Flow cytometry enumeration of T-lymphocyte subsets was performed on a FACSCalibur system (BD Biosciences, San Jose, CA, USA), and the percentages of T-lymphocyte subsets were analyzed using multiphase software (BD Biosciences, San Jose, CA). For laboratory measurements, blood was available from each patient early in the morning after an overnight fast. Laboratory data, including serum hemoglobin, albumin, creatinine, uric acid, urea nitrogen, calcium, phosphorus, intact parathyroid hormone (iPTH), and cystatin C, were analyzed by a Beckman AU5800 automatic biochemical analyzer (Beckman Coulter, Inc., Brea, CA) according to the manufacturer's protocol. The estimated glomerular filtration rate (eGFR) was calculated by the CKD Epidemiology Collaboration (CKD-EPI) creatinine equation (22 (link)).
Beckman au5800 automatic biochemical analyzer
Manufactured by Beckman Coulter
Sourced in United States
The Beckman AU5800 is an automatic biochemical analyzer designed for clinical laboratory testing. It performs a variety of biochemical assays, including colorimetric, enzyme, and immunoassay tests, to analyze samples and provide quantitative results. The AU5800 is capable of high-throughput testing with a large sample capacity and efficient workflow.
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6 protocols using beckman au5800 automatic biochemical analyzer
1
Lymphocyte Subset Analysis and Kidney Function
2
Monitoring HBV Viral Load and Markers
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The HBV viral load and HBsAg and ALT levels of patients were determined at the baseline and during treatment (weeks 12, 24 and 48) in the sera. Serum HBV DNA was measured by Roche COBAS AmpliPrep/COBAS TaqMan HBV Quantitative Test, Version 2.0 (Roche Diagnostics, Indianapolis, IN), with dynamic range 20-1.7 E8 IU/ml. The HBsAg level was measured by Abbott Architect Plus i2000 analyzer (Abbott, Abbott Park, IL), with a lower limit of detection of 0.05 IU/ml. The ALT level was determined by Beckman AU5800 automatic biochemical analyzer (Beckman Coulter Inc., Atlanta, GA).
3
Hyperhomocysteinemia induction in rats
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Male Sprague-Dawley rats (12 weeks old) were purchased from Sino-British SIPPR/BK Laboratory Animal Ltd. (Shanghai, China) in the whole study. All of the procedures were guided and approved by the Animal Care and Use Committee of the Second Military Medical University and conducted specifically to the principles of the Institutional Animal Care.
Animals were divided into two groups. The HCY group at the age of 12 weeks was given by L-methionine (1 g/kg body weight per day) in tap water for a period of 8 weeks, as previously described [18 (link)]. The amount of water drank by animals every day was divided into two parts. The first part was mixed with L-methionine according to body weight of the individual, while the second one was the normal potable water without L-methionine. The control group was given equal volume of water. Eight weeks after HCY treatment, plasma HCY levels in rats were measured. Blood samples were taken from rats and were centrifuged for 10 min at 1500 rpm at 4°C, followed by collection of plasma for the evaluation of HCY. HCY reagent (Beijing Strong Biotechnologies Inc., Beijing, China) was used to detect the levels of HCY by an enzymatic cycling method with Beckman AU 5800 automatic biochemical analyzer (Beckman, USA).
Animals were divided into two groups. The HCY group at the age of 12 weeks was given by L-methionine (1 g/kg body weight per day) in tap water for a period of 8 weeks, as previously described [18 (link)]. The amount of water drank by animals every day was divided into two parts. The first part was mixed with L-methionine according to body weight of the individual, while the second one was the normal potable water without L-methionine. The control group was given equal volume of water. Eight weeks after HCY treatment, plasma HCY levels in rats were measured. Blood samples were taken from rats and were centrifuged for 10 min at 1500 rpm at 4°C, followed by collection of plasma for the evaluation of HCY. HCY reagent (Beijing Strong Biotechnologies Inc., Beijing, China) was used to detect the levels of HCY by an enzymatic cycling method with Beckman AU 5800 automatic biochemical analyzer (Beckman, USA).
