Objective To construct and identify the transgenic plant vector recombinant pBI-Eg95 plasmid of Echinococcus granulosus. Methods Total RNA was extracted from hydatid cyst protoscoleces of Echinococcus granulosus after sonication. A couple of specific primers were designed on the basis of known sequences of Eg95 gene. The desired gene was amplified by PCR technique from the cDNA, and then was cloned into the plant expression vector pBI121 to construct the recombinant pBI-Eg95 plasmid. The recombinant plasmid was electroporated into Agrobocterium tumefaciens (At) LBA4404 strain. The positive recombinant clones were confirmed by restriction endonuclease digestion and characterized by PCR. Results For RT-PCR, a specific band around 471 bp was amplified. The result of DNA sequencing of Eg95 showed the identity with the published sequence. The same band was obtained by restriction endonuclease digestion and PER from the plasmids of positive recombinant At(rAt). Conclusions The recombinant pBI-Eg95 plasmid was successfully constructed, and it provides the basis to further research of the transgenic plant vaccine of Echinococcus granulosus.

Objective To investigate the effect of stigma maydis aqueous extract on glucolipid metabolism and oxidative stress in type 2 diabetic rats.Methods Male Sprague-Dawley rats were randomly divided into normal control group,model control group,positive control group(Metformin) and stigma maydis aqueous extract low,middle,high dose group.Type 2 diabetic rats model was induced by high-fat and high-sugar dietary feeds and streptozotocin(STZ) in all groups except the normal control group.After treatment of 8 weeks,automatic biochemistry instrument was applied to detecting the levels of fast blood glucose(FBG),triglyceride(TG),total cholesterol(TC) and low density lipoprotein cholesterol(LDL-C) in serum.The levels of free fat acid(FFA),malonaldehyde(MDA) and superoxide dismutase(SOD) in serum were assayed by uv-vis spectrophotometer.Results Compared with model control group,after stigma maydis aqueous extract treatment,middle and high dose groups remarkably lowered the levels of FBG,FFA,TG and TC in dose-dependent manner(P

Recombinant expression vector pcDNA3-MCPCD59DP containing human membrane complement regulatory proteins(hCRPs) MCP and CD59 cDNA was constructed successfully by using two independent promoters.After transfected into NIH3T3 cells with calcium phosphate-DNA precipitate method,NIH3T3 pcDNA3-MCPCD59-DP transfectants were obtained by G418 selection.Extraneous genes integration was identified by PCR.The co-expression of human CD59 and MCP at both mRNA and protein levels was confirmed by using RT-PCR and Western blot analysis.Human MCP and CD59 cDNA were integrated in NIH3T3 pcDNA3-MCPCD59-DP genomic DNA after continuous 30 times passages,indicating that NIH3T3 pcDNA3-MCPCD59-DP were stable cell lines.Human complement-mediated cytolysis assays showed that NIH3T3 cells transfected stably with pcDNA3-CD59,pcDNA3-MCP,and pcDNA3-MCPCD59-DP were protected from C-mediated damage and co-expressed human MCP and CD59 provided more excellent protection against C-mediated attack as compared with either CD59 or MCP expressed alone.The dicistronic vector represents an effective and efficacy strategy to overcome C-mediated damage and has potential therapeutic value for effectively controlling complement activation and finally for preventing hyperacute rejection in clinical gene therapy.

Humans ; Hypertension, Portal ; etiology ; surgery ; Liver Cirrhosis ; complications ; surgery ; Liver Transplantation ; Portasystemic Shunt, Surgical ; methods

Anticonvulsants ; adverse effects ; Body Weight ; drug effects ; Epilepsy ; drug therapy ; Female ; Fructose ; adverse effects ; analogs & derivatives ; Humans ; Infant ; Male

Humans ; Hypertension, Portal ; physiopathology ; surgery ; Liver ; physiopathology ; Liver Cirrhosis ; complications ; Liver Transplantation ; Portasystemic Shunt, Transjugular Intrahepatic

