To compare the difference on knowledge, attitudes, willingness of health workers and patients with their families towards organ donation and to explore the independent factors influencing the willing towards organ donation. During Dec. 2017 to Feb. 2018, we surveyed 241 hospitalized patients and their families in Hepatobiliary Center, the First Affiliated Hospital of Nanjing Medical University, with the self-defined questionnaire. Meanwhile, 202 health workers were investigated as a control group. Health workers’ knowledge towards organ donation was significantly higher than that of patients, but there were no statistical differences of attitudes and willingness to donate their own organs between the two groups. Multivariate logistic regression showed that factors contributing to the willingness to donate organs were age less than 35 years old, considering organ donation to be a noble thing, and people who supported organ donation. We recommended to incorporate the related knowledge and laws into medical courses, to improve the willingness of health workers to donate, and then to spread the knowledge to social population, ultimately to promote the development of organ donation in China.