[Analysis of hepatic pathological inflammation and fibrosis condition and its influencing factors in 721 patients with chronic hepatitis B with normal ALT].

Objective: To analyze the hepatic pathological inflammation and fibrosis condition in order to explore the relationship with related clinical indicators in patients with chronic hepatitis B patients with normal alanine aminotransferase (ALT). Methods: 721 cases of chronic hepatitis B with normal ALT who were initially diagnosed in the Department of Infectious Diseases of Henan Provincial People's Hospital from August 2016 to December 2019 were retrospectively collected. Liver biopsy was performed in all patients. General data of patients such as gender, age, liver function indexes, blood routine indexes, HBsAg level, HBeAg status, HBV DNA level, spleen thickness and prothrombin time were collected. Univariate and multivariate analysis methods were used to determine the influencing factors of inflammation and fibrosis degree with liver biopsy. A receiver operating characteristic curve (ROC) was used to evaluate the established multi-factor prediction model. Alpha=0.05 was considered as a standard orientation of test. Results: The average age of 721 cases with chronic hepatitis B was 36.1±9.7 years, and the male to female ratio was 1.28/1, with inflammation and fibrosis grade mainly concentrated in G1S1 (349 cases), G1S2 (132 cases), G2S2 (119 cases), and G2S1 (57 cases). Among them, there were 349 (48.4%) cases of G1S1, and 372 (51.6%) cases of G/S≥2. The main manifestations were mild to moderate inflammation and fibrosis, and only 64 (8.88%) cases had severe G/S≥3. HBsAg level (stratified with 4 log10 IU/ml as the boundary) analyzed in 721 cases were correlated with the relevant clinical indicators stratification and liver pathological inflammation and fibrosis, and the difference was statistically significant (inflammation grade, χ2=6.182, P=0.013; Fibrosis grade, χ2=36.534, P=0.001). Univariate analysis of the relevant clinical indicators that may influence the patient's liver pathological G/S ≥2 showed the patient's age, albumin, γ- glutamyltransferase (GGT), platelet, prothrombin time (PT), spleen thickness and HBsAg level were all statistically significant (P<0.05), while multivariate analysis showed that age, GGT, PT, and spleen thickness had statistical differences (P<0.05). The prediction model was established in accordance to multivariate analysis, and the area under the ROC curve was 0.642. Maximization of the sum of sensitivity and specificity as cut-off value of Logit P=0.497, the diagnostic sensitivity, specificity, and Youden's index were 60.6%, 64.5%, and 0.252, respectively. Conclusion: More than half of patients with chronic hepatitis B with normal ALT have significant inflammation and fibrosis and require timely antiviral therapy. Age, GGT, PT and spleen thickness can help comprehensively evaluate the liver inflammation and fibrosis status among patients, but the lack of accurate prediction models suggests that more effective indicators that can help predict the inflammation and fibrosis status of such patients have yet to be discovered. Therefore, liver biopsy should still be actively performed in patients with normal ALT to confirm the diagnosis and timely treatment.

[Immunotherapy the road of hope for patients with advanced liver cancer in the new era].

Liver cancer prevention has always been a key issue in the follow-up diagnosis and treatment of viral hepatitis. Insidious onset, high morbidity, monotherapy, short survival time, and high mortality are the outstanding problems encountered in the diagnosis and treatment of advanced liver cancer. In recent years, with the clinical application of targeted drugs and immune checkpoint inhibitors, phased progress has been made in the diagnosis and treatment of advanced liver cancer, especially the accessibility of drug prices under the new medical insurance has provided more and more patients the opportunity to achieve a longer survival time. In this paper, the hot issues in the diagnosis and treatment of patients with advanced liver cancer in the immunotherapy era are discussed.

[Analysis of bone mineral density and its influencing factors in 211 patients with chronic hepatitis B treated with long-term entecavir monotherapy].

