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The cycle threshold (CT) for the target genes ( SIRT-1 and CYP2E1 ) and the endogenous control GAPDH for each sample were detected and the relative quantitation of SIRT-1 and CYP2E1 mRNA expression levels was estimated by normalizing CT of each gene with the CT of GAPDH in each sample using the 2 CT method (Livak and Schmittgen, 2001)