Objective To screen a new schistosome vaccine candidate. \ Methods\ Schistosoma japonicum adult cDNA library was screened using sera from immune rabbits vaccinated with irradiated cercariae and monoclonal antibodies against membrane antigen of S.japonicum schistosomula. Three different fragments of S.japonicum cDNA genes were cloned into pGEM-T vector. The sequences of the inserts were determined using an automatic DNA sequencer and were analysed using Blast program. One of the unknown genes (B8) was selected and its ORF sequence (291 bp) was subcloned into eukaryotic expression vector. The recombinant plasmids were identified by restrictive enzymes and PCR amplification. The positive recombinant plasmids (pBK/SjB8) were transformed into host bacteria XL1-blue, and were then induced by IPTG for expression. SDS-PAGE and Western blotting analysis of total cellular protein from the bacteria were performed to detect the gene products. Results The results demonstrated that ORF of SjB8 gene was subcloned into the plasmid pBK-CMV and could express as fusion protein in XL1-blue. The results of SDS-PAGE and Western-blot also showed that the molecular weight of the fusion protein with 3 kDa ?-galactosidase was approximately 13\^6 kDa and the actual molecular weights of the SjB8 was 10\^6 kDa. The expressed fusion product of pBK/Sj-B8 could be recognized by immune serum and McAb. Conclusion A new gene of S.japonicum vaccine candidate (SjB8) was cloned into eukaryotic expression vector pBK-CMV and could express 10\^6 kDa schistosome protein. The results provide foundation for further study of the protein for its posibility as candidate vaccine.

Objective To clone and express a membrane protein(Tetraspanin 2) gene of Schistosoma japonicum (SjTsp2). Methods A pair of primers was designed to amplify the SjTsp2 gene which was subcloned into prokaryotic plasmid pET28a(+). The recombinant plasmid was transformed into E. coli BL21(D3) and followed by expression of the protein induced by IPTG. The protein was purified by affinity chromatography and used to immunize BALB/c mice. Dilution of antibody against SjTsp2 was determined by ELISA. The protein was also identified by Western blotting. Results Big loop of SjTsp2-A, 228 bp, was amplified in vitro by PCR. Its deduced amino sequence shared 52% similarity with SmTsp2. The soluble recombinant SjTsp2-A was expressed in the experiment and high dilution antibody against the recombinant (1 ∶ 32 000 in maximum) was produced in immunized mice. SjTsp2-A reacted positively with sera of acute and chronic schistosomiasis patients but not with sera from healthy persons by Western blotting. Conclusion SjTsp2 has been expressed and shows certain antigenicity.

Objective To investigate the morphological features of reproductive system of adult Schistosoma japonicum under an optical microscope.Methods Adult schistosomes were obtained from infected mice with cercariae shedding from Oncomelania snails.The adult worms fixed with 10% formalin,dehydrated,imbedded in paraffin,cut at 3 ?m thick,stained by HE staining and then observed under an optical microscope.Results The reproductive organs of adult Schistosoma japonicum such as testicle,ovary,fallopian tube,vitellarium,yolk duct and hystera were displayed distinctly and typically.Conclusions The morphological features of reproductive system of adult Schistosoma japonicum are distinct and typical by using routine pathological techniques preparing and HE staining,which establishes a morphological foundation for the morphological teaching of schistosomes and reproductive biology research.

Objective To obtain genes encoding the novel molecules for diagnosis of schistosomiasis. Methods The cDNA library of Schistosoma japonicum (Sj) schistosomula of day 15 post-infection was screened with positive patients sera,and the inserts of positive clones were sequenced. Then the sequences were compared with all sequences in GenBank database by Internet. The positive clones were analyzed by the software in bioinformatics. Results After three rounds of screening,15 positive clones were obtained,and among them 11 clones had the meaningful sequences,in which 5 genes coding for Sj mitochondrion,1 gene coding for Sj myosin and the others were SjCHGC05315,SjCHGC01371,SjCHGC04782,SjCHGC05166,SjCHGC09769,respectively. Conclusion Immunological screening by using positive patients sera can be used to discover Sj antigen genes with a potential value for diagnosis.

