Enterovirus type 71(EV71) causes severe hand-foot-and-mouth disease(HFMD) resulting in hundreds of deaths of children every year; However,currently,there is no effective treatment for EV71.In this study,the EV71 poly-protein(EV71-P1 protein) gene was processed and cloned into the eukaryotic expression vector pPIC9k and then expressed in Pichia pastoris strain GS115.The EV71 P1 pretein with a molecular weight of 100 kD was produced and secreted into the medium.The soluble EV71 P1 protein was purified by column chromatography with a recovery efficiency of 70％.The result of the immunological analysis showed that the EV71 P1 protein had excellent immunogenicity and could stimulate the production of EV71-VP1 IgG antibody in injected rabbits.We suggest that EV71-P1 protein is an ideal candidate for an EV71 vaccine to prevent EV71 infection.

Bipolar disorder(BD)is a serious mood disorder with high prevalence,morbidity and mortality rates. Glycogen synthase kinase-3β(GSK-3β) is a multifunctional serine/threonine protein kinase that is generally located in eukaryotic cells with such functions as the adjustment of the synthe?sis of glycogen metabolism,cell proliferation and differentiation and gene expression. It is involved in multiple signaling pathways and regulates cell signaling proteins,structural proteins and transcription factors through phosphorylation,affecting the survival of the neurons and plasticity. According to gene poly?morphism and clinical studies,GSK-3β may be associated with BD. As a GSK-3β inhibitor,lithium is an effective BD therapeutic drug,and the small molecule inhibitor targeting GSK-3βis also a hotspot of BD treatment. GSK-3βmay be a potential target in the treatment of BD.

Objective To examine the effects of IL-10 on cardiac fibroblasts ( CFBs) proliferation and phenotype transformation to myofibroblasts (MyoFbs) induced by transforming growth factor-β1 (TGF-β1);and to investigate the regulating pathways .Methods Cardiac fibroblasts were isolated from cardiac ventricles of neonatal SD rats . The passage 2~4 were used and divided into the following groups for treatment:1) control group, 2) IL-10 reac-tion group, 3) TGF-β1 reaction group, and 4) IL-10 plus TGF-β1 reaction group (TGF-β1 treatment followed with IL-10 pretreatment ) .Cells proliferation was assessed by MTT assay and immunocytochemistry staining for prolifera-ting cell nuclear antigen (PCNA);the phenotype transformation into MyoFbs was assessed by immunocytochemistry of α-smooth muscle actin (α-SMA);extracellular signal related kinase ( ERK1/2) and P38 kinase pathways were assessed by western-blot.Results TGF-β1 (10 μg/L) treatment boosted the proliferation and the expression ofα-SMA significantly (P<0.01), while IL-10 (10, 50 or 100 μg/L) plus TGF-β1 co-treatment induced lower cell proliferation and expression of α-SMA than treating with TGF-β1 alone ( P<0.05 ) , with the inhibitory effect of IL-10 being concentration dependent .TGF-β1 could significantly stimulate the ERK 1/2 and P38 kinase phospho-rylation ( P<0.01 ) , however IL-10 (100 μg/L) plus TGF-β1 co-treatment failed to down-regulated the phospho-rylation of ERK1/2 and P38 kinase compared with TGF-β1 alone ( ERK1/2:P<0.05;P38:P<0.01 ) .Conclu-sions IL-10 can attenuate TGF-β1-induced CFBs proliferation and phenotype transformation to MyoFbs .The in-hibitory effects may explained by a mechanism of inhibiting the activation of ERK 1/2 and P38 kinase .

