Both flipped classroom and peer assisted learning have their advantages on making learners more flexible, active and deeply involved in learning. This study described the design and implementation of flipped classroom combined with peer-assisted learning in ultrasound-guided puncture training course, taking standardized training resident physicians in department of anesthesiology, Tongji Hospital, Huazhong University of Science and Technology as an example. Through questionnaire and operation evaluation, the teaching effect is evaluated from three aspects: personal perception, academic achievements, and behavioral changes. The results showed that 71 students (100.0%) were satisfied with the classroom training in 71 effective classroom teaching feedback forms. After training, compared with before training, residents' self-confidence in completing ultrasound-guided puncture operation was improved [7(2) points vs. 4(3) points, P<0.001]. The operation time of successful puncture was shortened [37.5(35.5)s vs. 80.5(70.0)s, P<0.001], and the number of adjusting needles was reduced [1(1) vs. 3(2)], and the development score of puncture needle was increased [10(0) points vs. 6(3) points, P<0.001]. Of the 56 valid questionnaires collected 2 weeks after the course ended, 50 (89.3 %) trainees had used ultrasound-guided puncture technique in clinical practice, of which 49 (98.0 %) trainees had successful performance. The flipped classroom and peer assisted learning used in the ultrasound guided puncture course for residential trainees can help to improve their teaching satisfaction and perceived self-confidence, and help to improve their performance of clinical skills.