Hlc 723g8

Levels of total cholesterol, LDL-C, complete blood cell count, serum triglycerides, HDL-C, albumin, blood urine nitrogen (BUN), uric acid, and glucose were measured from fasting blood samples 30 (link). The Jaffe method was used to calculate serum creatinine. Hemoglobin A1c was measured using an automated analyzer (HLC-723G8, Tosoh Corp., Tokyo, Japan). CK-MB and troponin I serum levels were measured using chemiluminescent microparticle immunoassays. Total leukocyte count and the lymphocyte, neutrophil and monocytes proportions were measured using an automated cell counter (XE-2100 Hematology Alpha Transportation System; Sysmex, Kobe, Japan). Absolute leukocyte subtype counts were calculated as the product of its proportion and total leukocyte count. The CKD-EPI formula was utilized to determine estimated glomerular filtration rate (eGFR) 31 (link). An immunochemistry system (Beckman Coulter IMMAGE) was used to evaluate plasma levels of high-sensitivity C-reactive protein (hs-CRP). The detection limit was 0.2 mg/L, and all measurements were made twice.

Fasting blood samples were drawn in the morning as part of routine clinical practice. Blood glucose was assayed using an enzymatic hexokinase method. Glycated hemoglobin (HbA1c) was measured with automated glycohemoglobin analyzers (Tosoh HLC-723G8, Tokyo, Japan). Lipid profiles were measured with automatic biochemistry analyzers (Hitachi 7150, Tokyo, Japan). HsCRP was measured by rate turbidimetry with immunoassay analyzers (Beckman Assay, Brea, CA, USA).

In the physical examination center, a questionnaire was administered among participants by well-trained investigators via face-to-face interviews. The following data were collected: characteristics of social demography, medical history, and information about lifestyle including physical activity, cigarette smoking and alcohol consumption.
After the interview, participants were taken through a medical examination. Anthropometric measurements (body weight, height and waist circumference (WC)) were measured by trained nurses, according to standardized procedures [18 (link)]. BMI was then calculated as body weight (unit: kg) divided by the square of height (unit: m) using Quetlet’s index. After a period of 5-min rest in a quiet room, the systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured on the right upper arm in sitting position, using an automatic machine (AND TM-2655P, Japan). Venous blood samples were collected after an overnight fast of at least 10 h. All measurements were performed in air-conditioned room (22–26 °C) of the examination center, between 8:00 and 11:00 in the morning. Fasting plasma glucose (FPG), TC, TG, HDL-C and LDL-C levels were assayed by using a fully automated biochemistry analyzer (Roche Cobas 6000, Germany). Hemoglobin A_1c (HbA_1c) was assayed with an automated glycohemoglobin analyzer (TOSOH HLC-723G8, Japan).

Blood samples were collected from the tail or facial vein without anesthesia and centrifuged at 1500 g for 10 min at 4 °C to isolate the plasma. To prevent degradation of incretin hormone, blood samples were treated with not only heparin/aprotinin but also a dipeptidyl peptidase‐4 (DPP‐4) inhibitor. Plasma alanine aminotransferase (AST) and aspartate aminotransferase (ALT) levels were measured by an Autoanalyzer 7180 (Hitachi High‐Technologies Corporation, Tokyo, Japan). GHb was measured with an automated GHb analyzer (HLC‐723G8; Tosoh, Tokyo, Japan). Plasma insulin was measured with an ELISA kit (Shibayagi Co., Ltd., Gunma, Japan). Plasma levels of total GLP‐1 and total PYY were also measured with an ELISA kit (Wako Pure Chemicals).

Clinical measurements of the individuals were collected by well-trained investigators, including sex, weight, height, the age at T1D onset and some laboratory examination results, such as the FPG level, 2-h PPG level according to the OGTT test, fasting C-peptide (FCP) level and 2-h postprandial C-peptide (PCP) level. The FCP and 2-h PCP levels were detected by a chemiluminescence method (ADVIA Centaur XP Immunoassay System, Siemens, Germany) in the laboratory of the Second Xiangya Hospital, and the HbA1c level was detected by automated liquid chromatography (HLC-723G8, Tosoh, Japan). The radioligand binding assay was used in our laboratory to detect islet autoantibodies such as GADA, IA-2A and ZnT8A (21 (link)–23 (link)).

Blood samples of all participants were collected after a 10–12 h overnight fasting immediately after their admission. Plasma samples were prepared by centrifugation at 3500 × g twice for 10 min each at 15–18 °C and were stored at −80 °C until analysis according to our previous studies [20 (link)–22 (link)].
Plasma PCSK9 levels were measured using a high-sensitivity, quantitative sandwich enzyme immunoassay (Quantikine ELISA, R&D Systems Europe Ltd) according to our previous study [22 (link)]. The lower limit of detection was 0.096 ng/mL. PCSK9 levels of each patient were determined twice and the mean value of the two samples was used in the final analysis. Plasma levels of fibrinogen were quantitatively measured by the method of Clauss and a Stago autoanalyzer with STA Fibrinogen kit (Diagnostic Stago, Taverny, France). The levels of plasma triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), LDL-C, apolipoprotein (apo) A1, apoB and fasting plasma glucose (FPG) were determined by automatic biochemistry analyser (Hitachi 7150, Japan). Hemoglobin A_1C (HbA_1C) was determined using Tosoh Automated Glycohemoglobin Analyser (HLC-723G8, Tokyo, Japan).