Lh 780

Serobiochemical variables including glucose, bilirubin, protein, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), total cholesterol (Tchol), LDL cholesterol (LDLc), HDLc, triglycerides, calcium, sodium, phosphorus, potassium, chlorine, albumin, urea, and creatinine were measured in serum samples using automated biochemical auto-analyzers (Advia Systems, Siemens Healthcare Diagnostic Inc., Deerfield, IL, USA). Insulin was measured with the IMMULITE 2000 analyzer (DPC, LA, USA), and insulin resistance was calculated with the Homeostatic Model Assessment for Insulin Resistance (HOMA-IR). Whole blood was used to determine red and white cell series by an automated hematological analyzer (LH 780; Beckman Coulter, Fullerton, CA, USA). We determined plasma lipopolysaccharide-binding protein (LBP), a surrogate marker of metabolic endotoxemia, since plasma lipopolyshaccharide (LPS) determination shows several limitations, mainly due to the presence of endogenous inhibitors [33 (link)]. LBP was determined using a commercial ELISA kit (HycultBiotech, Uden, The Netherlands) as previously reported [34 (link)]. All samples were analyzed in triplicate, with intra- and inter-assay coefficients of variations (CVs) <10% for all parameters.

Independent variables Participants’ TLC was measured on study recruitment in a haematology analyzer (Beckman Coulter LH 780). Besides measured TLCs, assumed TLCs ranging from 4,000–11,000/μL by an increment of 500/μL was used to calculate the on-recruitment PIs.
Dependent variable PI on recruitment was expressed as absolute number of asexual and/or sexual parasites present in 1 μL of peripheral blood. It was calculated as: $\mathit{PI}\left(/\mathit{\mu L}\right)=\frac{PC}{WBC}\times TLC$
Where, PI is parasite index, PC is the number of parasites counted in the blood smear, WBC is the number of white blood cells counted in the blood smear, TLC is the patients’ total leucocyte count (either ‘assumed’ or ‘measured’). Leishman’s stained peripheral blood smears were examined under Olympus CH20i microscope and PIs were determined independently by three microscopists. Using a manual tally counter, number of parasites and WBCs in corresponding microscopic fields were counted. If ten or more parasites were noted in up to 200 WBCs counts, further counting was stopped, otherwise counting continued up until 500 WBCs [8 ]. A mean of three consecutive PIs was used for further analysis.

Upon admission of the patients, venous blood was collected to determine specific laboratory parameters. RDW was measured using an automated blood cell counter (LH780, Beckman Coulter, Brea, CA, United States), with a normal range of 11–16%. Clinical data were collected through chart review and electronic case reports entered into EpiData software 3.1 (The EpiData Association, Odense, Denmark) and processed using a consistency check on two copies. The estimated glomerular filtration rate (eGFR) in Chinese patients was evaluated using the 4-variable Modification of Diet in Renal Disease (MDRD) equation [(12 (link)), p. 2937–44].

Blood samples were obtained from participants in the morning after fasting for 12 hr and analysed by the Beckman Coulter LH 780 haematology analyser (Beckman, Coulter, FL, USA) in Tongji Hospital. Plasma FFA was measured with the FFA assay kit (Pulilai, China). Malondialdehyde (MDA) and SOD activity were assayed with the commercial kits (Nanjing Jiancheng, China). Triglyceride (TG) and hydroxynonenal (HNE) measurement in tissue extracts and cells were processed with the TG assay kit (Pulilai, China) and HNE elisa kit (Cayman, USA), respectively.