Objective To investigate the liver disease phenotypes and clinical features of patients with different genotypes of Wilson's disease(WD).Methods A retrospective analysis was performed for 163 patients with WD who were diagnosed and underwent genetic testing in The Fifth Medical Center of Chinese PLA General Hospital from August 2008 to June 2023,and clinical manifestations,laboratory examination,pathological examination,imaging examination,and ATP7B genetic testing results were collected.According to ATP7B gene mutation,the patients were divided into groups as follows:R778L mutation group and non-R778L mutation group;P992L mutation group and non-P992L mutation group;truncation mutation group and non-truncation mutation group.Liver disease phenotypes and clinical features were analyzed for the patients with c.2333G>T/p.R778L mutation(R778L mutation),c.2975C>T/p.P992L mutation(P992L mutation),and truncation mutation of the ATP7B gene.The Mann-Whitney U test or the Kruskal-Wallis H test was used for comparison of continuous data between groups,and the chi-square test or the Fisher's exact test was used for comparison of categorical data between groups.Results The 163 patients with WD had varying severities of liver disease phenotypes,among whom 121(74.23%)were diagnosed with chronic liver disease,36(22.09%)were diagnosed with decompensated cirrhosis,and 6(3.68%)were diagnosed with fulminant WD,and in addition,there were 5 patients(2 with chronic liver disease and 3 with decompensated cirrhosis)with neurological abnormalities.For the 163 patients with WD,R778L mutation(with an allele frequency of 28.2%)was the most common mutation in the ATP7B gene,followed by P992L mutation(with an allele frequency of 12.6%),and truncation mutation showed an allele frequency of 11.0%.There was no significant difference in the distribution of the three mutations across different liver disease phenotypes(P>0.05).The R778L mutation group had a significantly lower level of ceruloplasmin(CP)than the non-R778L mutation group[0.04(0.02-0.08)g/L vs 0.08(0.03-0.13)g/L,Z=-2.889,P=0.004].Compared with the non-P992L mutation group,the P992L mutation group had significantly higher levels of alanine aminotransferase[135.0(80.5-237.0)U/L vs 80.5(36.0-173.3)U/L,Z=2.684,P=0.007]and aspartate aminotransferase[121.4(77.0-195.0)U/L vs 84.0(39.0-123.3)U/L,Z=3.388,P<0.001].Compared with the non-truncation mutation group,the truncation mutation group had significantly lower levels of CP[0.03(0.02-0.08)g/L vs 0.06(0.03-0.11)g/L,Z=-3.136,P=0.002]and serum copper[3.20(2.15-5.00)mg/L vs 4.20(2.60-7.50)mg/L,Z=-2.296,P=0.025].Conclusion R778L mutation,P992L mutation and truncation mutation are not associated with liver disease phenotype in WD patients;however,R778L mutation is associated with a lower level of CP,P992L mutation is associated with higher levels of ALT and AST,and truncation mutation is associated with lower levels of CP and serum copper.

