Biochemical assessment
TNF-α
-20
Morning blood samples of 5mL were collected via venous puncture from participants who had fasted overnight, specifically between the hours of 8:00 a.m. and 10:00 a.m. Following collection, centrifugation was performed using an Eppendorf Centrifuge 5430R. The resulting samples were divided into aliquots and stored at -20°C until analysis. All biochemical tests were conducted using a fully automated clinical chemistry analyzer (EM 360; Transasia, ERBA Diagnostics [Transasia]) with ERBA diagnostics kits (ERBA Diagnostics Mannheim GmbH).The levels of TNF-α, IL-6, IL-1β, and CRP were quantified utilizing a two-step sandwich-type immunoassay known as the Human Leptin Quantikine ELISA Kit, which incorporates enzymatic amplification.