Objective To investigate the immunogenicity of immunodominant cytotoxic T lymphocyte epitope E749-57 of human papilloma virus (HPV) 16 oncoprotein E7 chaperoned by heat shock protein (HSP)110. Methods Mouse HSP110 gene was cloned into prokaryotic expression vector pQE-80L for the expression of HSP110 protein, which was purified using Ni-NTA column. SDS-PAGE and Western-blot were conducted to confirm the purified mHSP110 protein, which was subsequently incubated with E749-57 peptide under heat shock condition, and high-performance liquid chromatography (HPLC) was used to evaluate the binding efficiency of the recombinant protein and E749-57 peptide. Twenty mice were divided into 4 groups to be immunized with mHSP110 protein, E749-57 peptide, mHSP110-E749-57 complex and phosphate buffered saline (PBS),respectively. Two weeks after the last immunization, spleen cells were collected from the immunized mice and divided into 2 parts: one were stimulated by E749-57 peptide followed by the detection of CD8+ INF-γ+ T cells with flow cytometry; the other one were subjected to MTT analysis for the estimation of cell proliferation. The mHSP110-E749-57 complex was also used to immunize TC-1 tumor bearing mice to observe its anti-tumor effect.Results The full-length 2577 bp-sized mHSP110 gene was amplified from mouse liver cDNA and cloned into pQE-80L vector. Direct sequencing confirmed the correctness of the cloning. SDS-PAGE and Western-blot demonstrated the successful purification of mHSP110. HPLC assay showed that the purified mHSP110 protein could bind with E749-57 to form a relatively stable protein complex. The percentage of IFN-γ+ CD8+ T cells in and proliferation index of spleen cells from the complex-immunized mice were statistically higher than those from the other 3 groups of mice. Moreover, the complex could obviously inhibit the growth of TC-1 tumor in mice. Conclusion The mHSP110-E749-57 complex could enhance the generation of specific cytotoxic T lymphocytes and exert anti-tumor effects in mice.

OBJECTIVETo observe the change in IL-13 mRNA expression in rat lungs with acute lung injury (ALI) induced by lipopolysaccharide (LPS), so as to investigate the significance of anti-inflammatory mechanism in the development of ALI.

METHODS(1) Different doses (2 approximately 8 mg/kg) of LPS were injected via the tail vein to the rats to establish ALI model. (2) The IL-13 mRNA content in pulmonary tissue was determined by RT-PCR.

RESULTS(1) Different degrees of ALI was induced in the rats by different doses of LPS. ARDS was produced in the rats by LPS of 6 mg/kg. (2) The IL-13 mRNA expression in the pulmonary tissue could be enhanced by the induction of LPS, especially when its dose was larger than 6 mg/kg.

CONCLUSION(1) Rat ALI model could be produced by LPS injection via the tail vein. (2) LPS in dose of 6 mg/kg was the threshold for the production of rat ARDS. (3) The incidence of rat ARDS might be closely related to the enhanced expression of IL-13 mRNA in rat pulmonary tissue.

Acute Disease ; Animals ; Disease Models, Animal ; Female ; Interleukin-13 ; biosynthesis ; genetics ; Lipopolysaccharides ; Male ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Wistar ; Respiratory Function Tests ; Respiratory Insufficiency ; chemically induced ; metabolism ; physiopathology

Objective To compare the antibacterial activity in vitro of Haizhengmeite and Mepem. Methods Four hundreds and eighteen bacteria isolated were collected from clinical settings in different area,including 104 strains of Escherichia coli(52 strains of ESBLs +and 52 strains of ESBLs -), 104 strains of Klebsiella pneumonia(52 strains of ESBLs +and 52 strains of ESBLs -),56 strains of Proteus spp. (28 strains of ESBLs +and 28 strains of ESBLs -), 52 strains of other Enterobacteriaceae, 51 strains of Acinetobacter baumanii and 51 strains of Pseudomonas aeruginosa.Two pharmaceutical products of meropenem injection were Mepem from Japan Sumitomo Pharmaceuticals Co., Ltd and Haizhengmeite from Zhejiang Haizheng Pfizer pharmaceuticals Co.Ltd in China, respectively.Minimum inhibitory concentrations(MIC)of two products of meropenem were determined by broth microdilution method and agar dilution method according to the Clinical and Laboratory Standards Institute(CLSI,2016).Results The sensitive rates of Escherichia coli, Klebsiella pneumoniae, and Proteus spp.to Haizhengmeite and Mepem were >85%,while the rates of the sensitivity to Acinetobacter baumanii and Pseudomonas aeruginosa were lower,with the rates of 33.3%,31.4% and 58.8%,52.9%,respectively.Conclusions Haizhengmeite and Mepem both show good antibacterial activity against Enterobacteriaceae, but lower activity against Acinetobacter baumanii and Pseudomonas aeruginosa.Both products are stable to ESBLs,and no significant difference is observed between the two products in antibacterial activity in vitro.

Objective: To investigate the effect of nucleophosmin 1 (NPM1) mutant A on TGF-β1-induced K562 cell proliferation and AKT phosphorylation. Methods: K562 cells were infected with Ad5-NPM1 to create an NPM1 over-expression cell model. NPM1 levels were determined by ELISA and Western blot analysis. The levels of AKT and P-AKT were determined by Western blot. MTT was used to measure the proliferation of K562 cells. Results: NPM1 protein levels in K562 cells increased in an Ad5-NPM1-MOI-dependent manner. Cell proliferation and NPM1 levels in the supernatant were significantly increased in K562 cells infected with Ad5-NPM1-30 and Ad5-NPM1-100 compared to those infected with Ad5-vector-100 (P<0.01). Treatment with (10 ng/mL) TGF-β1 increased P-AKT levels, but not total AKT levels in K562 cells. TGF-β1-induced phosphorylation of AKT was significantly increased by infection of K562 cells with Ad5-NPM1-100. No significant differences were found in total AKT levels among all groups. TGF-β1 (10 ng/mL) treatment also in-creased the proliferation of K562 cells. TGF-β1-induced K562 cell proliferation was significantly increased by infection with Ad5-NPM1-100 (P<0.01). Conclusions: NPM1 improves TGF-β1-induced cell proliferation by up-regulating AKT phosphorylation levels.