1.Correlation between renin-angiotensin-aldosterone system gene polymorphisms and large artery atherosclerotic stroke: a study in a southem Chinese Han population
Xiaomu WU ; Xinghua ZHOU ; Weizhen WANG ; Chaodong WANG ; Guozhu HU ; Yingqiong XIONG ; Lijuan LIU ; Kunnan ZHANG
International Journal of Cerebrovascular Diseases 2012;20(1):35-41
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.
2.The significance in diagnosing tuberculous meningitis by detecting specific antigen in cerebrospinal fluid
Kunnan ZHANG ; Ziqiang XU ; Chaodong WANG ; Xinhui QU ; Xufang XIE ; Xiaomu WU
Chinese Journal of Neurology 2011;44(2):86-90
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.
3.Effect and mechanism of miR-3188 on malignant biological behavior of gastric cancer cell
Kunnan Wang ; Jinming Zhang ; Yun Zhang ; Junze Zhang ; Xinpu Yuan ; Guijun Zou ; Chaojun Zhang
Acta Universitatis Medicinalis Anhui 2022;57(4):545-551
Objective:
To explore the effect of miR-3188 on the proliferation, apoptosis, invasion and migration of gastric cancer cells and the underlying molecular mechanism.
Methods:
The expression level of miR-3188 was examined in normal human gastric mucosal cells(GES-1) and human gastric cancer cell lines(HGC-27,MGC-803,BGC-823,MKN-45) by qRT-PCR. It was determined by bioinformatic analysis and dual luciferase reporter assay whether NRAGE was the target gene of miR-3188.miR-3188 mimic, miR-3188 inhibitor and negative control miR-NC were transfected into gastric cancer cell line HGC-27 respectively, and the expression of NRAGE on protein and mRNA levels was detected by Western blot and qRT-PCR. miR-3188 mimic, NRAGE overexpression plasmid(pc-NRAGE) and its negative control(miR-NC,pc-control) were separately or co-transfected into gastric cancer cell line HGC-27. CCK-8, flow cytometry and Transwell experiment were used to detect cell proliferation, apoptosis, invasion and migration in each group. The expressions of epithelial-mesenchymal transition(EMT), proliferation, apoptosis, invasion and migration related proteins [CyclinD 1,matrix metalloproteinase 9(MMP 9),Bcl-2,N-cadherin, Vimentin, E-cadherin] in each group were detected by Western blot.
Results:
The expression of miR-3188 in human gastric cancer cell lines was lower than that of human normal gastric mucosal cells. Bioinformatics analysis and dual luciferase reporter assay confirmed that miR-3188 could target and bind to NRAGE 3′UTR.Western blot and qRT-PCR confirmed that miR-3188 negatively regulated the expression of NRAGE on the protein level in HGC-27 cells, but not on the mRNA level. Compared with the miR-NC group, Transfected miR-3188 mimic reduced the proliferation, invasion and migration ability of HGC-27 cells, and increased the apoptosis rate, and the co-transfection of pc-NRAGE could reverse the above effects. Compared with the miR-NC group, when miR-3188 was overexpressed in HGC-27 cell, the expression of cyclinD 1, MMP 9, Bcl-2, N-cadherin, and Vimentin was down regulated, while E-cadherin expression was up-regulated, and the above effects could be reversed by co-transfection with pc-NRAGE.
Conclusion
The miR-3188/NRAGE axis may play important roles in the progression of gastric cancer, and miR-3188 may be a potential therapeutic target for gastric cancer.