4
Serum Calcium Homeostasis Assessment
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Serum biochemical indices, including total calcium (Ca), phosphorus (P), alkaline phosphatase, creatinine, and alanine aminotransferase, were measured using a Beckman automatic biochemical analyzer (AU5800; Beckman Coulter, CA, USA). The serum PTH level and total 25-hydroxyvitamin D (25OHD) level were measured using an electrochemiluminescence immunoassay (e601; Roche Cobas, Germany). Calcium homeostasis can be divided into three conditions based on the changes in serum calcium and drug dosage, including improved, worsened, and stable. Given that the serum total calcium level can be slightly decreased due to the physiological reduction of the serum albumin level, patients whose serum calcium level declined obviously (defined as a decline of >0.20 mmol/L) with stable medicine dosage were described as worsened. Patients who had increased serum calcium level with stable/reduced medicine requirement were described as improved. Patients with stable serum calcium levels (including patients with a decline in the serum total calcium level <0.2 mmol/L) and unchanged drug dosage were described as stable (10 (link)).
5
Serum Biomarkers Assessment Protocol
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Blood sample were harvested in the morning after at least 8 hours fasting. Serum was gathered after centrifugation at 3,000 r/min for 10 minutes. Serum ucOC were assayed by ELISA (MK118, Takara Bio Inc, Japan), with the intra assay coe cient of variation of 4.58%-6.66% and the inter assay coe cient of variation of 5.67%-9.87%. Serum cOC were measured by ELISA (MK111, Takara Bio Inc, Japan), with the intra assay coe cient of variation of 3.3%-4.8%, the inter assay coe cient of variation of 0.7%-2.4%.
Serum calcium (Ca), phosphorus (Pi), and alkaline phosphatase (ALP) measurements were performed by a Beckman Automatic Biochemical Analyzer (AU5800, Beckman Coulter, Indianapolis, IN, USA). Serum OC, β-isomer of C-terminal telopeptides of type I collagen (β-CTX), N-terminal prepeptide of type 1 procollagen (P1NP) were measured by electrochemiluminescence immunoassay (Roche Cobas e601, Mannheim, Germany). Parathyroid hormone (PTH) was measured using chemiluminescence (Siemens ADVIA Centaur, Munich, Germany). 25hydroxyvitamin D (25OHD) was measured by electrochemiluminescence immunoassay (Roche Cobas e601, Mannheim, Germany). Creatinine (Cr) was measured by an automated Roche electrochemiluminescence system (E170; Roche Diagnostics, Basel, Switzerland). All measurements were completed by central laboratory of PUMCH.
Serum calcium (Ca), phosphorus (Pi), and alkaline phosphatase (ALP) measurements were performed by a Beckman Automatic Biochemical Analyzer (AU5800, Beckman Coulter, Indianapolis, IN, USA). Serum OC, β-isomer of C-terminal telopeptides of type I collagen (β-CTX), N-terminal prepeptide of type 1 procollagen (P1NP) were measured by electrochemiluminescence immunoassay (Roche Cobas e601, Mannheim, Germany). Parathyroid hormone (PTH) was measured using chemiluminescence (Siemens ADVIA Centaur, Munich, Germany). 25hydroxyvitamin D (25OHD) was measured by electrochemiluminescence immunoassay (Roche Cobas e601, Mannheim, Germany). Creatinine (Cr) was measured by an automated Roche electrochemiluminescence system (E170; Roche Diagnostics, Basel, Switzerland). All measurements were completed by central laboratory of PUMCH.
6
Serum and Urinary Biochemical Indices in Metabolic Studies
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Biochemical indices including serum Ca, phosphorus (P), alkaline phosphatase (ALP), and 24-h urine Ca (24-hUCa) were measured with the Beckman Automatic Biochemical Analyzer (AU5800; Beckman Coulter). Ionized-Ca (iCa) levels were measured with a blood-gas analyzer radiometer (ABL800 FLEX; Denmark). The serum parathyroid hormone (PTH) level was measured via chemiluminescence (ADVIA Centaur; Siemens, Germany). Serum 25-dihydroxyvitamin D (25OHD) value was measured using an electrochemiluminescence immunoassay (e601; Roche Cobas, Germany). Normal reference ranges for indices: serum Ca: 2.13–2.70 mmol/L. serum iCa: 1.08–1.28 mmol/L, serum P: 0.81–1.45 mmol/L, ALP: 30–120 U/L, PTH: 13–65 pg/mL, 25OHD: 30–50 ng/mL, 24hUCa: <7.5 mmol.
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