Objective To evaluate the immunoprotective effect of Schistosoma japonicum (Chinese strain) recombinant signaling protein 14-3-3(rSj14-3-3), and to observe the synergism of rSj14-3-3 and rSjGST proteins as candidate vaccine and the effect of ??-T cells activated by Mtb against Schistosoma japonicum. Methods BALB/c mice immunized with rSj14-3-3 and rSjGST purified through SDS-PAGE, electroelution and dialysis were challenged by cercaria infection. Six weeks after challenging infection, the mice were killed and the worm and egg reduction rates were calculated. Results Worm reduction rate was found to be 32.20% in rSj14-3-3+Freund adjuvant group, 31.10% in rSj14-3-3+rSjGST+Freund adjuvant group, 27.96% in rSj14-3-3+Mtb group, 26.00% in rSj14-3-3+rSjGST+Mtb group, and 27.10 % in rSjGST+Mtb group, respectively, number of eggs in liver tissue was reduced by 50.40%, 53.30%, 51.10%, 58.60% and 51.30%, respectively. Conclusion rSj14-3-3 could induce partial immunity against Schistosoma japonicum in BALB/c mice, and might serve as a candidate vaccine; ??-T cell activated by Mtb played a role in anti-Schistosoma japonicum similar to the immune reactions induced by Freund adjuvant, but no synergistic effect combined with rSjGST was observed.

Objective To study the localization of the signaling protein 14-3-3 of Schistosoma japonicum (Sj14-3-3)in the parasite. Methods Cercariae were collected from the infected Oncomelania hupensis for the infection of rabbits. Fifteen-day-old schistosomula and adult worms obtained from infected rabbits 15 and 42 days post-infection were used for frozen sections and indirect immunofluorescence staining with monoclonal antibody to rSj14-3-3. Results The results showed that the Sj14-3-3 distributed mainly in the tegument, subtegument, muscle, and parenchyma of both adult worms and 15-day-old schistosomula. Conclusion The wide distribution and large sites of Sj14-3-3 in the parasite were clearly demonstrated, which established a significant clue for further studies of biologic actions and application of 14-3-3 protein.

Objective To investigate expression of 14-3-3 sigma gene in patients with breast cancer and its clinical significance.Methods Expression of 14-3-3 sigma gene was semi-quantitated by using RT-PCR and western-blot in 40 specimens of breast cancer and 18 specimens benign breast disease tissue.Results Of 40 cases of breast cancer 35(87.5%)were negative for 14-3-3 sigma gene in RT-PCR,32(80%)were negative in Western-blot,and 31(77.5%)were negative in both RT-PCR and western-blot.Besides,the expression in 2 cases was down-regulation in both the 2 method.In 18 specimens with benign breast disease tissue the expression of 14-3-3 sigma gene was detectable,which was demonstrated by RT-PCR or western-blot.Conclusion Inactivation and down-regulation of 14-3-3 sigma gene is a frequent event in breast cancer,and it may contribute to diagnosis of breast cancer.

Liver fibrosis is characterized by an abnormal hepatic accumulation of extracellular matrix (ECM) that results from both increased deposition and reduced degradation of collagen fibres. Some studies show that transforming growth factor ?1(TGF-?1), alternatively activated macrophage (aaM) and interleukin 13(IL-13) play a key role in the evolution of fibrosis, of which TGF-?1 and IL-13 become research hotspots. TGF-?1 mainly activates hepatic stellate cells (HSC) through TGF-?1/Smad signal pathway, while IL-13 seems to play a rather crucial role through JAK-STAT6 signal pathway. aaM is an important source of TGF-?1 and activated with Il-13. This paper reviews the role of those signaling molecules in cellular signal transduction of hepatic fibrosis of schistosomiasis japonica, and provides some targets for future drug development.

Objective:To establish the model of apoptosis of activated human ??T cells induced by restimulating with Mycobacterium tuberculosis antigen(Mtb-Ag). To investigate the roles of p38 and phosphatidylinositol 3 kinase(PI3K) pathways in the apoptosis of activated human ??T cells induced by restimualting wih Mtb-Ag.Methods:Mtb-Ag activated human T cells(MtbAT) were cultured for 15 days to 25 days and restimulated with three concentrations of Mtb-Ag for 24 hours, and the apoptosis of ??T cells were measured by flowcytometry(FCM) using Annexin-V-FITC/PI staining. Mtb-AT were restimulating with Mtb-Ag(10 ?g/ml) for 3, 6, 12 and 24 hours, the apoptosis of ??T cells were detected. Mtb-AT cells were pretreated with SB203580(an inhibitor for p38 pathway), or LY294002(an inhibitor for PI3K pathway) for 60 minutes, and restimulating with Mtb-Ag for 3 hours, the apoptosis of ??T cells were detected.Results:Both 10.0 and 20.0 ?g/ml Mtb-Ag significantly induced the apoptosis of ??T cells(P0.05). Compared with control, the apoptosis of ??T cells could be significantly induced by restimulating MtbAT with Mtb-Ag(10.0 ?g/ml) for 3, 6, 12 and 24 hours(P0.05) in the percentages of apoptosis of ??T cells restimulated by Mtb-Ag(10.0 ?g/ml) between for 3 hours and for 24 hours, the percentages of apoptosis of the latter is higher than the former about 7.55%. The apoptosis of ??T cells induced by restimualting wih Mtb-Ag could be inhibited by SB203580(80.0 ?mol/L) or LY294002(10.0 ?mol/L), the inhibition rate of apoptosis was 91.6% and 43.1%, respectively.Conclusion:We established the model of apoptosis of activated human ??T cells by means of using Mtb-Ag(10.0 ?g/ml) to restimulate activated ??T cells for 3 hours. The test of inhibitors of signalling molecule suggested the signalling pathways including p38 and PI3K, participated in the apoptosis of activated human ??T cells restimulated by Mtb-Ag.

