AIM: To investigate a specific association between hepatic steatosis and hepatitis C virus (HCV) core. METHODS: HeLa cells and primary mouse hepatocytes were transfected with HCV core plasmid, and conditional transgen...AIM: To investigate a specific association between hepatic steatosis and hepatitis C virus (HCV) core. METHODS: HeLa cells and primary mouse hepatocytes were transfected with HCV core plasmid, and conditional transgenics in which hepatic over-expression of HCV core is regulated by the tetracycline-off system, were developed. The expression of the HCV core was assessed over one to six months after withdrawal of doxycycline (dox) by immunohistochemistry (IHC) and Western blotting and by sequential liver biopsy. Hepatic steatosis was evaluated using oil red stain. 8-hydroxydeoxyguanosine (8-OHdG) stains and caspase levels were conducted to clarify hepatic oxidative stress and apoptosis rate. Serum aminotransferase was checked. RESULTS: The transfected hepatocytes had globular cores under the lipid vesicles. In transgenic mice on control diet, core expression was robust, localized to the cytoplasmic vesicle membrane and strongly associatedwith microvesicular steatosis, which was gradually replaced by macrovesicular steatosis. However, both steatosis and core positive hepatocytes diminished with time. Increases in aminotransferase, caspase and 8-OHdG were associated with peak core expression. CONCLUSION: The core protein was readily detected and morphologically associated with steatosis in individual hepatocytes both in vitro and in vivo. In vivo, oxidative stress caused by the core potentially reduced the number of core positive hepatocytes and in parallel the level of steatosis. To our knowledge, this is the f irst animal model that directly shows topological relationship between HCV core and hepatic lipid vesicles.展开更多
AIM To explore factors associated with persistent hepatitis B virus (HBV) infection in a cohort of hepatocellular carcinoma (HCC)-affected families and then investigate factors that correlate with individual viral loa...AIM To explore factors associated with persistent hepatitis B virus (HBV) infection in a cohort of hepatocellular carcinoma (HCC)-affected families and then investigate factors that correlate with individual viral load among hepatitis B surface antigen (HBsAg)-positive relatives. METHODS questionnaire. Demographics, relationship to index case, HBsAg status of mothers and index cases were evaluated for association with the HBV persistent infection or viral load by generalized estimating equation analysis. RESULTS Among 729 relatives enrolled, parent generation (P = 0.0076), index generation (P = 0.0044), mothers positive for HBsAg (P = 0.0007), and HBsAg-positive index cases (P = 5.98 x 10(-8)) were associated with persistent HBV infection. Factors associated with HBV viral load were evaluated among 303 HBsAg-positive relatives. Parent generation (P = 0.0359) and sex (P = 0.0007) were independent factors associated with HBV viral load. The intra-family HBV viral load was evaluated in families clustered with HBsAg-positive siblings. An intra-family trend of similar HBV viral load was found for 27 of 46 (58.7%) families. Male offspring of HBsAg-positive mothers (P = 0.024) and older siblings were associated with high viral load. CONCLUSION Sex and generation play important roles on HBV viral load. Maternal birth age and nutritional changes could be the reasons of viral load difference between generations.展开更多
BACKGROUND Hepatic steatosis is a major cause of chronic liver disease.Two-dimensional(2D)ultrasound is the most widely used non-invasive tool for screening and monitoring,but associated diagnoses are highly subjectiv...BACKGROUND Hepatic steatosis is a major cause of chronic liver disease.Two-dimensional(2D)ultrasound is the most widely used non-invasive tool for screening and monitoring,but associated diagnoses are highly subjective.AIM To develop a scalable deep learning(DL)algorithm for quantitative scoring of liver steatosis from 2D ultrasound images.METHODS Using multi-view ultrasound data from 3310 patients,19513 studies,and 228075 images from a retrospective cohort of patients received elastography,we trained a DL algorithm to diagnose steatosis