PubMed Health⌕ Search

Biomedical subjects

D S Chen

Publications and source records attributed to D S Chen.

At least 163 records · Page 9Linked to original sources
PubMed1995

Genotypes of hepatitis C virus in Taiwan and the progression of liver disease.

The existence of four genotypes of hepatitis C virus (HCV)--types 1a, 1b, 2a, and 2b--has been suggested based on variations in nucleotide sequences of the core region. The aim of this study was to investigate the prevalence of HCV genotypes in chronic type C liver disease in Taiwan and correlate distinct genotypes to severity of liver disease. The genotypes of 175 patients with chronic type C liver disease were determined by a polymerase chain reaction with type-specific primers. The prevalence of each genotype in Taiwan was as follows: type 1a, n = 1 (0.6%); 1b, n = 125 (71.4%); 2a, n = 21 (12%); 2b, n = 6 (3.4%); mixed types, n = 18 (10.3%); and unclassified, n = 4 (2.3%). The demographic and clinical features were comparable between patients with different genotypes, except that the mean peak serum transaminase levels of patients with double viruses and type 1b HCV infections were significantly higher than were those of patients with type 2a virus. Moreover, type 1b HCV was more prevalent in patients with liver cirrhosis alone or with hepatocellular carcinoma. In conclusion, type 1b virus is the predominant genotype in chronic hepatitis C in Taiwan, and type 1b virus and mixed infection may trigger more severe liver disease.

Adult↗
PubMed1995

Prominent proliferative response of peripheral blood mononuclear cells to a recombinant non-structural (NS3) protein of hepatitis C virus in patients with chronic hepatitis C.

The proliferative response of peripheral blood mononuclear cells (PBMC) to a recombinant non-structural (NS3) protein of hepatitis C virus (HCV) was studied in 41 patients with chronic hepatitis C. Of them, 28 had chronic persistent hepatitis (CPH) and 13 chronic active hepatitis (CAH). The positive proliferation rate of PBMC to the recombinant NS3 protein, T9Ag, was 66% in the 41 patients (77% in CAH versus 61% in CPH; P > 0.05) when stimulation index (SI) = 4 was set as the cut-off value. However, mean SI of CAH patients was significantly higher than that of CPH patients (8.3 +/- 5.2 versus 5.1 +/- 3.6; P < 0.05). Six other chronic hepatitis patients who were repeatedly negative for anti-HCV antibody but positive for serum HCV RNA also had an SI of > or = 4.0. The frequency of cellular immune response to the T9Ag is among the highest results obtained by using HCV antigens tested so far. Our studies thus indicate that NS3 is an immunologically important region of HCV for T cells. Moreover, the proliferative response to T9Ag may help to establish hepatitis C etiology in chronic hepatitis patients who are seronegative with currently available anti-HCV assays.

Adult↗
PubMed1995

Prevalence, genotypes and antibody titer of hepatitis C virus in pregnant women in Taiwan.

OBJECTIVES: To investigate the prevalence and genotypes of hepatitis C virus (HCV) in pregnant women in Taiwan, as well as to examine whether any correlation occurs between HCV genotype and anti-HCV titer. METHODS: Forty-three pregnant women with positive anti-HCV and HCV-RNA were selected among 3,400 cases screened from January 1992 to March 1994. Each blood specimen was assayed for HCV genotypes by PCR method to detect HCV I, II, III and IV. Anti-HCV titer was determined by a second-generation EIA kit with serial dilutions. RESULTS: Twenty-eight cases (65%) belonged to HCV II, 11 (26%) HCV III, 3 (7%) HCV IV and 1 (2%) HCV II+III. Besides, the anti-HCV titers in HCV II and HCV III groups were similar, ranging from 8x to 40,000x and revealing no statistical significance (p = 0.75). CONCLUSIONS: Our data have verified that the prevalence rate of HCV infection in pregnant women is around 1.3% and no significant difference of anti-HCV titer occurs between HCV II and HCV III pregnant women.

