OBJECTIVE: To investigate the effect of Aurora kinase B (AURKB) silencing-induced autophagy on apoptosis of osteosarcoma 143B cells and the underlying molecular mechanisms. METHODS: Human osteosarcoma 143B cells were transfected with Lv/shAURKB or the negative control vector Lv/shScrambled followed by treatment with chloroquine (CQ) for 24 h. Western blotting was performed to detect the protein expression levels of AURKB, P62, LC3, cleaved caspase-3, Bcl-2, and P-ULK1. Transmission electron microscopy and LC3 dual-label fluorescence method were used to trace the autophagosomes in 143B cells to assess cell autophagy, and the cell apoptosis was detected using flow cytometry and TUNEL assay. Co-immunoprecipitation assay was used to detect the interaction between AURKB and ULK1. RESULTS: The ratio of autophagy-related proteins LC3 II/I and the number of autophagosomes were significantly increased in 143B cells after transfection with Lv/shAURKB ( < 0.05), which significantly increased the expression of cleaved caspase-3 and reduced the expression of Bcl-2 ( < 0.05). Combined treatment of the cells with Lv/shAURKB and the autophagy inhibitor chloroquine obviously restored the expressions of caspase-3 and Bcl-2 ( < 0.05). Transfection with Lv/shAURKB significantly increased the apoptosis rate of 143B cells ( < 0.05), and this effect was significantly antagonized by combined treatment with chloroquine ( < 0.05). AURKB silencing strongly activated the phosphorylation of the autophagy-initiating protein ULK1 in 143B cells ( < 0.05). The results of co-immunoprecipitation assay confirmed when AURKB was immunoprecipitated, ULK1 also precipitated. CONCLUSIONS Silencing AURKB can induce autophagy by activating ULK1 phosphorylation to promote apoptosis in 143B cells.

Objective: To evaluate the efficacy and safety of Xiaoaiping combined with chemotherapy in the treatment of advanced esophageal cancer. Methods: This is a multi-center, randomized, open label and parallel controlled study. A total of 124 advanced esophageal cancer patients with Karnofsky Performance Status (KPS) score ≥60 and expected survival time≥3 months were enrolled. We adopted design and divided the patients into study and control group. The patients in study group received Xiaoaiping combined with S-1 and cisplatin. The control group received S-1 and cisplatin. Each group included 62 patients and 21 days as a treatment cycle. The efficacy and adverse events in patients of the two groups were observed and compared. Results: 57 patients in the study group and 55 in the control group were included in efficacy assessment. The response rate was 54.4% and 34.5% in the study group and control group, respectively(P<0.05). Disease control rates were 86.0% and 69.1%, respectively(P<0.05). The median progression-free survival (PFS) was 7.97 in the study group and 6.43 months in the control group(P<0.05). The median overall survival(OS) was 12.93 in the study group and 10.93 months in the control group(P<0.05). The most common adverse events in the two groups were nausea and vomiting, thrombocytopenia, anemia, neutropenia, liver damage, pigmentation, oral mucositis, renal impairment and diarrhea. The incidences of nausea, vomiting, thrombocytopenia, leukopenia, neutropenia and diarrhea in the study group were significantly higher than those in the control group(P<0.05). Conclusion Xiaoaiping combined with S-1 and cisplatin significantly increased response rate, and prolongedpatients′ survival in patients with advanced esophageal cancer.

Gas chromatography with mass spectrum detector ( GC-MSD) coupled with purge-and-trap system was set up to analyze the concentration of isoprene in natural waters. The best experimental conditions were established, including purge gas flow rate ( 50 mL/min ) , purge time ( 15 min ) , the optimum capillary column ( Rt-Alumina BOND/KCl) and the appropriate condition of temperature programming. When analyzing isoprene in natural waters, the precision was <4% (n=6), the detection limit was 0. 5 pmol/L and the recovery was 91%-102%. The preservative experiment showed that there was no obvious variation in sample concentrations of isoprene within 60 days. The concentrations of isoprene measured with the method ranged from 60 . 8 to 278 . 7 pmol/L in the Jiaozhou Bay and its adjacent river estuaries and from 44 . 7 to 77 . 2 pmol/L in Yellow River estuary, which was in good accord with those results reported in literatures in other coastal waters. In conclusion, the analytical method could meet the requirements of the analysis of concentration of isoprene in natural waters.

Objective To construct a recombinant Bacillus Calmette-Guerin ( BCG ) vaccine strain, rBCG::Rv3478-pMV261, expressing the Rv3478 protein of Mycobacterium tuberculosis and to inves-tigate its immunogenicity.Methods The gene fragments encoding Rv3478 antigen were amplified by PCR and then respectively cloned into pMV261 and pET-28a vectors to construct the recombinant expression plas-mids (Rv3478-pMV261 and Rv3478-pET-28a).The Rv3478-pMV261 plasmids were transformed into the BCG cells to construct the rBCG vaccine strains, while the Rv3478-pET-28a plasmids were transformed into Escherichia coli BL21 strains for the expression of Rv3478 protein.Polyclonal antibodies were induced in mice upon the immunization with Rv3478 protein.The rBCG vaccine strains overexpressing Rv3478 protein were screened out with Western blot assay.The C57BL/6 mice were divided into four groups including the PBS treated group, BCG treated group, rBCG::pMV261 ( R0) treated group and rBCG::Rv3478-pMV261 ( R3) treated group.All mice were sacrificed in 4 or 12 weeks after immunization.Enzyme-linked immunos-pot assay ( ELISPOT) , ELISA and flow cytometry analysis were performed to evaluate the induced humoral and cell-mediated immune responses in mice.Results The Rv3478 protein was successfully expressed and could induce polyclonal antibodies in mice.High levels of IFN-γand TNF-αwere detected in mice treated with R3, indicating that the immunization with R3 enhanced the cellular immunity.Moreover, the ratios of CD4+to CD8+T cells and the percentages of CD44+CD62L+T cells were increased in mice upon the immuni-zation with R3.Conclusion The recombinant BCG vaccine strain overexpressing Rv3478 protein could in-duce stronger cell-mediated immune responses in mice.It might be have a great significance as a new tuber-culosis( TB) vaccine strain against TB infection in the future.