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 analyze the association between β2-AK 27 locus genetic polymorphisms and asthma, and the protective effect in airway hyperreactivity. Methods The allel polymerase chain reaction were used to determine β2-AR 27 locus genetic polymorphisms in 149 patients with cough variant asthma who have the airway hyperreactivity. To observe these people for two years in order to know the proportion of changed to typical asthma. And compare with 90 people in healthy group. Results (1) The distribution frequency of β2-AR 27 locus genetic polymorphisms is major in heterozygote (57 % ) , and the Glu/Glu homozygote has the least ( 20% ) , (2) There was a significant decrease in the frequency of Glu/Glu genotype in asthmatics compared with healthy group(9% VS 20% ) ,OR = 0.4(P<0.05) ,95% CI (0.2 ～0. 9) ,but there was no significant difference in the allele frequency of asthmatics compared with healthy group,(3)The frequency of Glu/Glu genotype in severe asthma was lower than stable asthma group(P<0. 05). Conclusion These results suggesteded that β2-AR 27 locus genetic polymorphisms is correlated with asthma,and the Glu27 could have the protective effect to the airway.

Objective To study the FMS-like tyrosine kinase-3 (FLT3) gene, NPM1 gene and c-kit gene mutations in acute myeloid leukemia (AML) by extracting DNA from the storage of bone marrow slides, and to investigate the relationship between the three gene mutations and clinical features in AML. Methods The bone marrow slides of 55 patients diagnosed with AML were enrolled in this study. The PCR, DNA sequencing and molecular cloning were used to detect and analyse the FLT3-ITD, NPM1 and c-kit gene mutations. Patients' remission, progression and survival time were also recorded. Results The DNA was successfully extracted from the bone marrow slides with -20 ℃ frozen storage without Wright stained, chemically fixed, and room temperature storage Wright stained discoloured by phenol ∶ chloroform ∶ isoamyl alcohol method, which can be used in PCR, direct sequencing and molecular cloning sequencing analysis. 10 of the 55 cases (18.2 %) were FLT3-ITD positive, including 9 cases with heterozygous mutations and 1 case with homozygous mutation. FLT3-ITD positive group had lower complete remission (CR) rate, shorter event-free survival (EFS) time and overall survival (OS) time than the negative group (P< 0.05). 9 of the 55 cases (16.4 %) had NPM1 heterozygous gene mutations, all belonging to type A. The EFS rate of the patients with NPM1 mutation was higher in 10 months and the OS rate was higher in 19 months (P< 0.05). 3 of 9 NPM1 mutations patients were FLT3-ITD positive. The CR rates of the four groups after initial remission induction therapy in order were NPM1+FLT3-ITD-, NPM1-FLT3-ITD-, NPM1-FLT3-ITD+, NPM1+FLT3-ITD+(P<0.05). Besides, NPM1-FLT3-ITD+was a risk factor affecting the OS (RR=1.250, P=0.005). 2 of the 55 cases (3.6 %) had c-kit gene mutations, namely mutant D816H and mutant D816V. The c-kit gene mutations were not found in patients with FLT3-ITD and NPM1 mutations. Conclusions The FLT3-ITD mutation is a poor prognosis molecular marker in AML, and NPM1 mutation is a good factor for the prognosis. NPM1-FLT3-ITD+is a risk factor affecting OS. The rate of c-kit gene mutation is low in AML, without the overlap of FLT3 and NPM1 mutations.

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

P1B-ATPases are a group of proteins that can transport heavy metal ions across membranes by hydrolyzing ATP and they are a subclass of the P-type ATPase family. It was found that P1B-ATPases are mainly responsible for the active transport of heavy metal ions in plants and play an important role in the regulation of heavy metal homeostasis in plants. In this paper, we dissusses the mechanism of P1B-ATPases from the structure and classification of P1B-ATPases, and review the current research progress in the function of P1B-ATPases, in order to provide reference for future research and application of P1B-ATPases in improving crop quality and ecological environment management.
Adenosine Triphosphatases/metabolism* ; Biological Transport ; Metals, Heavy ; Plants/enzymology*