Cd86 13395 1 ap

FZHY was obtained from Huanghai Pharmaceutical Co. (Shanghai, China). RPMI 1640 medium, fetal bovine serum (FBS), and 0.25% trypsin were purchased from Gibco (Rockville, MD, USA). BCA protein assay kit and TRIzol reagent were purchased from Invitrogen (Carlsbad, CA, USA). Special antibodies including p-JNK(#9251; 1 : 1000), T-JNK(#9252; 1 : 1000), p-ERK(#9101; 1 : 1000), T-ERK(#9102; 1 : 1000), p-P38(#9211; 1 : 1000), T-P38(#9212; 1 : 1000), p-STAT-1(#7649; 1 : 1000), T-STAT-1(#14994; 1 : 1000), and iNOS(#13120; 1 : 1000) were purchased from cell signaling technology (Beverly, MA, USA). Anti-HO-1 (ab68477; 1 : 10000), CD40 (ab252428; 1 : 1000), IL-6 (ab259341; 1 : 1000), and horseradish peroxidase-labeled goat antirabbit IgG were purchased from Abcam (Cambridge, UK). CD86 (13395-1-AP; 1 : 500) was purchased from ProteinTech Group (Chicago, IL, USA). Polyvinylidene difluoride (PVDF) membranes, western blotting detection reagent ECL, and murine recombinant IFN-γ were purchased from Millipore (Bedford, MA, USA). LPS, 1400W, Griess reagent, and MTT were purchased from Sigma-Aldrich (St. Louis, MO, USA). MicroRNA primers, transcription kits, and universal PCR master mix were obtained from GenePharma Company (Shanghai, China). Standard compounds sodium danshensu, salvianolic acid B, and adenosine were purchased from the National Institutes for Food and Drug Control (Beijing, China).

The diabetic rats were euthanized, and skin tissues were removed from the full‐thickness skin defects and burns of the rats. The tissues were fixed in neutral formaldehyde for 1 d. Then, the tissues had been embedded in paraffin and sectioned before being subjected to H&E, DHE, and immunofluorescent staining. Immunofluorescence single staining: VEGF (bs‐1313R, Bioss), CD86 (13395‐1‐AP, Proteintech), CD206 (60143‐1, Proteintech), TNF‐α (bs‐10802R, Bioss), HIF‐1α (bs‐1407R, Bioss), MMP‐9 (bs‐4593R, Bioss), IL‐6 (bs‐0782R, Bioss) and IL‐10 (bs‐6761R, Bioss). Immunofluorescence double staining: α‐SMA (bsm‐33188 M, Bioss)/CD31 (GB11063‐2, Servicebio) and Col I (14695‐1‐AP, Proteintech)/Vimentin (GB12192, Servicebio)). The cell nucleus of the tissues were labeled with DAPI, and the photographs were analyzed for intensities of fluorescent signals using ImageJ software. The intensities of the fluorescent signals in the full‐thickness skin defects and burns of the diabetic rats in the control groups were set to 100%. Laser Speckle Contrast Imaging (RFLSI Pro) was used to examine blood perfusion in the full thickness skin defects and burns.