Objective To investigate the current status of traditional Chinese medical (TCM) cultural literacy among non-medical students in TCM universities, analyze key issues in classic literature reading, knowledge acquisition channels, and curriculum-practice transformation, and propose targeted improvement strategies for interdisciplinary talent cultivation. Methods In December 2024, stratified sampling was employed to recruit non-medical students from 8 TCM universities across four economic zones in China: eastern region (Beijing University of Chinese Medicine, Nanjing University of Chinese Medicine, Zhejiang Chinese Medical University), central region (Henan University of Chinese Medicine, Jiangxi University of Chinese Medicine), northeastern region (Heilongjiang University of Chinese Medicine), and western region (Chengdu University of Traditional Chinese Medicine, Chongqing University of Chinese Medicine). Electronic questionnaires were distributed via an online platform. Descriptive statistical analysis was used to analyze relevant data. Results A total of 320 questionnaires were collected, of which 280 were valid. TCM theoretical cognition and classic reading: 88.2% (247/280) of students understood basic TCM theories, but only 32.1% (90/280) “had carefully read multiple TCM classics ” or “read parts of one or two classics”, with 31.1% (87/280) indicating “almost no understanding” of classics. Interest and knowledge acquisition channels: 91.4% (256/280) of students were “ interested” in TCM traditional culture, and 95.7% (268/280) acknowledged its modern value. 78.6% (220/280) accessed TCM culture primarily through online media and 76.8% ( 215/280) through school courses. However, the usage rate of health preservation methods including “frequently” and “occasionally” was only 65.7% (184/280). Curriculum evaluation and practice transformation: 83.9% (235/280) of students recognized the richness of course content, but only 31.4% (88/280) were very satisfied with the courses, and 65.0%(182/280) thought there were few opportunities for practical teaching. Conclusions The non-medical students have good cognition foundation of traditional Chinese medicine (TCM) theory but insufficient classic reading, high interest and recognition but limited channels for relevant knowledge acquisition, and rich TCM culture courses but insufficient practical transformation. These can be improved by increasing the proportion of TCM classics and traditional culture-related courses, integrating TCM traditional cultural elements into professional curriculum teaching, carrying out diverse campus cultural activities related to TCM culture as a beneficial supplement to school curriculum teaching, and strengthening the combination of practical teaching links with TCM traditional culture.