At every visit, all subjects underwent blood tests and oral glucose tolerance test (OGTT) after an overnight fast. After a fasting blood sample was collected, glucose load of 75 g was ingested within 5 min. Blood samples were collected at 30, 60, 90, and 120 min after the glucose load. Blood glucose concentrations were measured using the ADVIA® 2400 chemistry system (Siemens, Bayern, Germany). Total glucose incremental area under the curve (iAUC) during OGTT was determined using the trapezoidal method [10 (link)]. Insulin concentration was measured using a Cobas e 601 module (Hitachi High-Technologies Corporation, Tokyo, Japan). C-peptide concentration was measured using a Cobas 8000 chemistry autoanalyzer (Roche Diagnostics System, Basel, Switzerland). Hemoglobin A1c (HbA1c) concentration was measured using an ADAMS A1C HA-8180 (Arkray Factory, Kyoto, Japan). Total cholesterol, triglyceride, HDL-cholesterol, and LDL-cholesterol concentrations were measured using a Hitachi 7600–110 analyzer (Hitachi, Tokyo, Japan).
7600 110 analyzer
Manufactured by Hitachi
Sourced in Japan
The Hitachi 7600-110 analyzer is a versatile instrument designed for clinical laboratory use. It is capable of performing a range of analytical tests and measurements. The core function of the 7600-110 is to provide accurate and reliable results for various clinical samples.
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9 protocols using 7600 110 analyzer
1
Comprehensive Metabolic Assessment Protocol
2
Blood Glucose Measurement Protocols
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Accu-chek Active blood glucometer and test strips (Roche Diagnostics, Mannheim, Germany) are used in the entire institution. The meter used glucose dehydrogenase method and measurement is done from the bottom of the strip on the color comparison field by reflectance photometry. Measurable range was 10 to 600 mg/dL. Independent menu (which is then named blood sugar test (BST menu) for recording manual POC blood glucose test results was provided in the electronic health record system. The data analysis included only the data entered on this menu. In the preliminary analysis of the BST menu data, results out of measurable range, results containing decimal points, or symbols were analyzed.
In the central laboratory, glucose was measured by the hexokinase glucose 6-phosphate dehydrogenase method using L-type Glu 2 (Wako Pure Chemical Industries, Osaka, Japan) in a Hitachi 7600-110 analyzer (Hitachi Ltd., Tokyo, Japan). In POC gas analyzer, GEM Premier 5000 (Instrumentation Laboratory, Bedford, MA, USA) provides quantitative measurements of whole blood glucose. Glucose determination is accomplished by enzymatic reaction of glucose or lactate with oxygen in the presence of glucose oxidase or lactate oxidase and the electrochemical oxidation of the resulting hydrogen peroxide at the platinum electrode.
In the central laboratory, glucose was measured by the hexokinase glucose 6-phosphate dehydrogenase method using L-type Glu 2 (Wako Pure Chemical Industries, Osaka, Japan) in a Hitachi 7600-110 analyzer (Hitachi Ltd., Tokyo, Japan). In POC gas analyzer, GEM Premier 5000 (Instrumentation Laboratory, Bedford, MA, USA) provides quantitative measurements of whole blood glucose. Glucose determination is accomplished by enzymatic reaction of glucose or lactate with oxygen in the presence of glucose oxidase or lactate oxidase and the electrochemical oxidation of the resulting hydrogen peroxide at the platinum electrode.
3
Plasma Lp-PLA2 and Cardiovascular Biomarkers
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For measurement of Lp-PLA2 concentration in plasma, 5 mL of fasting blood samples were drawn before coronary angiography was performed. The blood samples, collected in EDTA, were centrifuged at 1000×g for 15 minutes at -20℃ within 30 minutes of collection, and the separated samples were stored at -80℃. Plasma concentrations of Lp-PLA2, which was the mass level of Lp-PLA2, was determined using a commercially available Lp-PLA2 enzyme-linked immunosorbent assay (ELISA) kit (CUSABIO Biotech, Wuhan, China), an ELISA test with two specific monoclonal antibodies.16 (link),17 (link) Each sample was measured twice and the mean of the two values was calculated to minimize error. The interassay coefficient of variation was 16.11%. hs-CRP levels were measured using the latex-enhanced immunoturbidimetric method (Denka Seiken Co., Ltd., Tokyo, Japan) with a detection limit of 0.09 mg/L in serum, using a Hitachi 7600-110 automatic analyzer (Hitachi Co., Tokyo, Japan). Total cholesterol, LDL cholesterol, HDL cholesterol, and triglyceride levels in serum were measured using the standard enzymatic method (Daiichi Sankyo Co., Ltd., Tokyo, Japan for total, LDL and HDL cholesterol; Roche Diagnostics, Indianapolis, IN, USA for triglycerides) with a Hitachi 7600-110 analyzer (Hitachi Co., Tokyo, Japan).