Objective:To investigate the clinical manifestations, pathological, and gene mutation characteristics of ABCB4 gene variant-associated cholestatic liver disease in adults. Methods:Eight adult cases of ABCB4 gene variant-associated cholestatic liver disease who were hospitalized in the Department of Hepatology, Fifth Medical Center of the People's Liberation Army General Hospital from May 2010 to December 2022 were enrolled in this study. The clinical manifestations, pathological features, gene variant features, and prognostic conditions were analyzed. Patient gene testing and biological information analysis were performed using whole-exome next-generation sequencing. SPSS 19.0 software was used to conduct descriptive analysis. Results:Among the eight adult cases of the ABCB4 gene variant, there were three males and five females, with a median age of onset of 24 (20, 37) years. There were three cases with a compound heterozygous variant in ABCB4, and the clinical phenotypes included two cases of progressive familial intrahepatic cholestasis type 3 and one case of intrahepatic cholestasis of pregnancy overlapping with low-phospholipid-associated cholelithiasis syndrome. There were five cases with a single heterozygous variant in ABCB4, and the clinical phenotypes included two cases of intrahepatic cholestasis of pregnancy overlapping with drug-induced liver injury and three cases of low-phospholipid-associated cholelithiasis syndrome. Imaging of all eight cases showed liver fibrosis, and six cases already had cirrhosis. All patients underwent liver histopathological examination, which mainly showed cholestasis and portal fibrosis in eight cases, small bile duct hyperplasia in seven cases, copper deposition in three cases, and cirrhosis in five cases. ABCB4 screening revealed 11 different mutations, including eight new mutations. The pathogenicity assessment showed that c.2394+82C>T (intron) was a benign mutation, and the rest were deleterious mutations. Ursodeoxycholic acid was the treatment for all patients, with a follow-up time of 7.5 (0.5, 12.7) years. One case died of end-stage liver disease, two cases developed cholestatic cirrhosis, and five cases were in stable condition. Conclusion:The adult ABCB4 gene variant-associated cholestatic liver disease are mostly single heterozygous mutations, the clinical phenotypes are diverse and overlapping, the disease is more severe in those who carried non-functional mutations.

Objective:To analyze the clinical manifestations, pathology, and gene variant characteristics in children with progressive familial intrahepatic cholestasis type 3 (PFIC3).Methods:This retrospective study assessed the clinical manifestations, pathological features, gene variants, and prognosis data of 11 children with PFIC3 hospitalized in the Department of Hepatology, Fifth Medical Center, PLA General Hospital, from January 2015 to December 2022. Panel or whole exome sequencing was performed on the probands, followed by Sanger sequencing for verification within the family. Detected pathogenic variants were compared with known disease databases. Additionally, the new variants were predicted the deleteriousness and protein structure using relevant software to evaluate their pathogenicity.Results:Among the 11 PFIC3 children, 8 were boys and 3 were girls. The age of onset was 3.1 (0.2, 15.6) years. The main complaint of onset was different in the 11 patients;5 of them were abnormal liver function, 3 of them were liver and spleen enlargement, 2 of them were abdominal distension, and 1 of them was jaundice. Alanine aminotransferase, asparate aminotransferase and γ-glutamyltransferase increased in all the patients, which were(113±40), (150±44) and (270±156) U/L respectively. Moreover, direct bilirubin increased in 9 patients, and cholestasis was showed in 8 patients. All patients showed liver fibrosis on imaging, and 8 patients had cirrhosis. The pathological features of 8 cases by liver biopsy were as follows: 8 cases of fibrosis in the portal area, 7 cases of small bile duct hyperplasia, 4 cases of positive copper staining, and 5 cases of cirrhosis. A total of 17 ABCB4 gene variants were detected, including 9 new variants: c.589C>T(p.Q197X), c.1230+1G>A(Splicing), c.2914G>A(P.D972N), c.1058G>A(p.C353Y), c.956G>T(p.G319V), c.473T>A(p.L158Q), c.164T>C(p.L55S), c.2493G>C(p.R831S), and c.1150G>C(p.G384R). All 11 patients were treated with ursodeoxycholic acid and followed up for 5.1(0.6, 7.4) years. Among them, 4 cases of cirrhosis progressed continuously, 3 cases had liver transplantations, and the remaining 4 cases were stable after medical treatment.Conclusions:Children with PFIC3 have early onset, diverse clinical manifestations, rapid progression of fibrotic and cholestasis, as well as poor prognosis. Genetic testing helps to confirm the diagnosis.