Objective: To investigate the changes of bone mineral density and its related influencing factors in chronic hepatitis B patients treated with long-term entecavir monotherapy. Methods: 211 cases with chronic hepatitis B treated with entecavir monotherapy in the Department of Infectious Diseases of Henan Provincial People's Hospital from June 2018 to September 2019 were retrospectively collected. Age, gender, body mass index, number of years of medication use, presence or absence of liver cirrhosis and current bone mineral density level (using dual-energy X-ray detection, taking lumbar L1 ~ 4 and left femur as observation region) and other related data were collected. 211 cases general situation was descriptively analyzed by case-control study design. Two independent sample t-tests were used to compare the differences in serum calcium, phosphorus, and renal function levels in patients with different medication durations. Univariate logistic regression was used to screen the influencing factors of bone mineral density level. Significant variables of univariate analysis were included in multivariate logistic regression to obtain the independent influencing factors leading to the decrease of bone mineral density level. The test level was set as α = 0.05. Results: The average age of 211 cases with chronic hepatitis B was (42.36 ± 11.10) years. The average medication time use was (2.52 ± 1.94) years. The body mass index (23.95 ± 3.11), and male-to-female ratio was 2.25/1. The incidence of liver cirrhosis was 35.5%. The incidence of low bone mass in the two observation sites (lumbar spine L1~4 and left femur) was 24.6% and 29.4%, respectively. There were statistically significant differences in serum calcium, phosphorus and renal function levels among patients with different entecavir treatment duration (≥3 years and < 3 years) (P < 0.05). Univariate analysis result showed that the influencing factors of BMD were age, the number of years of medication use, gender, liver cirrhosis (L1~4 of the lumbar spine region) and age, the number of years of medication, and gender (left femoral region). The variables that entered the two models after the multivariate analysis were age (L1~4 region of lumbar spine: OR = 2.225, left femur OR = 1.660), gender (L1~4 region of lumbar spine: OR = 3.048, left femur OR = 2.496), number of years of medication use (L1~4 region of lumbar spine: OR = 1.387, left femur OR = 1.276). Conclusion: Age, gender, and the number of years of medication use are independent factors that influence the bone mineral density of patients with chronic hepatitis B treated with long-term entecavir. Low bone mass risk at the two observation sites is 2.225 and 1.66 times the normal level for every 10 years of age increase. Compared with men, the risk of low bone mass at the two observation sites is 3.048 and 2.496 times for women, and for every additional year of medication use, the risk of low bone mass at the two observation sites is 1.387 and 1.276 times the normal level. Female patients with older age and prolonged medication use are at high risk of developing bone mineral density reduction.

[Differential proteomic screening of plasma exosomes before and after magnesium isoglycyrrhizinate treatment in chronic hepatitis B].

Objective: To screen the differential proteomic of plasma exosomes before and after magnesium isoglycyrrhizinate (MgIG) treatment in chronic hepatitis B patients. Methods: Plasma samples were collected from 36 cases with chronic hepatitis B before and after MgIG treatment (2 ml/case). Plasma exosomes were extracted by ultracentrifugation. Exosomal particles concentration and inner diameter were detected by Nanosight NS300 particle size analyzer. Three cases of plasma exosomes were randomly selected before and after MgIG treatment. Proteins were extracted after lysis and digested with trypsin. Label-free differential proteomics analysis was performed by liquid chromatography-tandem mass spectrometry to screen out differential proteins that changed more than 1.5 times. Enzyme linked immunosorbent assay (ELISA) was used to verify the quantitative differential protein expression (n = 30). Measurement data were compared by paired sample t-test. Results: The average particle concentration of the extracted exosomes was 2.2×10(9)/ml, and the average size was (107 ± 52) nm, which was consistent with the theoretical value of plasma exosome size, proving that the plasma exosomes were successfully extracted. Proteomics results showed that before and after MgIG treatment in chronic hepatitis B patients, a total of 153 differentially expressed proteins were screened, including 85 up-regulated and 68 down-regulated proteins. Enzyme-linked immunosorbent assay results showed that compared with the MgIG before and after treatment group of chronic hepatitis B patients, the differences in the concentrations of hepatocyte growth factor activator and hepatocyte growth factor like protein in plasma exosomes were statistically significant (P < 0.05). Hepatocyte growth factor activator concentration in the plasma exosomes before and after MgIG treatment group was (45.9 ± 9.4) µg/ml and (13.9 ± 2.0) µg/ml, respectively, and it was down-regulated by about 3 times. Hepatocyte growth factor-like protein concentration in the plasma exosomes before and after MgIG treatment group was (23.4 ± 4.9) µg/ml and (13.8 ± 2.2) µg/ml, respectively, and it was down-regulated by about 2 times. Enzyme-linked immunosorbent assay results had consistency with the proteomics results. Conclusion: This study successfully screened the differential proteomic of plasma exosomes before and after MgIG treatment in chronic hepatitis B, and provided experimental basis for studying the molecular mechanism of MgIG treatment for chronic hepatitis B.

