[Association between inflammatory cytokine CD40 gene polymorphism and risk of acute coronary syndrome].

OBJECTIVE: To investigate the expression levels of CD40, sCD40L, hs-CRP, WBC in acute coronary syndrome (ACS) patients and the association between CD40-1C/T single nucleotide polymorphism and risk of ACS in Han Chinese, moreover, the regulatory effects of IFN-gamma and fluvastatin on the expression of CD40 in peripheral blood mononuclear cell (PBMNC) were also observed. METHODS: (1) 160 ACS patients and 92 control patients diagnosed by coronary angiography were recruited. Enzyme linked immunosorbent assay, particle enhanced immunoturbidimetric assay, flow cytometry were used to detect the levels of soluble CD40L, hs-CRP, and WBC count. (2) The CD40 genotype and allele frequency were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing technology. (3) PBMNC were separated by density gradient centrifugation heparinized venous blood from 40 ACS patients, cultured for 24 h with or without 100 ng/ml IFN-gamma in the absence or present of 10 micromol/L fluvastatin. The CD40 expression levels were then detected by flow cytometry. RESULTS: Inflammatory cytokine CD40, sCD40L, hs-CRP levels were significantly higher in ACS patients than in controls. The CD40-1C allele frequency was 0.606 in ACS group and 0.489 in controls, while T allele frequency was 0.394 in ACS group and 0.511 in controls. The frequency of CC genotype was significantly higher in ACS group than in controls (P < 0.01). C allele carriers had significantly higher risk of ACS (OR = 1.608, 95%CI: 1.12 - 2.32, P = 0.011). CD40 production increased after 24 h culturing and the CD40 levels were significantly higher in subjects with CC genotype than that in subjects with CT or TT genotype [CC: (14.78 +/- 4.56) MFI, CT: (11.98 +/- 4.12) MFI, TT: (9.86 +/- 3.83) MFI, P < 0.05]. IFN-gamma further increased PBMNC CD40 expressions in all subjects after culturing for 24 h and fluvastatin equally inhibited IFN-gamma induced PBMNC CD40 expression from various genotypes (CC, CT, TT was 30.3%, 26.3%, 29.3% respectively, all P > 0.05). CONCLUSION: Inflammatory cytokines were increased in ACS patients and CD40-1C/T polymorphism is associated with higher risk for ACS in Han Chinese.

[Expression of WT1 gene and its isomer ratio changes during phorbol ester induced differentiation of K562 cell line].

OBJECTIVE: To explore the changes in expression of WT1 gene and ration of its isomers during phorbol ester (TPA) induced differentiation of leukemia cell line K562 by fluorescence quantitative RT-PCR and analysis the relationship between different isomers and hematogenic cell differentiation. METHODS: The degree of cellular maturation were verified by NBT reduction test and immunophenotyping. Expression of WT1 gene was determined by fluorescence quantitative RT-PCR during differentiation of K562 cell line. The relative ratio of the four splicing variants WT1 ( + / + ), WT1 ( + / - ), WT1 ( - / + ), WT1 ( - / - ) were calculated. RESULTS: During the differentiation of K562 cell, the NBT reduction rate and the CD9 positive rate both increased significantly (P < 0. 05). The expression of WT1 gene decreased immediately to (1.67 +/- 0.45) x 10(-3) from (4.67 +/- 1.11) x 10(-3), and then increased again to (4.64 +/- 1.53) x 10(-3) at 96 hours. The ratio of WT1 ( + / + ) was decreased gradually, from 0 hour (39.65 +/- 19.46)% to 96 hour (15.25 +/- 7.27)%. While the ratio of WT1( - / - ) was increased, from 0 hour (15.38 +/- 11.34)%, to 96 hour (37.60 +/- 11.90)%. The other two isomers ratios did not change significantly. CONCLUSION: During the TPA induced differentiation of K562 cell, there are two high expression levels of WT1 gene. Before differentiation, the majority is WT1 ( + / + ), and after differentiation, is WT1 ( - / - ). It indicates that WT1 gene may activate or inhibit cell differentiation by regulating the ratio of its four splicing variants.

[Correlation of CD40 gene polymorphisms with acute coronary syndrome, hypertension and diabetes].

OBJECTIVE: To investigate the correlation of CD40-E1SNP (-1C/T) and CD40-E4SNP with acute coronary syndrome (ACS), hypertension and diabetes in Chinese Han population. METHODS: The allele frequencies of CD40-E1SNP (-1C/T) and CD40-E4SNP were assayed by polymerase chain reaction-restriction fragment length polymorphism and DNA sequence technology in 160 definite ACS patients and 92 controls with negatively coronary arteriography. Multivariate logistic regression models were used to analyzed the interaction between the CD40 gene polymorphisms and hypertension and diabetes. RESULTS: CD40-1C allele frequency of the ACS group was 0.606, significantly higher than that of the control group (0.489, P < 0.01), while the T allele frequency of the ACS group was 0.394, significantly lower than that of the control group (0.511, P < 0.01). The frequency of CC genotype was much higher in the ACS group than in the control group (P < 0.01). C allele increased the risk of ACS (OR = 1.608, 95% CI: 1.12 - 2.32, P = 0.011). After adjustment of confounding variables, such as age, sex, and body mass index, the binary logistic analysis showed a significant gene-environment interaction (P < 0.05). The OR value were: 1.608 for C allele (95% CI: 1.12 - 2.32, P < 0.05), 5.71 for C allele-with hypertension (95% CI: 1.12 - 29.08, P < 0.05), 1.44 for C allele-without diabetes (95% CI: 1.12 - 5.13, P < 0.01), and 2.35 for C allele-with diabetes (95% CI: 1.47 - 4.82, P < 0.01). Expression of CD40-E4SNP was not found in these subjects. CONCLUSION: CD40-1C/T polymorphism is associated with ACS in Chinese people. The -1C allele carriers have higher risk of ACS if they get hypertension or diabetes; CD40-E4SNP may not exist in Chinese people.