Ere described by the median and interquartile range. When a normal

Ere described by the median and interquartile range. When a normal distribution and equal variance were found between the two groups, a t-test was used. When equal variances were not assumed, a t9-test was used. Skewed data were log transformed before analysis to achieve a normal distribution. Hardy-Weinberg Equilibrium, genotype and allele frequency Title Loaded From File distributions were analyzed by the chi-square test. Logistic regression analysis was also applied, with adjustment for confounders. Haploview software was used to evaluate the linkage disequilibrium of SNPs. One-way ANOVA analysis or KIndependent non-parametric analysis was used to compare the fatty acid levels among the genotype groups. P values less than 0.05 were considered statistically significant.Materials and Methods Study population and blood collectionCAD patients without cancer or diabetes were recruited from Wuhan Asia Heart Hospital. CAD was defined as follows: (1) angiography showed 50 stenosis in one or more major coronary artery; (2) myocardial infarction Tubastatin A site diagnosis according to the WHO criteria issued in 1979 [13], including 18325633 clinical symptoms, enzyme elevation or ECG changes; (3) absence of atherosclerosis, or occlusion of vascular stenosis and spasm; and (4) no clinical or pathological changes or any diagnosis of the diseases mentioned above. Control participants were recruited from Zhongnan Hospital of Wuhan University. Finally, 1015 genetically unrelated Chinese subjects (33?5 years) were included in this study (505 CAD, 510 controls). The participation rates in case and control subjects from recruitment were 49.8 and 50.2 , respectively. Written informed consent was obtained from each participant, and the study protocol was approved by the ethics committees of Zhongnan Hospital of Wuhan University and Asia Heart Hospital. After an overnight fast, samples of venous blood were drawn from each subject into EDTA tubes. The tubes were immediately placed on ice until they arrived at the laboratory. Then, the blood specimens were separated into plasma, and stored at 280uC until analysis.Results General characteristics, plasma fatty acids and desaturase activity of the control and CAD patientsThe general characteristics of the control and CAD patients are summarized in Table 2. Except for gender, age and diastolic, all the characteristics were different between the two groups (p,0.01). Figure S1 shows the Chromatograms of plasma fatty acids. The plasma fatty acid concentration differed between controls and CAD patients in several instances (Table 3). After adjustment for gender, age, body mass index (BMI), blood pressure, Total-cholesterol (TC), Triglyceride (TG), HDL-cholesterol (HDL-C) and LDL-cholesterol (LDL-C), CAD patients had higher concentrations of C16:0, C16:1, C18:1n-9, AA, total Table 1. Characteristics of SNPs in FADS gene cluster.Position1 61552680 61629122 61627881 61657110 61641542 Minor allele Major allele MAF2 T T C C C G C T T A 0.333 0.454 0.143 0.474 0.Measurement of fatty acid levels and desaturase activityThe fatty acids were extracted from 200 ml of plasma and converted into their methyl esters by transesterification using the methods described previously [14].The fatty acid methyl esters were analyzed using gas chromatography (Varian 450-GC, varian Inc., USA) on a 10 m60.1 mm60.1 mm polyethylene glycol column (DB-WAX, Agilent Technologies, USA). Peaks were identified by comparison with fatty acid methyl ester standards (Sigma-Aldrich, USA) using a mass.Ere described by the median and interquartile range. When a normal distribution and equal variance were found between the two groups, a t-test was used. When equal variances were not assumed, a t9-test was used. Skewed data were log transformed before analysis to achieve a normal distribution. Hardy-Weinberg Equilibrium, genotype and allele frequency distributions were analyzed by the chi-square test. Logistic regression analysis was also applied, with adjustment for confounders. Haploview software was used to evaluate the linkage disequilibrium of SNPs. One-way ANOVA analysis or KIndependent non-parametric analysis was used to compare the fatty acid levels among the genotype groups. P values less than 0.05 were considered statistically significant.Materials and Methods Study population and blood collectionCAD patients without cancer or diabetes were recruited from Wuhan Asia Heart Hospital. CAD was defined as follows: (1) angiography showed 50 stenosis in one or more major coronary artery; (2) myocardial infarction diagnosis according to the WHO criteria issued in 1979 [13], including 18325633 clinical symptoms, enzyme elevation or ECG changes; (3) absence of atherosclerosis, or occlusion of vascular stenosis and spasm; and (4) no clinical or pathological changes or any diagnosis of the diseases mentioned above. Control participants were recruited from Zhongnan Hospital of Wuhan University. Finally, 1015 genetically unrelated Chinese subjects (33?5 years) were included in this study (505 CAD, 510 controls). The participation rates in case and control subjects from recruitment were 49.8 and 50.2 , respectively. Written informed consent was obtained from each participant, and the study protocol was approved by the ethics committees of Zhongnan Hospital of Wuhan University and Asia Heart Hospital. After an overnight fast, samples of venous blood were drawn from each subject into EDTA tubes. The tubes were immediately placed on ice until they arrived at the laboratory. Then, the blood specimens were separated into plasma, and stored at 280uC until analysis.Results General characteristics, plasma fatty acids and desaturase activity of the control and CAD patientsThe general characteristics of the control and CAD patients are summarized in Table 2. Except for gender, age and diastolic, all the characteristics were different between the two groups (p,0.01). Figure S1 shows the Chromatograms of plasma fatty acids. The plasma fatty acid concentration differed between controls and CAD patients in several instances (Table 3). After adjustment for gender, age, body mass index (BMI), blood pressure, Total-cholesterol (TC), Triglyceride (TG), HDL-cholesterol (HDL-C) and LDL-cholesterol (LDL-C), CAD patients had higher concentrations of C16:0, C16:1, C18:1n-9, AA, total Table 1. Characteristics of SNPs in FADS gene cluster.Position1 61552680 61629122 61627881 61657110 61641542 Minor allele Major allele MAF2 T T C C C G C T T A 0.333 0.454 0.143 0.474 0.Measurement of fatty acid levels and desaturase activityThe fatty acids were extracted from 200 ml of plasma and converted into their methyl esters by transesterification using the methods described previously [14].The fatty acid methyl esters were analyzed using gas chromatography (Varian 450-GC, varian Inc., USA) on a 10 m60.1 mm60.1 mm polyethylene glycol column (DB-WAX, Agilent Technologies, USA). Peaks were identified by comparison with fatty acid methyl ester standards (Sigma-Aldrich, USA) using a mass.