In this research, the positive role of interface visual design in digital safety education was verified taking COVID-19 prevention and control knowledge as the content of public health safety education, where interface emotion (positive, negative, and neutral) and interface layout (waterfall typed and juxtaposition typed) were regarded as independent variables, and readers’ understanding, course evaluation and system usability score were dependent variables. As revealed in the results of a 3 × 2 two-factor experiment in which 252 college students participated: first, different interface emotion can cause significantly different understanding, where negative emotion has the best learning transfer effect; second, due to the difference in interface emotion, participants may give certain courses significantly different evaluation scores, while positive emotional interface contributes to the obviously high scores of three course-evaluation items, “appeal of the lesson”, “enjoyment of the lesson” and “interface quality”; third, significantly different system usability can be caused by different interface layout, where waterfall-type layout enjoys higher appraisal from users; fourth, interface emotion and interface layout have a similar interactive effects in terms of “effort of the lesson” and “interface quality”, where waterfall-type layout is favored in terms of positive emotional interface, and juxtaposition-type layout is more advantageous in terms of negative emotional interface. These results are of vital significance for interface design and safety education. Further, the visual design method for interface emotion and interface layout were analyzed to determine the most suitable design principles so as to improve the effect of digital public health safety education and provide constructive ideas for fighting against COVID-19 at the educational level.