OBJECTIVETo prepare sodium alginate-nanohydroxyapatite composite material and to explore its feasibility as a bone repair material.

METHODSSodium alginate-nanohydroxyapatite composite material was prepared using chemical cross-linking and freeze-drying technology. The composite was characterized by X-ray diffraction (XRD) and scanning electron microscope (SEM) and its porosity was measured by liquid displacement method. The fifth passage of bone marrow stromal stem cells (BMSCs) were incubated on the composite material and then growth was observed by inverted microscope and SEM. BMSCs were cultured with liquid extracts of the material, methyl thiazolyl tetrazolium (MTT) assay was used to calculate the relative growth rate (RGR) on 1, 3, 5 d and to evaluate the cytotoxicity. Fresh dog blood was added into the liquid extracts to conduct hemolysis test, the spectrophotometer was used to determine the optical density (OD) and to calculate the hemolysis rate.

RESULTSSodium alginate-nanohydroxyapatite composite material displayed porosity, the porous pore rate was (88.6 +/- 4.5)%. BMSCs showed full stretching and vigorous growth under inverted microscope and SEM. BMSCs cultured with liquid extracts of the material had good activities. The toxicity of composite material was graded as 1. Hemolysis test results showed that the hemolysis rate of the composite material was 1.28%, thus meeting the requirement of medical biomaterials.

CONCLUSIONThe composite material fabricated in this study has high porosity and good biocompatibility.

Alginates ; Biocompatible Materials ; Cells, Cultured ; Glucuronic Acid ; Hexuronic Acids ; Humans ; Mesenchymal Stromal Cells ; Porosity ; Tissue Engineering ; Tissue Scaffolds

Objective To investigate clinical effect and mechanism of Yudanrongxin pills in the treatment of patients with acute myocardial infarction ( AMI) .Methods 86 cases of AMI in our hospital from February 2014 to February 2015 were selected and divided into the observation group and the control group, with 43 cases in each group.The patients in the control group were treated with conventional therapy, while patients in observation group were treated with Yudanrongxin pills on the basis of the control group.The related inflammation factors, indicators of myocardial injury and heart function index were compared between two groups before and after treatment.Results Compared with before treatment, in both two groups after treatment, the serum inflammatory factors including high-sensitivity C-reactive protein(hs-CRP), IL-6,soluble vascular cellular adhesion molecule-1 (sVCAM-1), TNF-αand P-selectin decreased, the myocardial injury criterion including creatine kinase-MB (CK-MB), lactate dehydrogenase (LD), myocardial troponin I (cTnI) and myoglobin (Mb) decreased,the cardiac function indexes of left ventricular end systolic volume (LVESV) and left ventricular end diastolic volume (LVEDV) decreased and left ventricular ejection fraction (LVEF) increased (P<0.05).Compared with control group, the hs-CRP,IL-6,sVCAM-1,TNF-α,P-selectin,CK-MB,LD,cTnI and Mb in observation group were lower(P<0.05).The degree of improvement at cardiac function was better than that in control group (P<0.05).Conclusion Yudanrongxin pills could better improve cardiac function in treatment with AMI, its role was relative to inhibition of inflammatory factors and myocardial protection against injury.