stages(healthy,mild,moderate,or severe)from clinical ultrasound diagnoses.Performance was validated on two multiscanner unblinded and blinded(initially to DL developer)histology-proven cohorts(147 and 112 patients)with histopathology fatty cell percentage diagnoses and a subset with FibroScan diagnoses.We also quantified reliability across scanners and viewpoints.Results were evaluated using Bland-Altman and receiver operating characteristic(ROC)analysis.RESULTS The DL algorithm demonstrated repeatable measurements with a moderate number of images(three for each viewpoint)and high agreement across three premium ultrasound scanners.High diagnostic performance was observed across all viewpoints:Areas under the curve of the ROC to classify mild,moderate,and severe steatosis grades were 0.85,0.91,and 0.93,respectively.The DL algorithm outperformed or performed at least comparably to FibroScan control attenuation parameter(CAP)with statistically significant improvements for all levels on the unblinded histology-proven cohort and for“=severe”steatosis on the blinded histology-proven cohort.CONCLUSION The DL algorithm provides a reliable quantitative steatosis assessment across view and scanners on two multi-scanner cohorts.Diagnostic performance was high with comparable or better performance than the CAP.展开更多
BACKGROUND Acoustic radiation force impulse(ARFI)is used to measure liver fibrosis and predict outcomes.The performance of elastography in assessment of fibrosis is poorer in hepatitis B virus(HBV)than in other etiolo...BACKGROUND Acoustic radiation force impulse(ARFI)is used to measure liver fibrosis and predict outcomes.The performance of elastography in assessment of fibrosis is poorer in hepatitis B virus(HBV)than in other etiologies of chronic liver disease.AIM To evaluate the performance of ARFI in long-term outcome prediction among different etiologies of chronic liver disease.METHODS Consecutive patients who received an ARFI study between 2011 and 2018 were enrolled.After excluding dual infection,alcoholism,autoimmune hepatitis,and others with incomplete data,this retrospective cohort were divided into hepatitis B(HBV,n=1064),hepatitis C(HCV,n=507),and non-HBV,non-HCV(NBNC,n=391)groups.The indexed cases were linked to cancer registration(1987-2020)and national mortality databases.The differences in morbidity and mortality among the groups were analyzed.RESULTS At the enrollment,the HBV group showed more males(77.5%),a higher prevalence of prediagnosed hepatocellular carcinoma(HCC),and a lower prevalence of comorbidities than the other groups(P<0.001).The HCV group was older and had a lower platelet count and higher ARFI score than the other groups(P<0.001).The NBNC group showed a higher body mass index and platelet count,a higher prevalence of pre-diagnosed non-HCC cancers(P<0.001),especially breast cancer,and a lower prevalence of cirrhosis.Male gender,ARFI score,and HBV were independent predictors of HCC.The 5-year risk of HCC was 5.9%and 9.8%for those ARFI-graded with severe fibrosis and cirrhosis.ARFI alone had an area under the receiver operating characteristic curve(AUROC)of 0.742 for prediction of HCC in 5 years.AUROC increased to 0.828 after adding etiology,gender,age,and platelet score.No difference was found in mortality rate among the groups.CONCLUSION The HBV group showed a higher prevalence of HCC but lower comorbidity that made mortality similar among the groups.Those patients with ARFI-graded severe fibrosis or cirrhosis should receive regular surveillance.展开更多
BACKGROUND Genome-wide association studies from Asia indicate that HLA-DP and HLA-DQ loci are important in persistent hepatitis B virus(HBV)infections.One of the key elements for HBV-related carcinogenesis is persiste...BACKGROUND Genome-wide association studies from Asia indicate that HLA-DP and HLA-DQ loci are important in persistent hepatitis B virus(HBV)infections.One of the key elements for HBV-related carcinogenesis is persistent viral replication and inflammation.AIM To examine genetic and nongenetic factors with persistent HBV infection and viral load in families with hepatocellular carcinoma(HCC).METHODS The HCC families included 301 hepatitis B surface antigen(HBsAg)carriers and 424 noncarriers born before the nationwide vaccination