[Gene mutation pattern of Gilbert's syndrome combined with viral hepatitis and its relationship with the exploration of clinical data].

Objective: To study whether gene mutation pattern of Gilbert's syndrome (GS) is combined with viral hepatitis and its relationship with relevant clinical data. Methods: Clinical data of GS patients combined with viral hepatitis who was admitted to the Department of Infectious Diseases of Henan Provincial People's Hospital from August 2013 to December 2018 was retrospectively analyzed. The relationship between gene mutation pattern, general data (age, gender, etc.) and liver biochemical indexes was analyzed. The differences of the above data in patients with or without combined viral hepatitis were analyzed. The measurement data were compared by t-test. The categorical data was compared by the χ (2) test. The median and interquartile range of non-normally distributed data was used to indicate the central and discrete tendency. Results: A total of 107 GS eligible cases data were collected. The male to female ratio was 4.94:1 (89:18). The average age of onset was (36.36 ± 12.51) years. Alanine aminotransferase and total bilirubin levels were normal or slightly elevated, while aspartate aminotransferase, alkaline phosphatase, and γ-glutamyltransferase were all within the normal range. There were 49 cases in the combined viral hepatitis group (36 cases with HBV and 13 cases with HCV), and 58 cases in the GS alone group. Total bilirubin level in GS alone group was higher than the combined viral hepatitis group (z = 0.035, P < 0.05), and there were no statistically significant differences in gender, age, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and gamma glutamyltransferase (P > 0.05). Uridine diphosphate glucuronide transferase 1A1 (UGT1A1), specifically encoded by GS was detected in all 107 cases. Mutations was mainly occurred in the upstream promoter PBREM-3263 (-3279) (86 cases) and TATA box TA insertion mutation (71 cases), and GGA-AGA Gly71Arg (57 cases) mutation in EXON1 of the coding region. All mutation forms had manifestations of homozygous and heterozygous abnormalities. The combined incidence of main mutation forms in the genetic testing data were sequenced as: A2 + B2 + C2 (17 cases, 25.23%), A1 + B1 (17 cases, 15.89%), A2 (11 cases, 10.28%), C2 (10 Cases, 9.34%), A2 + B2 (7 cases, 6.54%), A1 + B2 (7 cases, 6.54%), C1 (7 cases, 6.54%), and there was no statistically significant difference between different mutation combinations in patients with or without hepatitis (P > 0.05). The results of total data analysis showed that the total bilirubin level in the single-site mutation group was higher than the multi-site mutation group (Z=2.019, P = 0.043), and other biochemical indicators had no effect (P > 0.05) and the differences were not statistically significant. Further analysis showed that the total bilirubin level of the single-site mutation subgroup in the GS alone group was higher than the multi-site mutation subgroup (Z = 1.999, P = 0.046), and the statistical difference was similar to the combined viral hepatitis group (P > 0.05). Different mutation combinations had no effect on biochemical indexes, and had no relationship with combined viral hepatitis (P > 0.05). Conclusion: GS is common in patients with combined viral hepatitis, and there is no significant difference between the incidence of gene mutation, mutation forms, biochemical indexes, and non-hepatitis group. The increase in the number of GS mutation sites does not aggravate the deterioration of bilirubin levels due to the decrease in the content and activity of uridine diphosphate glucuronosyltransferase, and the combination of different mutation sites does not affect the changes of various biochemical indexes, and at the same time it is not related to hepatitis.

[Advances in combination therapy of ombitasvir and dasabuvir for chronic hepatitis C virus genotype 1 infection].

Since 2014, the United States and Europe has approved all oral, interferon free- regimens that combine with direct-acting antiviral agents. Hence, the sustained virological response rate of patients with chronic HCV genotype 1 infection has improved over 90%, and the treatment modalities has introduced a new era. These drugs, ombitasvir and dasabuvir, received customary authorization of Food and Drug Administration in 2015 and are the first combined direct-acting antiviral agents for treating HCV genotype 1 infection. It has superior application prospects in China because of its high-sustained virological response rate and safety profile. This article reviews the pharmacokinetics, drug interactions, efficacy and safety of this therapeutic regimen.