Objective:To investigate the role of serum hepatitis B virus RNA (HBV RNA) in predicting HBeAg serological conversion in children with chronic hepatitis B.Methods:175 children aged 1~17 years with chronic hepatitis B who received interferon α (IFNα) for 48 weeks were selected. Patients were divided into HBeAg seroconversion and non-conversion based on whether HBeAg seroconversion occurred at 48 weeks of treatment.T-test and Mann-Whitney U test were used to compare between groups; chisquare test or Fisher exact probability method was used to compare the frequency between groups of classified variables; and Pearson correlation was used to analyze the correlation between indicators. Univariate and multivariate logistic regression analyses were used to identify influencing factors associated with HBeAg serological conversion. The predictive effect of HBV RNA, HBV DNA, and HBsAg on HBeAg serological conversion was compared and analyzed by the receiver operating characteristic curve (ROC).Results:The seroconversion rate of HBeAg at 48 weeks was 36.0% (63/175). The reduction in HBVRNA levels from baseline to the 12th, 24th, 36th, and 48th weeks of antiviral therapy was significantly greater in the HBeAg serological conversion group than that in the non-conversion group, and the difference was statistically significant between the two groups (P < 0.05). Univariate and multivariate regression analyses showed that age and a decline in HBV RNA levels at week 12 were independent predictors of HBeAg serological conversion. The area under the ROC curve (AUROC) of HBV RNA decline at week 12 was 0.677(95% CI∶0.549-0.806, P = 0.012), which was significantly better than the same period of AUROC of HBV DNA (0.657, 95% CI∶0.527-0.788, P = 0.025) and HBsAg (0.660, 95% CI∶0.526-0.795, P = 0.023) decline. HBV RNA levels decreased (>1.385 log10 copies/ml) at week 12, with a positive predictive value of 53.2%, a negative predictive value of 72.2%, a sensitivity of 77.4%, and a specificity of 57.9% for HBeAg seroconversion. Conclusion:HBV RNA level lowering during the 12th week of antiviral therapy can serve as an early predictor marker for HBeAg serological conversion in children with chronic hepatitis B.

Objective:To evaluate the efficacy and safety of direct antiviral drugs based on sofosbuvir in the treatment of chronic hepatitis C in children.Methods:Data of children diagnosed with chronic hepatitis C from May 2017 to December 2019 and received sofosbuvir (SOF, body weight ≥ 17 kg 200 mg/d, <17 kg 150 mg/d)/ledipasvir (LDV, body mass ≥ 17 kg 45 mg/d, <17 kg 33.75 mg/d) or velpatasvir (VEL, body weight ≥ 17 kg 50 mg/d), were collected. The patients were treated with ribavirin (RBV, 15 mg/kg·d) for 12 weeks and followed up for 12 weeks. The main outcome measures were sustained virological response (SVR) at 12 weeks follow-up after drug discontinuation, and the occurrence of adverse events during the treatment period was observed to evaluate the safety of the drug.Results:HCV RNA turned negative in 10 of the 13 children (76.92%), within 2 weeks after antiviral treatment, in 2 cases (15.38%) HCV RNA turned negative in 2-4 weeks, in 1 case (7.69%) RBV was added when HCV RNA did not turn negative in 8 weeks, but turned negative at 10 weeks. At 12 and 24 weeks after discontinuation of antiviral drugs (SVR12), HCV RNA continued to be negative, and at 24 weeks (SVR24), it was also negative. Adverse reactions: headache occurred in 2 cases (15.38%), fatigue in 3 cases (23.08%), no serious adverse reactions occurred in any of the patients.Conclusions:The efficacy and safety of direct antiviral drugs based on SOF in the treatment of children with chronic hepatitis C were good.

