Objective: To realize the status of junior college students’ health literacy in Nanjing Medical University. Methods: A questionnaire survey was conducted to investigate 197 medical college students by using the random cluster sampling method. Results: The proportion of students who had held health literacy was 16.24%. The holding rate of health knowledge and concept, the forming rate of health behavior and the mastering rate of health skill were 45.17%, 5.58% and 55.84%, respectively. The holding rate of health literacy among male students was higher than that of female students. The freshman had higher health literacy than sophomore students. The holding rate of health literacy among students from urban areas was higher than that of students from rural areas. Parental educational level and monthly living expenses were relate to health literacy level. Conclusion: The health behavior and lifestyle literacy of the subjects in this university need to be improved. Targeted health education for different groups of college students should be carried out according to the survey results. The work of school health education in multi-ways should be improved.