β?carbonic anhydrases(β?CAs)are ubiquitous metalloenzymes which active site contains a zinc ion(Zn2+),and they could catalyze the hydration of carbon dioxide to bicarbonate and protons efficiently and are involved in many biological pro?cesses,such as respiration,pH and CO2 homeostasis,biosynthetic reactions,virulence regulation and so on,and may play a critical role in the life activity of many organisms which contain these enzymes. β?CAs are widely distributed in fungi,bacteria, algae,plants and a small number of protozoan and metazoan except vertebrates. Therefore,as potential drug targets for design?ing and developing antibacterial and anti?parasitic drugs,β?CAs promise a broad application prospect. This paper focuses on the distribution,physiological function and the progress of researches on β?CAs in parasites and their vectors.

Objective To evaluate the imaging features,pathologic characteristics and diagnostic methods of gastrointestinal stromal tumor(GIST).Methods A retrospective study was performed to analyze the clinical pathological and imaging data of 14 GIST patients diagnosed by surgery and pathology.Results 14 cases of GIST were all solitary.10 cases were located in stomach and 4 cases in small intestine.Tumors were mostly round or oval,and a few of them were irregular lobulated.Immunohistochemistry analysis:12 cases were CDll7 positive and 9 cases were CD34 positive.Conclusion The imaging examination was important to the diagnosis and localization of GIST,but the final diagnosis of GIST depended on histopathological and immunohistochemical examination.

Objective:To identify the influence of occupational stress on occurrence of hypertension.Methods:A prevalent cohort of 964 male workers employed for at least 3 years with different occupational stress were investigated with Occupational Stress Index, hypertension was diagnosed by WHO criteria of 1996. Results:The occupational stress was a risk factor for hypertension with RR=1.70 and in a dose-effect pattern after adjusted for other factors. In high exposure group, the adjusted incidence density was 12.47/per 1000 person-years with RR=2.99, in the medium exposure group, that was 8.81/per 1000 person-years with RR=2.11, that of low exposure group was 4.17/per 1000 person-years, the differences between different groups had statistic significance. When the length of exposure was more than 10 years, the adjusted incidence of hypertension increased significantly in high and medium exposure groups.Conclusions:Causality between occupational stress and hypertension exist, but it needs long exposure of at least 10 years for occurrence of hypertension.

Objective To develop and identify the monoclonal antibodies (McAbs) against Region II~+ motif in circumsporozoite protein of Plasmodium falciparum. Methods BALB/c mice were immunized with 12 peptides within Region Ⅱ~+ in circumsporozoite protein of P. falciparum. Spleen cells isolated from the immunized mice were fused with myeloma cell. After three times screening with ELISA, 3 positive hybridoma cell lines were obtained. Results ELISA test indicated that the McAbs reacted with recombinant circumsporozoite protein fragment containing tandemly repeat region and conserved Region II~+. IFA test showed that the McAbs recognized not only the sporozoites of P. falciparum, but also the sporozoifes of P. yoelii. Conclusion McAbs obtained can probe the Region II~+ motif in circumsporozoite protein of P. falciparum, which might also recognize that of other Plasmodium species.

Objective To identify the recombinant aldolase (ALD) of Plasmodium falciparum, and to develop monoclonal antibodies (McAbs) against the recombinant ALD. Methods ALD gene was amplified by PCR from genomic DNA of FCC1/HN strain, and expressed in E.coli DH5?. BALB/c mice were immunized with the recombinant ALD of P. falciparum via celiac injection for 3 times with 2 weeks interval. Three days after a booster injection, spleen cells of the immunized mice were used for producing McAbs. The immune serum was tested by IFAT and Western blotting. Results BALB/c mice immunized with purified aldolase protein developed strong immune response to the antigen, and the titer of specific antibody reached 1∶105 in all immune sera after the third immunization. Moreover, immune sera specifically recognized the cultured P. falciparum. Western blotting showed that the immune sera recognized specifically a Mr 41 000 band of crude malaria antigen. No cross-reaction with human red cells was detected. Seven positive hybridoma cell lines were obtained after 3 rows of selection. All the McAbs′subclasses belong to IgG1. IFAT showed that only 4 McAbs could recognize the cultured P.falciparum. Conclusion Plasmodial aldolase has been successfully expressed and purified, and the established hybridoma cell lines can secrete McAbs specific to the aldolase of P. falciparum.

