Surveys showed that vaccine hesitancy may influence the acceptance of COVID-19 vaccines in healthcare workers (HCWs) and the general population. Currently, the actual acceptance of COVID-19 vaccination in HCWs has rarely been reported. In the present survey, we investigated the real-world acceptance of COVID-19 vaccination in HCWs in perinatal medicine during the first three-month period of vaccination in China and to identify the main reason for the decline of vaccination. HCWs (1087) who participated in a Chinese national symposium on perinatal medicine during 16-18 April 2021 were invited to answer a 27-question questionnaire online. A total of 1051 HCWs completed the questionnaire. Of them, 86.2% (906/1051) accepted the COVID-19 vaccination and 13.8% (145/1051) declined the vaccination. Because of the vaccine hesitancy, one-fourth of the vaccinated participants did not accept the vaccination until consulted with others or requested by employers. The main reason for the decline of vaccination in 145 unvaccinated HCWs was the concern about vaccine safety. The results indicate that vaccination request by employers may promote vaccine acceptance. More convincing data on the safety of COVID-19 vaccines appears to be important to increase the acceptance of vaccination.