Test kits for human IL-6, IL-10, Il-12 and IL-1b were purchased from Ruiboao Biotech, Guangzhou, China. Test kits for human CCL-5 and MIP-2 were products of Hengyuan Biotech, Shanghai, China. ELISA was performed according to the manual instructions. In brief, total proteins were extracted from both CD68 high infiltration and low infiltration groups. Both standard samples and tissue samples were added into 96-well plates, followed by the addition of 0.1 mL biotin-labelled antibody and 60-min incubation at 37℃. After gentle washing in 0.01 M TBS, ABC reagents were added into each well for further 30-min ncubation. The color was developed in the dark by TMB reagents. After stopping, a microplate reader quantified the optical density value at 470 nm for further analysis.
