Objective To evaluate the general situation of occupational exposure to antineoplastic drugs (ADs) in medical institutions in Hunan Province, and to provide basis for formulating prevention and control measures of ADs occupational exposure.Methods A self-designed questionnaire was used to investigate doctors, nurses, pharmacists, administrators and workers in Ads-related departments of 19 grade A hospitals in Hunan Province. The survey contents included the general information, ADs exposure and protection, cognition and training, and health status.Results A total of 588 questionnaires were sent out, and 539 were effectively recovered, with an effective recovery of 91.67%. Among the 539 respondents, 46.75% were nurses, and most of them were female staff members (80.15%). Respondents aged from 30 to 39 (46.94%), from oncology department (31.17%), with bachelor degree (63.82%), with intermediate professional titles (47.31%), working for 6~10 years (29.87%) accounted for the highest proportion. ADs of more exposure were cisplatin (78.11%), cyclophosphamide (73.10%). There were significant differences in the frequency of ADs exposure among staffs in different departments and occupations (P<0.001). Hematology department staff (72.97%) and nurses (55.16%) had the highest frequency of ADs exposure. 8.01% of the respondents were not equipped with any protective devices in their department. 1.91% of the respondents did not use any personal protective equipment when exposed to Ads. There were significant differences in cognition of ADs and occupational exposure among respondents from different departments (P<0.001), and the cognition degree of respondents from oncology department, pharmacy intravenous admixture service (PIVAS) and hematology department was relatively higher. There were significant differences in cognition of the mechanism, absorption pathway and post-exposure treatment of ADs among the respondents with different occupations (P≤0.001), and doctors, nurses and pharmacists had relatively higher degree of cognition. 25.97% of the respondents received no relevant training. There were significant differences in health problems of the respondents in different departments and occupations after exposure to ADs (P<0.001), and those in hematology department (54.05%) and nurses (35.71%) had the highest proportion of health problems. The most common health problems were hair loss (34.60%), stomatitis and canker ulcer (13.50%), abnormal menstruation (13.08%). Among the 316 married females, 43 cases (13.61%) had infertility, 14 cases (4.43%) had habitual abortion, 94 cases (29.75%) had induced abortion, and 32 cases (34.04%) had induced abortion because of fetal development problems.Conclusion Medical institutions have the risk of occupational exposure to ADs, so medical staff should improve their awareness of occupational exposure to ADs. Medical institutions should strengthen management and training, and attach great importance to the health of staff. Administrative departments should formulate relevant laws and regulations according to the prevention and control situation of occupational exposure in the region occupational exposure , so as to reduce the occurrence of occupational exposure.