Base Sequence↗
PubMed1995

Large hepatitis delta antigen in packaging and replication inhibition: role of the carboxyl-terminal 19 amino acids and amino-terminal sequences.

Hepatitis delta virus (HDV) encodes two proteins, the small delta antigen (SHDAg) and large delta antigen (LHDAg). The latter is identical to the former except for the presence of additional 19 amino acids at the C terminus. While SHDAg is required for HDV replication, LHDAg inhibits replication and, together with hepatitis B surface antigen (HBsAg), is required for the assembly of HDV. The last 19 C-terminal amino acids of LHDAg are essential for HDV assembly. Most of LHDAg (amino acids 19 to 146 and 163 to 195) had been shown to be dispensable for packaging with HBsAg. To discern whether the last 19 C-terminal amino acids solely constitute the signal for packaging with HBsAg, we constructed two LHDAg deletion mutants and tested their abilities to be packaged with HBsAg in cotransfection experiments. We found that deletion of amino acids 2 to 21 and 142 to 165 did not affect LHDAg packaging. This result suggested that only the last 19 C-terminal amino acids of LHDAg are required for packaging. We further constructed two plasmids which expressed c-H-ras with or without additional 19 C-terminal amino acids identical to those in LHDAg. Only c-H-ras with additional 19 amino acids could be cosecreted with HBsAg in the cotransfection experiment. This result confirmed that the C-terminal 19 amino acids are the packaging signal for HBsAg. We also tested the trans activation activity and trans-dominant inhibitory activity of the deletion mutants of SHDAg and LHDAg, respectively. In contrast to deletion of amino acids 142 to 165, deletion of amino acids 2 to 21 impaired the trans-dominant inhibitory activity of LHDAg. Deletion of amino acids 2 to 21 and 142 to 165 did not affect the trans activation activity of SHDAg. This result suggested that a functional domain which is important for the trans-dominant inhibitory activity of LHDAg exists in the amino terminus of HDAg.

Antigens, Viral↗
PubMed1995

The significance of spontaneous hepatitis B e antigen seroconversion in childhood: with special emphasis on the clearance of hepatitis B e antigen before 3 years of age.

To investigate the significance of spontaneous hepatitis B e antigen (HBeAg) seroconversion during childhood, 415 hepatitis B surface antigen (HBsAg) carrier children (ages 0 to 15 years) were prospectively followed for 7.1 +/- 2.9 years. Hepatitis B virus (HBV) markers and liver function profiles of each child were tested at least once every 6 months. Among them, 50 were initially anti-HBe positive and 140 seroconverted from HBeAg to anti-HBe during follow-up. Before HBeAg seroconversion, jaundice occurred in 9 and alanine transaminase (ALT) activities elevated in 99 of the 140 seroconverters. Serum ALT returned to normal in all patients within 1 to 5 years of seroconversion. Six had reelevated ALT later after seroconversion. Only 7 (9.7%) of the 72 carrier infants seroconverted before 3 years of age. The peak ALT levels in five of them exceeded 100 IU/L, and two had jaundice before HBeAg seroconversion. One of the early seroconverters developed hepatocellular carcinoma (HCC) at 11 years of age, although his liver function profiles remained normal after HBeAg seroconversion. Liver biopsy specimens from 30 children during the anti-HBe-positive stag e showed inactive cirrhosis in 2 (including one with HCC), chronic hepatitis with marked fibrosis in 1, mild activity and moderate fibrosis in 2, mild activity and mild fibrosis in 9, and minimal histologic changes in the remaining 16. Although most will achieve a normalization of ALT and inactive liver histologic changes, the seroconversion of HBsAg carrier children from HBeAg to anti-HBe is not necessarily an indicator of favorable prognosis; a small proportion of children will develop cirrhosis or even HCC.

Adolescent↗
PubMed1995

Hepatitis C virus in chronic liver disease and hepatocellular carcinoma in Taiwan.

