Objective:To study the immunological characteristics of recombinant Rv3425 protein,to evaluate its diagnosis value and the role in the pathogenicity. Methods: Rv3425 gene was cloned into pET28a vector,the recombinant protein was induced and purified;we analyzed the antigenicity and specificity of Rv3425 by ELISA ( Enzyme-linked immuno sorbent assay) and Western blot methods,apoptosis effect of Rv3425 to macrophage was deteced by FCM ( Flow cytometry method ) . Results: Purified prokaryotic expressed protein Rv3425 was acquired,we found Rv3425 could elicit high level of IFN-γin spleen cells and combine with the serum of iH37Rv (inactivated mycobacterium tuberculosis) immunized mice;the specific IgG and IgM antibodies of TB (Tuberculosis) patients were significantly higher than healthy donors;Rv3425 also could induce the necrosis of macrophage. Conclusion:Our study found that Rv3425 had strong antigenicity and could induce the apoptosis of macrophage,these findings were very important for the research of TB diagnosis and pathogenicity.

Objective To obtain the antigen and antibody of JC virus(JCV)VP2.Methods The JCV VP2 gene were amplified from a cerebrospinal fluid sample by polymerase chain reaction (PCR)and confirmed by sequencing.Then,the gene was cloned into plasmid pET32a(+)to construct recombinant prokaryotic expression vector pET-32a(+)-VP2.The recombinant plasmid was transformed into the competent E.coli BL21.Induced with isopropyl-β-D-1-1 thiogalactopyranoside (IPTG),E.coli BL21 were subsequently crushed by ultrasound.The gene expression in the supernatant was analyzed by Western blot.Thereafter,the expressed protein was purified by isoeleetric point method.The polyclonal antibody against JCV VP2 protein was obtained from the BALB/c mouse immunized with the purified protein.Results The VP2 fusion protein was expressed in the E.coli BL21.The recombinant fusion protein was expressed by IPTG induetion with relative molecular mass of 58.5×10~3.Sodium dodecyl sulphate-polyacrylamide gel electrophoresis(SDSPAGE)analysis showed that the expression level was highter after 6-10 h of IPTG induction.The recombinant protein had good antigenicity which was confirmed by BALB/c mice immunized with the protein.Conclusions The successful expression and purification of VP2 fusion protein and the antibody will be valuable for the study on the biological function of VP2 and JCV epidemiologieal investigation.

Objective To construct prokaryotic expression vector carrying jc virus(JCV)t-antigen gene,express and purify this fusion protein.Methods The JCV t-antigen gene from a cerebrospinal fluid sample was amplified using polymerase chain reaction(PCR)method.After sequencing.the gene was cloned into plasmid pET32a(+)to construct recombinant prokaryotic expression vector pET32a(+)-t.The t-antigen fusion protein was expressed by isopropy-~D-thiogalactoside(IPTG)induction and prepared in large scale,then purified by Ni+affinity column chromatography.The polyclonal antibody was obtained from the BAI.B/C mouse immunity by the purified protein.Results The relative molecular nlass of recombinant protein expressed by pET32a(+)-t was about 41 000.Sodium dodeeylsulfate-polyaerylamide gel electrophoresis(SDS-PAGE)showed that the fusion protein W&S highly expressed after 3.5～20.Oh of IPTG induction.The antigenicity of the purified protein Was well confirmed by Western blot.The anti-mousepolyclonal antibody was obtained successfully from immunized BALB/c mice.Conclusions The prokaryotic expression vector pET32a(+)-t is successful constructed and the fusion protein is expressed and purified.Furthermore,the antibody of JCV small envelop protein t is successfully prepared.This work provides vMuable information for further study on epidemiology and biological function of t antigen.

Objective To evaluate the validity and reliability of simplified Chinese version Scoliosis Research Society-22 (SRS-22) questionnaire on assessment of life quality of adolescent idiopathic scoliosis (AIS) patients at different time points pre-and post-operation. Methods Translation and cross-cultural adaptation of the English version of SRS-22 questionnaire was done. 177 AIS patients were involved in present study and the life quality before and 6-and 12-months after operation was assessed by SRA-22 and Short Form-36 (SF-36) questionnaire, of them 145 cases (26 males and 119 females; 11.1-17.9 years old, averaged 15.8; Cobb angle 41?-109?, averaged 63?) completed all the 3 steps. To evaluate the reproducibility, 65 patients were assessed twice in an interval time of 4-6 days (averaged 4.6 days) before operation and 55 valid questionnaires were received, 65 patients were assessed twice in an interval time of 7-21 days (averaged 14.7 days) at 12 months following up and 57 valid questionnaires were received. The two measures of reliability as internal consistency and reproducibility were determined by Cronbach's ? coefficients and intraclass correlation coefficient (ICC), respectively. Concurrent validity was measured by comparing with SF-36 questionnaire. Measurement was made using the Pearson correlation coefficient (r). Results The Chinese version SRS-22 questionnaire had a satisfactory validity and reliability, with average Cronbach's ? coefficients of 0.79-0.85 and average ICC of 0.77-0.87. Considering concurrent validity, 2 domains had excellent correlation (r=0.75-1.00), while 12 had moderate correlation (r=0.50-0.75). The function/activity (SRS-22) and role-physical or physical functioning (SF-36), pain (SRS-22) and pain index (SF-36), mental health (SRS-22) and mental health (SF-36) had good correlation (r=0.70-1.00). For the 145 patients, there were some differences in 5 various domains between the three measurements (pre-operation and 6 and 12 months post-operation). The scores of all domains 12 months after operation increased significantly than those of pre-operation and 6 months post-operation (P

