Tissue sections were deparaffinized with xylene and rehydrated through graded alcohols into water. Tissues were placed in EDTA (pH 9.0) solution in a microwave in a short period of time for antigen retrieval, before washing for three times in pH 7.4 PBS solution. The endogenous peroxidase was blocked with 3% H2O2 for 25 min and then washed three times in pH 7.4 PBS solution. Protein blockage was applied using albumin from BSA (Solarbio) for 30 min before incubation with primary antibody (α-SMA; 1:100 dilution) at 4°C overnight. Sections were then incubated for 50 min with secondary antibodies (Dako, Denmark) at room temperature. Diaminobenzidine chromogen (Dako) was used for color development followed by counterstaining with Harris hematoxylin. TUNEL staining was performed using cell death detection kit (Roche, Germany). Quantification data were performed by calculating the mean optical density value of positive-staining areas versus the whole areas in five randomly selected fields using Image-Pro Plus software (Media Cybernetics). Antibodies used are as follows: anti-rabbit Ki-67 (1:400; Bioss, China), anti-rabbit P53 (1:400; Bioss, China), anti-rabbit Cyt C (1:400; Bioss, China), anti-rabbit Bcl-2 (1:400; Bioss, China), anti-rabbit Bax (1:100; Proteintech, USA), anti-rabbit NOS2 (1:100, Santa Cruz Biotechnology, CA, USA), anti-rabbit Arginase 1 (1:100; Santa Cruz Biotechnology, CA, USA), anti-rabbit CD31 (1:100;Abcam, UK), and anti-rabbit α-SMA (1:100; Abcam, UK).

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