4
Comprehensive Metabolic Assessment in Diabetes
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Efficacy assessments included the following: glucose [FPG, plasma glucose during OGTT (postprandial plasma glucose: PPG), glucose incremental area under the curve (iAUC), and glucose maximum concentration (Cmax)], insulin [fasting plasma insulin (FPI), plasma insulin during OGTT (postprandial plasma insulin: PPI), insulin iAUC, and insulin Cmax], homeostatic model assessment (HOMA)–insulin resistance (IR), and HOMA-β, glycated albumin, fructosamine, and HbA1c. The glucose and insulin iAUCs during the OGTT were determined by the trapezoidal method.
Safety assessments included the following: electrocardiogram, hematology, and laboratory tests (white blood cells, red blood cells, hemoglobin, hematocrit, platelet count, total protein, albumin, alanine aminotransferase, aspartate aminotransferase, blood urea nitrogen, creatinine, etc.), pulse rate, and blood pressure, along with a personal report and were recorded at every visits.
Blood samples were analyzed on a Hitachi 7600-110 analyzer (Hitachi High-Technologies Corporation, Tokyo, Japan).
Safety assessments included the following: electrocardiogram, hematology, and laboratory tests (white blood cells, red blood cells, hemoglobin, hematocrit, platelet count, total protein, albumin, alanine aminotransferase, aspartate aminotransferase, blood urea nitrogen, creatinine, etc.), pulse rate, and blood pressure, along with a personal report and were recorded at every visits.
Blood samples were analyzed on a Hitachi 7600-110 analyzer (Hitachi High-Technologies Corporation, Tokyo, Japan).
5
Anthropometric and Metabolic Assessments
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Anthropometric parameters (weight, height, and body mass index [BMI]) were measured using Inbody 720 (BioSPACE Co., Seoul, Korea) at each visit. We measured waist circumference and hip circumference using a tape measure and were expressed in centimeters, rounded to three times to the first decimal place. The waist-to-hip circumference ratio was calculated by dividing the waist circumference by the hip circumference.
After a 12 h fast, the blood samples were collected from the cubital vein in ethylenediaminetetraacetic acid-coated tubes. Blood samples were centrifuged at 3,000 rpm for 20 min (Hanil Science Industrial Co. Ltd., Seoul, Korea) and stored frozen at -80°C until the analysis. A Hitachi 7600-110 analyzer (Hitachi High-Technologies, Tokyo, Japan) was used to analyze blood glucose and homeostasis model assessment of insulin resistance (HOMA-IR) was performed [24 (link)].
After a 12 h fast, the blood samples were collected from the cubital vein in ethylenediaminetetraacetic acid-coated tubes. Blood samples were centrifuged at 3,000 rpm for 20 min (Hanil Science Industrial Co. Ltd., Seoul, Korea) and stored frozen at -80°C until the analysis. A Hitachi 7600-110 analyzer (Hitachi High-Technologies, Tokyo, Japan) was used to analyze blood glucose and homeostasis model assessment of insulin resistance (HOMA-IR) was performed [24 (link)].
6
Anthropometric and Metabolic Measurements
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Anthropometric parameters (weight, height, BMI) were measured using Inbody 720 (BioSPACE Co., Seoul, Korea). Waist circumference (WC) and hip circumference (HC) were measured three times in centimeters to the first decimal place with a tape measure. The waist–hip circumference ratio (WHR) was obtained by dividing WC by HC.
Blood samples were collected from the cubital vein in ethylenediaminetetraacetic acid (EDTA)-coated tubes after a 12 h fast to minimize the effect of circadian rhythm at baseline and endpoint. Blood was centrifuged at 3000 rpm for 20 min (Hanil Science Industrial Co. Ltd., Seoul, Korea) and stored frozen at −80 °C until the analysis. Blood lipid levels were analyzed using an automatic blood analyzer (COBAS NIRA, Roche, Switzerland), and liver function and blood sugar tests were analyzed using a Hitachi 7600–110 analyzer (Hitachi High-Technologies, Japan). The low-density lipoprotein cholesterol (LDL-C) level was calculated according to the Friedewald formula [20 (link)]. A homeostatic model assessment for insulin resistance (HOMA-IR) was performed [21 (link)].
Blood samples were collected from the cubital vein in ethylenediaminetetraacetic acid (EDTA)-coated tubes after a 12 h fast to minimize the effect of circadian rhythm at baseline and endpoint. Blood was centrifuged at 3000 rpm for 20 min (Hanil Science Industrial Co. Ltd., Seoul, Korea) and stored frozen at −80 °C until the analysis. Blood lipid levels were analyzed using an automatic blood analyzer (COBAS NIRA, Roche, Switzerland), and liver function and blood sugar tests were analyzed using a Hitachi 7600–110 analyzer (Hitachi High-Technologies, Japan). The low-density lipoprotein cholesterol (LDL-C) level was calculated according to the Friedewald formula [20 (link)]. A homeostatic model assessment for insulin resistance (HOMA-IR) was performed [21 (link)].