program was initiated in 1984.Five HBV-related single nucleotide polymorphisms(SNPs)—rs477515,rs9272105,rs9276370,rs7756516,and rs9277535—were genotyped.Factors associated with persistent HBV infection and viral load were analyzed by a generalized estimating equation.RESULTS In the first-stage persistent HBV study,all SNPs except rs9272105 were associated with persistent infection.A significantly higher area under the reciprocal operating characteristic curve for nongenetic factors vs genetic factors(P<0.001)suggests that the former play a major role in persistent HBV infection.In the second-stage viral load study,we added 8 HBsAg carriers born after 1984.The 309 HBsAg carriers were divided into low(n=162)and high viral load(n=147)groups with an HBV DNA cutoff of 105 cps/mL.Sex,relationship to the index case,rs477515,rs9272105,and rs7756516 were associated with viral load.Based on the receiver operating characteristic curve analysis,genetic and nongenetic factors affected viral load equally in the HCC family cohort(P=0.3117).CONCLUSION In these east Asian adults,the mechanism of persistent HBV infection-related SNPs was a prolonged viral replication phase.展开更多
Aim:Chronic persistent hepatitis B virus carriers are generally asymptomatic until the advanced stage of the disease.The"Hepatitis B-Carrier Clinics"of Chang Gung Memorial Hospital has been using alpha-fetop...Aim:Chronic persistent hepatitis B virus carriers are generally asymptomatic until the advanced stage of the disease.The"Hepatitis B-Carrier Clinics"of Chang Gung Memorial Hospital has been using alpha-fetoprotein(AFP)and liver ultrasound for early detection of hepatocellular carcinoma(HCC)in hepatitis B surface antigen(HBsAg)carriers since 1980.Methods:We evaluated the results of surveillance between 1980 and 2012 by collecting clinic data,matched cancer registry status,and national mortality database status.Results:Of 15,235 HBsAg carriers,238 instances of HCC(1.5%or 156.2/100,000 person-years)were detected over a mean follow-up period of 10.0±7.6 years.There were more men(89.1%)and patients with liver cirrhosis(70.2%)in the HCC group(P<0.001),and both the initial and maximal alanine aminotransferase(ALT)levels were higher in this group(P<0.001).One hundred and thirty cases of HCC(54.6%)were identified during regular follow-up sessions,55(23.1%)were detected after the regular schedule had lapsed("out-of-schedule"),and 53(22.3%)were lost to follow-up completely.The mean tumor size was smaller in the regular group than in the out-of-schedule group(2.72 cm vs.4.59 cm,P<0.001),and the survival rate was higher(43.8%vs.30.9%,P<0.001).Conclusion:The incidence of HCC was relatively low in the HBsAg-Carrier Clinics cohort.Surveillance for early diagnosis of HCC improved the survival of high-risk HBsAg carriers.To ensure cost-effectiveness,we suggest using different screening strategies according to the individual risk of hepatocarcinogenesis.展开更多
Familial clustering of hepatitis B surface antigen carriers(HBsAg)and hepatocellular carcinoma(HCC)has led to the evaluation of the role of genetics in hepatitis B-related diseases.Consistent reports indicate that the...Familial clustering of hepatitis B surface antigen carriers(HBsAg)and hepatocellular carcinoma(HCC)has led to the evaluation of the role of genetics in hepatitis B-related diseases.Consistent reports indicate that the HLA-DP and-DQ loci are associated with persistent hepatitis B virus(HBV)infection.However,for hepatocarcinogenesis,existing studies have low power and conflicting data.Global single nucleotide polymorphism(SNP)data was collected from the 1000 Genomes Project and correlated with local epidemiological information.Southeastern Asia has a higher prevalence of HBsAg than Northeastern Asia;this was used in the evaluation of persistent HBV infection.The higher incidence of HCC in West Africa compared with East Africa was used in the evaluation of hepatocarcinogenesis.The allele frequencies for SNPs were significantly different between East Asians and Africans.Therefore,SNPs that have been identified in persistent HBV infections in East Asia may not be completely applicable in Africa.SNPs in NTCP,CTF19,and the HLA-DQ and-DP loci showed North-to-South allele frequency changes in East Asia.These