1.The development of competency model for undergraduate medical students based on factor analysis
Yifu RU ; Shimeng BAI ; Zhou XING ; Hai XIAO ; Tao SUN ; Libin YANG
Chinese Journal of Medical Education Research 2016;15(6):541-547
Objective To explore the abilities and personal qualities that medical undergraduates need for their future clinical work, and develop their competency model. Methods The self-made ques-tionnaire survey on medical undergraduates' competencies was conducted among the 1326 medical under-graduates in a medical university by using the cluster sampling method, and the 1099 effective question-naires were collected with effective rates of 82.88%. The exploratory factor analysis was used to analyze the internal structure of competency item and to build the competency model. The rating method was used to calculate the weight of each competency item. Results The overall Cronbach's alpha coefficient of the competency questionnaire was 0.965, and the coefficient of each dimension was above 0.832. The KMO statistic value was 0.970, and the probability associated with Bartlett test statistic was P=0.000, showing that the questionnaire has high reliability and validity. The competency model of medical undergraduates covers six aspects, namely, craftsmanship, personal qualities, professional learning, interpersonal communication, psychological adaptation and the pursuit of excellence, which contain 49 factors. The cumulative variance contribution rate was 54.729%. The weights of these six aspects in the model were 0.204 4, 0.202 1, 0.175 3, 0.158 8, 0.137 6 and 0.121 8 respectively. Conclusion The competency model of undergraduate medi-cal students has a certain scientific and practical value, which can provide new evaluation methods and ideas for medical education objectives, quality assurance, teaching evaluation, medical personnel selection and training.
2.Deletion of D8L region reducing the immunogenicity of recombinant vaccinia virus vector
Ziling ZHANG ; Kangli CAO ; Shimeng BAI ; Lanlan DONG ; Tianhan YANG ; Chen ZHAO ; Jianqing XU ; Xiaoyan ZHANG
Chinese Journal of Microbiology and Immunology 2023;43(11):836-842
Objective:To reduce the immunogenicity of vaccinia virus vector by replacing the D8L region, which is a neutralizing antibody epitope in vaccinia virus, with an exogenous gene.Methods:A gene fragment encoding influenza virus hemagglutinin (HA) was inserted into the D8L region to replace it using homologous recombination technique. Then, a recombinant vaccinia virus influenza vaccine was constricted. A recombinant vaccinia virus vaccine with the TK region expressing HA was used as a control. The expression of HA was validated by Western blot. BALB/c mice were immunized with the vaccines and the serum antibody titers two weeks after each immunization were evaluated by ELISA and hemagglutination inhibition assay. The protective efficacy of the recombinant vaccinia virus was assessed through a challenge experiment.Results:Western blot confirmed the successful expression of HAD8L protein in the constructed recombinant vaccines. ELISA and hemagglutination inhibition assay showed that after the primary immunization, the anti-HA antibody titer induced by the recombinant vaccinia virus with D8L region mutation was slightly higher than that induced by the vaccine with TK region mutation, and the difference was statistically significant with the increase of immunization times ( P<0.05). The recombinant vaccinia virus with D8L region mutation showed significantly lower immunogenicity than the recombinant virus with TK region mutation after the primary immunization, but there was no significant difference between them with the increase of immunization times ( P>0.05). After H1N1pdm challenge, no virus was detected in the mice immunized with the recombinant vaccinia virus with D8L region mutation and the mice showed mild lung inflammation and less tissue damage. Conclusions:This study indicated that inserting exogenous genes into the D8L region of the neutralizing antibody epitope in the vaccinia virus vector could help to reduce the immunogenicity of the vector itself and enhance the immunogenicity of the exogenous genes. This provided a reference for the use of the vaccinia virus vector as a delivery tool in the field of vaccines or gene therapy.
3.Expression and self-assembly of HIV-1 CAP2NC protein.
Shimeng BAI ; Zhiqing ZHANG ; Jiaming QIAO ; Honglin SHEN ; Fang HUANG ; Shuangquan GAO ; Shaoyong LI ; Shaowei LI ; Ningshao XIA ; Ying GU
Chinese Journal of Biotechnology 2018;34(4):586-593
We constructed the CAP2NC prokaryotic expression vector of HIV-1 NL4-3 strain and obtained relatively pure CAP2NC protein by optimizing its purification conditions to explore its in vitro self-assembly conditions. Primers were designed according to the CAP2NC DNA sequence of HIV-1 NL4-3 strain. The target gene was amplified by PCR and cloned into prokaryotic expression vector pTO-T7. Then the recombinant strain was transformed into Escherichia coli BL21 (DE3). IPTG induced protein expression, then the protein was purified by hydrophobic chromatography. SDS-PAGE and Western blotting were performed to analyze the target protein, and the biological activity of the antigen was identified through ELISA. The self-assembly of CAP2NC protein was analyzed by transmission electron microscopy and gel filtration chromatography. The protein had good reaction with the specific antibodies of p24 and formed different structures in various conditions. When 10% yeast RNA was added to the protein complex, the recombinant protein only formed into a tubular structure, which was similar to the self-assembled structure of the HIV-1 virus capsid. The results showed that the HIV-1 CAP2NC protein had in vitro self-assembly activity, and the RNA affected the structure of CAP2NC protein assembly. The protein can be used as a simple and effective molecular model to study its structure, and then it can provide a reference for the study of HIV immature virus particles.