Hepatitis B virus (HBV) has long been known to be the major etiologic factor of chronic liver diseases and hepatocellular carcinoma (HCC), and in Taiwan 80-90% of chronic liver diseases and HCC are caused by HBV. Assays for antibody to hepatitis C virus (HCV) and to detect its viral genome (HCV-RNA) have revealed HCV as the next most common cause of these diseases in Taiwan. The prevalence of antibody to HCV (anti-HCV) in hepatitis B surface antigen (HBsAg)-negative patients is around 70-80%, and most of the patients are viremic. Anti-HCV is found in 0.5-1.0% of healthy adults. The epidemiology of HCV infection in Taiwan is similar to other areas of the world, with horizontal transmission as the major route of infection. Blood transfusion was an important route of transmission, accounting for 30-40% of chronic HCV infection. After screening for anti-HCV in blood donors was instituted, this infection route was effectively controlled. The nucleotide sequences of a Taiwanese isolate of HCV are similar to Japanese isolates (homology of > 90%) and less similar to the prototype U.S. isolate (78% homology). The predominant genotype is type II/lb, being detected in 66-71% of patients with chronic hepatitis C and in 83% of those with cirrhosis or HCC. Analysis of serum HCV cDNA levels by competitive polymerase chain reaction showed that the levels ranged from 10(1) to 10(7) copies/ml and did not correlate with the gender of the patients, past blood transfusion, serum aminotransferase activities, or histologic severity. However, the serum HCV levels were higher in patients with genotype II/lb than those with type III/2a or type IV/2b (p < 0.005), indicating genotype as an important determinant of levels of HCV viremia. Mixed infections of multiple genotypes of HCV may contribute to the acute exacerbations of chronic hepatitis C; among 20 patients with exacerbations, 11 (55%) had evidence indicating the emergence of a different predominant HCV genotype; among 26 without exacerbations, this was found in only 2 (8%) (p < 0.005). The incidence of HCC was studied by prospective follow-up of patients with cirrhosis by regular hepatic ultrasound examinations and serum alpha-fetoprotein surveillance in the following four groups: i) HBsAg-positive, 300 patients; ii) anti-HCV positive, 151 patients; iii) both positive, 144 patients; and iv) both negative, 62 patients. Each year, 3-5% developed HCC, and the difference in incidence between the four groups was not statistically significant. The mean age when HCC was detected was 56 +/- 10, 63 +/- 9, 55 +/- 11 and 60 +/- 14 years in each group, respectively. The results indicate a high incidence of HCC in cirrhotic patients in Taiwan, whether the cirrhosis was related to HBV or HCV; dual infections of both viruses did not accelerate the occurrence of HCC. Although most anti-HCV-positive patients with HCCs had cirrhosis, HCC did occur in some patients without cirrhosis. Studies of these two groups of HBsAg-negative, anti-HCV-positive HCC patients revealed less frequent detection of HCV-RNA in serum and lower titers of HCV RNA in HCC without cirrhosis. In fact, 10/20 (50%) non-cirrhotic HCC patients were actually positive for serum HBV DNA by PCR, indicating the possible role of HBV in the etiology of HCC in these patients.

Carcinoma, Hepatocellular↗
PubMed1994

Frequent genetic alterations at the distal region of chromosome 1p in human hepatocellular carcinomas.

Cytogenetic analysis of hepatocellular carcinoma (HCC) cell lines and primary HCC tissues has demonstrated chromosome 1p to be the region most commonly affected. To refine the altered locus, genetic abnormalities of this region were surveyed systemically by microsatellite polymorphism analysis. Twelve sets of primers evenly distributed on chromosome 1p which can amplify di- or tetranucleotide repeat length polymorphism by polymerase chain reaction were selected. The results were then supplemented by the conventional restriction fragment length polymorphism study. A comparison of the allele patterns between 30 pairs of HCC and their corresponding nontumor DNAs discovered chromosome 1p aberrations in 15 of 30 tumors (50%). The abnormalities can be classified into three groups. The first aberration was typical loss of heterozygosity that was found in 9 HCCs (30%). The second aberration was a 2-3-fold increase of allelic dosage, which was detected in 6 HCCs (20%). The third aberration was the novel microsatellite polymorphism, which was detected in 3 cases (10%). These abnormalities seemed to cluster at the distal part of chromosome 1p, with a common region mapped to 1p35-36, which is also the region with frequent loss of heterozygosity in neuroblastoma and colorectal and breast cancers. Therefore, loss of putative tumor suppressor gene(s) in this locus may participate in the development of hepatocellular carcinoma and a wide range of human cancers.

