Objective To observe the application effect of layered cooperation teaching in clinical teaching in intensive care unit (ICU).Methods The master graduate students who practiced in ICU of Fangcun branch of Guangzhou Provincial Hospital of Traditional Chinese Medicine form September 2013 to July 2014 were divided into two groups according to the rotation cycle.Diagnostic test was conducted to all the above students.The new teaching group (21 people) was determined according to the diagnostic test results and students' majoring to frm study group and the layered cooperation teaching was adopted,while the traditional teaching was used in the traditional clinical education group (19 cases).In the end of the rotation training,a theoretical examination was conducted among the two groups of students,and the questionnaire of teachers' teaching quality was issued.The related data were processed by SPSS 17.0,and the data between groups were compared by t test.Results The examination result in the layered cooperation teaching group (78.35 ± 3.13) were better than those in the traditional clinical education group (70.21 ± 4.58) and the difference was statistically significant (P=0.041).Survey results showed that the new teaching group students' evaluation to teaching content,teaching methods and teaching effectiveness was higher than traditional teaching group,and the score difference was statistically significant (P＜0.05).Conclusion The layered cooperation teaching in the clinical teaching in intensive care unit (ICU) to mobilize the students' subjective initiative,so that students of different knowledge structure can be integrated into the clinical practice of ICU,and enhance their ability of clinical analysis of ICU disease.