Anti cd66 fitc

Ischemic Preconditioning Therapy Apparatus (Model: IPC-906; Beijing Renqiao Cardiovascular and Cerebrovascular Disease Prevention Research Nantong Company, China). BD FACS Calibur flow cytometer (BD Company, USA); Low-temperature high-speed centrifuge (Heraeus, Germany); Anti-CD11b-BB700 (Clone Number: M1/70), Anti-CD66-FITC (Clone Number: B1.1/CD66) and Anti-IL-1β-Alexa Fluor 647 (Clone Number: JK1B-1), (BD Company, USA); Multi-function microplate reader: (BioTek Company, USA); PCR instrument: (Bio-Rad Company, USA); QuantStudio 3 Real-Time PCR System: (Thermo Fisher Scientific Company, China); Various peripheral animal blood neutrophil isolation liquid kits: (Number: Solarbio.P9040. Beijing Soleibao Technology Company, China); MonScript™ RTIII All-in-One Mix with dsDNase Kit: (Wuhan Mona Biotechnology Company, China); MonAmp™ ChemoHS qPCR Mix Kit: (Wuhan Mona Biotechnology Company, China); Primers: (Shanghai Sangon Bioengineering Company, China).

Heparin anticoagulated blood was collected within 24 h of hospital admission, and one month thereafter. Polymorphonuclear cells were isolated from the lowest fraction of Ficoll-Paque separated heparin tubes, and erythrocytes were lysed with erythrocyte lysis buffer (Sigma-Aldrich, Saint Louis, Missouri). Cell purity was 94.1% [IQR, 86.0–97.3] as determined by flow cytometry (FACS Canto II with FACSDiva Software; BD Biosciences, Heidelberg, Germany) using anti-CD14-APC, anti-CD3-PECy7, and anti-CD66-FITC (BD Bioscience, San Jose, CA). See Figure S1 for the gating strategy.