In order to find a teaching mode that can meet the needs of the teaching in large class, we integrate various theories and teaching methods, construct grouping micro-video competition teaching mode and implement it in the biochemistry course in 6 undergraduate classes of 2016 nursing major of Youjiang Medical University for Nationalities to test its application effect in large class. After surveying to one experimental class (n = 103) and 2 control classes (n = 210) in the form of questionnaire and analyzing the final examine scores of experimental and control contents, it is shown that students' average scores in experimental class in which the micro-video competition teaching mode is adopted and the scores of objective and subjective questions are significantly higher than those in controls (P < 0.05), while there is no significant difference in the scores of contents between experimental class and controls (P > 0.05). In particular, there is a statistical difference in the scoring rate of objective questions between experimental content and control content in the experimental class (P < 0.05), while there is no significant difference in the scoring rate of subjective questions (P > 0.05). Students prefer to affirm the micro-video competition teaching mode (P < 0.05), while they are generally uncertain towards traditional lecturing teaching mode in questionnaire (P < 0.05). The research results show that the micro-video competition teaching mode has promoted students' memory and mastery on the experiment content. Especially, in the aspect of training students' comprehensive analysis and the ability to apply knowledge, the teaching effect of the mode is better that of traditional mode. It not only promotes the ability of students' comprehensive analysis and its application to the experiment content, but also the ability of their comprehensive analysis to other teaching content and relevant skills.