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Background and Aims: Hepatitis C virus (HCV) infection is reported to be associated with or to cause type 2 diabetes mellitus (T2DM). Our study aimed to elucidate the role of triglyceride (TG) and cholesterol (CHOL) levels in the association between anti-HCV seropositivity and T2DM in an HCV-endemic area., Methods: We analyzed a computerized dataset of 56 338 residents from a community-based comprehensive screening program in Tainan County in southern Taiwan. Fasting glucose, anti-HCV status, hepatitis B surface antigen (HBsAg) status, platelet counts, TG levels, CHOL levels, age, gender, and body mass index were included in the analyses. Multivariate logistic analysis was used to identify factors independently associated with T2DM., Results: Older age, being overweight, thrombocytopenia, hypertriglyceridemia, hypercholesterolemia, anti-HCV seropositivity, and HBsAg seronegativity were common factors independently associated with diabetes. Among all models of multiple logistic regression analysis used for identifying factors independently associated with T2DM, anti-HCV seropositivity was only identified in the models that included either hypertriglyceridemia or hypercholesterolemia. When subjects were divided into hyperlipidemia (CHOL, > 200 or TG, > 150 mg/dL; n = 33 393) or non-hyperlipidemia subgroups (CHOL, < 200 and TG, < 150 mg/dL; n = 22 945), anti-HCV seropositivity was identified as an independent factor only in the non-hyperlipidemia subgroup. The odds ratio was 1.35, with a 95% confidence interval of 1.17-1.55., Conclusions: This study demonstrates that the lipid level is associated with the relationship between T2DM and anti-HCV seropositivity in non-hyperlipidemic individuals. However, the relationship between HCV and T2DM did not exist when the lipid level was not included in the analysis., (© 2012 Journal of Gastroenterology and Hepatology Foundation and Wiley Publishing Asia Pty Ltd.)

Background and Aim: To evaluate the association of chronic hepatitis B virus (HBV) and hepatitis C virus (HCV) infection with hypercholesterolemia and hypertriglyceridemia., Methods: We analyzed the computerized health datasets of 56,336 residents from a community-based comprehensive screening in Tainan County in southern Taiwan. The overall prevalence rates of HBV surface antigen (HBsAg) and anti-HCV were 10.9% and 10.2%, respectively. Anti-HCV, HBsAg, platelet counts, albumin/globulin ratio (A/G ratio), fasting glucose, triglyceride, cholesterol levels, and body mass index (BMI) were abstracted for analyses. Multivariate logistic analysis was used for identification of the independent factors of hypercholesterolemia and hypertriglyceridemia., Results: The prevalence of hypercholesterolemia and hypertriglyceridemia were 48.9% and 28.0%, respectively. Hypercholesterolemia and hypertriglyceridemia were associated with each other. Older age, negativity for HBsAg and anti-HCV, normal platelet counts, A/G ratio ≥ 1, higher BMI, and being diagnosed as diabetic were common independently associated factors of both hypercholesterolemia and hypertriglyceridemia. Men had higher risk for hypertriglyceridemia, while women had higher risk for hypercholesterolemia., Conclusions: This large scale community-based study demonstrated that subjects with seropositivity for Hepatitis C not only had lower prevalence of hypercholesterolemia and hypertriglyceridemia but subjects with seropositivity for Hepatitis B had the same trend., (© 2010 The Authors. Journal of Gastroenterology and Hepatology © 2010 Journal of Gastroenterology and Hepatology Foundation and Blackwell Publishing Asia Pty Ltd.)

Background and Aim: The early detection of hepatocellular carcinoma (HCC) and opportunity to select appropriate treatment are important benefits of HCC screening. Our aim in the present study was to investigate the survival rate, prognostic factors and treatment effects in HCC patients of community-based screening., Methods: Community-based ultrasound (US) screening for HCC in adults with platelet counts (< 150 x 10(3)/mm(3)) and/or alpha fetoprotein (AFP) > 20 ng/mL was conducted in 2002 and 2004. As per the Barcelona Clinic Liver Cancer (BCLC) stage, 90 cases of intermediate or earlier stage HCC were detected and 88 cases had sufficient information for analysis (49 men and 39 women, aged 65.8 +/- 9.6 years). The tumor diameter was mostly less than 5 cm (76.1%). The follow up was continued until June 2008., Results: The 4-year overall survival rate was 46.8%. Old age (> or = 70 years) (P = 0.046), later stage of HCC (intermediate vs earlier) (P = 0.012), low platelet count (< 100 x 10(3)/mm(3)) (P = 0.013) and refusal of modern treatment (P = 0.026) were independent poor prognostic factors. Curative treatment increased survival in patients of all ages. Both curative treatment and transcatheter arterial embolization (TAE) increased survival in cases of intermediate HCC. However, treatment benefits were not found for patients with (very) early stage HCC., Conclusions: Early detection and prompt treatment of HCC leads to increased survival. For elderly patients this benefit was seen only for early stage cases receiving curative treatment. Differences between treatment types for patients with (very) early stage HCC might emerge with a longer follow-up period.