Objective To investigate the protective immunity in mice immunized with recombinantBifidobacteria bifidum(Bb)-Eg95 vaccine of Echinococcus granulosus (Eg) and challenged with Eg protoscoleces.Methods Fifty-six female BALB/c mice 12-14 weeks old and weighed 20-25 g were vaccinated with the recombinant Bb-Eg95 vaccine subcutaneously,intramuscularly,intranasally and orally,respectively,with blank vector,Bb and medium of solution(MRS) as control,8 mice in each group.Mice were challenged with Eg protoscoleces on week 8 after immunization and killed on week 25 after infection.The weight of hydatid cyst was measured and the decreased larva rate was calculated.Sera were collected to determine the levels of IgE,IgG and its subclasses by enzyme linked immunosorbent assay(ELISA).Splenocytes were collected and cultivated to test the proliferation of splenocytes using methyltetrazolium (MTT) assay under EgAg and concanavalin A (ConA) stimulation.The results were compared with analysis of variance and the comparison between two groups was performed with LSD-t test.Results There was significant difference in the weight of hydatid cyst between groups (F =11.062,P ＜ 0.05).Compared with MRS control group[(0.075 ± 0.019)g],the hydatid cyst weight decreased in subcutaneous group [(0.050 ± 0.013)g],intramuscular group[(0.050 ± 0.019)g],intranasal group[(0.028 ± 0.016)g] and oral group [(0.031 ± 0.018)g,all P ＜ 0.01).Compared with subcutaneous and intramuscular groups,the hydatid cyst weight decreased in intranasal and oral groups(all P ＜ 0.05).The decreased larva rate was inversely proportional to the weight of hydatid cyst.There was significant difference in the levels(obsorbancy,A) of IgG,IgG2a,IgG2b,IgG1,IgG3 and IgE between these groups(F =21.774,36.977,27.071,14.746,10.131,9.444,P ＜ 0.05 or P ＜ 0.01).Compared with MRS control group (0.015 ± 0.002,0.002 ± 0.001,0.003 ± 0.001),the levels of IgG,IgG2a and IgG2b increased in subcutaneous group(0.022 ± 0.004,0.007 ± 0.002,0.008 ± 0.002),intramuscular group (0.023 ± 0.003,0.008 ± 0.002,0.007 ± 0.002),intranasal group(0.032 ± 0.007,0.012 ± 0.002,0.013 ± 0.004)and oral group(0.028 ± 0.006,0.010 ± 0.003,0.010 ± 0.002,P ＜ 0.05 or P ＜ 0.01).Compared with subcutaneous and intramuscular groups,the levels of IgG,IgG2a and IgG2b increased in intranasal and oral groups(P ＜ 0.05 or P ＜ 0.01).Compared with MRS control group(0.009 ± 0.001,0.009 ± 0.002,0.009 ± 0.001),the levels of IgG1,IgG3 and IgE decreased in subcutaneous group(0.022 ± 0.004,0.007 ± 0.002,0.008 0.002),intramuscular group(0.004 ± 0.001,0.004 ± 0.001,0.004 ± 0.002),intranasal group(0.005 ± 0.002,0.005 ± 0.003,0.005 ± 0.002)and oral group(0.005 ± 0.001,0.004 ± 0.002,0.004 ± 0.003,all P ＜ 0.01).There was significant difference in the proliferation of splenocytes in the supernatant of cultured splenocyte,of cultured splenocyte + EgAg and of cultured splenocyte + ConA(F =63.975,359.833,167.399,P ＜ 0.01).There was significant difference in the proliferation of splenocytes inside groups(F =6741.955,4953.667,869.320,201.235,175.413,139.653,169.994,all P ＜0.01).Compared with the cultured splenocyte the proliferation of splenocytes increased in the cultured splenocyte +EgAg and splenocyte + ConA (all P ＜ 0.01).Compared with the cultured splenocyte + EgAg,the proliferation of splenocytes increased in the cultured splenocyte + ConA(P ＜ 0.01).Conclusion An effective and protective immunity is induced by the recombinant Bb-Eg95 vaccine of Eg in mice.