Objective To investigate the relationship of matrix metalloproteinases(MMP?3 and MMP?9)and the tissue inhibitor TIMP?1 with left ventricular hypertrophy(LVH)in patients with primary hypertension. Methods Totally 140 patients with primary hypertension and 132 healthy controls were included. Matrix metalloproteinase?3(MMP?3),matrix metalloproteinase?9(MMP?9)and tissue inhibitor metalloproteinase?1 (TIMP?1)were measured. All subjects were taken echocardiography examination ,then left ventricular mass index(LVMI)was calculated. Re?sults MMP?3,MMP?9,TIMP?1(488.32±100.32 vs 314.59±99.78;340.56±43.21 vs 290.15±33.98;389.16±57.53 vs 243.45±62.31;P<0.001) and LVMI(113.7±9.9 vs 88.3±10.4,P<0.001)in patients with primary hypertension were significantly higher than those in controls. In a multiple stepwise regression analysis with LVMI as the variable,it was found that age,SBP,MMP?3,MMP?9 and TIMP?1 were main determinants for LVMI (r2=0.78,P<0.001). 3. Patients with primary hypertension were divided into two subgroups according to LVMI,i.e.,hypertension with LVH (group A)and hypertension without LVH(group B). SBP,MMP?3,MMP?9 and TIMP?1(178±31 vs 166±25;490.14±99.13 vs 405.56±53.12;340.56±43.21 vs 290.15±33.98;393.45±47.69 vs 301.58±39.57;P<0.05)of group A were significantly higher than those of group B. Conclusion MMP?3,MMP?9 and TIMP?1 are influencing factors for LVH in patients with primary hypertension.

This paper reported a new type of amylase (neoamylase) secreted by a Bacillus strain ZX99. The enzyme was a kind of ectoenzyme that could catalyze starch into isomalto-oligosaccharide effectively, but could not act on pullulan as substrate. The strain Bacillus ZX99 was mutated by ultraviolet ray and a mutant strain BS3.232 was screened. The activity of the neoamylase produced from BS3.232 increased by 60% over that from ZX99 under the same conditions. The results of thin-layer chromatography of products from starch and pullulan catalyzed by the enzyme demonstrated that the enzyme was different from neopullulanase and can be used to produce isomaltooligosaccharide from starch, including isomaltose, panose, isomaltotriose, isomaltotetose.

Objective To observe the regulatory effect of recombinant human DCN on the expression of HLA class Ⅰ, Ⅱ molecule on hepatoma carcinoma cell HepG2 and investigate the relatively immune mechanism of human DCN on enhancing anti-tumor effect. Methods After transfected HepG2 cells with pcDNA3.1 (+)-DCN by liposome transfection, the expression of HLA class Ⅰ , Ⅱ molecule and the apoptotic indexes were analyzed by the flow cytometer. Results The expression of HLA class Ⅰ , Ⅱ molecule on the HepG2 cells transfected with pcDNA3.1 (+)-DCN was up-regulated with the apoptosis indexes being significantly higher than that of other control groups. Conclusion The human DCN can induce HepG2 apoptosis and up-regulate the expressions of HLA class Ⅰ , Ⅱ molecule which may contribute to its antitumor effect.

Objective To investigate the relations between serum adiponectin and coronary artery calcification score (CACS) and to find the risk factors for coronary artery calcification in maintenance hemodialysis(MHD) patients.Methods Twenty-seven MHD patients(MHD group) and 13 healthy persons (control group) were enrolled in this study.The serum adiponectin was measured by enzyme-linked immunosorbent assay method.CACS was calculated by multi-row spiral CT.The circulating parameters such as hemoglobin (Hb),calcium,phosphate,calcium-phosphate product,intact parathyroid hormone (iPTH),total cholesterol (TC),low density lipoprotein cholesterol (LDL-C),albumin (Alb),high sensitive C-reactive protein (hs-CRP),and so on were detected.Results The level of serum adiponectin in MHD group [(15.00 ± 7.47) mg/L] was significantly higher than that in control group [(2.07 ± 0.83) mg/L],and there was significant difference (P＜ 0.01).Coronary artery calcification(CACS ＞ 0 score) was observed in 88.9％ (24/27) in MHD group and 10/13 in control group.The mean CACS in MHD group was significantly higher than that in control group [655 (0-3 570) scores vs.126 (0-731)scores],and there was significant difference (P ＜ 0.05).Eleven MHD patients and 1 healthy person had severe coronary artery calcifications,(CACS ≥ 400 scores).There was significan t difference in dialysis duration,diastolic pressure,phosphate calcium-phosphate product and adiponectin (P ＜ 0.05 or ＜ 0.01).Spearman analysis showed that CACS of MHD patients was positively correlated with dialysis duration,phosphate,calcium-phosphate product,serum creatinine and adiponectin (P ＜ 0.01 or ＜ 0.05).Only calcium-phosphate product remained as independent predictor of CACS in multivariate analysis (P ＜0.01).Conclusion Coronary artery calcification is common in MHD patients and which is correlated with dialysis duration,serum phosphate,calcium-phosphate product,serum creatinine and adiponectin.