Carcinoma, Hepatocellular↗
PubMed1994

Transfusion-transmitted human T-cell lymphotropic virus type I infection in Taiwan: a true risk and occasional coinfection with hepatitis C virus shown in a prospective study.

To study the incidence of human T-cell lymphotropic virus (HTLV) after blood transfusion in Taiwan, serum samples from 699 patients in a prospective study were examined for seroreactivity of anti-HTLV. By an enzyme immunoassay, 9 of the 699 recipients were repeatedly positive. Serial serum samples of these 9 patients were then confirmed with a Western blot analysis and with a polymerase chain reaction (PCR) assay for HTLV-I genome. Four were already positive for anti-HTLV before transfusion, 1 carried antibodies to HTLV-I transiently after transfusion, and only 4 cases had de nova seroconversions. These patients and their family members were called back and tested for HTLV-I genome in the peripheral blood mononuclear cell (PBMC) and plasma. All the serologically positive patients, except the "transient one," were positive for HTLV sequences in the PBMCs. Viral sequences could also be detected in several serum or plasma samples. In the family members, only the spouse of a pretransfusion-positive patient was infected. These results suggested that approximately 0.6% of the blood recipients were infected by HTLV-I through transfusion in Taiwan, and that the frequency of intrafamilial HTLV-I transmission is low. We also observed the unusual coinfection by both HTLV-I and hepatitis C virus in 2 patients, and superinfection of hepatitis C virus after blood transfusion in 1 HTLV-I carrier. Cases of coinfection suggest a prevalence of both viruses in blood donors and warrant further screening.

Adolescent↗
PubMed1994

Isoprenylation of large hepatitis delta antigen is necessary but not sufficient for hepatitis delta virus assembly.

Hepatitis delta virus (HDV) encodes two proteins, the small hepatitis delta antigen (SHDAg) and large hepatitis delta antigen (LHDAg). Both proteins are identical except for the presence of additional 19 amino acids at the C terminus of LHDAg. While SHDAg is required for HDV RNA replication, LHDAg inhibits replication and is required together with hepatitis B surface antigen for the assembly of HDV. The C-terminal last 4 amino acids of LHDAg (Cys-Arg-Pro-Gln) is an isoprenylation motif. It has previously been shown that the mutation of the Cys inhibited the assembly of HDV. In order to discern whether this effect is due to change of amino acid residue or abolition of isoprenylation, we constructed several LHDAg mutants of the terminal three amino acid residues and tested their abilities to be packaged with HBsAg by cotransfection experiments. We also made GST-fusion proteins of these mutants and tested their abilities to be isoprenylated in rabbit reticulocyte lysate system. We found that some, but not all, of the substitutions of the amino acid residues other than the Cys also inhibited isoprenylation and that the status of isoprenylation of these mutant proteins correlated well with their abilities to be packaged with HBsAg into virions. This result indicates that isoprenylation, rather than the primary amino acid sequence, is required for LHDAg packaging. Furthermore, we found that the attachment of an isoprenylation motif to SHDAg did not enable it to be packaged with HBsAg and that the deletions of any 5 amino acids in the last 15 amino acids (amino acids 196 to 210) unique to the LHDAg abolished the packaging ability. In contrast, the deletion of 33 amino acids (amino acids 163 to 195) upstream of the last C-terminal 19 amino acids of LHDAg did not interfere with its packaging ability. Therefore, we conclude that the 15 amino acids upstream of the isoprenylation site of LHDAg are also essential for HDV assembly, and a large portion of the alleged C-terminal Pro/Gly-rich region (amino acids 146 to 195) is not required for the assembly process.