ObjectiveTo investigate mechanisms for IL-27 induced proliferation and differentiation of peripheral blood CD4+ T cells in primary biliary cirrhosis (PBC).MethodsPeriperal blood CD4+ T cells were isolated from patients with PBC,chonic hepatitis B (CHB) and health controls (HCs).After IL-27 stimulation,proliferation ability of CD4+ T cells was evaluated by CCK-8 kit,and cytokines were analyzed by ELISA.Real-time PCR was employed to assay mRNA expression of T-bet and GATA3 in CD4+ T cells.p-STAT-1 and pSTAT-3 expression in CD4+ T cells were detected by Western blot.ResultsEnhanced proliferation of CD4+ T cells was found in all subjects after IL-27 stimulation.However,the proliferation ability in patients with PBC was greater than that in CHB and HCs ( P＜0.001 ).Levels of IL-2 and IFN-γ in supernatant from IL-27-incubated PBC blood CD4+ T cells were higher than that from CHB and HCs (P＜0.001 ).In normal situation,T-bet mRNA of CD4+ T cells in PBC group was higher than that in CHB group (P=0.007).Furthermore,after IL-27 stimulation,elevated T-bet mRNA expression and GATA3 inhibition were found in patients with PBC.High expression of p-STAT-1 and p-STAT-3 in blood CD4+ T cells were found in PBC,CHB and HCs after stimulation by IL-27.But their expression in patients with PBC were higher than those in patients with CHB and HCs.ConclusionProliferation of blood CD4+ T cells could be induced by IL-27 in patients with PBC.The signaling pathways of p-STAT-1,p-STAT-3 were involved to induce Th1 immune response and related cytokines expression.This study implicated that IL-27 may play important roles in early inflammation damage in PBC.

Objective To investigate the correlation between immune inflammation and overactivity of T helper cells in childhood asthma by cell proliferation assay and activation induced cell death in vitro.Methods Th1/Th2/Th17 cytokines were determined by cytometric bead array.Cell proliferation and activation induced cell death were detected when CD4+ T cells were purified by magnetic beads and stimulated by PHA and antiCD3.At last,mRNA of Fas,FasL and Bcl-2 were mesured by real-time PCR.Results Cytokines of IL-4(2.451± 1.052ng/L vs 1.796 ±0.615 ng/L,P =0.018),IL-10( 1.920 ±0.813ng/L vs 1.390 ±0.162ng/L,P =0.006)and TNF(5.112 ±5.842 ng/L vs 1.506 ±0.551 ng/L,P =0.009) in sera of asthma group were higher than those in control group.Compared to control group,proliferation ability of CD4 + T cells in asthma group was greater ( OD450:0.498 ± 0.052 vs 0.274 ± 0.032,P ＜ 0.001 ) and apoptosis rate was lower( 35.62 ± 0.05 ％ vs 65.28±3.85％,P ＜0.001 ).mRNA expression of Fas in asthma group was lower but Bcl-2 was higher than those in control group.Conclusion It is implicated that defective expression of Fas and over expression of Bcl-2 in CD4+ T cells may contribute to apoptosis inhibition and cell proliferation,which could explain overeactivity of CD4 + T cells and lvmphocvte infiltration in childhood asthma.

Objective To investigate a new therapeutic pathway for primary biliary cirrhosis (PBC) by immune tolerance reestablishment in a PBC mouse model. Methods Spleenic cells from naive mice were incubated with M2 in the presence of ECDI and the ceils were injected into caudal vein of the mice which would be used for development of PBC model. Spleenic cells incubated with bovine serum albumin (BSA) were injected as controls. 16 weeks later, anti-mitoehondrial antibody (AMA) , alkaline phosphatase(AKP) and portal inflammation were assayed for evaluating the prevention effect. Results AMA positive rate in tolerance group was lower than that in BSA and PBC groups ( P = 0. 007, P = 0. 003 ). The difference between BSA and PBC was not significantly. Serum AKP levels in tolerance, BSA and PBC group were (80.5 ±9.8) U/L, (93.8 ±15.7) U/L and (92.5 ±17.7) U/L, separately. The level in tolerance group was lower than that in BSA and PBC groups (P =0.0095, P =0.029). The rates of portal areas with cell infiltration were 42. 67% ± 12. 30% , 57. 07% ± 11. 35% and 51. 53% ± 9. 96% , separately. The number of infiltrated portal tracts in tolerance group was less than that in PBC group (P = 0.039) and BSA group (P = 0. 0024). Conclusion PBC was prevented to some extent by reestablishing immune tolerance to M2 autoantigen. This provides clues for finding a better treatment proposal.

