Donor motivation in Xi'an, China: comparison with Canadian donors.

BACKGROUND AND OBJECTIVES: In China, paid donation is prohibited by law. There is little literature assessing donor motivation in China, and comparison with western countries such as Canada is important in understanding the application of Western literature. We compared motivational factors in donors from the city of Xi'an, China, with Canadian donors matched for age, sex and donation status. MATERIALS AND METHODS: A total of 218 donors in Xi'an completed an interview about motivation as did 218 Canadian donors matched for age, sex and donation status. Frequencies and percentages of responses to questions were tabulated and compared using the Chi-squared test. RESULTS: Donors in Xi'an and Canada felt a personal responsibility to donate blood (81·2% vs. 78·0%, P = 0·2057), but Xi'an donors were more likely to consider blood donation a social responsibility (81·7% vs. 45·2%, P < 0·0001). Xi'an donors more often believed that society views donation as a normal activity (98·6% vs. 48·4%, P < 0·0001) and that the social atmosphere promotes donation (90·3% vs. 53·5%, P < 0·0001) and saw greater health benefit (52·3% vs. 12·5%, P < 0·0001). Most Xi'an donors believed in balance between their life force (Qi) and blood (86·7% vs. 49·8%, P < 0·0001) but did not believe blood lost from donating would affect this (0·5% vs. 3·8%, P = 0·01). CONCLUSION: While traditional Chinese beliefs may not be seen as a barrier among people in Xi'an who donate blood, blood donation is seen differently than by Canadian donors. There is a need for more research specific to China to tailor recruitment strategies.

Properties of hepatitis C virus minigenome containing mutated 5'UTR region and luciferase transgene.

UNLABELLED: Sequences at the 3'UTR of Hepatitis C virus (HCV) negative-strand (-)RNA play an important role in the initiation of positive-strand (+)RNA synthesis. However, the underlying mechanism in cellular context is still unclear. In this report, we designed several cDNA-based HCV-like minigenomes containing different mutations at the 5'UTR of (+)RNA. These (+)RNAs transcribed from the minigenomes in vitro were transfected into HCV replicon cells for producing (-)RNAs with deletions of different stem loops (SL) at the 3'-end. The results showed that expression of the antisense transgene from minigenome increased, when the minigenome containing deletion of SL-C1+D1+E1 at the 3'-end of (-)RNA was transfected into the HCV replicon cells compared to that of the full minigenome. The expression of the transgene from minigenome decreased using other mutant minigenomes containing deletions SL-A1, SL-A1+B1, and SL-A1+B1+C1 at the 3'-end of (-)RNA. Finally, the transgene from SL-C1+D1+E1 of (-)RNA using CMV promoter-driven minigenome was expressed at higher level than full minigenome in HCV replicon cell lines. These results indicated that the region of (-)RNA interacting with HCV replicase may locate in the SL-C1+D1+E1 region of (-)RNA. KEYWORDS: Hepatitis C virus; minigenome; RNA dependent RNA polymerase; replication.

Development of a cell-based assay for monitoring hepatitis C virus ns3/4a protease activity.

Hepatitis C virus (HCV) NS3/4A (non-structural 3 and 4 B) protease plays a key role in the processing of polyprotein precursor and it becomes an attractive target for antiviral drug discovery. We developed a cell-based assay for monitoring of the NS3/4A protease activity in mammalian cells that is an important step in screening of specific drugs against the protease. The recombinant caspase 3 (rCasp3) was used as the specific substrate for NS3/4A protease. The endogenous cleavage sites in the procaspase 3 molecule were substituted by decapeptides specific for NS3/4A protease. The activation of rCasp3 depended on its specific cleavage by NS3/4A protease and resulted in an apoptosis of stable cells expressing the protease. The difference in cell viability between the cells expressing NS3/4A protease transfected with rCasp3 and the counterparts pretreated with NS3/4A protease inhibitors could be estimated by a spectrophotometry based on 3-(4,5-dimethylthioazol- 2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) staining of cells in microplates. Thus, we developed a simple and cost-effective colorimetric assay for evaluating NS3/4A protease activity enabling the screening of candidate NS3/4A protease inhibitors.

A novel hepatitis C virus vaccine approach using recombinant Bacillus Calmette-Guerin expressing multi-epitope antigen.

Hepatitis C virus (HCV) is a major cause of liver disease worldwide. HCV infection is associated with high morbidity and has become a major problem in public health. Until now, there has been no effective prophylactic or therapeutic vaccine. BCG, a live vaccine typically used for tuberculosis prevention, has been increasingly utilized as a vector for the expression of recombinant proteins that will induce specific humoral and cellular immune responses. In this study, recombinant BCG (rBCG) was engineered to express a HCV multi-epitope antigen CtEm, and HLA-A2.1 transgenic mice were immunized with rBCG-CtEm. High levels of specific anti-HCV antibodies targeted to mimotopes of HVR1 were detected in the serum. HCV-specific lymphocyte proliferation assay, cytokine determination and cytotoxicity assay indicated that HCV epitope-specific cellular immune responses were elicited in vitro. The rBCG-CtEm immunization conferred protection against infection with the recombinant vaccinia virus (rVV-HCV-CNS) in vivo. These results suggest that rBCG expressing multi-epitope antigen may serve as an effective vaccine against HCV infection.