Amino Acid Sequence↗
PubMed1994

Allelotype and loss of heterozygosity of p53 in primary and recurrent hepatocellular carcinomas. A study of 150 patients.

BACKGROUND: The allelotype and loss of heterozygosity (LOH) of the p53 gene in human hepatocellular carcinoma (HCC) were studied in 150 patients with resected primary HCC and 18 with recurrent HCC. METHODS: DNA samples of paired HCC and livers were cut with BanII enzyme for the study of p53 allelotype and allele loss. The medical records of the patients were carefully reviewed. RESULTS: Sixty-four (42.7%) patients were heterozygous for the p53 gene, 69 (46%) were homozygous for the 1.5/1.4 kb small (S) allele, and 17 (11.3%) were homozygous for the 2.9 kb large (L) allele. The frequencies of the minor L allele (0.323) and of the major S allele (0.677) in this population of Chinese patients differed from the frequencies previously reported for North American Caucasians (0.13 and 0.87, respectively). The heterozygous patients tended to have lower serum hepatitis B surface- and e antigens (HBsAg and HBeAg) and higher diabetes mellitus (DM) than did homozygous patients (SS and LL). Thirty-seven (57.8%) of the 64 heterozygous patients had a tumor-specific p53 allele LOH, being two times more common in HCC tumors larger than 8 cm than in HCC tumors 2 cm or smaller. The frequency of DM was four times higher in the heterozygous patients who had p53 LOH than in those who retained both alleles. LOH of p53 did not correlate with tumor invasiveness or differentiation, hepatitis B or C virus infection, or prognosis. CONCLUSION: The allelotype of p53 gene in HCC correlates with HBsAg and HBeAg seropositivities and DM. LOH of the p53 gene is a common event in HCC, correlates with DM, and occurs less often in familial HCC. LOH can identify the clonal origin of recurrent HCC but is not a critical prognostic factor.

Adult↗
PubMed1994

Genotypes of hepatitis C virus in chronic liver disease in Taiwan.

The genotypes of hepatitis C virus (HCV) infection in 81 patients with liver cirrhosis (LC) or hepatocellular carcinoma (HCC) were investigated by the polymerase chain reaction using type-specific primers. All the patients were positive for HCV RNA in the serum. Forty-two patients had LC with HCC, while the remaining 39 patients had LC without HCC. Genotype II was detected in 47 samples (58.0%), type III in 6 samples (7.4%), and type IV in 4 (6.2%). No evidence of genotype I was found. Mixed infection was detected in 11 samples (13.6%). The prevalence of genotype II in LC with HCC patients (69.0%) was significantly higher (P < 0.05) than in the LC without HCC patients (46.2%). It is concluded that genotype II is the most predominant type in patients with LC or HCC in Taiwan, and is found more frequently in patients who had LC with HCC than in those who had LC alone.

Adult↗
PubMed1994

Amniocentesis in mothers who are hepatitis B virus carriers does not expose the infant to an increased risk of hepatitis B virus infection.

Sixty-seven pairs of mothers with hepatitis B virus (HBV) surface antigen (HBsAg) and their infants were divided into two study groups to determine the effect of amniocentesis on intrauterine HBV infection. In the first study group (35 pairs), the infant's HBsAg status in cord blood was studied and the results were compared with those obtained in the cord blood from 65 infants born to HBsAg-positive women who did not have an amniocentesis. In the second study group (32 pairs), the HBV status of the infants was studied at the age of three months to five years and compared with the HBV status of 3,454 infants in the National HBV Prevention Program. In the first study group, one sample (2.9%) was weakly positive for HBsAg; while in the first control group, two (3.1%) were positive. In the second study group, three (10%) infants were positive for HBsAg. The failure rates of immunoprophylaxis in the second study and control groups were similar (9.4% vs 11% for HBsAg carrier mothers; 30% vs 14% for HBe antigen-positive carrier mothers). This suggested that genetic amniocentesis did not increase the risk of intrauterine HBV infection.

