OBJECTIVETo explore the involvement of intron A into eukaryotic expression vector to improve antigen expression efficiency and enhance immunogenicity of DNA vaccine in mice.

METHODSAs model antigen, the coding gene of mycobacterial Hsp65 was cloned into eukaryotic expression vector pCMV4.0 with intron A involved and pVAX1 without intron A involved, respectively. The resulted recombinant expression vectors were transfected into 293T cells and were then injected into BALB/c mice as DNA vaccines. Anti-Hsp65 specific IgG and isotype were detected by ELISA and T cell immune response was analyzed by enzyme-linked immunosorbent spot (ELISPOT) assay and intracellular cytokine staining.

RESULTSCompared with non-intron A pVAX1hsp65, the recombinant plasmid pCMV4.0hsp65 involved with intron A pVAX1hsp65 caused higher expression level of Hsp65 in 293T cells, and enhanced Th1 type immune response, which was defined as higher level of anti-Hsp65 specific total IgG level (3.76 ± 0.23 vs 3.15 ± 0.22, P < 0.01) and IgG2a/IgG1 ratio (4.08 ± 0.04 vs 2.23 ± 0.12, P < 0.01) and more IFN-γ-secreting CD4(+) ((2.0 ± 0.058)% vs (1.5 ± 0.087)%, t = 4.804, P < 0.01) and CD8(+) ((0.6 ± 0.058)% vs (1.0 ± 0.115)%, t = 3.098, P < 0.05) T lymphocytes. The difference showed statistical significance.

CONCLUSIONIntron A can improve the expression efficiency of mycobacterial Hsp65 antigen and enhance immunogenicity of DNA vaccine in mice when involved into eukaryotic expression vector.

Animals ; Bacterial Proteins ; genetics ; immunology ; Chaperonin 60 ; genetics ; immunology ; Female ; Genetic Vectors ; Introns ; immunology ; Mice ; Mice, Inbred BALB C ; Plasmids ; Vaccines, DNA ; genetics ; immunology

H7N9 avian influenza is the latest subtype of influenza virus to emerge in the world. By April 17, 2013 in Shanghai, a total of 31 confirmed cases were reported, and 11 of these patients died. The epidemiological characteristics and the clinical progress of this new human flu infection are still not clear. Thirteen confirmed patients have now been treated in Shanghai Public Health Clinical Center. Among the first batch of patients, hospitalised at the beginning of April 2013, two who were admitted with the same estimated date of onset of disease had very different outcomes. After active treatment at the Centre, one recovered by April 18, 2013, but one patient entered critical condition and died on April 11, 2013. The clinical and laboratory characteristics in hospital are here analysed and compared to learn more about H7N9 avian influenza. Confirmation that the observed differences are valuable for prognosis and treatment decisions for H7N9 patients awaits authentication by analysis of more patients.
Influenza in Birds ; Influenza A Virus, H7N9 Subtype ; Communicable Diseases ; Laboratories

Aged, 80 and over ; Animals ; Antiviral Agents ; therapeutic use ; Humans ; Influenza, Human ; diagnosis ; drug therapy ; virology ; Male

Objective: To investigate patients' perception of service quality at hospitals in nine Chinese cities and propose some measures for improvement. Methods: The ServQual scale method was used in a survey involving patients at outpatient and in-patient facilities in Shanghai, Chongqing, Chengdu, Nanning, Guilin and Laibin of Guangxi, Honghezhou of Yunnan, Wulumuqi of Xinjiang and Zhongshan of Guangdong. The data collected were entered and analyzed using SPSS 20.0. Statistical analyses included descriptive statistics, factor analyses, reliability analyses, productmoment correlations, independent-sample t-tests, One-way ANOVA and regression analyses. Results: The Kaiser-Meyer-Olkin value for the factor analysis of the scale was 0.979. The Cronbach's α for the reliability analysis was 0.978. All the Pearson correlation coefficients were positive and statistically significant. Visitors to out-patient facilities reported more positive perception than visitors to in-patient facilities on tangibles (t = 4.168, P < 0.001) and reliability (t = 1.979, P < 0.05). Patients of 60 years of age and above reported more positive perception than those between 40 and 49 on reliability (F = 3.311, P = 0.010), assurances (F = 2.751, P < 0.05) and empathy (F = 4.009, P = 0.003). For the five dimensions of the scale, patients in Laibin, Guangxi reported the most positive perceived service quality, followed by patients in Shanghai. On the other hand, patients in Chongqing and Nanning and Guilin of Guangxi reported relatively poor perceptions of service quality. Standardized regression coefficients showed statistically significant (P < 0.001) positive values for all ServQual dimensions. Empathy (β = 0.267) and reliability (β = 0.239) most strongly predicted perception of service quality. Conclusions: Chinese patients perceived service quality as satisfactory. Hospitals in various regions of China should enhance their awareness and ability to serve their patients.