Objective To determine the effects of Sodium Nitroprusside ( SNP) , superoxide dismutase ( SOD) and catalase ( CAT ) on the expression of catalytic subunit of the DNA-dependent protein kinase ( DNA-PKcs ) and Ku70/80 heterodimer in cardiomycyte H9C2, as well as their expression in the myocardial tissues of SD rats . Methods H9C2 cells were co-cultured with SNP at concentreations of 10, 20 and 40 mmol/L for 6 hours, SD rats were injected with normal saline , SNP, SNP+SOD, SNP+CAT or SNP+SOD+CAT.Western blot and immuno-histochemistry assay were used to examine DNA-PKcs and Ku70/80 protein expression respectively .Results The expression of DNA-PKcs and Ku70/80 increased in H9C2 cells co-cultured with SNP when compared with control group, but they were be decreased when treated with SOD or/and CAT.The expression of DNA-Pkcs and Ku70/80 in myocardial tissues of experimental groups were higher than the control .Conclusions Radical scavengers may play a role as a protective effect for sodium nitroprusside related injury in cardiac myocytes .

Objective To discuss the effect and clinical value of serum sialic acid (SA) and lactadherin(MFG-E8) in the coronary atherosclerotic heart disease(CHD).Methods Selected 295 patients who visited to Wuhan University Renmin's Hospital cardiovascular medicine (from January to June, 2014), and then according to the results of coronary angiography divide the subjects into control group (n =78, the result of coronary angiography completely normal)and coronary heart disease group(n =217, the results of coronary angiography showed that one major coronary stenosis was more than 30％).Detected their serum level of lipid, SA, MFG-E8, hs-CRP index.At last, analyze the differences of SA and MFG-E8 between these groups, and the correlation with the severity of coronary stenosis (symbolize with Gensini score), use the binary logistic regression analysis to research the relationship among the serum level of SA, MFG-E8 and the risk of CHD.Results Compared with the control group, the patients with CHD have a higher level of serum SA and a lower level of MFG-E8 (U =4 402.5, 2 736.0 respectively,P ＜0.001);the AMI group has a higher level of serum SA than the StA and UA groups (U =1 329.0,957.5 respectively, P ＜0.001);the AMI group has a lower level of serum MFG-E8 than the StA groups (U =1517.0, P ＜ 0.001).The spearman correlation analysis showed that the serum SA has a positively correlation with the level of TG, hsCRP and Gensini score, and has a negatively correlated with the level of MFG-E8 (r =0.214, 0.462, 0.378,-0.322 respectively,P ＜0.01);the MFG-E8 has a negatively correlated with the level of TG, hsCRP and Gensini score (r =-0.18,-0.425,-0.564 respectively, P ＜0.01).Binary logistic regression analysis showed that there was a positive correlation between the serum SA and the risk of CHD (OD =4.782, 5.319, 4.179 3.832, 3.648 respectively, P ＜0.001), and the was a negative correlation between the serum MFG-E8 and the risk of CHD (OD =O.118, 0.112, 0.127, 0.135, 0.134 respectively, P ＜ 0.001).Conclusion With the serum SA rised and the MFG-E8 reduced, the risk of coronary heart disease was higher, combined detection these two indexes may be able to diagnose the CHD and assess the degree of stenosis and the risk of CHD.

Objective To explore the effect of chimera on acute graft-versus-host disease (aGVHD) after xenogeneic bone marrow transplantation. Methods Chimera was produced by exposing rats to sublethal total body irradiation and mouse bone marrow transplantation plus intraperitoneal injection of CTX. Mice were divided into three groups. Mice in group A were transplanted with marrow cells of rats without chimera, mice in group B with marrow cells with chimera, and mice in group C with C57BL/6 of rat marrow cells with chimera. Prophylactic effect on the model of aGVHD in the mice induced after xenogeneic bone marrow transplantation was observed. Results All transplantated mice showed typical aGVHD signs. Animals in group A developed typical aGVHD, and death occurred in 10-12 days after transplantation. The mean survival time of group B were longer than that of group A and C, and the signs and histopathological changes of aGVHD in group B were less severe. Conclusion Xenogeneic chimera can prevent aGVHD, alleviate its symptoms and histopathological changes, and prolong mean survival time. The prophylactic effect is specific.