Objective:To explore the status of HBV infection, immune response to hepatitis B vaccine and its influencing factors of one-year old children born to HBV infected mothers in real world.Methods:In this cross-sectional study, eligible mothers infected with chronic HBV and children who completed standard vaccination against hepatitis B vaccine and hepatitis B immunoglobulin injection were selected. Clinical biochemical, virological measurements, data of antiviral therapy and complications during pregnancy and childbirth were collected by HIS(Hospital Information System)system and LIS (Laboratoty Information Management System) system. At one year of age, the children were tested for HBsAg, HBsAb and HBV DNA in venous blood specimens.Results:A total of 1 302 eligible mothers and children were collected, including 600 in high viral load group (mothers’ HBV DNA≥2×10 5 IU/ml) and 702 in low viral load group (mothers’ HBV DNA<2×10 5 IU/ml). In high viral load group, 587 patients received antiviral drugs in the middle or late trimester (Treated group) and 13 patients did not receive antiviral drugs (Untreated group). No chronic HBV infection occurred in children of low viral load group, and in 5 cases (0.83%) the infection occurred in high viral load group. In the five HBV infected children, 3 cases (0.51%) were in the treated group, and 2 cases (15.38%) in untreated group. The failure rate of mother-to-child blocking in low viral load group was significantly lower than that in high viral load group ( χ2=5.87, P=0.015), and it was significantly lower in treated group than that in untreated group ( χ2=29.195, P=0.001). The percentages of HBsAb level <10 mIU/ml, 10- <100 mIU/ml and ≥100 mIU/ml in 1 297 children without HBV infection were 1.15%, 15.65% and 83.19%, respectively. Univariate and multivariate logistic regression analysis showed that maternal total bilirubin level and hypothyroidism during pregnancy were correlated with HBsAb level at 1 year of age ( χ2=29.003, P <0.05). Conclusions:Antiviral drugs taken during pregnancy by pregnant women with high HBV viral load can significantly reduce mother-to-child transmission of HBV. Follow-up on the response of children born to HBV-infected mothers to hepatitis B vaccine should be enhanced after birth.

Objective: To investigate the curative effect of antiviral therapy and related factors influencing the curative affect in children with immune-tolerant phase chronic hepatitis B. Methods: From May 2014 to April 2015, 46 children with chronic hepatitis B, aged 1 to 16 years with immune-tolerant phase were enrolled as the treatment group. All cases in the treated group either received interferon alpha (3-5 MIU/m², once daily) in lamivudine combination (if HBV DNA decreased < 2 log₁₀) or repeatedly received interferon-alpha alone (if HBV DNA decreased >2 log₁₀) for 12 weeks. Interferon was discontinued at 72 weeks and followed-up period was continued with lamivudine for 24 weeks. At the same time, data of 23 cases of untreated children with immune-tolerant phase chronic hepatitis B were collected as the control group. The treatment group and the control group were divided into two age groups: 1-7 years old and 7-15 years old. Data measurements were compared using t-test, analysis of variance and single factor analysis methods, and the count data were analyzed by χ ² test. Multiple logistic regression analysis was used to analyze the effects of different factors on response. Results: (1) There were 22 cases aged 1-7 years in the treatment group (47.8%) and 12 cases aged 1-7 years in the control group (52.2%). The cases of mother-to-child transmission (MTCT) in treatment and control group were 34 (73.9%) and 17 (73.9%), while children with normal baseline ALT in the treatment and control group were 18 (39.1%) and 10 (43.5%). (2) At the end of follow-up, 15 cases in the treatment group (32.6%) had HBeAg serological conversion. Among them, nine (19.6%) cases had HBsAg clearance or HB-Ag seroconversion with anti-HBs, and one (2.2%) case had HBsAg clearance, but both HBeAg and anti-HBe were positive. In the control group, one case had HBV DNA lower than the lower limit of detection level, and one case had HBeAg seroconversion without HBsAg clearance. (3) At the end of follow-up, the seroconversion rates of HBeAg in patients aged 1 to 7 years and patients aged 7 to 15 years were 45.5% and 20.8%, respectively (P = 0.078) and the clearance rates of HBsAg were 36.4% and 8.3% (P = 0.023). The serum conversion rates of normal and abnormal baseline alanine aminotransferase levels were 5.6% and 50.0% (P = 0.005), and the clearance rates of HBsAg were 5.6% and 32.1% (P = 0.077), respectively. There was no statistically significant difference in gender, mother-to-child transmission, HBV DNA genotyping and baseline HBsAg level in antiviral efficacy among children (P > 0.05). (4) HBsAg and HBeAg clearance occurred in 100% of patients at the end of follow-up who had HBsAg < 3 000 IU/ml at 24 weeks of treatment. (5) Multivariate logistic regression analysis showed that serum HBeAg conversion rate had relation with non-MTCT transmission and abnormal baseline alanine aminotransferase. Furthermore, HBsAg clearance rate was associated with the age of children. Conclusion Sequential combination of interferon and lamivudine with a prolonged course can improve the HBV DNA negative conversion rate, HBeAg seroconversion rate, HBsAg loss rate and mild ALT abnormalities at baseline in children under the age of 7 years with immune-tolerant phase chronic hepatitis B.

