Beckman reagents

Fasting blood samples (10 mL) were taken from IS patients and NCs within 72 h of recruitment, separated by centrifugation, and frozen at −80°C immediately after processing. Samples were shipped in packaging incorporating dry ice-cooling agents by courier from every site to a blood storage site, where they were stored at −80°C. Total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), apolipoprotein B (ApoB), ApoA1, fasting plasma glucose (FPG), high-sensitive C-reactive protein (hs-CRP), and homocysteine (Hcy) concentrations were measured in the laboratory of the Dian Diagnostics Group Co. Ltd with Beckman Coulter AU680 Clinical Chemistry Analyzer and Beckman reagents (Beckman Coulter, Brea, CA, USA). All the participants have not taken antibiotics, probiotics, and prebiotics in the 3 months prior to the feces collection. Feces were collected at hospital or home. The feces were collected according to the instruction and delivered immediately at low temperatures. The frozen feces were shipped using dry ice to Xiangxi Center for Disease Prevention and Control. Once received, fecal samples were divided into three parts of 200 mg and stored at −80°C until extraction.

The contact thermometer, TFD-04 (Fudan University, Shanghai, China), CZF ultrasonic therapeutic apparatus (output power, 1–5 W; Chongqing Haifu Technology Co. Ltd., Chongqing, China) and Hitachi H-600 transmission electron microscope (S-4000, Hitachi, Ltd., Tokyo, Japan) were provided by the Institute of Ultrasonic Engineering in Medicine located at the Chongqing Medical University (Chongqing, China). The mixtures to suspend Nano-HA within the various solvents were sonicated using an XL2020 sonicator microtip (Heat Systems, Farmingdale, NY, USA), and the biochemical indicators of multiple organ function were analyzed using a Beckman CX7 analyzer and Beckman reagents (Beckman Coulter, High Wycombe, UK).