Background and Aim: This study attempted to clarify accuracy of acute hepatitis C (AHC) and its clinical characteristics., Methods: We reviewed 632 reported cases from national surveillance data of the Taiwan Center for Disease Control between 1995 and 2004, and reclassified diagnoses. A definite case was defined as alanine aminotransferase (ALT) > 10 x the upper limit of normal (ULN) with seroconversion of anti-hepatitis C virus antibody (anti-HCV). A probable case was defined as (i) seroconversion of anti-HCV and/or elevated ALT levels; or (ii) anti-HCV(+) but titers increased (from < 40 S/CO to >or= 40 S/CO) and ALT > 10 x the ULN. A suspected case was defined as initial anti-HCV(+) and ALT level > 10 x the ULN and/or jaundice. Excluded cases were defined as ALT levels less than 10 x ULN with initial positive anti-HCV Ab., Results: A total of 310 (49%) cases were confirmed as AHC; these included 95 (15%) definite and 215 (34%) probable cases. Higher incidence rates and accuracy of AHC were demonstrated in the southern area significantly if compared with northern, eastern and central areas, respectively (all P < 0.05). On comparison between blood centers and hospitals, more AHC cases were found in Southern Taiwan than in other areas (157/73 vs 24/40, P < 0.001), younger mean age (33.3 +/- 11.1 vs 49.3 +/- 16.4, P < 0.001), lower ALT levels (263.1 +/- 200.9 vs 1264.2 +/- 706.8, P < 0.001) and male predominance (191/39 vs 46/18, P = 0.046)., Conclusions: This study showed our reporting system over-estimated the AHC diagnosis, which is also a common issue worldwide. Greater efforts are needed to establish appropriate reporting systems, as well as more supplemental methods to distinguish between prevalent and incident cases.

Aim: To evaluate the potential therapeutic effect of liposomal gene delivery, genes encoding for human thymosin alpha1 (Talpha1) and interferon omega1 were injected via the tail vein into mice bearing a Hep-A-22 liver tumor., Methods: The cDNA of human Talpha1 and interferon omega1 were obtained by synthesis or reverse transcription-polymerase chain reaction (RT-PCR), respectively. Eukaryotic expressing vectors pIRES2, encoding Talpha1 and/or interferon omega1, were constructed and injected with liposome via the tail vein into ICR mice bearing a Hep-A-22 tumor. The potency of tumor inhibition was evaluated when three treated groups were compared with the group receiving the empty vector. Apoptosis of tumor cells was investigated by analyzing DNA fragmentation., Results: Only the group treated with dual-gene plasmid reached an eligible level of tumor inhibition (43%). The difference in tumor weight was statistically significant between the Talpha1 gene or the interferon omega1 gene treated groups and the control (P<0.05), and highly significant between the dual-gene treated group and the control (P<0.01). DNA ladder was observed in the tumor cells from the purpose gene treated groups but not from the control., Conclusion: The dual-gene plasmid-liposome complex showed more potent inhibition than the single gene constructs on the growth of Hep-A-22 tumor cells in mice, which may be attributed to indirect and additive induction of apoptosis in tumor cells by increased expression of Talpha1 and interferon omega1.

To investigate whether there are preferential VJC gene rearrangements of the gamma-chain of the human T-cell antigen receptor (TCR), we amplified and sequenced gamma-chain TCR transcripts from peripheral blood lymphocytes from adult normal donors. cDNA was synthesized from total RNA and amplified by the polymerase chain reaction (PCR) using 5' primers specific for either the V gamma I or the V gamma II subgroups of the gamma-chain of the TCR. The amplified cDNAs were then cloned and sequenced. The majority (approximately 83%) of the cDNAs employing V-I subgroup gene segments rearranged to J gamma 2 (J gamma 2.1 or J gamma 2.3) C gamma 2 gene segments. This was in contrast to the predominant rearrangement of the V gamma II subgroup (V gamma 9) to J gamma 1.2C gamma 1. The remaining 13% of the cDNAs employing V gamma I subgroup gene segments rearranged to J gamma 1.1C gamma 1 or J gamma 1.3C gamma 1. There was significant N diversity as well as imprecise joining at the VJ junction. gamma delta TCR utilizing the C gamma 1 gene segment are disulfide-linked, whereas those utilizing the C gamma 2 gene segment are non-disulfide-linked. These results demonstrate that peripheral blood gamma-chain transcripts exhibit preferential rearrangements of V gamma I subgroup gene segments to J gamma 2(2.1,2.3)C gamma 2 gene segments. By contrast, V gamma II subgroup (V gamma 9) transcripts exhibit rearrangements to J gamma 1.2C gamma 1.

