7600 clinical analyzer

For each patient, demographic and baseline clinical data (including age; sex; complications related to liver disease such as ascites, HE, and HRS; and clinical course in the hospital) were obtained from medical records and were recorded in a specific liver disease pro forma. Laboratory variables that were evaluated included levels of total protein, serum albumin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin, creatinine, and blood urea nitrogen (BUN) as well as the international normalized ratio (INR). All biochemical values were measured using a Hitachi 7600 clinical analyzer (Hitachi) and a Sysmex CA1500 fully automatic analyzer (Sysmex Corp.). Hematological parameters including white blood cell (WBC) count and the relevant subpopulations (ie, lymphocyte, neutrophil, and monocyte counts), RDW, platelet counts, and hemoglobin levels were analyzed using an automated analyzer (Sysmex XN‐9000). The MLR and NLR were calculated by dividing the number of monocytes by lymphocytes and by dividing the neutrophil count by lymphocyte count, respectively. Additionally, hepatic function was evaluated using the Model for End‐Stage Liver Disease (MELD) score, which was calculated as previously described.¹³ The 28‐day patient survival rate was determined. Date of death was obtained from medical records.

Blood and urine samples were obtained on admission and every 24 hours up to 72 hours for measuring the NGAL, Cys-C, and sTREM-1 levels. Blood was centrifuged at 3,000 revolutions per minute (rpm) for 15 minutes, and urine was centrifuged at 2,000 rpm for 5 minutes. The supernatants were transferred to Eppendorf tubes and stored at −80°C. All of the specimens were renumbered before the experiment. The plasma NGAL level was determined by using a Triage NGAL Assay (Alere Inc., San Diego, CA, USA), and the measurable range was 15 to 1,300 ng/mL. The urine NGAL level was analyzed by using a NORMAN-2 scattering turbidimetry analyzer with an NGAL Assay (Norman Inc., Nanjing, China), and the measurable range was 0 to 4,000 ng/mL. The Cys-C level was measured by using an automated chemistry analyzer (Hitachi 7600 Clinical Analyzer; Hitachi, Tokyo, Japan) with a latex immunoturbidimetry assay. The level of sTREM-1 was determined by using a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) (R&D Systems Inc., Minneapolis, MN, USA) with a measurable range of 0 to 4,000 pg/mL. ELISA was performed in duplicate, and other assays were performed in strict accordance with the instructions of the manufacturers. Laboratory investigators were blinded to the clinical information throughout the study.

Fasting blood samples were drawn from all the adolescents after a 12-hour fast. The samples were added to an ethylenediaminetetraacetic acid (EDTA) anticoagulation tube, mixed upside down 8–10 times, and centrifuged at 2,500 rpm for 10 minutes at room temperature for plasma separation. The extracted plasma sample (40 μL) was transferred to a 2 mL Eppendorf (EP) tube. Ice-cold methanol was added (160 μL) to remove protein. The centrifuge tube was vortexed at maximum speed for 10 s and centrifuged at 13,500 rpm for 15 min. Supernatants (80 μL) were transferred to liquid chromatography (LC) vials and stored at -80°C until the ultra-performance liquid chromatography (UPLC)–mass spectrometry (MS) analysis. In addition, triglycerides (TG), cholesterol (TC), high-density lipoprotein cholesterol (HDLC), and low-density lipoprotein cholesterol (LDLC) were measured by the Shengjing Hospital of China Medical University using a Hitachi 7600 clinical analyzer (Hitachi, Tokyo, Japan).

Clinical examinations included questionnaires, medical history, anthropometric measurements, and biochemical analysis. During the examination, the physician recorded the medical history (including previous diseases and drug prescriptions) and drinking frequency and amount. The smoking history was also recorded and distinguished as yes or no.
The anthropometric measurements were performed as previously described, including body weight, standing height, waist circumference, and blood pressure [25 (link), 26 (link)]. Weight and height were measured when the patient was wearing light clothing and no shoes. Waist circumference was measured after the patient exhaled, with the tape measure placed between the lowest rib and the upper edge of the iliac crest. Blood pressure was measured after resting for 5 min. Body mass index (BMI) was calculated as the weight (kg) divided by height (m) squared.
Fasting blood samples were taken from the anterior cubital vein and were used for biochemical analysis. Measurements included liver enzymes, blood lipids, glucose, and uric acid. All of the biochemical values were measured by a Hitachi 7600 clinical analyzer (Hitachi, Tokyo, Japan) using standard methods. Serum 25-hydroxyvitamin D levels were measured with the electro-chemiluminescence immunoassay (ECLIA) platform using the Roche cobas e602 analyzer (Roche Diagnostics GmbH, Germany).

A peripheral venous blood sample (6 mL) was collected from each
patient within the first 24 hours after admission to the ER. Blood
samples were used to analyze the hematological index and biochemical values.
Laboratory parameters measured included: white blood cells (WBC), platelets,
hemoglobin, red blood cell distribution width (RDW), neutrophil-lymphocyte
ratio (NLR), prothrombin time, total protein, albumin, alanine aminotransferase
(ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), creatine
kinase (CK), creatinine, potassium, pH, PaCO₂, and PaO₂.
All biochemical analyses were conducted using a Hitachi 7600 Clinical Analyzer
(Hitachi, Tokyo, Japan), Sysmex CA-7000 System (Sysmex, Kobe, Japan), and
Sysmex XE-2100 Automated Analyzer (Sysmex) using standard methods.

Each patient provided a peripheral venous blood sample (5 mL) during the first 24 hours after admission to the ER. Sera were isolated, frozen, and stored at −80 °C until use. Serum HMGB1 levels were measured using an enzyme-linked immunosorbent assay kit according to the manufacturer’s instructions (Yanhui Biotech, Shanghai, China).
Baseline hematological and laboratory parameters, including white blood cell (WBC), neutrophil, lymphocyte, and platelet counts, hemoglobin, prothrombin time, total bilirubin, albumin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine, serum amylase, troponin I (cTnI), pH, partial pressure of carbon dioxide (PaCO2), and partial pressure of oxygen (PaO2) were measured using a Hitachi 7600 Clinical Analyzer (Hitachi, Tokyo, Japan), Sysmex CA-7000 System (Sysmex, Kobe, Japan), and Sysmex XE-5000 (Sysmex)^{8 (link)}, according to standard protocols.