[A real-world study of paritaprevir/ritonavir-ombitasvir combined with dasabuvir in the treatment of genotype 1b chronic hepatitis C].

Objective: To recognize the efficacy and safety of paritaprevir/ritonavir-ombitasvir combined with dasabuvir (OBV/PTV/RTV+DSV) in the treatment of genotype 1b chronic hepatitis C. Methods: Patients with genotype 1b chronic hepatitis C who were admitted to the People's Hospital of Henan Province, Huashan Hospital of Shanghai and the Fifth Medical Center of the General Hospital of the People's Liberation Army of China between November 2017 to August 2018 were enlisted. All patients received OBV/PTV/RTV+DSV antiviral therapy. HCV RNA levels were measured at baseline, weeks 1, 2, 3, 4, 8, 12, and 24, then 12 weeks, and 24 weeks after completion of treatment; patients' comorbidity, concomitant medications, and clinical adverse events were recorded. Results: 108 patients were enrolled in the study, with an average age of 49.1 years, 44 patients were male (40.8%), 96.3% (104/108) were newly diagnosed, and four patients had previous treatment history, of whom three were treated with IFN and one with IFN + DAA. Ninety-eight cases completed 12 weeks treatment and 89 cases were in follow up for 12 weeks, after discontinuation of the drug. Overall, 89 cases (100%) achieved SVR12.One patient treated with PR and DAA had HCV RNA level of 869175 IU/mL at 4 weeks of treatment, which was significantly higher than the baseline HCV RNA level (301776IU/ML), and was judged as failure of treatment; and follow-up was discontinued. Of all enrolled patients, 19 (17.6%) had underlying diseases and 15 (13.9%) had combined medications. During treatment, adverse events (AE) occurred in 11 patients (10.1%). The main adverse events were pruritus and elevated bilirubin. Conclusion: Combined antiviral therapy (OBV/PTV/RTV+DSV) of 12 weeks are highly effective with good safety profile in the treatment of Chinese patients with genotype 1b chronic hepatitis C.

Levels of HBx, VEGF, and CEACAM1 in HBV-related hepatocellular carcinoma and their correlation with cancer prognosis.

OBJECTIVE: Hepatitis B virus X protein (HBx), vascular endothelial growth factor (VEGF) and carcinoembryonic antigen related cell adhesion molecule 1 (CEACAM1), are related to HBV associated hepatocellular carcinoma (HCC). This study recruited HCC patients and employed the SMMC-7721 and L02 liver cell lines, to analyze the expression levels of HBx, VEGF and CEACAM1 in liver cancer and their correlation with the cancer prognosis. PATIENTS AND METHODS: HBV-related HCC patients were recruited from our hospital. Immunohistochemistry (IHC) and Western blotting assay were used to detect the expression of HBx, VEGF and CEACAM1 in liver tissues. Multi-variant analysis and the correlation analysis between HBx, VEGF, CEACAM1 expression and clinical/pathological features of HCC were performed by using the Cox regression analysis. RESULTS: In HBV-related HCC tissues, positive expression rates of HBx, CEACAM1, and VEGF, were 80%, 50%, and 65%, respectively. In HBx-positive group, positive rate for CEACAM1 and VEGF were 56.25% and 75%, while in HBx-negative group such figures were 75% and 25% (p<0.05). HCC cells had lower expression of CEACAM1 and higher VEGF levels compared to normal hepatocytes. Those HCC cells transfected with HBx had even lower CEACAM1 and higher VEGF levels compared to un-transfected cells. HBx was negatively correlated with CEACAM1 and positively correlated with VEGF. Expressions of these three factors were all independent risk factors as they were correlated with lesion size, venous infiltration, metastasis, and capsule. CONCLUSIONS: HBx, VEGF and CEACAM1 were widely expressed in HBV-related HCC. HBx may facilitate occurrence and progression of HBV-related HCC via down-regulating CEACAM1 and up-regulating VEGF.

[Influence of hepatitis B virus X gene on apoptosis of hepatic cells mediated by Fas].

