Abstract: Objective To evaluate effect of health literacy intervention in occupational groups, explore suitable ways and provide suggestions for workplace health promotion. Methods In application of convenience sampling, Minhang District, Hongkou District and Qingpu District each selected 2 enterprises respectively according to their type, scale and location, setting one as intervention enterprise and the other as control. Then a six-month comprehensive intervention was conducted among intervention groups. Two hundred employees in each enterprise were randomly investigated before and after intervention to evaluate the effect. Results The overall health literacy level of participants in the intervention group rose from 16.7% to 33.4%, with improvement on the literacy levels of scientific view, infectious diseases prevention and treatment, chronic disease control and prevention and the three aspects of health literacy content (P<0.05), while such improvement was not reflected in the control group, and health skill literacy, safety and first aid literacy declined (P<0.05). Conclusion Comprehensive intervention based on health needs could improve the level of health literacy of occupational groups. Intervention in future should focus on improving the literacy of chronic disease control and prevention and basic medical care in occupational population.