Objective To investigate the population and role of IL-10+ CD19+ regulatory B cell (Breg) in patients with chronic hepatitis B.Methods Patients with acute hepatitis B (AHB) (n =28),chronic hepatitis B (CHB) (n =31) and normal subjects (n =25) were collected from Changshu No.2 People's Hospital between 2011 June and 2012 October.Peripheral blood mononuclear cells (PBMC) were isolated and stimulated with CpG ODN 2006 and PMA.Flow cytometry was used to analyze the population of IL-10-CD19 + Breg,CD4 + CD25high Treg,and ELISA was used to analyze the concentration of IL-10 in culture supernatant.Results The population of Breg in Peripheral blood of the CHB group [1.28％ (1.05％-2.20％)] was higher than that in the AHB group [0.87％(0.55％-1.22％)] and the HCs group [0.89％ (0.51％-1.37％)] (P =0.001,0.006),and the difference between the AHB group and the HCs group was not statistically significant (P=0.669).Breg in the CHB group [14.30％ (10.70％-16.70％)] was higher than that in the AHB group [10.30％ (7.05％-13.30％)] and the HCs group [10.40％(6.85％-12.60％)] (P =0.003,0.001),treg in the CHB group [5.80％ (4.20％-9.10％)] was also higher than that in the AHB group [4.05％ (2.53％-5.40％)] and the HCs group [4.50％ (2.55％-5.50％)] (P ＜0.001,P =0.005),and there was no significantly difference between the AHB group and the HCs group (Breg:P =0.796 ; Treg:P =0.227).Spearman correlation analysis showed that Breg was positively correlated with Treg in the CHB group (r =0.50,P =0.004),however there was no significantly correlation in the AHB group and the HCs group (r =-0.15,P =0.462; r =0.09,P =0.669).The concentration of IL-10 in the CHB group was higher than that in the AHB group and the HCs group (P ＜ 0.001),and the difference between the AHB group and the HCs group was not statistically significant (P=0.341).Spearman correlation analysis showed that IL-10 were positively correlated with the population of Breg in the CHB group (r =0.409,P =0.022).Conclusion The poluations of regulatory B cell and regulatory T cell increased in patients with chronic hepatitis B,and Breg cell might play the immune regulation role through secreting IL-10 in chronic HBV infection.

Objective To investigate the roles of TLR3 pathway activiated by polyI:C in proliferation and apoptosis of multiple myeloma (MM) RPMI8226 cell line.Methods RPMI8226 cells were cultured in RPMI 1640 with different dose of polyl:C.Cells were collected in different time.Proliferation and apoptosis were detected by CCK-8 kit and flow cytometry,separately.Results The proliferation of RPM18226 was inhibited by polyI:C,and it was dose and time dependent,24 h:12.30％ ±2.04％,22.50％±2.20％,37.90％ ±1.30％ ; 48 h:17.80％ ±1.52％,29.60％ ±0.85％,45.80％ ±1.68％ ;72 h:25.10％±1.01％,34.60％±1.27％,60.50％±2.08％,P＜0.05.RPMI8226 cells were incubated with 50 μg/ml,100 μg/ml and 200 μg/ml polyI:C for 48 h.Apoptotic rate were 5.60％ ±1.06％,8.71％ ±1.06％ and 13.93％ ±1.17％,P＜0.05.TLR3 and TRIF mRNA expression increased obviously and dose dependent,TLR3:1.41±0.10,2.24±0.16,4.08±0.13; TRIF:1.07±0.16,1.97±0.13,3.56±0.19,P＜0.05.Conclusion The proliferation of MM cells were inhibited by TLR3 pathway obviously,and apoptosis was induced by polyI:C.