The BALB/c mice were immunized with rMS-Sj26GST and rBCG-Sj26GST vaccine in Schistosoma japonicum by subcutaneous injection. After they were immunized for 8 weeks, the eyeballs were removed to get blood and macrophages of abdominal cavity and spleen cells were harvested. The lymphocytic stimulating index (SI) was used to measure the cellular proliferating ability and NO release was used to measure the phagocytic activity of the macrophages. By using ELISA kit, the levels of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) in serum and the splenic lymphocytic cultured supernatant were detected. The results showed that after the mice were immunized with 10(6) CFU of rMS-Sj26GST and rBCG-Sj26GST vaccine separately by subcutaneous injection, proliferating ability of splenic lymphocytes in the mice showed no difference (P > 0.05), but both were significantly increased as compared with that in the control group (P < 0.05); The contents of NO in the intraperitoneal macrophages of rMS-Sj26GST vaccine group were significantly lower than in the control group (P < 0.001) and rBCG-Sj26GST vaccine group (P < 0.01); The levels of serum IL-2 in the rMS-Sj26GST vaccine group were significantly increased as compared with that in the control group (P < 0.001), vector group (P < 0.01) and rBCG-Sj26GST vaccine group (P < 0.05); The contents of serum IFN-gamma in the rMS-Sj26GST vaccine group were significantly increased as compared with that in the control group (P < 0.01) and rBCG-Sj26GST vaccine group (P < 0.05), The contents of IFN-gamma in the cultured supernatant were significantly lower than those of rBCG-Sj26GST vaccine group (P < 0.001), but were significantly increased as compared with that in the control group (P < 0.01). It was indicated that both vaccines could enhance the immune response of the mice, but rMS-Sj26GST vaccine had stronger immunogenicity than rBCG-Sj26GST vaccine.

Objective To investigate the correlation of clinicopathological parameters and prognosis with serum and pancreatic cancer tissue klotho. Methods Immunohistochemistry EnVision two step method was used to assess klotho protein expression of a tissue microarray ( TMA) of 79 pairs of pancreatic tissue and normal surrounding tissue. The serum klotho levels in 39 pancreatic cancer patients and 39 healthy controls who had matched clinical data were measured by ELISA. The relationships between the expression of klotho and the clinicopathological features and survival were analyzed. Results Klotho expression positivity in pancreatic cancer tissue was significantly higher than that in adjacent normal tissues (59. 5％ vs 96. 3％);serum level of klotho was markedly higher in pancreatic cancer patients than that in control group [(670. 30 ± 82. 24)pg/ml vs (310.35 ± 34.65) pg/ml], and both the difference was statistically significant (P<0.001). Klotho expression was negatively associated with tumor clinical stage and lymph node metastasis (P<0. 05), and the expression of klotho did not correlate with patients' gender, age, tumor size, location, local invasion depth and the like. The median survival time in pancreatic cancer patients with positive klotho expression were longer than that in in pancreatic cancer patients with negative klotho expression [(48. 31 ± 6. 94) months vs (19. 50 ±6. 78)months], and the difference was statistically significant (P<0. 01). ROC analysis on serum klotho gave a cutoff value of 376. 51 pg/ml to diagnosis pancreatic cancer with a sensitivity of 84. 6％ and specificity of 87. 2％. Conclusions Klotho level in serum and tissue of pancreatic cancer patients was closely correlated with clinicopathological parameters and prognosis, which may be a potential biomarker for pancreatic cancer.

Heat shock protein 65 (HSP65) is one of the most important protective immunogens against the tuberculosis infection. The signal sequence of antigen 85B and the whole HSP65 DNA sequence of human Mycobacterium tuberculosis (M. tuberculosis) were amplified from BCG genome and plasmid pCMV-MTHSP65 respectively by polymerase chain reactions (PCR). These two sequences were cloned into the plasmid pBCG-2100 under the control of the promoter of heat shock protein 70 (HSP70) from human M. tuberculosis, yielding the prokaryotic shuttle expression plasmid pBCG-SP-HSP65. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis showed that the two cloned DNA sequences were consistent with those previously reported, and the direction of their inserting into the recombinant was correct and the reading frame had been maintained. The recombinants were electroporated into BCG to construct the recombinant BCG vaccine and induced by heating. The induced expression detected by SDS-PAGE showed that the content of 65 kD protein expressed in recombinant BCG was 35.69% in total bacterial protein and 74.09% in the cell lysate supernatants, suggesting that the recombinant HSP65 gene could express in BCG with high efficiency and the expressed proteins were mainly soluble. Western-blot showed that the secretive recombinant proteins could specifically combine with antibody against M. tuberculosis HSP65, indicating that the recombinant proteins possess the biological activity of HSP65.
BCG Vaccine ; biosynthesis ; immunology ; Bacterial Proteins ; biosynthesis ; immunology ; Chaperonin 60 ; Chaperonins ; biosynthesis ; immunology ; Cloning, Molecular ; Escherichia coli ; metabolism ; Genetic Vectors ; Humans ; Mycobacterium tuberculosis ; genetics ; immunology ; Plasmids ; genetics ; Sequence Analysis, DNA ; Vaccines, Synthetic ; biosynthesis ; immunology

