Objective To compare the immune protective effect of different antigens of Trichinella spiralis:excretory-secretory(ES) antigen of adult worms, ES antigen of muscle larva and their mixed antigens in mice. Methods ES antigens of Trichinella spiralis adult worms and muscle larva were produced by the physiological saline culture methods. The ES antigens of Trichinella spiralis adult worms, muscle larva and the mix were used to immunize mice 3 times at 7 day interval, adjuvant control and normal control were set up. Seven day after the final immunization, each mouse was orally challenged with 200 Trichinella spiralis larva. Intestinal adult worms and muscle larva of Trichinella spiralis of each group were recoveried and examined on Day 7 and Day 30 post-challenge,respectively. Results The intestinal adult worms of Trichinella spiralis reduced by 87.95%, 69.48% and 84.34% in the adult worm ES antigen group, muscle larval ES antigen group and their mixed antigen group, respectively, while muscle larva reduced by 74.79%, 87.97% and 86.87% respectively. Adult worm reduction rates of the adult worm ES antigen group and mixed antigen group were higher than those of muscle larval ES antigen group (P

Objective:To explore the regulatory effect of Hsf 1 on PLC/PRF5 hepatoma cells proliferation.Methods: By shRNA gene silencing technology ,constructed PLC/PRF5 hepatoma cell line of Hsf 1 gene silencing.To detect the expression of Hsf 1, p53 and Rb proteins in PLC/PRF5 hepatoma cells by Western blot.The proliferation of PLC/PRF5 cell line was observed by methylthiazolyl tetrazolium assay ( MTT ) , plate clone formation assay ( PCFA ) and cell cycle assay.Results: shRNA-Hsf1 could significantly inhibit the expression of Hsf 1 in PLC/PRF5 cells.It could induce PLC/PRF5 cells stopping at G1 phase of cell cycle , inhibit cell proliferation and colonal formation;silencing Hsf1 caused up-regulation of p53 and Rb proteins expression in PLC/PRF5 cells.Conclusion: Silencing Hsf1 is involved in up-regulation of p53 and Rb proteins expression , which results in inhibiting proliferation of PLC/PRF5 hepatoma cells.

Objective:To study the role of DR5 in TRAIL apoptotic signal transduction. Methods:BALB/C mice were immunized with recombinant DR5 that contains the full-length extracellular domain of the human DR5. Anti-DR5 mAb was generated by hybridoma. The level of DR5 expression on Jurkat cell line was examined by flow cytometry. The rates of TRAIL-induced apoptosis and anti-DR5 mAb blocking on Jurkat cells were tested by flow cytometry with TRAIL apoptosis kit.Results:The percentage of DR5 expression on Jurkat cells was 94 83%. TRAIL and anti-TRAIL mAb could kill Jurkat cells on dose-dependent, and the killing rate was 90% in the concentration of 50～100 ng/ml. The killing role of TRAIL could be blocked on Jurkat cells pretreated with anti-DR5 mAb. The average percentage of blocking was 90 49%.Conclusion:DR5 plays a very key role in TRAIL induced apoptosis in Jurkat cells.

Objective:To determine the sensitivity of SMMC-7721 cells to mDRA-6,the synergistic damage effect of mDRA-6 and adriamycin on human hepatocellular carcimoma cell lines and its possible mechanism.Methods:SMMC-7721 cells were cultured with RPMI1640 medium in regular condition.The morphology was observed by microscope.Cytotoxicity was examined by MTT assay. Apoptosis were detected by flow cytometry.Results:(1)The apoptosis of SMMC-7721 cells could be induced by mDRA-6,1.89 ?g/ml of mDRA-6 cound kill 36% of the cells;(2)Concentration-dependent cytotoxicity of adriamycin was exhibited in SMMC-7721 cells;(3)The combination of mDRA-6 and adriamycin exhibited synergistic effect on SMMC-7721 cells.2 ?g/ml of mDRA6 and 40 ng/ml of adriamycin killed 60%SMMC-7721.Conclusion:MDRA-6 can induce SMMC-7721 cell apoptosis.The combination of mDRA-6 and adriamycin exhibit synergistic effect on SMMC-7721 cells.

Objective:To observe anti-DR5 monoclonal antibody on apoptosis and CDC effect in EC109 cells.Methods:The anti-DR5 monoclonal antibody was prepared by hybridoma technique.Tumoricidal effects and complement-dependent cytotoxicity of the McAb to EC109 cells was screened by MTT assay.The apoptosis of EC109 cells was detected by flow cytometry with annexin Ⅴ-FITC/PI staining.Morphological change of EC109 was observed by microphotograph.Results:An anti-DR5 monoclonal antibody was obtained.It induced apoptosis of EC109 dose dependently.The cytotoxic action was notably enhanced by addition of complement.The cells growth inhibition ratios reached 83.04%.The apoptotic body and cathepsis were seen in microphotograph.Conclusion:The anti-DR5 monoclonal antibody could induce EC109 cells apoptosis and cause the complement dependent cytotoxic (CDC) effects powerfully.

