Fully automatic biochemical analyzer

The lipid profiles, including serum TC, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol LDL-C, and triglyceride (TG), were detected by Fully Automatic Biochemical Analyzer (Hitachi, Tokyo, Japan). The apolipoprotein A, B, E contents in serum; the TC, TG and free cholesterol (FC) contents in liver; the TC and total bile acid contents in feces were measured using Elisa kits from Applygen Technologies Co., Ltd. (Beijing, China). The liver cholesterol ester (CE) was calculated by liver TC minus liver FC. The concentrations of 3-hydroxy-3-methyl glutaryl coenzyme A reductase (HMG-CoA reductase), cholesterol 7α-hydroxylase (CYP7A1), fatty acid synthase (FAS), cholesterol acyltransferase (ACAT), lipoprotein lipase (LPL), and low-density lipoprotein receptor (LDLR) in liver were measured with Elisa kits (Shanghai Enzyme-linked Biotechnology Co., Ltd. Shanghai, China) following manufacturer's instructions.

To evaluate the biocompatibility in vivo toxicity of the ALP-(MIs)n/DOX. PBS, free DOX, ALP-(MIs)25/DOX and ALP-(MIs)48/DOX were administered intravenously to healthy tumor-free ICR mice at a single dose of PBS, 3 mg kg^-1 of free DOX, 14.1 mg kg^-1 of P-(MIs)25 and P-(MIs)48 (n = 3). The hemanalysis and biochemical analyses were performed on blood withdrawn from the mice on 24 h post drug treatment. The blood samples were centrifuged at 5000 rpm for 10 min to separate the plasma. The plasma was then submitted to the Nanjing Biomedical Research Institute Affiliated to Nanjing University for analysis of ALT, AST, TBIL, BUN and CREA levels, which studied the samples by 7020 Japan Hitachi fully automatic biochemical analyzer.

Whole blood (K2EDTA tube) was collected from the wing vein before slaughter for routine hematological parameters using an automatic hematology analyzer (Sysmex, Japan), including white blood cells (WBCs), red blood cells (RBCs), hemoglobin (HGB), and platelets (PLTs). Serum was isolated from non-anticoagulant blood via centrifugation at 3,000 rpm/min for 10 min at 4°C for serum biochemical indicator determination. Serum total cholesterol (TC) and triglyceride (TG) levels were analyzed using a Fully Automatic Biochemical Analyzer (Hitachi, Japan).

The whole blood (K₂EDTA-tube) was collected from the wing vein of NW, UAW, and BAW phenotype of HW geese (n = 9–10) for routine hematological parameter using the automatic hematology analyzer (Sysmex, Japan), including white blood cells (WBC), red blood cells (RBC), hemoglobin (HGB), neutrophils (NEUT), lymphocyte (LYMPH), monocytes (MONO), eosinophilic cell (EO), basophilic cell (BASO), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MHC), mean corpuscular hemoglobin concentration (MCHC), platelet (PLT), thrombocytocrit (PCT), mean platelet volume (MPV), platelet ratio (P-LCR), and platelet distribution width (PDW). Serum was isolated from non-anticoagulant blood via centrifuged at 3,000 rpm/min for 10 min at 4°C for serum biochemical indicators determination. Serum total cholesterol (TC), high density lipoprotein (HDL), low density lipoprotein (LDL), triglycerides (TG), total protein (TP), albumin (ALB), globulin (GLB), Ca, P, creatine kinase (CK), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) were analyzed using Fully Automatic Biochemical Analyzer (Hitachi, Japan).

Height, weight, systolic blood pressure (SBP), and diastolic blood pressure (DBP) were measured by trained staff, and the BMI (kg/m²) was calculated. Fasting venous blood from the eligible participants was drawn the next morning after they were water fasted for 8 h. A fully automatic biochemical analyzer (Hitachi) was used to detect the total cholesterol (TC), triglyceride, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol (LDL-C) levels. For the OGTT, 75 g of anhydrous glucose was taken orally under the fasting state. It was dissolved in 250–300 mL of water and consumed within 5–10 min. The venous blood was separately collected under the fasting state 2 h after the administration of the glucose. The fasting plasma glucose (FPG), 2-h plasma glucose (2hPG), fasting insulin (FINS), and 2-h insulin (2hINS) levels were detected. In addition, 4 mL of blood was drawn under the fasting state. The serum was separated and stored in a refrigerator at −80°C for testing. An ELISA was used for detecting the FGF21. The kit was provided by BIM Company (USA), and the detection was performed according to the manufacturer’s instructions. The serum insulin level was detected using a chemiluminescence microparticle immunoassay. The kit was provided by Abbott Laboratories. The homeostasis model was used to assess the insulin resistance index (HOMA-IR = FPG × FINS/22.5).