Au480 chemical analyzer

The levels of plasma glucose and creatinine were measured on an AU480 Chemical Analyzer (Beckman Coulter, Brea, CA, USA). The levels of fructosamine and glycated albumin in the plasma were determined by ELISA using Mouse Fructosamine (FTA) ELISA kit and Mouse Glycated Albumin (GALB) ELISA kit (MyBioSource, San Diego, CA, USA, catalog number MBS9371486 and MBS733522 respectively). The plasma concentrations of leptin, insulin, ghrelin and PAI-1 were assessed by Multiplex analysis (Bio-Plex Pro™ Mouse Diabetes Multiplex Metabolic Biomarkers Assay, #171F7001M, Bio-Rad Laboratories, Hercules, CA, USA). Urinary albumin and creatinine were measured with a Mouse Albumin ELISA kit and a Mouse Creatinine kit (Crystal Chem, Elk Grove Village, IL, USA, catalog number 80630, 80350); albuminuria was estimated by UACR.

Routine laboratory measurements, including glycated hemoglobin A_1C (HbA_1C), serum creatinine, UACR, and urinary excretion of total protein, were performed on AU480 Chemical Analyzer (Beckman Coulter, Brea, CA, USA). The eGFR was calculated by the CKD-EPI formula (2009).
The concentrationsof RBP-4, GST-α_1, GST-π, TGF-β, Col1, Col4, BMP-7, and HGF were assessed in the morning urine samples by ELISA. The commercially available kits: SEA929Hu for RBP-4; SEA609Hu for GST-α₁; SEB090Hu for GST-π; SEU124Hu for TGF-β;HEA571Hu for Col1; HEA180Hu for Col4; SEA799Hu for BMP-7;and EA047Hu for HGF(Cloud Clone Corp., Katy, TX, USA) were used in this study. The activation reagent IS044 (Cloud Clone Corp., Katy, TX, USA) was applied additionally for the measurement of TGF-β concentration. None of the received results were out of the upper limit of the range defined by the kit manufacturer. The results which were below the lower limit of the range defined by the manufacturer were set as zero. The results were adjusted to the urinary creatinine.