Seven-wk-old male Balb/c mice(National Laboraory Animal Breeding and Research Center, Taipei, Tawan)were housed individually in solid-bottomed plastic cages with wood shavings for bedding, in a room maintained on a 12-h light dark cycle(0800-2000),24+- 1C and 50% humidity. All animals were allowed to have free access to water and food during the study Animal Care and Use Committee in the Chung Shan Medical University. Mice(n=8 per group) were randomly divided to three groups and were fed modified AIN-93G diets that differed only in the source of nondigestible carbohydrates, i.e. cellulose, chicory inulin(Santisa Co., Taiwan, active components 85% w/w) or synthetic OFS (Taiwn Sugar Co., Taiwan ,active components 67% w/w), for 3weeks. Chicory inulin used in this study has the mean degree of polymerization of eight. The active components of OFS were fructosylnystose, nystose, andl-kestose, of which the degree of polymerization was as follows(g/kg):casein, 200.0; cornstarch, 529.5; sucrose, 100.0; soybean oil, 70.0; AIN-93G mineral mix, 35;AIN-93G vitamin mix, 1.0; L-cystine 3.0;choline bitartrate, 2.5;BHT, 0.014; and nondigestible carbohydrates, 50(on the dry weight basis of active components).Mice were housed individually in metabolic cages during days 18-20 in order to collect feces without the interference of coprophagy and wood bedding. Feces voided were collected immediately into ice-bathed tubes in order to prevent the loss of SCFA and the deterioration of fecal fat. The feces collected during the 3-d were mixed together to be a 3-d fecal composite and stored at -20C until analysis .On day 21, each mouse was decapitated after light anesthesia from 0900 to 1200 after an overnight fast. Bloods were collected into heparinized tubes. The liver and colon were immediately removed, cleaned with saline, blotted dry and then weighed, The livers were frozen using liquid nitrogen and stored in -20C until analysis.
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