[Objectives] The Chinese Soft-shelled Turtle Pelodiscus sinensis is an important species in China’s aquaculture. The intestine serves as the primary organ for the digestion, absorption, and transport of nutrients, while also functioning as a barrier against harmful external substances. However, the structural characteristics of the intestinal tissue in P. sinensis remain unclear. [Methods] In this study, the complete intestinal tract of P. sinensis was divided into 20 intestinal segments from fundus gastris to genital pores. Histological analysis was performed by hematoxylin and eosin (H.E) and Alcian Blue-Periodic Acid-Schiff (AB-PAS) staining. SPSS 22.0 was used for one-way analysis of variance of quantitative analysis data, and P < 0.05 was defined as significant difference. [Results] Segments I﹣IV are adjacent to the liver and gallbladder. Segments V﹣VIII are close to the pancreas and near the liver and gallbladder. Segments IX﹣XII and XIII﹣XVI are near the spleen. Segments XVII﹣XX are located adjacent to the kidneys and ovaries (Fig. 1). H.E staining showed that segments I﹣IV had larger diameter, thicker muscular layer, pronounced and numerous intestinal folds, and nearly no intestinal villi. Segments V﹣VIII had significantly smaller diameter and thinner muscular layer than segments I﹣IV, with large but singular folds and well-developed villi. Segments IX﹣XII had smaller diameter and thinner muscular layer than segments V﹣VIII, with shorter and numerous folds and almost no villi. Segments XIII﹣XVI had smaller diameter and thinner muscular layer than segments IX﹣XII, with short singular folds and relatively well-developed villi. Lastly, segments XVII﹣XX showed no significant differences in diameter or muscular layer thickness compared with segments IX﹣XII, with large and numerous folds and almost no villi (Figs. 2，3). AB-PAS staining revealed that segments I﹣IV predominantly contained type I and type III goblet cells, while segments V﹣XVI primarily had type I and type II goblet cells. No goblet cell was observed in segments XVII﹣XX. [Conclusion] These results indicate distinct classification characteristics among the intestinal segments of P. sinensis, allowing the intestine to be categorized into five regions:initial of small intestine (ISI), anterior of small intestine (ASI), middle of small intestine (MSI), posterior of small intestine (PSI), and finale of small intestine (FSI).