Automatic analyzer

Serum activity levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and γ- glutamyltransferase (γ-GT) were measured using HITACHI 7170S biochemistry analysis equipment. Hepatitis B e antigen (HBeAg) was measured by an automatic analyzer (Roche, Basel, Switzerland). The HBV DNA was determined with PCR System (Applied Biosystem, Foster City, USA). Serum concentration of A20 and TNF-α were measured by ELISA kit (West Tang biological technology, Shanghai, China) according to the manufacturer's instructions.

The mice were fasted overnight and then blood was sampled from the eyes of the mice. Serum triglycerides (TG), total cholesterol (TC), blood urea nitrogen (BUN), and serum creatinine (Scr) levels were determined using an automatic analyzer (Roche, Basel, Switzerland). Fasting blood glucose (FBG) was measured using a glucose meter (Roche Diagnostics GmbH, Mannheim, Germany). Fasting insulin (FINS) was measured by an ELISA assay (Jiancheng, Nanjing, China).

The body weight, blood pressure, plasma glucose, insulin, cholesterol, and triglyceride levels were measured before and after the 6-month feeding period. Systolic blood pressure was measured with a noninvasive tail-cuff monitor (MK2000; Muromachi Kikai, Tokyo, Japan). Eight-hour fasting plasma glucose was measured with a Beckman Glucose Analyzer. Plasma insulin was determined with a radioimmunoassay kit (Linco Research, Inc., St. Charles, MO). The plasma cholesterol and triglyceride levels were measured using enzymatic methods (Roche, Pleasanton, CA, USA) through an automatic analyzer (Roche).

Male Sprague-Dawley rats, initially weighing 330-350 g, were housed in cages in a temperature- and light-controlled room, with free access to water. All procedures were approved by the Dong-A University Institutional Animal Care Committee (DIACUC-approval-16-9).
Thirty-two adenine-induced uremic rats were fed diets containing 0.75% adenine and 2.5% protein (Envigo Teklad, Madson, WI, USA) for three weeks. After three weeks, rats were randomly divided into four groups, which were supplemented and fed diets containing 2.5% protein for four weeks. These groups included an adenine control group (0.9% saline by gastric gavage), an omega-3 FA-supplemented group (300 mg/kg/day by gastric gavage), an MK-7-supplemented group (50 µg/kg/day by gastric gavage), and a combined omega-3 FA and MK-7-supplemented group [12 (link),13 (link)]. Six normal control rats were fed diets containing 2.5% protein for seven weeks. All animals were fed evenly and had access to water ad libitum. Rats were anesthetized with diethyl ether, and blood samples were obtained from the heart. Serum creatinine, blood urea nitrogen (BUN), and phosphorus levels were measured by an automatic analyzer (Roche, Germany).

Serum content of total TG, total cholesterol (TC), total bilirubin, high-density lipoprotein (HDL), low-density lipoprotein (LDL), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) was detected using an automatic analyzer (Roche, Basel, Switzerland) according to the manufacturer's introduction.