Objective To identify the expression of long non-coding RNAs(lncRNAs) and their putatively modulated cytokines in human immunodeficiency virus(HIV)-1 infected monocytes and to explore the role of lncRNAs in immunomodulation of HIV-affected host cells.Methods RNAs arrays were used to screen the lncRNAs differentially expressed in HIV-infected monocytes and subsequently confirmed by quantitative real time PCR (qRT-PCR).A siRNA was designed and transfected to the human monocytes, followed by interleukin(IL)-1β, IL-10 and tumor necrosis factor(TNF)-α detection with ELISA.Results The results of the screening assays and qRT-PCR showed that the expression of lncRNA-n266623 was up-regulated in monocytes after HIV-1 infection;the secretions of IL-1β,IL-10 and TNF-α in human monocytes were significantly enhanced following transfection of siRNA targeting lncRNA-n266623 in the monocytes.Conclusion The HIV-1 infection can promote the expression of lncRNA-n266623 which may inhibit the expressions of IL-1β, IL-10 and TNF-α in monocytes and negatively regulate the immune response to HIV-1 infection.

Objective To explore the effect of salvianolate combined with Qumei trimetazidine on cardiac function in patients with chronic heart failure.Methods Seventy-four patients with chronic heart failure were randomly divided into treatment group and control group (37 cases per group).Patients in control group were treated with the regular treatment scheme including digitalis,diuretics,vasodilators,angiotensin converting enzyme inhibitor(ACEI),angiotensin receptor blockers (ARB) or β blocker therapy for 24 weeks treatment.Patients in treatment group were given the regular treatment scheme plus salvianolic acid and Qumei trimetazidine treatment,of which,the dose of salvianolic was 0.2 g into 5％ glucose injection 250 ml or 0.9％ sodium chloride injection 250 ml by intravenous injection,1 times/day,and Qumei trimetazidine for 20 mg,3 times/day,for 24 weeks.Cardiac function was observed in patients of two groups before and after treatment.The level of brain natriuretic peptide (BNP) was measured.Results Heart function were improved,the total effective rate in treatment group was 91.9％ (34/37),higher than that of control group (70.3％ (26/37),x2 =5.638,P ＜ 0.05).In treatment group,left ventricular ejection fraction (LVEF),stroke volume (SV),cardiac output (CO) of patients after treatment were (52 ± 7) ％,(65.10 ± 12.87) ml,(5.65 ± 1.18) L/min respectively,significant different from that before treatment ((39 ±5)％,(46.53 ± 12.14) ml,(4.79 ± 1.02) L/min,and the differences were statistic significant (t =9.192,6.384,3.352,P ＜ 0.05).Meanwhile,in treatment group,systolic pressure,diastolic pressure,heart rate,left ventricular end diastolic diameter (Dd),left ventricular diastolic posterior wall thickness(PWT),interventricular septal thickness (IVST),left ventricular mass (LVMW),plasma brain natriuretic peptide of patients after treatment were (105 ± 8) mmHg,(75 ± 9) mmHg,(76±8) time/min,(48.7 ±3.7) mm,(9.1 ±1.4) mm,(8.7 ±1.2) mm,(170±59) g,(104.1 ±19.5) ng/L respectively,significant different from that of before treatment((134 ± 12) mmHg,(84 ±8) mmHg,(118 ±11) time/min,(55.2 ±7.8) mm,(11.7 ±2.3) mm,(10.5 ±2.4) mm,(228 ± 111) g,(568.7±179.5) ng/L t=-12.231,-4.546,-18.782,-4.579,-5.874,-4.080,-2.806,15.652,P ＜ 0.01).The same trend was seen in control group in terms of LVEF,SV,systolic blood pressure,heart rate,PWT,plasma BNP before and after treatment(LVEF:(38 ±6)％ vs.(43 ± 8)％ ;:(46.76 ± 11.80) ml vs.(58.69 ± 11.58) ml; systolic blood pressure:(132 ± 10) mmHg vs.(116 ± 11) mmHg; heart rate:(116 ± 10) time/min vs.(77 ±9) time/min;PWT:(11.5 ±2.6) mm vs.(10.4 ±2.0) mm;plasma BNP:(570.2 ± 177.3) ng/L vs.(211.6 ± 21.2) ng/L;t =3.041,4.389;-6.546,-17.632,-2.039,12.21 ;P ＜ 0.05 or P ＜ 0.01).Moreover,after treatment,systolic pressure,diastolic pressure,LVEF,SV,CO,Dd,PWT,IVST,LVMW,plasma brain natriureticpeptide in treatment group were significantly better than that of control grouo (t =-4.919,-2.867,5.510,2.252,2.581,-2.319,-3.238,-3.628,-2.231,-22.701,P ＜0.01 or P ＜ 0.05).Conclusion The effect of salvianolate combined Qumei trimetazidine on treating chronic heart failure is significant,and there is a reverse effect on the left ventricle.

