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Objective To explore the effect of effective questioning on the teaching of “Toxicology” based on case-based teaching, and to accumulate experience for further promoting the reform of Toxicology. Methods We selected 317 undergraduates of preventive medicine in Grade 2012, 2013 and 2014, from Shaanxi University of Chinese Medicine as research objects. Two teaching cases were planned in the instructional design for Grade 2012, while six teaching cases were planned for Grade 2013. The instructional design for Grade 2014 implemented effective questioning on the basis of Grade 2013. The difference in test paper scores, grade point average (GPA), the difficulty level of the test paper and grade were analyzed before and after the increase of case-based teaching and effective questioning. Results The test paper scores and grade point averages increased gradually from Grades 2012, 2013 and 2014, and the difference was statistically significant (F=99.722, P |