AIM and METHODS: To study the culture met ho d of human neural stem/progenitor cells, these cells derived from human fetal fo rebrains were maintained and expanded in serum-free defined medium containing ba sic fibroblast growth factors (bFGF), epidermal growth factor (EGF) and B27. Whe n they formed neurosphere, these three factors and supplemented FBS were removed to induce differentiation. Cell were cultured for 12-14 d, then fixed for immun ocytochemistry examination. RESULTS: This period of expansion resulted in a 107-fold incre ase in this heterogeneous population of cells. Upon differentiation, they form n eurons, astrocytes and oligodendrocytes, the three main phenotypes in the CNS. CONCLUSION: These results demonstrate the feasibility of long-t erm in vitro expansion of human neural progenitor cells. The advantages of s uch a population of neural precursors for allogeneic transplantation, including t he ability to provide an expandable, well-characterized, defined cell source, ca n form specific neuronal or glial subtypes.

Objective To evaluate the value of echocardiography on the diagnosis of arrhythmogenic right ventricular cardiomyopathy (ARVC),and to improve the diagnositic accuracy of ARVC by echocardiography.Methods According to the 2010 European Heart Association guideline,twenty-one patients with ARVC were diagnosed from September 2003 to June 2014.The patients were divided into four groups (confirmed,suspiciously diagnosis,miss diagonisis,misdiagnosis) and the echocardiographic features were retrospectively analyzed including the right ventricular (RV) movement,the diameter of RV outflow tract (RVOTd),fractional area change of RV (RVFAC),the severity of tricuspid regurgitation (TR) and peak pulmonary artery systolic pressure (PASP).Results Of 21 patients,15 (71.4％) were confirmed by echocardiography,which had the typical ARVC echocardiographic features including the hypokinetic,akinetic or aneurysm of RV,dilation of RVOTd [mean RVOTd (40 ± 3)mm],and RV FAC＜33 ％ [mean (21 ± 7)％].TR were noticed in all the 15 patients but the PASP were normal [mean (27 ± 9)mmHg,1 mmHg =0.133 kPa].Three (14.3％) were suspiciously diagnosed which had the RV wall hypoakinetic,1 with pure RVOTd dilation and 2 with RV and RVOTd dilation,all 3 patients had mild TR,33％＜RVFAC ≤40％ and PASP were in normal range.Two patients had normal echocardiography which was miss diagnosed,one patient was misdiagnosed as dilated cardiomyopathy.Conclusions The different stages of ARVC patients had different echocardiographic features,the patients were easily diagnosed when the ARVC patients in RV failure stage.But for the early and late stage,the diagnosis should combine the clinical manifestation and other imaging facilities to avoid miss diagnosis and misdiagnosis.

AIM: To observe the clinical effects of intravitreal injection of triamcinolone acetonide ( TA ) before vitrectomy for retinal detachment associated with choroidal detachment. · METHODS: Totally 23 cases ( 23 eyes ) of retinal detachment associated with choroidal detachment in our hospital were treated by intravitreal injection of TA 4-5d before 23-Gauge micro-invasive vitrectomy combined with silicone oil injection. All the cases were followed up between 6 to 9mo. The anatomic retinal reattachment, visual acuity, intraocular pressure and postoperative complications were observed and analyzed. ·RESULTS: After the surgery, the visual acuity of all patients were improved, with 9 eyes better than 0. 3 (39%), and 18 eyes better than 0. 05 (78%). The BCVA at 1wk, 1 and 3mo and last follow up were different compared with before operations (P<0. 05). The mean intraocular pressure was 4. 02±1. 47mmHg before injection, 13.69±4. 68mmHg before operation (P<0. 05), and17.72±5.87 mmHg after operation (P<0.05). The retina of all patients treated were reattached 2wk post-operatively. The retinal reattachment rate after the primary surgery and the secondary surgery was 87% and 100%, respectively. Post-operative complications included 7 eyes of transient high intraocular pressure, occurred during 12-14d after operations and returned to normal after less glucocorticoid eye drops and giving IOP lowering drugs. There were no intraocular hemorrhage, iatrogenic retinal breaks, infections, or lens injuries. · CONCLUSION: Intravitreal injection of TA before vitrectomy for retinal detachment associated with choroidal could improve the clinical effects, and decrease the difficulty of surgery while the injection itself is pretty safe.