Vacuette vacutainer

All specimens for blood indicators and PAF-AH activity measurements were collected by venipuncture into 5-mL drying Vacuette vacutainers (Greiner Bio-One GmbH, Kremsmunster, Austria) in the morning after a 12 h fast on the second day after admission. The samples were sent to the laboratory, and serum was isolated by centrifugation (10 min, 3000 × g) and preserved at −80°C.

All specimens for blood indicators and coagulation measurements were collected by venipuncture into 5 mL drying VACUETTE vacutainers (Greiner Bio-One GmbH, Kremsmunster, Austria) and 5 mL Becton Dickinson vacutainers containing 0.5 mL 0.109 M sodium citrate (Becton Dickinson, Franklin Lakes, NJ, USA) in the morning after a 12-hour fast. The samples were sent to the laboratory, and the serum and plasma were isolated by centrifugation (10 min, 3000 g).

Peripheral venous blood was collected from all participants in the morning between 9 and 11 am. For RNA extraction, blood was collected in 9 mL acid citrate dextrose (ACD-B) tubes (BD Biosciences, North Ryde, New South Wales, Australia). Total RNA was extracted using the Trizol method (Invitrogen, Carlsbad, CA, USA) and RNA quality and concentration were assessed on the Agilent Technologies 2100 Bioanalyzer and Nanodrop ND-1000 spectrophotometer. Four control and 3 patient samples had low RIN values < 4 and were excluded from RNA analysis. RIN values were not significantly different between diagnostic groups (Table 1). cDNA was synthesized from 1 μg total RNA per case with the Invitrogen Superscript IV kit (Invitrogen, Carlsbad, CA, USA) and random hexamers following manufacturer instructions (Life Technologies). For plasma assays, blood was collected in 9 mL ethylenediaminetetraacetic acid (EDTA) tubes (Vacuette Vacutainer, Greiner Bio-One, Kremsmünster, Austria). Plasma was collected from EDTA tubes via centrifugation for 15 min at 2000 x g and aliquoted into protein low-binding tubes (Eppendorf, Hamburg, Germany) and stored at −80°C until the day of the assay.