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With the development of medical technique, many medical challenges have been solved. This cross-sectional study was aimed to assess the reliability and validity of a new developed scale for assessing the professionalism of young health care workers.This cross-sectional population-based study was conducted based on cluster sampling method in 2015. The participants were from 2 medical centers of Nantong city of Jiangsu province in China. The internal consistency reliability was assessed using Cronbach's alpha and split-half reliability coefficients. Construct validity was analyzed using exploratory factor analysis (EFA) and confirmatory factor analysis (CFA). Content, convergent, and discriminant validities were also assessed in this study.Total 749 individuals aged 29.09 ± 3.91 years old participated in this investigation. There were 745 valid questionnaires and 730 were complete. Cronbach's alpha value (0.944) and the split-half reliability coefficient (0.873) reflected satisfactory internal consistency reliability of this new professionalism assessment scale. EFA extracted a 7-factor model. About 63.4% of the total variance was explained by these factors. However, CFA showed a good model fit after excluding the items with factor loading lower than 0.5. Good discriminant validity of this new developed professionalism assessment scale was also shown (P < .05). However, the evidences for content and convergent validity were not enough in this study.The results showed the satisfactory reliability of this new developed professionalism assessment scale. However, this scale should be modified to improve the validity in further studies.