ObjectiveHere, we develop a seven-layer gastric wall stratification theory based on the physical basis of ultrasound and histology, and further discuss its potential clinical application. Methods1. Experimental methods: Ex vivo human gastric specimens were immersed in normal saline and examined with a high-frequency probe to study the relationship between the sonograms and the corresponding anatomy of the gastric wall. 2. The study enrolled 136 patients admitted to our hospital with gastric diseases who underwent gastric contrast ultrasonography supplemented with the pathological examination. The seven-layer stratification theory was adopted during the analysis to profile sonogram characteristics with lesions originating from various layers. ResultsAll the sonograms of the in vitro human gastric specimens could be divided into seven intervals of strong and weak echoes. The pathological examinations were performed on 136 patient-derived samples as the golden criteria of diagnosis: 29 cases of gastric polyps, 10 cases of lymphomas, 5 cases of neuroendocrine tumors, 11 cases of ectopic pancreas, 22 cases of gastric stromal tumor, 19 cases of leiomyomas, 29 cases of chronic inflammation, 9 cases of diffuse invasive cancer, and 2 cases of neurilemmoma. The ultrasound and pathological examination results were consistent in 110 cases, showing a coincidence rate of 80.9%. ConclusionBy adopting the seven-layer stratification theory of the gastric wall, the ultrasound can accurately locate the position of mucosal muscularis, which is of great significance for accurate measurement of the thickness of each anatomical layer and the correct judgment of the origin and the classification of the space-occupying lesions. Keywords Gastric wall; ultrasound; seven-layer stratification; clinical application