findings confirm the role of genetics in persistent HBV infection.Some of the SNPs in the HLA loci show a trend of West-to-East allele frequency changes in Africa,indicating they may participate in hepatocarcinogenesis.Among the non-HLA related SNPs,rs2596542 in MICA shows a strong trend of allele frequency changes and is correlated with HCC incidence in Africa.SNPs in KIF1,IL-1A,and STAT4 also show,albeit with low statistical power,allele frequency trends compatible with HCC incidence.Taken together,there are strong correlations between background genetics in HLA-DP and-DQ loci with persistent HBV infection and hepatocarcinogenesis.The correlations were weak-positive in non-HLA loci.展开更多
基金grants from National Science Council, Taiwan, China. No. NSC 93-2314-B-182A-148, 94-2314-B-182A-185 and 95-3112-B-182A-002 Chang Gung Memorial Hospital, Taoyuan, Taiwan, China. No. CMRPG 33014, CMRPG 340341 and SMRPG350081
文摘AIM: To investigate a specific association between hepatic steatosis and hepatitis C virus (HCV) core. METHODS: HeLa cells and primary mouse hepatocytes were transfected with HCV core plasmid, and conditional transgenics in which hepatic over-expression of HCV core is regulated by the tetracycline-off system, were developed. The expression of the HCV core was assessed over one to six months after withdrawal of doxycycline (dox) by immunohistochemistry (IHC) and Western blotting and by sequential liver biopsy. Hepatic steatosis was evaluated using oil red stain. 8-hydroxydeoxyguanosine (8-OHdG) stains and caspase levels were conducted to clarify hepatic oxidative stress and apoptosis rate. Serum aminotransferase was checked. RESULTS: The transfected hepatocytes had globular cores under the lipid vesicles. In transgenic mice on control diet, core expression was robust, localized to the cytoplasmic vesicle membrane and strongly associatedwith microvesicular steatosis, which was gradually replaced by macrovesicular steatosis. However, both steatosis and core positive hepatocytes diminished with time. Increases in aminotransferase, caspase and 8-OHdG were associated with peak core expression. CONCLUSION: The core protein was readily detected and morphologically associated with steatosis in individual hepatocytes both in vitro and in vivo. In vivo, oxidative stress caused by the core potentially reduced the number of core positive hepatocytes and in parallel the level of steatosis. To our knowledge, this is the f irst animal model that directly shows topological relationship between HCV core and hepatic lipid vesicles.
基金Supported by grants from the Chang Gung Memorial Hospital(No.CMRPG3C0701)the National Science Council(No.NSC101-2314-B-182A-025-MY3)China Medical University(No.CMU103-N-15)
文摘AIM To explore factors associated with persistent hepatitis B virus (HBV) infection in a cohort of hepatocellular carcinoma (HCC)-affected families and then investigate factors that correlate with individual viral load among hepatitis B surface antigen (HBsAg)-positive relatives. METHODS questionnaire. Demographics, relationship to index case, HBsAg status of mothers and index cases were evaluated for association with the HBV persistent infection or viral load by generalized estimating equation analysis. RESULTS Among 729 relatives enrolled, parent generation (P = 0.0076), index generation (P = 0.0044), mothers positive for HBsAg (P = 0.0007), and HBsAg-positive index cases (P = 5.98 x 10(-8)) were associated with persistent HBV infection. Factors associated with HBV viral load were evaluated among 303 HBsAg-positive relatives. Parent generation (P = 0.0359) and sex (P = 0.0007) were independent factors associated with HBV viral load. The intra-family HBV viral load was evaluated in families clustered with HBsAg-positive siblings. An intra-family trend of similar HBV viral load was found for 27 of 46 (58.7%) families. Male offspring of HBsAg-positive mothers (P = 0.024) and older siblings were associated with high viral load. CONCLUSION Sex and generation play important roles on HBV viral load. Maternal birth age and nutritional changes could be the reasons of viral load difference between generations.
基金Supported by the Maintenance Project of the Center for Artificial Intelligence,No.CLRPG3H0012 and No.SMRPG3I0011.
