosin staining. Results Here we developed a murine model of ADLI that recapitulates liver injury in the human disease. Using this model, we investigated the potential involvement of the enhancer of zeste homolog 2 methyltransferase (EZH2), a histone methyltransferase which inhibits the transcriptional activation of the Nrf2-ARE oxidative stress pathway. Compared to controls, mice livers with ADLI showed decreased expression of EZH2 together with reduced H3K27me3 marks in the Nrf2 promoter. This was accompanied by increased expression of Nrf2 and its target genes NQO1 and HO-1. Liver injury in the mice with ADLI could be alleviated to an extent by in vivo delivery of siRNAs targeting EZH2, which further downregulated EZH2 expression and H3K27me3 levels in the Nrf2 promoter along with accompanying increases in Nrf2, NQO1 and HO-1 expression. Conclusions Therefore, inhibiting EZH2 likely reduced liver damage in ADLI by enhancing this key anti-oxidative stress pathway. Our results establish a role for EZH2 in a mouse model of ADLI and furthermore provides valuable mechanistic insights into the development of ADLI pathology.

目的:探讨异烟肼(INH)诱导肝细胞CYP2E1启动子区组蛋白乙酰化对肝细胞凋亡的影响.方法:INH浓度1000μg/mL,Garcinol为5μmol/L,药物作用6 h.ELISA检测乙酰化酶HAT与去乙酰化酶HDAC活性.Chip检测H3K56、H4K5水平.RT-PCR检测CYP2E1、JNK、Bax、Bcl2表达.结果:INH组与对照比,HAT活性降低,HDAC及CYP2E1启动子区acH3K56和acH4K5水平升高,JNK、Bax的mRNA增加,Bcl2下降.与INH组比,INH+Garcinol组HAT活性、acH3K56、acH4K5表达及JNK、Bax降低,Bcl2升高.结论:抑制肝细胞CYP2E1基因启动子乙酰化可减少INH诱导的肝细胞凋亡.

目的 初步探讨利福平诱导小鼠肝损伤过程中,炎症因子及相关指标的变化情况.方法 48只昆明小鼠随机分为基线组和利福平组,灌胃3、5、7、10、15天后处死.酶联免疫吸附(ELISA)法检测血清中丙二醛(MDA)、超氧化物歧化酶(SOD)含量.RT-PCR和ELISA法检测肝组织中核因子κB(NF-κB)和肿瘤坏死因子-α(TNF-α)的mRNA及蛋白表达水平.结果 肝组织病理学及MDA、SOD水平变化,提示肝脏损伤进展.与基线组相比,利福平组NF-κB或TNF-α的mRNA及蛋白表达水平均在第7天和第10天出现显著升高(P<0.05).结论 利福平可能通过激活NF-κB信号通路诱导小鼠肝脏发生炎性损伤.

目的:研究一线抗结核药物诱导小鼠肝损伤发生过程中NF-κB变化情况.方法:48只昆明小鼠随机分为基线组和HRZ组(INH+RFP+PZA),于灌胃3d、5d、7d、10d、15d后处死.ELISA检测血清中ALT、AST含量.ELISA和RT-PCR检测肝组织中NF-κB、IκB mRNA及蛋白表达水平.结果:肝组织病理学及ALT、AST水平发生改变,提示肝损伤发生.与基线组相比,HRZ组NF-κB mRNA和蛋白水平呈上升趋势,IκB mRNA和蛋白水平逐渐下降.结论:NF-κB通路在抗结核药物性肝损伤小鼠模型中被激活.