Objective To investigate the effect and mechanism of Yiqi Shuxin pills in the treatment with viral myocarditis(VMC).Methods 56 cases of VMC in our hospital were randomly divided into the experimental group and the control group,28 cases in each group.The patients in the control group were with routine treatment,and these in the experimental group were treated with Yiqi Shuxin pills based on the control group.The clinical effects between the two groups were compared, and the changes of miR, myocardial enzyme spectrum and immunity were compared before and after treatment.Results The total effective rate was 92.86% in the experimental group and was 72.57% in the control group,there was statistical significance between the two groups(P<0.05);There was no significant difference between the two groups before treatment at miR-1,miR-133,miR-21,CK-MB, cTnI,LDH,CD3 +,CD4 +,CD8 +,NK in serum;The content of miR-1,miR-133,CD3 +,CD4 +,CD8 +,NK after treatment in the two groups were higher than before treatment(P <0.05),miR-21,CK-MB,cTnI and LDH were lower than before(P <0.05);The improvement of these indicators in the experimental group was superior to that in the control group(P<0.05).Conclusion Yiqi Shuxin pills could improve the therapeutic effect of VMC,the mechanism maybe related to improving the circulation of blood miR and enhance cellular immune function.

The protective effect and mechanism of Oroxylin A, a naturally occurring compound in Scutellaria baicalensis Georgi, was investigated in this study. Isoproterenol administration to rats triggered classic cardiac failure, as demonstrated by objective parameters of cardiac dysfunction. Intragastric administration of oroxylin A at the dose of 25, 50 and 100 mg/(kg·d)significantly improved deranged cardiac parameters in the isoproterenol-induced heart failure model in a dose-dependent manner. At the same time, oroxylin A markedly ameliorated cardiac histological changes and down-regulated serum levels of various neuroendocrine factors including norepinephrine, aldosterone, brain natriuretic peptide, endothelin 1, angiotensin II and so on. Mechanistically, augmenting autophagy of myocardial cells via the inhibition of AKT1-RPS6KB1 signaling contributed to the improvement of isoproterenol-induced rat heart failure by oroxylin A. Taken together, these results suggest that oroxylin A ameliorates heart failure through promoting autophagy in myocardial cells.

Objective To prepare sodium alginate-nanohydroxyapatite composite material and to explore its feasibility as a bone repair material. Methods Sodium alginate-nanohydroxyapatite composite material was prepared using chemical cross-linking and freeze-drying technology. The composite was characterized by X-ray diffraction (XRD) and scanning electron microscope (SEM) and its porosity was measured by liquid displacement method. The fifth passage of bone marrow stromal stem cells (BMSCs) were incubated on the composite material and then growth was observed by inverted micros-cope and SEM. BMSCs were cultured with liquid extracts of the material, methyl thiazolyl tetrazolium (MTT) assay was used to calculate the relative growth rate (RGR) on 1, 3, 5 d and to evaluate the cytotoxicity. Fresh dog blood was added into the liquid extracts to conduct hemolysis test, the spectrophotometer was used to determine the optical density (OD) and to calculate the hemolysis rate. Results Sodium alginate-nanohydroxyapatite composite material displayed porosity, the porous pore rate was (88.6±4.5)%. BMSCs showed full stretching and vigorous growth under inverted microscope and SEM. BMSCs cultured with liquid extracts of the material had good activities. The toxicity of composite material was graded as 1. Hemolysis test results showed that the hemolysis rate of the composite material was 1.28%, thus meeting the requirement of medical biomate-rials. Conclusion The composite material fabricated in this study has high porosity and good biocompatibility.

Objective:To explore the design of an evaluation index system with feasibility and appropriateness, and to conduct cloud model evaluation, in order to provide reference for the high-quality development of public traditional Chinese medicine(TCM) medical institutions in China.Methods:The Delphi method and analytic hierarchy process were used to construct an evaluation index system for public TCM medical institutions and calculate the index weights. Then the cloud model theory was applied to comprehensively evaluate the index system.Results:The index system of TCM medical institutions including 5 primary indicators and 12 secondary indicators of " external governance" module and 9 primary indicators and 62 secondary indicators of " internal management" module was constructed. The cloud model of the index system was(4.608, 1.022, 0.151), and the cloud rank was in the range of " very good" .Conclusions:The indicator system constructed in this study is relatively objective, scientific, and reasonable. While ensuring the accuracy and credibility of the evaluation results, it also adds evaluation information, which has certain guiding significance for promoting the high-quality development of public TCM medical institutions.