Au5400 automatic biochemical analyzer

Clinical data were collected from the electronic medical record database of Xianyang Central Hospital. The FAR was calculated from serum fibrinogen and albumin concentrations on admission. Plasma fibrinogen levels were detected by the coagulation method using STA CompactMax automatic coagulation analyzer (Diagnostica Stago Ltd.) and high-sensitivity c-reactive protein (hs-CRP) was detected by immunoturbidimetry using an AU5400 automatic biochemical analyzer (Olympus Corporation). Albumin levels and myocardial injury markers, including cardiac troponin T (cTnT) as well as creatinine kinase (CK)-MB, creatinine, NT-proB-type natriuretic peptide (NT-proBNP), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were also tested using the Abbott Architect C16000 autoanalyzer (Abbott Laboratories) on admission.

Anticoagulant-free blood (5 ml) was centrifuged (1,700 x g, 18°C, 4 min), and the serum was collected for the measurement of the levels of aspartate transaminase (AST), lactate dehydrogenase (LDH), α-amylase (AMY), lipase (LPS) and P-type amylase (P-AMY) using an Olympus AU5400 automatic biochemical analyzer (Olympus Corporation, Tokyo, Japan).

Before any treatment, 5 mL blood was collected with a vacuum blood collection tube without anticoagulant. The serum was separated by centrifugation at 4°C and stored at −80C. The levels of serological markers of hepatitis B and antibodies against hepatitis C and hepatitis D virus were determined using an enzyme-linked immunosorbent assay kit (Wantai, Beijing, China). The hepatitis B viral loads were measured at the Institute of Hepatology of Xi’an Jiaotong University using the HBV fluorescence polymerase chain reaction diagnostic kit (Da An Gene Co., Ltd. of Sun Yat-Sen University, Guangzhou, China), which has a dynamic range of 10² to 10⁸ IU/mL according to the manufacturer's instructions. Biochemical liver function was assayed using the Olympus AU5400 automatic biochemical analyzer (Olympus Corporation, Mishima, Japan). Serum alpha fetal protein levels (ng/mL) were measured using the automated Eleceyes platform (Roche Diagnostics, Mannheim, Germany), and liver ultrasonography or computer tomography scans were performed at each visit.

Sputum and serum samples were collected on the day of admission (before treatment), during treatment (day 4, 7 and 14) and subsequent baseline (8 w after discharge). Spontaneous (97%) and induced (3%) sputa were collected, processed within 2 h of collection and divided into two aliquots. One sputum aliquot was processed with phosphate buffered saline24 (link)25 (link). Cytospins were prepared and cell-free supernatant was collected and stored in aliquots at −80°C pending analyses of soluble mediators. Differential cells were counted on May Grünwald Giemsa stained cytospins in a blinded fashion. Peripheral venous blood (7 mL) was collected into a Vacutainer tube (BD Diagnostics, NJ, USA) and centrifuged at 6716 G for 10 min at 4°C. Serum was then separated and stored at −80°C till further analysis.
Serum interleukin (IL)-6 and sputum IL-8 and myeloperoxidase (MPO) were quantified using commercial sandwich ELISA kits (R&D Systems, Abingdon, UK). Serum high sensitivity C-reactive protein (hs-CRP) was measured by a latex agglutination test, using an Olympus AU5400 automatic biochemical analyzer. All the samples from each patient were measured in the same assay to reduce inter-assay variability. The limit of detection was: 0.7 pg/mL for serum IL-6, 7.5 pg/mL for sputum IL-8, 0.062 ng/ml for sputum MPO, and 0.1 mg/L for serum hs-CRP, respectively.

Serum TSH level was determined using a mouse ELISA kit (MyBioSource), following the product manual. Serum FT4 concentrations were determined using specific radioimmunoassay kits (Jiuding Diagnostic). Serum triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), alanine transaminase (ALT) and aspartate aminotransferase (AST) levels were analyzed using an Olympus AU5400 automatic biochemical analyzer (Olympus Corporation) at Shandong Provincial Hospital.