文摘BACKGROUND Hepatic steatosis is a major cause of chronic liver disease.Two-dimensional(2D)ultrasound is the most widely used non-invasive tool for screening and monitoring,but associated diagnoses are highly subjective.AIM To develop a scalable deep learning(DL)algorithm for quantitative scoring of liver steatosis from 2D ultrasound images.METHODS Using multi-view ultrasound data from 3310 patients,19513 studies,and 228075 images from a retrospective cohort of patients received elastography,we trained a DL algorithm to diagnose steatosis stages(healthy,mild,moderate,or severe)from clinical ultrasound diagnoses.Performance was validated on two multiscanner unblinded and blinded(initially to DL developer)histology-proven cohorts(147 and 112 patients)with histopathology fatty cell percentage diagnoses and a subset with FibroScan diagnoses.We also quantified reliability across scanners and viewpoints.Results were evaluated using Bland-Altman and receiver operating characteristic(ROC)analysis.RESULTS The DL algorithm demonstrated repeatable measurements with a moderate number of images(three for each viewpoint)and high agreement across three premium ultrasound scanners.High diagnostic performance was observed across all viewpoints:Areas under the curve of the ROC to classify mild,moderate,and severe steatosis grades were 0.85,0.91,and 0.93,respectively.The DL algorithm outperformed or performed at least comparably to FibroScan control attenuation parameter(CAP)with statistically significant improvements for all levels on the unblinded histology-proven cohort and for“=severe”steatosis on the blinded histology-proven cohort.CONCLUSION The DL algorithm provides a reliable quantitative steatosis assessment across view and scanners on two multi-scanner cohorts.Diagnostic performance was high with comparable or better performance than the CAP.
基金Supported by the Chang Gung Memorial Hospital and PAII Inc.(a United States subsidiary company of Ping An Insurance Group),No.SMRPG3I0011.
文摘BACKGROUND Acoustic radiation force impulse(ARFI)is used to measure liver fibrosis and predict outcomes.The performance of elastography in assessment of fibrosis is poorer in hepatitis B virus(HBV)than in other etiologies of chronic liver disease.AIM To evaluate the performance of ARFI in long-term outcome prediction among different etiologies of chronic liver disease.METHODS Consecutive patients who received an ARFI study between 2011 and 2018 were enrolled.After excluding dual infection,alcoholism,autoimmune hepatitis,and others with incomplete data,this retrospective cohort were divided into hepatitis B(HBV,n=1064),hepatitis C(HCV,n=507),and non-HBV,non-HCV(NBNC,n=391)groups.The indexed cases were linked to cancer registration(1987-2020)and national mortality databases.The differences in morbidity and mortality among the groups were analyzed.RESULTS At the enrollment,the HBV group showed more males(77.5%),a higher prevalence of prediagnosed hepatocellular carcinoma(HCC),and a lower prevalence of comorbidities than the other groups(P<0.001).The HCV group was older and had a lower platelet count and higher ARFI score than the other groups(P<0.001).The NBNC group showed a higher body mass index and platelet count,a higher prevalence of pre-diagnosed non-HCC cancers(P<0.001),especially breast cancer,and a lower prevalence of cirrhosis.Male gender,ARFI score,and HBV were independent predictors of HCC.The 5-year risk of HCC was 5.9%and 9.8%for those ARFI-graded with severe fibrosis and cirrhosis.ARFI alone had an area under the receiver operating characteristic curve(AUROC)of 0.742 for prediction of HCC in 5 years.AUROC increased to 0.828 after adding etiology,gender,age,and platelet score.No difference was found in mortality rate among the groups.CONCLUSION The HBV group showed a higher prevalence of HCC but lower comorbidity that made mortality similar among the groups.Those patients with ARFI-graded severe fibrosis or cirrhosis should receive regular surveillance.
基金Supported by Chang Gung Memorial Hospital,No.CMRPG3C0701and National Science Council,No.NSC101-2314-B-182A-025-MY3 and No.MOST 107-2314-B-039-059.
文摘BACKGROUND Genome-wide association studies from Asia indicate that HLA-DP and HLA-DQ loci are important in persistent hepatitis B virus(HBV)infections.One of the key elements for HBV-related carcinogenesis is persistent viral replication and inflammation.AIM To examine genetic and nongenetic factors with persistent HBV infection and viral load in families with hepatocellular carcinoma(HCC).METHODS The HCC families included 301 hepatitis B surface antigen(HBsAg)carriers and 424 noncarriers born before the nationwide vaccination program was initiated in 1984.Five HBV-related single nucleotide polymorphisms(SNPs)—rs477515,rs9272105,rs9276370,rs7756516,and rs9277535—were genotyped.Factors associated with persistent HBV infection and viral load were analyzed by a generalized estimating equation.RESULTS In the first-stage persistent HBV study,all SNPs except rs9272105 were associated with persistent infection.A significantly higher area under the reciprocal operating characteristic curve for nongenetic factors vs genetic factors(P<0.001)suggests that the former play a major role in persistent HBV infection.In the second-stage viral load study,we added 8 HBsAg carriers born after 1984.The 309 HBsAg carriers were divided into low(n=162)and high viral load(n=147)groups with an HBV DNA cutoff of 105 cps/mL.Sex,relationship to the index case,rs477515,rs9272105,and rs7756516 were associated with viral load.Based on the receiver operating characteristic curve analysis,genetic and nongenetic factors affected viral load equally in the HCC family cohort(P=0.3117).CONCLUSION In these east Asian adults,the mechanism of persistent HBV infection-related SNPs was a prolonged viral replication phase.
