Stable replication of the EBNA1/OriP-mediated baculovirus vector and its application to anti-HCV gene therapy

<p>Abstract</p> <p>Background</p> <p>Hepatitis C virus (HCV) is one of the main causes of liver-related morbidity and mortality. Although combined interferon-α-ribavirin therapy is effective for about 50% of the patients with HCV, better therapies are needed and preventative vaccines have yet to be developed. Short-hairpin RNAs (shRNAs) inhibit gene expression by RNA interference. The application of transient shRNA expression is limited, however, due to the inability of the shRNA to replicate in mammalian cells and its inefficient transduction. The duration of transgene (shRNA) expression in mammalian cells can be significantly extended using baculovirus-based shRNA-expressing vectors that contain the latent viral protein Epstein-Barr nuclear antigen 1 (EBNA1) and the origin of latent viral DNA replication (OriP) sequences. These recombinant vectors contain compatible promoters and are highly effective for infecting primary hepatocyte and hepatoma cell lines, making them very useful tools for studies of hepatitis B and hepatitis C viruses. Here, we report the use of these baculovirus-based vector-derived shRNAs to inhibit core-protein expression in full-length hepatitis C virus (HCV) replicon cells.</p> <p>Results</p> <p>We constructed a long-term transgene shRNA expression vector that contains the EBV <it>EBNA1 </it>and <it>OriP </it>sequences. We also designed baculovirus vector-mediated shRNAs against the highly conserved core-protein region of HCV. HCV core protein expression was inhibited by the EBNA1/OriP baculovirus vector for at least 14 days, which was considerably longer than the 3 days of inhibition produced by the wild-type baculovirus vector.</p> <p>Conclusion</p> <p>These findings indicate that we successfully constructed a long-term transgene (shRNA) expression vector (Ac-EP-shRNA452) using the EBNA1/OriP system, which was propagated in <it>Escherichia coli </it>and converted into mammalian cells. The potential anti-HCV activity of the long-term transgene (shRNA) expression vector was evaluated with the view of establishing highly effective therapeutic agents that can be further developed for HCV gene therapy applications.</p

Supplemental Material - Can Computerized Simulation be Used to Assess Surgical Proficiency in Laparoscopic Colorectal Surgeries? A Systematic Review

Supplemental Material for Can Computerized Simulation be Used to Assess Surgical Proficiency in Laparoscopic Colorectal Surgeries? A Systematic Review by Si Yuan Chang, Kai Siang Chan and Aung Myint Oo in Surgical Innovation</p

Measuring health literacy in Asia: Validation of the HLS-EU-Q47 survey tool in six Asian countries

BACKGROUND: Health literacy has been increasingly recognized as one of the most important social determinants for health. However, an appropriate and comprehensive assessment tool is not available in many Asian countries. This study validates a comprehensive health literacy survey tool European health literacy questionnaire (HLS-EU-Q47) for the general public in several Asian countries. METHODS: A cross-sectional survey based on multistage random sampling in the target countries. A total of 10,024 participants aged ≥15 years were recruited during 2013-2014 in Indonesia, Kazakhstan, Malaysia, Myanmar, Taiwan, and Vietnam. The questionnaire was translated into local languages to measure general health literacy and its three domains. To evaluate the validity of the tool in these countries, data were analyzed by confirmatory factor analysis, internal consistency analysis, and regression analysis. RESULTS: The questionnaire was shown to have good construct validity, satisfactory goodness-of-fit of the data to the hypothetical model in three health literacy domains, high internal consistency (Cronbach's alpha >0.90), satisfactory item-scale convergent validity (item-scale correlation ≥0.40), and no floor/ceiling effects in these countries. General health literacy index score was significantly associated with level of education (P from <0.001 to 0.011) and perceived social status (P from <0.001 to 0.016), with evidence of known-group validity. CONCLUSIONS: The HLS-EU-Q47 was a satisfactory and comprehensive health literacy survey tool for use in Asia. Copyright © 2016 The Authors. Production and hosting by Elsevier B.V. All rights reserved