Automatic analyzer 7600

Venous blood samples were collected after 8 h overnight fasting. Fasting plasma concentrations of glucose, low-density lipoprotein cholesterol, HDL cholesterol, and TG were determined using a Hitachi Automatic Analyzer 7600 (Hitachi, Tokyo, Japan). Glycated hemoglobin (HbA1c) levels were determined using high-performance liquid chromatography with an automated HLC-723G7 analyzer (Tosoh Corporation, Tokyo, Japan). Serum creatinine levels were measured colorimetrically (Hitachi Automatic Analyzer 7600), and eGFR was calculated using the Chronic Kidney Disease Epidemiology Collaboration equation (CKD-EPI) [13 (link)]. To obtain the UACR, urinary albumin was measured in spot urine using the immunoturbidimetric method, and urinary creatinine was measured using the colorimetric method.

Venous blood samples were collected after 8 h of overnight fasting. Fasting plasma concentrations of glucose, TGs, high-density lipoprotein (HDL)-cholesterol, and low-density lipoprotein (LDL) cholesterol were determined by a Hitachi Automatic Analyzer 7600 (Hitachi, Tokyo, Japan). Glycated hemoglobin (HbA1c) levels were determined by high-performance liquid chromatography with an automated HLC-723G7 analyzer (Tosoh Corporation, Tokyo, Japan). Serum creatinine levels were measured colorimetrically (Hitachi Automatic Analyzer 7600), and eGFR was calculated using the Chronic Kidney Disease Epidemiology Collaboration equation [21 (link)]. To obtain the UACR, urinary albumin was measured in fasting morning spot urine using the immunoturbidimetric method, and urinary creatinine was measured using the colorimetric method. Urine sodium concentrations were measured using an ion-selective electrode, and 24-h sodium excretion was estimated based on concentrations of sodium and creatinine in fasting morning spot urine specimens according to 3 different methods (S1 Table).

Venous blood collection were carried out after 8 h of overnight fasting. Fasting plasma concentrations of glucose, triglyceride (TG), high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol were determined by a Hitachi Automatic Analyzer 7600 (Hitachi, Tokyo, Japan). Glycated hemoglobin levels were determined by high-pressure liquid chromatography with an analyzer HLC-723G7 (Tosoh Corporation, Tokyo, Japan). Serum creatinine (Cr) levels were measured colorimetrically (Hitachi 7600 automatic analyzer, Hitachi, Tokyo, Japan) and estimated glomerular filtration rate (eGFR) was measured using the Chronic Kidney Disease Epidemiology Collaboration equation (CKD-EPI) (Levey et al., 2009 (link)). To obtain the Urine albumin/Cr ratio (UACR), urine albumin was measured in spot urine by the immunoturbidimetric method and urinary Cr was measured using the colorimetric method.
Hepatic necro-inflammation and related liver fibrosis were estimated based on 11 different non-invasive scoring systems including Gholam’s model, BARD scores, and FIB-4 using routinely measured laboratory test and anthropometric parameters (Table S1).

Glomerular filtration rate (GFR) was estimated from the simplified equation developed using MDRD data: eGFR = 186.3 × (serum creatinine in mg/dl)^−1.154 × age^−0.203 × (0.742 for women) × (1.212 if African American).^{(18 (link))} Decreased eGFR was classified as eGFR <60 ml/min/1.73 m². Urine microAlbumin was measured with a turbidimetric assay (Albumin; Roche, Germany) using a Hitachi Automatic Analyzer 7600 (Hitachi, Japan). The urine creatinine was measured with a colorimetric assay (CREA; Roche, Indianapolis, IN) using a Hitachi Automatic Analyzer 7600. Elevated uACR was classified as uACR ≥30 mg/g. Serum 25(OH)D levels were measured with a radioimmunoassay (25-hydroxy-vitamin D ¹²⁵I RIA Kit; DiaSorin, Still Water, MN) using a 1470 Wizard Gamma Counter (Perkin Elmer, Turku, Finland). To minimize the analytical variation, serum 25(OH)D levels were analyzed by the same institute, which carried out a quality assurance program through the analysis period. Serum 25(OH)D levels were classified as either vitamin D deficiency [25(OH)D <15.0 ng/ml] and vitamin D sufficiency [25(OH)D ≥15.0 ng/ml].^{(19 (link))}

Venous blood samples were collected after 8 h overnight fasting. Serum ferritin was measured with an immunoradiometric assay using a 1470 WIZARD^® automatic gamma counter (PerkinElmer, Waltham, MA, USA) in the central laboratory. Fasting plasma concentrations of glucose and lipid were measured enzymatically using a Hitachi Automatic Analyzer 7600 (Hitachi, Tokyo, Japan). Glycated hemoglobin levels were determined by high-performance liquid chromatography with an automated HLC-723G7 analyzer (Tosoh Corporation, Tokyo, Japan). Serum creatinine levels were measured colorimetrically (Hitachi Automatic Analyzer 7600) and eGFR was calculated using the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation [17 (link)]. To obtain the UACR, urinary albumin was measured in spot urine using the immunoturbidimetric method and urinary creatinine was measured using the colorimetric method.

Participant blood samples were collected by trained nurses following overnight fasting. Drawn samples were adequately prepared and transported to the Central Laboratory after proper preparation and analyzed within 24 hours. Plasma glucose, total cholesterol, HDL-C, TG, aspartate aminotransferase (AST), ALT, blood urea nitrogen (BUN) and creatinine were measured using a Hitachi Automatic Analyzer 7600 (Hitachi, Tokyo, Japan). The level of HbA1c was measured using high performance liquid chromatography (HLC-723G7; Tosoh, Tokyo, Japan), which is the method that is certified by the National Glycohemoglobin Standardization Program.
A random 20–30 mL of midstream voided urine was collected in the morning, and then used to determine its level of urinary albumin and creatinine. Urinary albumin was measured using the turbidometric method (Roche, Germany) and urinary creatinine was measured using the Jaffe rate-blanked and compensated method (Wako, Japan) with a Hitachi Automatic Analyzer 7600 (Hitachi, Tokyo, Japan). The urinary albumin to urinary creatinine ratio (UACR) was reported as milligrams of urinary albumin per grams of urinary creatinine (mg/g). The eGFR was calculated using the Bedside Schwartz equation; eGFR (mL/min/1.73 m²) = 0.413 x height (cm) / serum creatinine (mg/dL) [17 (link)].