Objective To study the cyfluthrin resistance and potential mechanisms of Anopheles sinensis in Nanchang Chang-bei International Airport,Nanchang City,Jiangxi Province. Methods The resistance levels of the local An. sinensis were de-tected by WHO drug resistance bioassay. During the bioassay,the dying mosquitos were classed as sensitive mosquitos,and the survival ones were classed as resistant mosquitos. The P450 monooxygenase activity and glutathione S-transferase activity were detected and compared between the two groups. At the same time,the death time of each sensitive mosquito was recorded,and the correlations between the death time and the P450 monooxygenase activity and glutathione S-transferase activity were ana-lyzed,respectively. Results The bioassay mortality of the local An. sinensis was 59.5%. The differences of the P450 monooxy-genase activities among the resistant mosquitos,sensitive mosquitos and laboratory sensitive mosquitos had statistical signifi-cances(F=151.89,P<0.01),the resistant mosquitos>sensitive mosquitos>laboratory sensitive mosquitos. The differences of glutathione s-transferase activities among the three groups had no statistical significance(F=0.72,P=0.49). There existed positive correlation between the mosquito death time and the P450 monooxygenase activity,and the regression equation was y=79.479+1.512x with the correlation coefficient of 0.88,while there was no correlation between the mosquito death time and the glutathione S-transferaseactivity. Conclusion The An. sinensis in Nanchang Changbei International Airport has been resistant to cyfluthrin,and the promotion of P450 monooxygenase activity maybe one of the reasons for the resistance.

Transplantation acquired food allergy (TAFA) is a rare complication of solid organ transplantation. The pathogenesis of TAFA has not been fully elucidated. There are two possible mechanisms for its occurrence: food allergy mediated by IgE delivery of the donor and food allergy caused by food intolerance after transplantation. At present, there is still insufficient understanding of this complication among transplant physicians. Through systematic review of relevant literature, the authors summarized the research progress of TAFA, which mainly included the pathogenesis, clinical characteristics, treatment and prognosis of TAFA. In order to provide reference for the diagnosis and treatment of TAFA.

Objective : To investigate the effect of paederosidic acid methyl ester (PAME) on postoperative learning and memory impairment in mice and its mechanism. Methods : C57BL/6J male mice were randomly divided into Sham group , operation group , operation + PAME group ( PAME group) , operation + NADPH oxidase 4(Nox4) adeno⁃associated virus overexpression group (Nox4 overexpression group) , operation + Nox4 adeno⁃associated virus no⁃laden group ( AAV no⁃load group) , and operation + PAME + Nox4 overexpression group ( PN group) . Exploratory laparotomy was performed. PAME(20 mg/kg) was administered by continuous gavage for 7 days after operation , and adeno⁃associated virus was injected into the hippocampus 28 days before operation. Morris water maze test and conditioned fear test were used to detect the learning and memory ability of mice. The expression of Nox4 protein was observed by immunofluorescence. The protein expressions of Nox4 , long chain acyl CoA synthetase 4 (ACSL4) and glutathione peroxidase 4 (GPX4) were detected by Western blot. Reactive oxygen species (ROS) and iron content were determined by spectrophotometry. Results : Compared with the Sham group , the learning and memory ability of the operation group , the Nox4 overexpression group and the AAV no⁃load group decreased , the protein expression of Nox4 and ACSL4 increased , the protein expression of GPX4 decreased , and the ROS and iron content increased. After PAME treatment , the postoperative learning and memory ability of mice was improved , and Nox4 and ferroptosis in hippocampal neurons were alleviated. Conclusion Conclusion PAME treatment can improve the learning and memory ability of postoperative mice , which may be related to the inhibition of hippocampal Nox4⁃mediated ferroptosis.