Objective To explore the signal transduction mechanism of inhibitor kappa B kinase α( IKKα) , one of the catalytic subunits of IKK complex , for regulating p53 transactivation in the cellular ultraviolet radiation ( UVB) repsonse. Methods The transactivation of p53 was determined by dual-luciferase reporter gene analysis system while the expression and activation of IKKα, IKKβ, p53 and p38K was detected by Western blotting assay .Results UVB exposure induced activation and transactivation of p 53 in the wild type mouse fibroblasts ,but the effect was blocked by IKKa deficiency and recovered by reconstitution of IKKαexpression.Under the same conditions , IKKαregulated p38K activation, while inhibi-ting p38K activation down-regulated p53 transactivation under UVB exposure .Conclusion IKKαregulates UVB-induced phosphorylation and activation of p 53 in a p38K-dependent manner .

Objective To investigate the self-assembly and cellular localization patterns of fila-mentous temperature-sensitive protein Z (FtsZ) in Escherichia coli (E.coli) strains by using FtsZP74R, FtsZG77D and FtsZA81R mutants.Methods YFP or His labeled FtsZ proteins and the plasmids of FtsZ mu-tants were constructed by using molecular clone and site-directed mutagenesis methods.The targeted proteins were purified by affinity chromatography.FL37(△ftsZ-Cat) strains were constructed via linear DNA homol-ogous recombination.Living cell imaging was performed to observe the cellular localization patterns of FtsZ protein and its mutants in E.coli strains.The interactions between FtsZ-FtsZ/FtsZ mutants were examined by coi-mmunoprecipitation assay . The polymerization properties of FtsZ mutants were analyzed by light scattering.The activities of GTPase were monitored by using high performance liquid chromatography.Re-sults The P74, G77 and A81 amino acids were respectively replaced by different polar amino acids to change the amphipathicity of the helix within the domain of FtsZ ( 74-82 ) .The YFP-labeled FtsZP74R , FtsZG77D and FtsZA81R mutants failed to assemble into functional Z-ring structure in E.coli strains.The inter-actions between FtsZ protein and its mutants were weakened or completely disappeared.In addition, in vitro experiments showed that P74R, G77D and A81R mutations caused a decrease in the polymerization efficien-cy of FtsZ monomer.The activity of GTPase was significantly decreased in the FtsZA81R mutant. Conclusion The P74, G77 and A81 were critical amino acids in the function and assembly of FtsZ protein in E.coli strains.Moreover, A81 amino acid regulated the lateral interaction of FtsZ monomer and the activity of GTPase.

PD-1/PD-L1 signaling pathway as a T cell immune response co-stimulatory signaling pathway plays an important role in adaptive immunity. PD-1 is a major co-receptor expressing on T cells, binding with its ligands(PD-L1 and PD-L2), PD-1 can inhibit T cell activation and protect the body against the attacks from its own immune system. In addition to adjusting and maintaining autoimmune tolerance, in tumor cells PD-L1 expression is up-regulated, while in the virus-infected T cells PD-L1 expression is also upregulated. PD-1 / PD-L1 are involved in the tumor and infectious pathogen immune evasion, thus blocking the PD-1 / PD-L1 signaling pathway has become a hot research of cancer and chronic diseases. Currently, there are several anti-PD-1 or PD-L1 monoclonal antibodies approved by the FDA to enter clinical studies, which have shown significant anti-cancer effect.

AIM:To clarify the impact of heart shock factor 4b (Hsf4b) K65R mutation on the regulation of downstream protein expression .METHODS:Non-functional Lys mutant plasmid pWZL-blast-HA-Hsf4b/K65R was genera-ted by replacing single , homologous amino acids using KOD-Plus-Mutagenesis-Kit.Mouse lens epithelial mLEC stable cell lines expressing Hsf4b or Hsf4b/K65R were constructed by lentivirus infection .The expression of Hsf4b in the mutant and the wildtype mLEC cells was confirmed by Western blotting .The expression of Hsf4b downstream proteins such as heat shock protein ( Hsp)70, Hsp90, Hsp27 and CryAB was examined by Western blotting and real-time PCR.RESULTS:The results of PCR and DNA sequencing confirmed the successful construction of mLEC Hsf 4b/K65R mutant.The K65R mutation didn’t influence Hsf4b expression in the mLEC cells.After K65R mutation in Hsf4b, the expression levels of Hsp27 and CryAB were down-regulated and the expression of Hsp 70i and Hsp90a upregulated.CONCLUSION: pWZL-blast-HA-Hsf4b/K65R can be used to construct a stable cell line by infecting with lentivirus .Hsf4b/K65R mutation influ-ences the regulation of downstream heat shock proteins .

Objective:To determine the correlation of BCG concentration in Freund complete adjuvant ( FCA) and severity of arthritis during arthritis establishment in DBA 1/j male mice induced by bovine type Ⅱcollagen.Methods:CIA was induced by the im-munization of DBA1/j mice with bovine type Ⅱcollagen and BCG of various concentrations dissolved in FCA.To ascertain the effects administering the collagen booster,CIA-related features(including body weight,clinical scoring of arthritis),TNF-αin serum and the histopathological changes in the spleen and the joints regions were measured.Results:4 mg/ml BCG induced more serious arthritis than 1 mg/ml in DBA1/j mice after collagen exposure.Conclusion:There is a positive correlation of arthritis severity with BCG.Which indicates that selection of adjuvant with suitable BCG concentration could determine pathological outcome in CIA mouse model .