基金This research was supported by the grant from Chang Gung Memorial Hospital(CMRPG3E1121 and CIRPG3H0021).
文摘Aim:Chronic persistent hepatitis B virus carriers are generally asymptomatic until the advanced stage of the disease.The"Hepatitis B-Carrier Clinics"of Chang Gung Memorial Hospital has been using alpha-fetoprotein(AFP)and liver ultrasound for early detection of hepatocellular carcinoma(HCC)in hepatitis B surface antigen(HBsAg)carriers since 1980.Methods:We evaluated the results of surveillance between 1980 and 2012 by collecting clinic data,matched cancer registry status,and national mortality database status.Results:Of 15,235 HBsAg carriers,238 instances of HCC(1.5%or 156.2/100,000 person-years)were detected over a mean follow-up period of 10.0±7.6 years.There were more men(89.1%)and patients with liver cirrhosis(70.2%)in the HCC group(P<0.001),and both the initial and maximal alanine aminotransferase(ALT)levels were higher in this group(P<0.001).One hundred and thirty cases of HCC(54.6%)were identified during regular follow-up sessions,55(23.1%)were detected after the regular schedule had lapsed("out-of-schedule"),and 53(22.3%)were lost to follow-up completely.The mean tumor size was smaller in the regular group than in the out-of-schedule group(2.72 cm vs.4.59 cm,P<0.001),and the survival rate was higher(43.8%vs.30.9%,P<0.001).Conclusion:The incidence of HCC was relatively low in the HBsAg-Carrier Clinics cohort.Surveillance for early diagnosis of HCC improved the survival of high-risk HBsAg carriers.To ensure cost-effectiveness,we suggest using different screening strategies according to the individual risk of hepatocarcinogenesis.
基金This research was supported by the grant from Chang Gung Memorial Hospital(CMRPG3F0331).
文摘Familial clustering of hepatitis B surface antigen carriers(HBsAg)and hepatocellular carcinoma(HCC)has led to the evaluation of the role of genetics in hepatitis B-related diseases.Consistent reports indicate that the HLA-DP and-DQ loci are associated with persistent hepatitis B virus(HBV)infection.However,for hepatocarcinogenesis,existing studies have low power and conflicting data.Global single nucleotide polymorphism(SNP)data was collected from the 1000 Genomes Project and correlated with local epidemiological information.Southeastern Asia has a higher prevalence of HBsAg than Northeastern Asia;this was used in the evaluation of persistent HBV infection.The higher incidence of HCC in West Africa compared with East Africa was used in the evaluation of hepatocarcinogenesis.The allele frequencies for SNPs were significantly different between East Asians and Africans.Therefore,SNPs that have been identified in persistent HBV infections in East Asia may not be completely applicable in Africa.SNPs in NTCP,CTF19,and the HLA-DQ and-DP loci showed North-to-South allele frequency changes in East Asia.These findings confirm the role of genetics in persistent HBV infection.Some of the SNPs in the HLA loci show a trend of West-to-East allele frequency changes in Africa,indicating they may participate in hepatocarcinogenesis.Among the non-HLA related SNPs,rs2596542 in MICA shows a strong trend of allele frequency changes and is correlated with HCC incidence in Africa.SNPs in KIF1,IL-1A,and STAT4 also show,albeit with low statistical power,allele frequency trends compatible with HCC incidence.Taken together,there are strong correlations between background genetics in HLA-DP and-DQ loci with persistent HBV infection and hepatocarcinogenesis.The correlations were weak-positive in non-HLA loci.
