Dsassistant

To identify histological alterations, dorsal skin was obtained and fixed in 10% formalin. The fixed skins were embedded in paraffin and sliced into 4-μm sections. Sliced sections were stained with hematoxylin and eosin (H&E) for skin epidermis thickness analysis and Masson’s trichrome stain for collagen fiber analysis. To measure the expression of AGEs, immunohistochemistry (IHC) was performed on skin tissue. Images were obtained using a digital slide scanner (Motic Easyscan one, Motic, Hong Kong), and skin epidermis thickness was measured using image analysis software (Motic DS Assistant, Hong Kong).

Clinicopathological characteristics, including vascular and perineural invasion and mismatch repair (MMR) status, were obtained from pathology reports. The TNM stage was re-evaluated by the eighth edition TNM staging system. The patients’ demographic information included age, sex and tumor location. The tumor location was classified as left, right colon, and rectum. All patients were routinely followed up for five years after CRC resection. Censored Data was defined as patients alive at the last follow-up and those who died from non-CRC causes.
All available hematoxylin and eosin (H&E)-stained sections were reviewed and the representative sections were selected. These representative sections should include the tumor tissue and normal tissue surrounding the tumor tissue. The corresponding tissues were subsequently resected into 4-μm thick slices. All the representative H&E-stained sections were scanned using MoticEasyScan Infinity and analyzed using Motic DSAssistant.

Clinicopathological features included age, sex, tumor location (tumor distance from the anal verge), adjuvant chemotherapy, tumor grade, tumor stage, vascular invasion and perineural invasion. As systemic inflammation markers, the blood-based NLR was calculated based on pre-operative data. Blood-based NLR = blood-based total neutrophil count (10⁹/L)/ blood-based total lymphocyte count (10⁹/L). Following previously established procedures^{6 (link)}, we meticulously reviewed all hematoxylin and eosin-stained sections, selected representative sections, and cut sections into 4-μm-thick slices. The MoticEasyScan Infinity was used to scan all hematoxylin and eosin-stained sections, and Motic DSAssistant was used to analyze.