Nesfatin 1

Weight and height measurements and the body mass index (BMI) calculation were carried out meticulously for all subjects. Study nurses administered questionnaires through face-to-face interviews. After an overnight fast, blood samples were drawn in the morning at approximately 8 a.m. from a forearm vein of the participants. Tubes that had a 5-mL capacity and contained EDTA were used for collecting blood. After centrifugation of the blood at 3,000× g for 5 min at 4°C, the plasma was obtained. The separated plasma was stored in a −80°C freezer until the time of assay.
The concentrations of Nesfatin-1, corticosterone, and TSH were measured using commercially available enzyme-linked immunosorbent assay kits (Nesfatin-1: Cusabio Biotech. Co., Ltd, Wuhan, Hubei, People’s Republic of China; corticosterone: Enzo Life Sciences, Inc., New York, NY, USA; TSH: Cusabio Biotech. Co., Ltd) according to the manufacturer’s instructions. The plasma fT3 and fT4 levels were measured using a radioimmunoassay kit (North Institute of Biological Technology, Beijing, People’s Republic of China) and apparatus from USTC Zonkia (USTC Zonkia Scientific Instruments Co., Ltd, Anhui, People’s Republic of China).

The weight, height, and body mass index (BMI) of all subjects were measured. The blood sample was taken from the participants’ vein between 8:00 A.M. and 9:00 A.M. All subjects were fasting while giving blood. Tubes containing EDTA were used for collecting the samples. The blood samples were immediately centrifuged at 3,000 rpm for 5 minutes at 4°C. Extract supernatant as plasma sample was collected. The extracted plasma was stored at −80°C until detection. Commercially available ELISA kits were used to measure the concentrations of nesfatin-1 (Cusabio Biotech. Co., LTD, Wuhan, Hubei, China), cortisol (Enzo Life Sciences, Inc., Farmingdale, NY, USA), IL-6, CRP, and TNF-α (Yuanye Biotech.Co., LTD, Shanghai, China) according to the manufacturer’s instructions.