Objective: To investigate the influence of hepatitis B virus X gene (HBx) on apoptosis of hepatic cells mediated by Fas in HePG2 cells. Methods: HBx eukaryotic vector pcDNA3.1(+)-X was transfected into HEPG2 cells with lipofectamine, and the null vector pcDNA3.1(+) and untransfected HEPG2 were used as normal controls. The cells were collected 72 h after transfection, and the expression of HBx mRNA and protein was determined using RT-PCR and Western blot, respectively. The mRNA expression of apoptosis-related genes Bcl-2 and Bax mRNA was also determined using RT-PCR. Cytotoxicity and apoptosis were evaluated using CCK-8 and flow cytometry, respectively, after HepG2-HBx and HepG2-3.1 cells were treated with stimulatory monoclonal antibody anti-Fas CH11. The t test was used for pairwise comparison. Results: The cell line HepG2-HBx was successfully established, as confirmed by RT-PCR and Western blot, and RT-PCR results showed that HepG2-HBx cells had significantly higher expression of Bcl-2 mRNA than HepG2-3.1 and HepG2 cells (P < 0.05), but had significantly lower expression of Bax mRNA than HepG2-3.1 and HepG2 cells (P < 0.05); CCK-8 and flow cytometry showed that anti-Fas CH11 had a lower cytotoxicity to HepG2-HBx cells and allowed for a lower apoptosis rate of HepG2-HBx cells compared with HepG2-3.1 and HepG2 cells. Conclusions: HBx can inhibit apoptosis of hepatic cells mediated by the Fas pathway.

[Changes in serum ß2-microglobulin, retinol-binding protein, and cystatin C and their value in identifying early renal dysfunction in patients with chronic hepatitis B undergoing tenofovir or entecavir monotherapy: a comparative analysis].

Objective: To investigate the dynamic changes in serum ß2-microglobulin, retinol-binding protein, and cystatin C in chronic hepatitis B(CHB)patients treated with tenofovir or entecavir alone as the anti-HBV therapy, as well as their value in identifying early renal dysfunction. Methods: A total of 61 previously untreated CHB patients who were diagnosed and treated in the Department of Infectious Diseases in Henan Provincial People's Hospital from June 2013 to August 2015 were enrolled and divided into tenofovir group and entecavir group. The serum levels of ß2-microglobulin, retinol-binding protein, cystatin C, and creatinine and estimated glomerular filtration rate(eGFR)were compared between the two groups at baseline and 4, 8, 39, 52, 78, and 104 weeks after antiviral therapy. The independent samples t-test was used for comparison of continuous data, and the chi-square test was used for comparison of categorical data. P < 0.05 was considered statistically significant. Results: A total of 61 CHB patients were enrolled, with 31 in the tenofovir group and 30 in the entecavir group. The two groups had comparable serum levels of ß2-microglobulin, retinol-binding protein, and cystatin C at baseline, but there were significant differences in ß2-microglobulin and retinol-binding protein over time(both P < 0.05). There was a significant difference in cystatin C at 78 weeks(t = -2.062, P = 0.044), but there was no significant difference at 104 weeks(t = -1.544, P = 0.128). There were no significant differences in serum creatinine or eGFR at any time point between the two groups(P > 0.05). At 104 weeks, there were no significant differences in HBV-DNA clearance rate or the level of virologic breakthrough between the two groups(P > 0.05). Conclusion: Serum ß2-microglobulin, retinol binding protein, and cystatin C are more sensitive than eGFR in the monitoring of early renal dysfunction during the anti-HBV therapy with tenofovir or entecavir alone.

[Clinical features of malignant tumor liver infiltration with liver failure as initial clinical manifestation: a single-center retrospective analysis of 9 cases].

Objective: To investigate the clinical features of patients with liver failure caused by tumor diffuse liver infiltration. Methods: A retrospective analysis was performed for the clinical data of 1008 patients with liver failure who were admitted to our hospital from July 2009 to December 2015. Among these patients, 9 had acute liver failure caused by liver metastasis of malignant tumor. Their clinical manifestations, laboratory markers, clinical progress, and outcome were observed, and the clinical features were summarized. Results: Such patients were manifested as liver enlargement and rapid clinical progression, and imaging examination showed stenosis due to external compression in the inferior vena cava. The patients might be easily misdiagnosed with Budd-Chiari syndrome and had a poor prognosis, with a mortality rate as high as 100%. Conclusion: As for the liver failure patients with unexplained liver enlargement, the possibility of liver metastasis of malignant tumors should be considered. Liver biopsy should be performed as early as possible before the deterioration of liver function, in order to facilitate the targeted therapy for the primary tumor.