Objective To explore the possibility of heterogenous gene to express in juvenile Schistosoma japonicum and the application of electroporation in transformation of schistosomulae. Methods The plasmids of pEGFP-C1 were introduced into mechanically transformed schsitosomula with electroporation. The presence, transcription and translation of the transgene in electroporated schistosomula were confirmed by PCR, RT-PCR and Western blotting analysis respectively using the genomic DNA, total RNA and protein extracted and isolated from schistosomula cultured in vitro for 48 hours. Meanwhile, localization of EGFP within electroporated schistosomula was performed with confocal laser scanning microscope. Results 760 bp and 276 bp amplified products by PCR and RT-PCR were found coincident with the expected size and expression of EGFP gene in elctroporated schistosomula was confirmed by Western blotting. Fluorescence of EGFP was localized in tegument and subtegument of the electroporated schistosomula with confocal microscopy, especially in the anterior part of the worm. Conclusion The heterogenous gene of EGFP has been successfully introduced into juvenile S. japonicum by electroporation and the expression of transgene was confirmed with molecular and microscopical methods.

Objective To clone and express the cell signaling protein 14-3-3 gene from Toxoplasma gondii RH strain. Methods Toxoplasma RH strain tachyzoites, which maintained by mouse passage, were harvested from ascites of mice and genomic DNA was prepared. A pair of primers were designed and synthesized based on the sequence of Toxo 14-3-3 cDNA. A specific fragment of Toxo 14-3-3 gene was obtained by RT-PCR amplification from Toxoplasma genomic DNA. The PCR products were ligated to pGEM-T. The EcoRI / Xho I restricted fragments, confirmed by PCR and EcoRI / XhoI digestion, were cloned into expression vector pET28a and the recombinants were transformd into E.coli BL21. Fusion expression was induced by isopropyl-beta-D-thiogalactoside (IPTG) and confirmed by Western blotting with rabbit anti-Toxoplasma sera. Results The molecular size of Toxo 14-3-3 was 803 bp, which is highly homologous to the previous report cloned from the parasites of intestinal epithelial stage in cat. High expression was obtained in pET28a/ Toxo 14-3-3/E.coli BL21 when confirmed by Western blotting. Conclusion The recombinant construction of Toxo 14-3-3 was generated and expression was induced.

To probe the effect of paeoniflorin on periovular granuloma and liver fibrosis in mice infected with Schistosoma japonicum in different times of infection and the treatment with praziquantel (PZQ). The models of hepatic fibrosis induced by S.japonicum were established by exposure of BALB/c mice percutaneously through the tail to cercariae of S.japonicum. and mice with treatment were randomly divided into 3 groups: i.e. groups of pre-treatment (I), group of simultaneous treatment (Ⅱ) and group of post-treatment (III). All groups, except the normal control group, were orally introduced with PZQ. And mice in the paeoniflorin-treated group and control group were separately introduced with paeoniflorin and 0.5% sodium carboxymethycellulose respectively. The treatments in group I, II and III were started 30 days before PZQ usage, simultaneously with PZQ or 30days-after PZQ usage respectively. Mice in these groups were sacrificed on the 102, 132 or 162 days after infection. Then the serum levels of hyaluronic acid (HA), amino-terminal peptide of type III procollagen (PIIIP) and liver hydroxyproline (Hyp) were detected. The histopathology was examined by HE and Masson staining; the degree of hepatic fibrosis and the area of egg granuloma were analyzed. The expression of collagen I was examined by immunohistochemical method. It was found that the area of granuloma and degree of hepatic fibrosis in the paeoniflorin-treated groups in group I and III were significantly lower than those in the model control groups. Also, paeoniflorin could induce decreas expression of collagen I. Meanwhile the levels of serum HA, PIIIP and liver Hyp were all reduced in comparison with those in the control group (P<0.05 or P<0.01). However, in group Ⅱ, no significant difference was noted between the treated and the control group in most data. Paeoniflorin also showed the effects to reduce the size of periovular granuloma and to reduce the expression of type I collagen, thereby to resist the development of hepatic fibrosis caused by S. japonicum.-It is evident that PAE shows an efficaciously therapeutic effect on the development of liver fibrosis of shistosomiasis, whenever it is administered before or after the usage of schistosomicides.