7
Baseline Characteristics of Study Participants
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The study protocol was approved by the Institutional Review Board of Jeonbuk National University (IRB No. JBNU 2019-12-005). For the analysis of baseline laboratory values, blood samples after an overnight fast were collected from the antecubital vein of the subjects. Patients completed a demographic information questionnaire (including current treatment, medical history, smoking status, and alcohol intake) and a health check-up according to a standardized protocol that includes electrocardiogram, laboratory tests, pulse rate, and blood pressure. Alcohol intake was calculated based on the number and type of drinks consumed by each patient and then expressed in grams of absolute alcohol per day. Routine medical examination included measurements of weight, height, body mass index (BMI), electrocardiogram, and vital signs taken in a seated position after at least 5 min of rest. Lipid profiles (TC, TG, HDL cholesterol (HDL-C), and LDL cholesterol (LDL-C)) were analyzed on a Hitachi 7600-110® analyzer (Hitachi High-Technologies Corporation, Tokyo, Japan). Complete blood count was analyzed using a Sysmex XE-5000TM (Sysmex Corporation, Kobe, Japan), and other laboratory tests (liver enzymes, glucose, creatinine, etc.) were analyzed using the ADVIA® 2400 chemistry system (SIEMENS, Munich, Germany).
8
Abdominal Fat Reduction Intervention
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The 30 subjects who met the study criteria were asked to visit the clinic once every 6 weeks (weeks 0, 6 and 12 of the study period) for four clinical visits including the initial screening. During each visit, current supplementation use was reviewed and symptoms or side effects were recorded. During the screening visit, demographic and lifestyle information was collected (gender, age, alcohol consumption and smoking). A medical history was taken and a urine pregnancy test was conducted.
The following parameters were assessed: abdominal fat distribution was measured and analyzed using CT (Somatom Sensation 16 MDCT; Siemens, Forchheim, Germany) before (0 week) and after the 12 weeks intervention period. Body weight, BMI, body fat mass, percent body fat and lean body mass were measured using the Inbody 3.0 (Biospace, Seoul, Korea) during each visit. Blood samples were collected after a minimum 12-h fast during initial screening and at the 6 weeks and 12 weeks intervention period to obtain lipid profiles changes of total cholesterol, triglycerides, high-density lipoprotein (HDL), low-density lipoprotein (LDL), and free fatty acids. Blood samples were analyzed with a Hitachi 7600–110 analyzer (Hitachi High Technologies, Tokyo, Japan) using standard methods in the biochemical laboratory of Chonbuk National University Hospital.
The following parameters were assessed: abdominal fat distribution was measured and analyzed using CT (Somatom Sensation 16 MDCT; Siemens, Forchheim, Germany) before (0 week) and after the 12 weeks intervention period. Body weight, BMI, body fat mass, percent body fat and lean body mass were measured using the Inbody 3.0 (Biospace, Seoul, Korea) during each visit. Blood samples were collected after a minimum 12-h fast during initial screening and at the 6 weeks and 12 weeks intervention period to obtain lipid profiles changes of total cholesterol, triglycerides, high-density lipoprotein (HDL), low-density lipoprotein (LDL), and free fatty acids. Blood samples were analyzed with a Hitachi 7600–110 analyzer (Hitachi High Technologies, Tokyo, Japan) using standard methods in the biochemical laboratory of Chonbuk National University Hospital.
9
Comprehensive Metabolic Profiling of Blood Samples
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Blood samples were collected after an overnight fasting and centrifuged on the spot in 1 h after collection. Serum lipid profiles including total cholesterol, TG, HDL-C and LDL-C were measured by a Hitachi 7600-110 analyzer (Hitachi High-Technologies Corporation, Tokyo, Japan). Serum E2 and FSH were measured by Modular Analytics (E170) (Roche, Basel, Switzerland). Serum TSH was measured by ADVIA Centaur XP Immunoassay System (Siemens Healthcare GmbH, Erlangen, Germany). Hematology (CBC) and other laboratory tests (total protein, albumin, ALP, ALT, AST, γ-GTP, total bilirubin, BUN, and creatinine, uric acid, total calcium, calcium, glucose, urine specific gravity, urine pH,) were measured using the Sysmex XE-5000TM (Sysmex Corporation, Kobe, Japan) and ADVIA®2400 chemistry system (SIEMENS, Munich, Germany), respectively.
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