7600 biochemical analyzer

GH and IGF-1 were measured as described previously (78 (link)). Briefly, GH was measured by a two-site chemiluminescent immunometric assay (AutoDELFIA hGH, PerkinElmer Life and Analytical Sciences, Wallac Oy), with intraassay coefficient of variation (CV) of 5.3 to 6.5%, interassay CV of 5.7 to 6.2%, and sensitivity up to 0.01 μg/L (0.026 mU/L).
IGF-1 was measured with the Immulite 2000 solid-phase, enzyme-labeled chemiluminescent immunometric assay (Siemens Healthcare Diagnostic Products Limited, UK); normal age-appropriate ranges are as follows:1 to 6 years, 49 to 327 μg/L; 7 to 11 years, 57 to 551 μg/L; 12 to 13 years, 143 to 850 μg/L; 14 to 16 years: 220 to 996 μg/L; 17 to 18 years, 163 to 731 μg/L; 19 to 20 years, 127 to 483 μg/L; 21 to 35 years, 115 to 358 μg/L; 36 to 50 years, 94 to 284 μg/L; >50 years, 55 to 238 μg/L; intraassay CV, 2.3 to 3.5%; interassay CV, 7.0 to 7.1%; sensitivity, 20 μg/L. IGF-1 index = IGF-1/upper limit of normal range (ULN) (79 (link)).
Plasma glucose was measured by HITACHI 7600 Biochemical Analyzer (Tokyo, Japan).

All laboratory tests were performed at the Central Laboratory of Beijing Ditan Hospital. HBsAg, HBsAb, and HBeAg were detected by a chemiluminescent microparticle immunoassay (Architect i2000 analyzer; Abbott Diagnostics, Abbott Park, IL, USA). The detection range of HBsAg level was 0.05–250 IU/mL, the detection range of HBsAb level was 0–1000 mIU/mL, and the detection limit of HBeAg was 1.0 S/CO (sample/cut off). Serum HBV DNA load was quantified using a real-time fluorescent quantitative PCR amplification (Kehua Biological Company, Shanghai, China), with a lower limit of 500 copies/mL. Blood routines were run on a Sysmex XE-5000 system (Japan), while liver and kidney function was tested using a Hitachi 7600 biochemical analyzer (Hitachi 7600–020, Japan).
The auditory screening was performed using the Madsen Echo Screen (Germering, Germany). After 72 h of breastfeeding or artificial feeding, heel blood was collected on filter paper from the infants and tested by the Beijing Neonatal Diseases Screening Center to rule out congenital phenylketonuria and hypothyroidism. The same physician completed all GDS examinations from the Beijing Child Health Care Institute.

On the last day of the experiment, the mice were fasted for 6 h (with free access to drinking water), and then blood and tissues were sampled. Blood samples were collected. The blood sample was centrifuged at 5000 rpm for 10 min to collect the serum, and the serum was frozen at -40°C until use. According to standard clinical protocols, the levels of serum glucose, low-density lipoprotein (LDL), high-density lipoprotein (HDL), total cholesterol, and triglyceride (TG) were measured using Hitachi 7600 biochemical analyzer (Hitachi, Japan). In addition, serum insulin and corticosterone levels were evaluated using the ultrasensitive mouse insulin ELISA kit (CrystalChem, IL, USA) and the commercially available corticosterone ELISA kit (Abnova, Taiwan, China) according to the manufacturers’ instructions.

Fasting blood samples, collected from all of the study subjects, were used both as a hematological index and for biochemical analysis. Mid-stream urine was obtained in the morning for the URBP measurements. Commercial assay kits (Chongqing Zhongyuan Biotechnology Limited Company, Chongqing, China) were used to measure RBP levels, and the detection method was latex particle-enhanced immune-turbidimetry. The normal level of URBP ranged from 0.00 to 0.70 mg/L. All samples were analyzed using Hitachi 7600 biochemical analyzer (Hitachi, Tokyo, Japan) and Sysmex XE-2100 automatic analyzer (Sysmex, Kobe, Japan) by professional laboratory medical personnel.

After the 14-h fasting, animals were anesthetized with 10 mg/kg ketamine and weighted after complete sedation. Four-microliters of blood was obtained from the femoral vein either in the left or right leg with a 20-gauge syringe, and transferred into an EDTA anticoagulant tube for each animal by well-trained lab technicians. The blood was immediately centrifuged at 3,000 rpm for 15 min, and the plasma was then stored at -20C°, which was then analyzed within 48 h. After sampling, most animals were put back to their group colony however a few were taken to the primate facility of Wincon to avoid desertion and inapt to the original groups.
Fasting plasma glucose (FPG), total cholesterol (T-Cho), triglyceride (TG), HDL-C and LDL-C were measured using the Hitachi 7600 Biochemical Analyzer (Hitachi Ltd., Tokyo, Japan) in a clinical laboratory, which is an affiliated hospital of Guangxi Medical University. All assay kits were manufactured by Shanghai Zhicheng Biological Technology Co., Ltd., FPG concentrations were measured using GOD-PAP method; Fasting T-Cho concentrations were measured using CHOD-PAP method; Fasting TG concentrations were measured using GPO-PAP method and Fasting HDL-C and LDL-C concentrations were measured using direct clearance method.