Immunohistochemistry
From each group, six randomly selected samples underwent deparaffinization using xylene and were rehydrated with a graded ethanol series. Endogenous peroxidase was blocked with a 3% hydrogen peroxide solution after washing. It was then buffered with sodium citrate to facilitate antigen retrieval (10.2 mM, 20 min, 95°C). Using PBS containing 10% (w/v) bovine serum albumin, the sample was blocked for 10 minutes, and then at 4°C with a target of cadherin 5 (1: 200), SOD1 (1: 100), CD34 (1: 100), CASP3 (1: 200), or VEGF (1: 200). After that, the slide underwent incubation at room temperature for two hours with the corresponding HRP-conjugated secondary antibody (1:1000). Following washing, DAB dyeing, and counterstaining with hematoxylin was accomplished. A microscope was utilized to collect images for further analysis from at least six random fields of view (Olympus Corp., Japan).