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C677T (MTHFR) and G1691A (FVL) mutations were screened using PCR and restriction fragment length polymorphism analysis performed with previously published primers and restriction enzymes (21) (Table ?(Table1).1). PCR amplification conditions were as follows; 35 cycles of denaturation at 95��C for 5?min, followed by annealing at 68��C for 30?s (C677T) or 56��C for 30?s (G1691A), and extension at 72��C for 5?min. PCR products were digested with appropriate restriction enzymes in a total volume of 20?��L for 16?h at 37��C, and analyzed by 2.5% agarose gel electrophoresis with ethidium bromide staining. Table 1 Details for genotyping MTHFR and FVL genes. Statistical Resiquimod analysis Hardy�CWeinberg equilibrium was calculated in the cases and controls as described in our prior study (22). Differences between genotype/allele frequencies of GDM and non-GDM subjects were calculated using T test. The Yates correction was applied prior to analyzing genotype frequencies of MTHFR and FVL mutations. The odds ratio (OR) for genotype/phenotype relations and its 95% confidence interval (CI) was calculated performing chi-square test in the cross-tabs procedure for a 2?��?2 tables using SPSS software. All statistical analyses were performed using SPSS version 19.0 (SPSS Inc., Chicago, IL, USA). Results Characteristics of the study http://www.selleckchem.com/products/3-methyladenine.html population The clinical characteristics of GDM patients and non-GDM subjects are listed in Table ?Table2.2. GDM cases (n?=?137) were aged 22�C38?years with a mean age of 26.7?��?5.1?years, whereas the age and mean age for controls (n?=?137) were 17�C34 and 24.6?��?3.55?years. The pre-pregnancy BMI range was 19.8�C35.6?kg/m2 (mean: 26.8?��?3.93?kg/m2) for GDM and 19�C31.1?kg/m2 (mean: 24.1?��?3.55?kg/m2) for controls (p?=?0.28); 40.9% of individuals managed the diabetes with appropriate diet therapy and exercise, while 59.1% of patients required four to eight units of insulin therapy over the entire prenatal period. Age, weight, FBS, and PPBG values differed significantly between Selleckchem Compound Library GDM cases and controls (p?
