Identifying Qingkailing ingredients-dependent mesenchymal-epithelial transition factor-axiation "π" structuring module with angiogenesis and neurogenesis effects.

OBJECTIVE: To explore the functional role of the drug-dependent mesenchymal-epithelial transition (Met)-axiation "π" structural module of neurogenesis after processing by three components of Qingkailing injection in neurogenesis and angiogenesis in cerebral ischemia. METHODS: We used a Glutathione S-transferase (GST)-pull down assay, isothermal titration calorimetry assay, and other related methods to identify the relationships among Met, inositol polyphosphate phosphatase like 1 (Inppl1), and death associated protein kinase 3 (Dapk3) in this allosteric module. The biological effects of the modules of neurons generation composed of Met, Inppl1, and Dapk3 were measured through Western blot, apoptosis analysis, and double immunofluorescence labeling. RESULTS: The GST-pull down assay revealed that proline-serine-threonine rich domain of Met binds to the Src homology domain of Inppl1 to form a protein-protein complex; Dapk3 with a C-terminal domain interacts weakly with the protein kinase C domain of Met in the intracellular region. Thus, we obtained a "π" structuring module considered a neural regeneration module. The biological effects of angiogenesis and neurogenesis modules composed of Met, Inppl1, and Dapk3 were also verified. CONCLUSION: The study suggested that understanding the functional modules that contribute to pharmaceutics might provide novel signatures that can be used as endpoints to define disease processes under stroke or cerebral ischemia conditions.

[Investigation of Toxoplasma gondii infection among special populations in Nanjing City, Jiangsu Province].

OBJECTIVE: To understand the prevalence of Toxoplasma gondii infection among special populations in Nanjing City, so as to provide the reference for formulating the interventions for the prevention and control of T. gondii infections in this population. METHODS: The HIV/AIDS patients, tumor patients, pregnant women, and people with livestock and poultry breeding or processing works were selected as the study subjects from September to November 2015. The venous blood samples were collected from each participant for detecting IgG and IgM antibodies against T. gondii by ELISA. RESULTS: The overall prevalence of T. gondii infection was 10.2% in the study subjects in Nanjing City. The T. gondii infection rates were 12.2%, 11.3%, 4.0%, and 13.0% among the HIV/AIDS patients, tumor patients, pregnant women, and people with livestock and poultry breeding or processing works, respectively, and there was no statistically significant difference among the four groups (χ2 = 5.668, P = 0.130). The prevalence of T. gondii infection was higher in men than in women (15.3% vs. 5.8%; χ2 = 10.213, P = 0.001), and there were significant differences in the prevalence of T. gondii infection in terms of gender (χ2 = 9.501, P = 0.023), education levels (χ2 = 9.850, P = 0.043) or occupations (χ2 = 8.983, P = 0.062). CONCLUSIONS: The infection rate of T. gondii among the special population in Nanjing City is high. Therefore, the health education intervention should be strengthened in the follow-up work for the special population.

A DNA vaccine constructed with human papillomavirus type 16 (HPV16) E7 and E6 genes induced specific immune responses.

OBJECTIVE: Cervical cancer is found highly associated with human papillomaviruses type 16 (HPV16). HPV16 E6 and E7 oncogenes are important transforming genes which have become the main focus of anti-cervical cancer therapy. In this study, a recombinant DNA vaccine candidate, termed HPV16-DNA-E6E7, constructed with HPV16 E7 and E6 genes was generated and used to against HPV16-induced tumors. METHODS: We inserted an E7 DNA fragment into E6 gene to produce a recombinant gene (E6E7-DNA). The E6E7-DNA gene was inserted into a mammalian expression vector, pcDNA 3.1+, to construct the DNA vaccine candidate. Animals (C57BL/6 mice) were immunized with the vaccine candidate with various concentrations (50 microg, 100 microg or 200 microg, respectively), and cytotoxicity measurement and tumor protection assay were carried out to examine the immunological effects of the vaccine candidate. RESULTS: Immunization of with HPV16-E6E7-DNA induced HPV16-specific immune response and also conveyed protection against TC-1 induced tumor in vivo. A survival rate (90%) after 45 days of tumor challenge was observed. The animals injected with a higher dosage of the vaccine (200 microg) exhibited prolonged survival duration of more than 55 days. No transforming activity of the vaccine candidate was detected, as determined by focus formation and degradation of endogenous p53. CONCLUSION: Our results demonstrated that the HPV16-E6E7-DNA compound might become a candidate for HPV16 precautionary and immunotherapy.