The BALB/c mice were immunized with rMS-Sj26GST and rBCG-Sj26GST vaccine in Schistosoma japonicum by subcutaneous injection. After they were immunized for 8 weeks, the eyeballs were removed to get blood and macrophages of abdominal cavity and spleen cells were harvested. The lymphocytic stimulating index (SI) was used to measure the cellular proliferating ability and NO release was used to measure the phagocytic activity of the macrophages. By using ELISA kit, the levels of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) in serum and the splenic lymphocytic cultured supernatant were detected. The results showed that after the mice were immunized with 10(6) CFU of rMS-Sj26GST and rBCG-Sj26GST vaccine separately by subcutaneous injection, proliferating ability of splenic lymphocytes in the mice showed no difference (P > 0.05), but both were significantly increased as compared with that in the control group (P < 0.05); The contents of NO in the intraperitoneal macrophages of rMS-Sj26GST vaccine group were significantly lower than in the control group (P < 0.001) and rBCG-Sj26GST vaccine group (P < 0.01); The levels of serum IL-2 in the rMS-Sj26GST vaccine group were significantly increased as compared with that in the control group (P < 0.001), vector group (P < 0.01) and rBCG-Sj26GST vaccine group (P < 0.05); The contents of serum IFN-gamma in the rMS-Sj26GST vaccine group were significantly increased as compared with that in the control group (P < 0.01) and rBCG-Sj26GST vaccine group (P < 0.05), The contents of IFN-gamma in the cultured supernatant were significantly lower than those of rBCG-Sj26GST vaccine group (P < 0.001), but were significantly increased as compared with that in the control group (P < 0.01). It was indicated that both vaccines could enhance the immune response of the mice, but rMS-Sj26GST vaccine had stronger immunogenicity than rBCG-Sj26GST vaccine.
Animals ; Antigens, Helminth ; genetics ; immunology ; BCG Vaccine ; immunology ; Escherichia coli ; genetics ; Genetic Engineering ; methods ; Genetic Vectors ; immunology ; Glutathione Transferase ; genetics ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Mycobacterium smegmatis ; genetics ; immunology ; Polymerase Chain Reaction ; Random Allocation ; Recombinant Fusion Proteins ; genetics ; immunology ; Schistosoma japonicum ; enzymology ; genetics ; immunology ; Vaccines, Synthetic ; genetics ; immunology

Objective: This study explored the relationships among psychological stress, achievement motivation and psychological capital in college students to provide a reference for improving the level of psychological capital in college students. Methods: A multi stage stratified cluster sampling method was used to select 9 940 college students from ten universities in Anhui Province. The achievement motivation scale (AMS), psychological stress scale (SRQ-20) and psychological capital scale (PPQ) were applied. The moderating effect of the questionnaire was analyzed with χ 2 tests, Spearman rank correlation and stratified regression. Results: Statistical differences were found in psychological stress according to major, whether students leader, family economic status and whether students had left behind experience ( χ 2=15.50, 10.25, 28.61, 25.55, P <0.05). The rank correlation results indicated that psychological stress was negatively correlated with the pursuit of success ( r =-0.27) and four dimensions of self efficacy,optimism,hope and resilence in psychological capital ( r =-0.43, -0.41,-0.36,-0.45)( P <0.05), and was positively correlated with the avoidance of failure ( r =0.25, P <0.05). The stratified regression model indicated that psychological stress in the dimensions of college students achievement motivation (pursuit of success: β =0.02, Δ R 2=0.01, P <0.01; failure avoidance: β = 0.03 , Δ R 2=0.01, P <0.01) played a moderating role in the relationship between psychological capital and psychological capital. Conclusion Being female, senior students, low household economic status, and left behind experience are associated with more psychological stress among college students. Psychological stress is correlated with achievement motivation and psychological capital, and has a moderating effect on the relationship between achievement motivation and psychological capital.