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Objective To study the correlation between the renin-angiotensin-aldosterone system angiotensinogen (AGT) gene M235T,angiotensin Ⅱ type 1 receptor (AGTR1) gene Al166C,aldosterone synthase (CYP11B2) gene -344C/T polymorphisms and large-artery atherosclerotic (LAA) stroke in a southern Chinese Han population.Methods Polymerase chain reaction and gene sequencing technology were used for the genotyping in patients with LAA and normal controls with AGT gene M235T,AGTR1 gene A1166C,and CYP11B2 gene - 344C/T polymorphisms in a southern Chinese Han population,and to determine the correlation between the 3 gene polymorphisms and LAA by binary logistic regression analysis.Results A total of 107 patients with LAA and 142 healthy controls were included in the study.The frequencies of the AGT gene 253TT genotype (66.36％ vs.50.70％,x2 =6.122,P =0.047) and T allele (79.44％ vs.70.07％ ％,x2 =5.581,P =0.018) in the LAA group were significantly higher than those in the control group.The frequencies of the AGTR1 gene 1166CC genotype (0％ vs.0％,x2 =1.494,P =0.222) and C allele (7.48％ vs.4.93％,x2 =1.399,P =0.237) in the LAA group were no significantly differences with those in the control group.The frequencies of the CYP11B2 gene - 344CC genotype (9.35％ vs.4.23％,x2 =3.603,P =0.165) and C allele (27.10％ vs.26.06％,x2 =0.069,P =0.793) in the LAA group were no significant differences with those in the control group.Binary logistic regression analysis showed that there was no significant correlation between the three gene polymorphisms and the simple LAA diseases.The frequencies of AGT gene 235TT genotype (68.00％ vs.41.90％,x2 =12.446,P =0.002) and T allele (79.33％ vs.64.76％,x2 =8.993,P =0.003) in the LAA patients complicated with hypertension were significantly higher than those in the normotensive control group.Logistic regression analysis showed that the odds ratio (OR) exposed to TT genotype was 2.153 (95％ confidence interval [CI] 0.789-5.872).The OR of T allele was 2.089 (95％ CI 1.285-3.396).Conclusions The AGT gene M235T polymorphism is not associated with the simple LAA in the southern Chinese Han population,but it may be associated with the risk of LAA complicated with hypertension;CYP11B2 gene -344C/T polymorphism and AGTR1 gene A1166C polymorphism are not associated with the onset of LAA in the southern Chinese Han population.
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Objective To investigate the therapeutic effect and the detailed mechanisms of intraarterially delivery of bone marrow mesenchymal stem cells ( BMSCs) for treatment of middle cerebral artery occlusion (MCAO) in rats.Methods BMSCs were isolated,purified and amplified with the adherence culture method.BMSCs were labeled with 5-bromo-2-deoxyuridine ( BrdU ) (10 μmol/L) for 48 h before transplation.Surface antigens of CD90,CD29,CD106,CD34,CD45,CD11b were identified by flow cytometry.The MCAO model was established with suture emboli method.In this study,3×106 BMSCs were injected into rats with MCAO through intraarterial route at day 7 after stroke.The effects on functional and physical recovery were assessed with the behavioral tests (mNSS test and adhesive test) and body weight.Bielshowsky-Luxol Fast Blue double staining was used to demonstrate the reconstruction of axon and myelin.The Brdu-labeled BMSCs in vitro and in vivo were detected with direct immunofluorescent staining.The expression of neuron specific enolase ( NSE),neurite outgrowth inhibitor-A ( Nogo-A),synaptophysin (SYN),ki-67 nuclear antigen (Ki-67),glial fibrillary acid protein( GFAP),vascular endothelial growth factor ( VEGF) in brain were analyzed with immunohistochemical staining.Results Flow cytometry indicated that the positive rates of high expression of CD90,CD29,CD106 in BMSCs were respectively 91.70%,88.40% and 52.20%.Meanwhile,the positive rates of low expression of CD34,CD45,CD11b in BMSCs were 2.70%,5.65% and 7.82%,respectively.There was a significant difference in behavioral tests ( mNSS test and adhesive test) between BMSCs group and PBS group at day 21,28,35 after MCAO (mNSS:4.89 ±1.36,7.00 ±1.67,3.78 ±1.30 and 6.33 ±1.21,2.44 ±1.13,5.67 ± 1.51;t =2.69,3.83,4.75;adhesive test:54.00 ± 10.48,68.17 ± 11.09,36.89 ±9.80 and 59.33 ± 12.40,23.44 ± 9.04,46.50 ±9.38;t =2.51,3.92,4.77;P ＜0.05).Meanwhile,a significant difference in body weight was discovered between them at day 28,35 after MCAO.In BMSCs group,the area of corpus callosum in the ipsilateral hemisphere was significantly enlarged,the positive number of Brdu,SYN,Ki-67,GFAP,VEGF in brain was significantly increased,the expression of Nogo-A in brain was significantly decreased,nevertheless,the number of NSE-positive cells in brain and the infarct volume were not significant different from PBS group at day 35 after MCAO.Conclusions These results suggest that intra-arterial transplantation of BMSCs is an efficient treatment protocol for stroke.Treatment with BMSCs increases endogenous cells proliferation,angiogenesis,synaptogenesis,enhances axonal regeneration and the protective function of astrocytes,all of which may contribute to neurological functional recovery.