Objective To review and analyze the clinical and pathological data of children with autoimmune hepatitis (AIH).Methods Medical records of 46 patients hospitalized in Pediatric Liver Diseases Treatment and Research Center,Fifth Medical Center,General Hospital of People's Liberation Army(PLA) from April 2012 to April 2018 were extracted.Medical data included type of AIH,clinical manifestations,biochemical parameters,liver biopsy results,and outcomes of treatment were analyzed retrospectively.Among 46 children,19 were males and 27 were females.The age of onset was 10.1 (1.4-18.0) years old.Chi-Square test,Rank sum test or t test were used for inter-group comparison.Results There were 32 (70％)AIH-Ⅰ cases and 14 (30％)AIH-Ⅱ cases (x2=12.565,P=0.000).Among the 46 patients,there were 5 modes of onest:17 cases (37％) had acute viral hepatitis-like presentation,2 cases (4％)had fulminant hepatic failure,9 cases (20％) had insidious onset,5 cases (11％) showed cirrhosis and portal hypertension,and 13 cases (28％) were incidentally found to be due to elevated hepatic aminotransferases.Comorbidities including primary sclerotic cholangitis (n=3),primary biliary cholangitis (n=1),systemic lupus erythematosus (n=1) and inflammatory bowel disease (n=2),were all seen in AIH-Ⅰ cases.The elevated biochemical parameters of these patients were as follows:alanine aminotransferase (n=46),aspartate transminase (n=46),total bilirubin (n=35) γ-glutamyl transpeptadase (n=39),γ-globulin (n=32) and IgG (n=33).The γ-globulin and IgG levels were significantly higher in AIH-Ⅰ patients than those with AIH-Ⅱ ((32±9)％ vs.(23±8)％,t=3.217,P=0.002,(27± 10) vs.(18±8)g/L,t=3.193,P=0.003,respectively).Thirty-nine patients received liver biopsy,among whom 22 (56％) with inflammation grade (G)≥3,26(67％) with fibrosis stage (S) ≥3,and 7 with hepatic cirrhosis (S4) according to pathological analysis.Typical histopathological changes of AIH included:36 cases of interfacial hepatitis (92％),23 cases of lymphocyte/plasma cell infiltration (59％),3 cases of rosette (8％).Forty patients received prednisolone monotherapy or combined with azathioprine after diagnosis.Complete remission was seen in 29 (72％) patients,partial remission in 10 (25％) patients and no response in 1 (3％) patient.Among complete remission patients,15 (52％) had relapse in the process of prednisolone reduction.Repeated liver biopsy performed in 8 patients after treatment showed that hepatic inflammation and fibrosis were both improved in 6 patients,only inflammation was alleviated without fibrosis improvement in 1 patient,and neither inflammation nor fibrosis was improved in 1 case.The length of follow-up was 3.3 (0.3-10.5) years,and none of the 39 prednisolone-responded cases discontinued treatment successfully.Adverse effect of long-term prednisolone therapy included bilateral cataract (n=6),spinal fracture accompanied with delayed bone age development (n=1).Conclusions AIH-Ⅰ is more common than AIH-Ⅱ in children,with diverse clinical characteristics.Most cases have progressive liver inflammation and fibrosis when diagnosed.Prednisolone monotherapy or combined with azathioprine could achieve both biochemical and pathological improvement,but relapse is inevitable during drug tapering,hence long-term treatment is essential.