The levels of intact PTH, FGF23, 1,25-dihydroxy vitamin D [1,25D], and GDF15 in the plasma were determined using rat intact PTH ELISA kit (60-2500, Immutopics International, San Clemente, CA, USA), FGF23 ELISA kit (CY-4000, KINOS Inc., Tokyo, Japan), 1,25-(OH)2 Vitamin D EIA (AC-62F1, Immunodiagnostic Systems), and Mouse/Rat GDF15 Quantikine ELISA Kit (MGD150, R&D Systems), respectively. The phosphate levels in the plasma were determined using a Malachite Green Phosphate Assay Kit (POMG-25H, BioAssay Systems) or Autoanalyzer 7180 (Hitachi). The levels of creatinine in the plasma were determined using an Autoanalyzer 7180 (Hitachi).
7180 autoanalyzer
Manufactured by Hitachi
Sourced in Japan
The Hitachi 7180 autoanalyzer is a clinical chemistry analyzer designed for automated analysis of various biochemical parameters in medical laboratories. It is capable of performing multiple simultaneous tests on a range of biological samples, including blood, urine, and other bodily fluids. The core function of the Hitachi 7180 is to provide accurate and reliable results through its automated sample processing and analysis capabilities.
Automatically generated - may contain errors
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Xt 2000i hematology analyzer, by Sysmex (1 mentions)
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52 protocols using 7180 autoanalyzer
1
Plasma Biomarkers in Rats
2
Mice Fasting and Serum Analysis
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At necropsy day, mice were fasted for 16 hours and approximately 0.6~0.9 mL of blood sample was collected from abdominal vein. Samples were centrifuged and serum was transferred to Biotoxtech Co. Ltd., Republic of Korea, for liver function and lipid profiles analysis. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (T-Chol), triglycerides (TG), LDL-cholesterol (LDL-C), HDL-cholesterol (HDL-C), and glucose were measured using Hitachi autoanalyzer (7180, Hitachi, Japan).
3
Blood Collection and Liver Function Assessment
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At necropsy day, mice were fasted for 16 h and approximately 0.6 ~ 0.9 mL of blood sample were collected from the abdominal vein. Samples were centrifuged and serum was transferred to Biotoxtech Co., Ltd. for liver function and lipid profiles analysis. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (T-Chol), triglycerides (TG), LDL- cholesterol (LDL-C), HDL- cholesterol (HDL-C) and glucose were measured using a Hitachi auto-analyzer (7180, HITACHI, Japan).
4
Plasma Biomarker Measurement Protocol
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Blood samples were collected from the tail vein or superficial temporal vein under fed or 6 h fasting conditions and centrifuged at 13000 × g for 5 min at 4°C to obtain the plasma. PG, TG, plasma total cholesterol, and alanine aminotransferase levels were enzymatically measured using the Autoanalyzer 7180 (Hitachi High‐Technologies Corporation). Fasting PG was enzymatically measured using the Glucose CII Test Wako (FUJIFILM Wako Pure Chemical Corporation). Plasma insulin levels were measured using a sandwich enzyme‐linked immunosorbent assay (Shibayagi, Gunma, Japan, or Morinaga, Kanagawa, Japan). GHb was measured using an automated high‐performance liquid chromatography‐based GHb analyzer (HLC‐723 G8, TOSOH Corporation). The homeostasis model assessment of insulin resistance (HOMA‐IR) was calculated using the following formula38 : Further, if the fasting insulin level was lower than the detection limit (20.28 μU/ml), as seen in three and five mice in the WT and KO groups, respectively (WT; n = 3/40, KO; n = 5/40), HOMA‐IR was calculated using the following formula:
5
Plasma Biomarkers for Metabolic Evaluation
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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).
6
Podocan Expression in Diabetic Nephropathy
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Blood samples were obtained by facial vein puncture in male 28-week-old db/+ and uninephrectomized (UNx)-db/db mice that received unilateral nephrectomy at the age of 6 weeks. Plasma samples were separated by centrifugation, and PG levels were measured with the Autoanalyzer 7180 (Hitachi, Japan). Urine samples were collected over 8 hours using metabolic cages, and urine creatinine levels were measured with the Autoanalyzer 7180. After desalination of the urine samples using the PD MiniTrap G-25 column (GE Health Care, United Kingdom), urinary albumin levels were measured by ELISA (Shibayagi, Tokyo, Japan), and the urinary albumin-to-creatinine ratio (UACR) was calculated. These mice were then anesthetized using 2% isoflurane inhalation, and their remaining left kidneys were dissected, weighed, and processed for histopathological analysis using hematoxylin and eosin staining. The renal cortex was immersed in RNAlater® RNA stabilization reagent. The podocan mRNA levels were measured by quantitative RT-PCR. The plasma podocan levels were measured using the mouse podocan ELISA kit (Cloud-Clone Corp., Katy, TX).