Adult↗
PubMed1994

Fibrosing cholestatic hepatitis in a hepatitis B surface antigen carrier after renal transplantation.

A 45-year-old hepatitis B surface antigen carrier had an allograft kidney transplantation and maintenance immunosuppression with cyclosporin A and prednisolone. Six months later, she experienced a rapidly progressive hepatic failure manifested by elevation of serum bilirubin level, prolongation of prothrombin time, and mild to modest increase of serum aminotransferase levels. She died in 6 weeks. Postmortem liver histology showed canalicular and cellular cholestasis and ground-glass appearance and ballooning of most hepatocytes, but only mild inflammatory cell infiltration. Immunohistochemical staining showed massive loads of hepatitis B surface and core antigens in the hepatocytes and extensive periportal fibrosis. The whole picture was compatible with fibrosing cholestatic hepatitis described in hepatitis B virus-infected liver transplant. Sequencing of the hepatitis B virus genome amplified from the patient's serum indicated a precore mutant but few mutations in the core, pre-S, and S genes. Little inflammatory reaction was observed histologically despite HLA compatibility, a situation differing from that in liver transplant. This observation indicates that fibrosing cholestatic hepatitis may also occur in non-liver transplant setting.

Base Sequence↗
PubMed1994

Inverse polymerase chain reaction for cloning cellular sequences adjacent to integrated hepatitis B virus DNA in hepatocellular carcinomas.

Integration of hepatitis B virus (HBV) DNA is found in most HBV-related human hepatocellular carcinomas (HCCs). In the past, construction of genomic libraries was mainly employed to study the role of viral integration. However, large amounts of tissue DNA and a laborious screening procedures were required. Inverse polymerase chain reaction (IPCR) is based on the simple procedures of digestion of DNA with restriction enzymes and circularization of cleavage products before amplification using primers synthesized in the opposite orientations to those normally employed for PCR. This technique allows the in vitro amplification of DNA flanking a region of known sequence. By employing this method, starting from nanograms of hepatoma DNA, two adjacent cellular sequences were cloned from 11 HBV integrants in three HCCs. The original configurations in the chromosomes were further confirmed. One of the flanking cellular sequences was identified as the human 28S rRNA gene, the other was not found homologous to any known human sequences. This method appears to be practical and can be improved further to clone more flanking cellular sequences, especially in early and small HCCs.

Base Sequence↗
PubMed1994

Posttransfusion hepatitis C virus infection in children.

Hepatitis C virus (HCV) infection was investigated retrospectively and prospectively in children who had open heart surgery. The retrospective study included 196 patients who had open heart surgery 2 to 6 years before enrollment and were regularly followed. Antibody to HCV was detected in eight cases, and seven of these eight patients had viremia caused by HCV infection. The other patient's anti-HCV titer gradually decreased and HCV ribonucleic acid was absent throughout the course. Four of these eight had persistent viral replication without clinical evidence of hepatitis; the other three had viremia and chronic hepatitis. The HCV infection rate in the retrospective study group was about 4%. The HCV was of genotype 2 in five cases and of genotype 3 in two cases. The clinical outcome was not related to sex, age, amount of transfusion, or HCV genotype. For the prospective study, we enrolled 94 patients. Of the 56 enrolled after the initiation of anti-HCV screening in the blood bank, none was infected by HCV. Of the 94 patients, 38 were enrolled before screening; 4 had abnormal liver function 1 to 3 months after operation, and 2 were infected with HCV. One patient had an acute but resolving course; the other had persistent anti-HCV and HCV viremia, although the liver function test results returned to the normal range. The HCV in both cases belonged to genotype 2. We conclude that in Taiwan the seroconversion rate of HCV in children who underwent open heart surgery was 4% to 5%. Anti-HCV screening in donor blood significantly decreased the incidence of posttransfusion hepatitis C.

Acute Disease↗