Male C57BL/6 mice (BioLasco, Taipei, Taiwan) were housed in a specific-pathogen-free (SPF) facility, which is accredited by the Association for Assessment and Accreditation of Laboratory Animal Care International (AAALAC). Mid-grade sepsis was induced in the animal models by cecum ligation and perforation (CLP) 6. Briefly, mice were anesthetized with a combination of 0.1 mg/g ketamine and 0.01 mg/g xylazine. Via midline abdominal incision, the cecum was mobilized and ligated in the middle of the cecum, below the ileocecal valve, punctured once using a 21-G needle, and a small stool sample was squeezed out of the cecum to induce polymicrobial peritonitis. The abdominal wall was closed in two layers. Sham-operated mice underwent the same procedure, including opening of the peritoneum and exposing the bowel, but without ligation and needle perforation of the cecum. After surgery, the mice were resuscitated by subcutaneous injection of pre-warmed (37°C) normal saline (at 5 mL per 100 g of body weight). Total fifty-four mice were used in this study. All animal protocols were approved by the Institutional Animal Care and Use Committee of Chang Gung Memorial Hospital. All surgical procedures, including analgesia, were performed according to national and institutional guidelines.

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