We have developed a highly efficient new method for the amplification of alpha- and beta-chain human T-cell receptor (TCR) cDNAs. This method is designated non-palindromic adaptor polymerase chain reaction (NPA-PCR). cDNA was synthesized from total RNA isolated from mononuclear leucocytes, using either an oligo (dT)15-NotI or a C alpha-NotI or a C beta-NotI primer and RNase H-negative reverse transcriptase. The double-stranded cDNA was ligated with the non-palindromic adaptors EcoRI-XmnI [d(ATTCGAACCCCTTCG)] and XmnI G strand [d(pCGAAGGGGTTCG)] (phosphorylated), which resulted in the addition of the EcoRI-XmnI site in both 5' and 3' ends. These two non-palindromic adaptors, EcoRI-XmnI and XmnI G strand, are complementary to each other and both are required for ligation. The EcoRI-XmnI adaptor was removed from the 3' end by treatment with NotI restriction nuclease, whereas it was retained at the 5' end. The non-palindromic adaptor EcoRI-XmnI was used as the 5' amplification primer. C alpha or C beta constant region primers were used as 3' amplification primers. The amplified cDNAs were cloned and the plasmids were used to transform DH5 alpha competent cells. Over 1000 white colonies per 0.1-0.25 micrograms of total RNA or per 10,000 to 50,000 human peripheral blood mononuclear cells were obtained after amplification of either the alpha- or the beta-chain TCR cDNAs. Between 40 and 62% of the colonies (range from five donors) were positive after screening with either a C alpha or a C beta probe, located 5' to the C alpha and C beta amplification primers. A total of 50 amplified alpha- or beta-chain cDNA positive clones from two normal donors were randomly chosen and sequenced, and the sequences obtained were typical of alpha beta TCR. Two new J alpha gene segments were identified. Approximately 30% of the alpha-chain positive clones have 5' untranslated region, and most of the remaining alpha- or beta-chain TCR clones started from the initiation codon or near the 5' end. NPA-PCR has several advantages over existing PCR methods for the amplification of cDNAs with unknown or variable 5' end, such as the T-cell antigen receptors and the immunoglobulins. Among these advantages is that only one 5' end extension primer is required. Because of the large number of TCR V alpha and V beta families, a large number of different 5' end primers are required for amplification of alpha beta TCR cDNAs by conventional PCR.

We have developed two stable IL-2-dependent T-cell lines designated AKV-I and AKV-N from the enlarged spleens, respectively, of an AKV1 and an NFS mouse. Immunofluorescence staining with the appropriate monoclonal antibodies revealed that cells of the AKV-I cell line were alpha beta TCR-CD3+CD4-CD5-CD8+CD25+, whereas cells of the AKV-N cell line were alpha beta TCR-CD3+CD4-CD5+CD8-CD25+. A number of T-cell clones were developed from the AKV-I or AKV-N T-cell lines by limiting dilution and analysed by immunofluorescence. All clones tested were alpha beta TCR-CD3+CD4-CD25+. Certain T-cell clones expressed the CD5 antigen, whereas others expressed the CD8 antigen. The AKV-I cell line responded by proliferation to rIL2, rIL4, phorbol myristate acetate (PMA), PMA plus IL-4 and PMA plus PHA or Con A. In contrast, the AKV-N cell line did not respond to rIL-4 or rIL-4 plus PMA and exhibited only a modest proliferative response to PMA alone. Both AKV-I and AKV-N T-cell lines as well as a large number of T-cell clones examined were able to lyse cells of the PU5-IR murine cell line in the presence of the anti-CD3 (clone 145-2C11) MoAb, demonstrating their ability to mediate cytotoxicity in this system. Biochemical analysis of both AKV lines and a number of clones by immunoprecipitation with the anti-CD3 MoAb, followed by one-dimensional (either non-reducing or reducing) or two-dimensional (non-reducing/reducing) SDS-PAGE, revealed that the AKV lines and clones expressed a disulphide-linked dimer. Under non-reducing conditions, a band in the range of 75-85 kDa was observed and upon reduction it was resolved into two discrete polypeptide chains of 43-44 kDa and 48 kDa in certain AKV-I cells or 38 kDa and 42 kDa in certain AKV-N cells. In other T-cell clones or lines a broad band of 42-47 kDa was observed in AKV-I cells or 38-45 kDa in AKV-N cells. These results suggest the presence of different forms of disulphide-linked dimers on these cells. Northern blotting analysis using probes specific for the constant regions of the alpha-, beta-, gamma- and delta-chains of the T-cell antigen receptor revealed that all the AKV cell lines or clones tested expressed full-length alpha-, gamma- and delta-chain mRNA, whereas beta-chain mRNA was absent.(ABSTRACT TRUNCATED AT 250 WORDS)