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Objective To investigate the correlation between the glial fibrillary acidic protein(GFAP),neuron-specific enolase(NSE),synaptoghysin (SYN),neurite outgrowth inhibitor-A(Nogo-A)expression and neurological outcome in tissue surrounding the infarct during the recovery after cerebral ischemia-reperfusion injury in rats.Methods A 2-hour middle cerebral artery occlusion(MCAO)and reperfusion model in rats was induced by the intraluminal suture method.The modified neurological severity score(mNSS)was performed at day 28,35,42,and 49.Immunohistochemistry was used to detect the expressions of GFAP,NSE,SYN,and Noga-A in tissue surrounding the infarct.Results The mNSS score decreased gradually over time after cerdnal ischemia-reperfusion injury in rats.Except day 35(5.11±0.737)vs.day 42 (4.54±0.519),and day 42 vs.day 49(4.29±0.488),there were significant differences at all other time points(all P<0.05).The numbers of GFAP positive cells deergased gradually form day 28 to day 49,in which,the numbers of GFAP positive cells at day 42(51.00±13.59)vs.day 49(44.38±11.94) were significantly less than those at day 28(69.00±15.10)(P<0.05).There were no significant differences in the numbers of NSE positive cells at all time points,but their integrated optical density(IOD)increased gradually.There were significant differences between day 28(6 218.57±1 864.25)and day 42(9 414.00±2 491.12) or day 49(12 522.50±3 106.99),and between day 35(7 343.40±1 533.35)and day 49(all P< 0.05).There were no significant differences at all other time points.The SYN express (IOD)increased gradually.and it was significantly lower at day 49(66 503.00±12 834.61)than that at day 28(43 905.14±13 208.59)(P<0.05).The numbers of Nogo-A positive cells decreased gradually,and they were significantly less at day 49(42.13±14.45) than those at day 28(59.57±15.25)(P<0.05).The GFAP expression was positively correlated with the mNSS scores(r=0.993,P=0.007).The NSE(r=-0.954,P=0.044)and SYN(r=-0.992,P=0.008) expression was negatively correlated with the mNSS scores.Conclusion The neurological outcome was associated with the downregulation of GFAP expression and the upregalation of NSE and SYN expression during the recovery after cerebral ischemia-reperfusion injury in rats.
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Objective To evaluate the detection of culture filtrate protein 10 (CFP10) and 6000 early secretory antigenic target (ESAT-6) in cerebrospinal fluid to be used in diagnosing tuberculous meningitis. Methods Dot enzyme linked immunosorbent assay ( Dot ELISA) method that was improved by applying concentrated cerebrospinal fluid was used to detect CFP10 and ESAT-6 in cerebrospinal fluid to analyze small protein antigen secreted by M. tuberculosis. Cerebrospinal fluid of 111 subjects were collected,in which 58 specimens were clinically diagnosed as tuberculous meningitis and 53 as non-tuberculous.CFP10 and ESAT-6 were detected in cerebrospinal fluid using Dot ELISA method and the results were analyzed. Results The sensitivities of detecting CFP10 and ESAT-6 antigen were 93.1% and 91.4% respectively, and the specificities were 92. 5% and 94. 3% respectively. The sensitivities and specificities are generally higher compared with the other methods of detecting M. tuberculosis or materials of M. tuberculosis by acid-fast staining or mycobacterium tuberculosis culture and polymerase chain reaction.Conclusions Using Dot ELISA method to detect CFP10 and ESAT-6 in cerebrospinal fluid to diagnose tuberculous meningitis has a high sensitivity and specificity. Our study provided the evidence of detecting the specific antigen of M. tuberculosis to be used in diagnosing tuberculosis.