7
Oral Glucose Tolerance in Diabetic Rats
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Diabetic N-STZ-1.5 rats with impaired insulin secretion were used for oral glucose tolerance test [33 (link), 34 (link)] . Overnight fasted rats (23 weeks old) were randomized based on plasma glucose, triglyceride and BW, and were administered drugs 1 h before oral glucose loading (1.5 g/kg) (N = 6 per group, total N = 18). Blood samples were obtained from the tail vein at indicated time points and plasma parameters were determined. Plasma glucose and triglyceride were measured using an Autoanalyzer 7180 (Hitachi, Tokyo, Japan), and plasma insulin was measured using a radioimmunoassay kit (RI-13K, Merck Millipore).
8
Transwell Assay for B16BL6 Cell Migration
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Migration of B16BL6 cells was determined with the use of Transwell inserts with a pore size of 8 mm (Corning). The filters were precoated on the lower surface with fibronectin (5 mg/mL; Sigma-Aldrich) by incubation overnight at 4 C. Cells that had been cultured overnight in migration medium (RPMI 1640 supplemented with 0.5% FBS) were harvested and resuspended in migration medium containing either vehicle (0.1% DMSO) or D4-2 (1 or 10 mM). Cells (6 3 10 4 in 100 mL) were placed in the upper chamber of the Transwell apparatus, and the lower chamber was filled with migration medium. After incubation for 6 h at 37 C, the cells were fixed in methanol for 10 min at room temperature and then stained with crystal violet. After cells on the upper surface of the membrane had been removed with cotton swabs, the number of migrated cells on the lower surface was counted in five fields (2003) with a BX51 microscope (Olympus).
Hematologic and Blood Biochemical Analyses Female C57BL/6J mice at 6 weeks of age were injected intravenously with PBS, with vehicle (PBS containing 5% DMSO), or with 500 mg of D4-2 daily for 6 days. Two days after the last injection, hematologic and blood biochemical parameters were analyzed with the use of a hematology analyzer (ADVIA 2120; Siemens, Munich, Germany) or an Auto Analyzer 7180 (Hitachi, Tokyo, Japan), respectively.
Hematologic and Blood Biochemical Analyses Female C57BL/6J mice at 6 weeks of age were injected intravenously with PBS, with vehicle (PBS containing 5% DMSO), or with 500 mg of D4-2 daily for 6 days. Two days after the last injection, hematologic and blood biochemical parameters were analyzed with the use of a hematology analyzer (ADVIA 2120; Siemens, Munich, Germany) or an Auto Analyzer 7180 (Hitachi, Tokyo, Japan), respectively.
9
Cardiac Blood Collection and Analysis
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Mice were anesthetized with sustained isoflurane inhalation (3%), and blood was collected by cardiac puncture after rib removal. Freshly obtained EDTA blood was used to measure complete blood counts using the XT‐2000i hematology analyzer (Sysmex, Germany). To obtain plasma, blood was placed for 15 minutes at room temperature, then centrifuged at 1500g for 15 minutes at 4°C. Plasma levels of total cholesterol and triglycerides were measured with Hitachi 7180 autoanalyzer (Hitachi High‐Technologies Corp, Japan). Exsanguination was used as a means of euthanasia in this condition. Detailed method descriptions are available in Data S1 .
10
Unilateral Ureteral Obstruction in Mice
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UUO was performed as described previously[32 (link)]. Briefly, after induction of general anesthesia by intraperitoneal injection of pentobarbital (50 mg/kg body weight), the abdominal cavity was exposed via a midline incision, and the left ureter was ligated at two points with 4–0 silk. At the indicated times after UUO, mice were sacrificed, and the kidneys were removed for histological examination. Ureteral obstruction was confirmed by observing dilation of the pelvis and proximal ureter and collapse of the distal ureter. Sham-operated mice had their ureters exposed and manipulated but not ligated. Arterial blood pressure was measured at the indicated time points using a programmable apparatus by the tail cuff method. All mice were euthanized by cervical dislocation while under general anesthesia. Serum creatinine and blood urea nitrogen (BUN) levels were assessed by a Hitachi 7180 autoanalyzer (Hitachi High-Technologies, Tokyo, Japan).
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