Au5400 chemistry system

Blood samples were taken by cardiac aspiration after the mice were anesthetized with 2% isoflurane. Blood was collected in tubes and centrifuged at 3,000 rpm for 15 min at 4 °C by a refrigerated centrifuge (HEMA). Serum noradrenaline (NE) and urine adrenaline protein levels were assayed using ELISA kits (Catalog number: E-EL-0047c and ml002049) from Elabscience Biotechnology Co., Ltd. (Wuhan, China) and Shanghai Enzyme-linked Biotechnology Co., Ltd. (Shanghai, China) according to the provided guidelines. Serum concentrations of LDL, HDL, total cholesterol (TC), and free fatty acid (FFA) were measured by enzymatic method using AU5400 Chemistry System from Beckman Coulter (Brea, CA, USA).

Venous blood was drawn from fasting patients by venipuncture into vacutainer tubes with no additives (Becton–Dickinson, San Jose, CA). The GA assay (Lucica GA-L glycated albumin assay kit, Asahi Kasei Pharma Corporation, Japan) was performed in the clinical laboratory in Anzhen Hospital using an automated biochemical analyzer (Beckman AU5400 Chemistry System), and the results are expressed as the ratio of glycated albumin to albumin.

ALT, aspartate aminotransferase (AST), and total bilirubin in serum were measured by the Olympus AU5400 Chemistry System (Beckman Coulter, Switzerland). Viral markers, including hepatitis B surface antigen, hepatitis B e antigen (HBeAg), and anti-HBe, were tested by Chemiluminescent Assay (Architect i2000SR; Abbott Diagnostics, Abbott Park, IL, USA). HBV DNA was quantified by real-time PCR using a commercial detection kit (Kehua Bio-Engineering) with the aid of Light Cycler Detection System (ABI 7300 Realtime PCR system, USA). The lower limit of detection for this assay is 50 IU/mL.
HBV resistance mutations were identified by DNA sequencing. The fragments (nt130–nt1129), encompassing the reverse transcriptase (RT) domain, were amplified by nested PCR. The first pair of primers for the PCR were 5′-AAGCTCTGCTAGATCCCAGAGT-3′ (sense: nt18–nt40) and 5′-TTTCGCTCCAGACCGGCTGC-3′ (antisense: nt1320–nt1301). The second pair of primers for the PCR included 5′-GCGGGGTTTTTCTTGTTGAC-3′ (sense: nt56–nt75) and 5′-AGTATGGATCGGCAGAGGAG-3′ (antisense: nt1272–nt1253). PCR products were directly sequenced on an automated DNA sequencer (ABI 3730XL, USA) using the second sense primer. Substitutions at positions rt80, rt173, rt180, rt181, rt184, rt202, rt204, rt236, and rt250 were taken as resistance mutations for analysis [9 (link)].