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Influenza is an acute respiratory disease that seriously threatens public health. The occurrence of influenza has been proved to be related to a variety of meteorological factors. However, less attention has been paid to the effect of relative humidity (RH) on different types of influenza, especially in subtropical regions. Daily data on laboratory-confirmed influenza cases, weather variables, and air pollutants in Hefei covering the 2014–2019 period were collected. The seasonality and trend of daily influenza cases were explored by the time series seasonal decomposition method. Generalized linear model was fitted in conjunction with distributed lag nonlinear model to quantify the associations of RH with influenza A and influenza B. Subgroup analyses were conducted by sex, age (0–4, 5–17, and ≥18 years), and season (cold and warm seasons). A total of 5238 influenza cases including 2847 influenza A cases and 2391 influenza B cases were recorded. The epidemic of influenza presented a distinct seasonal pattern, and the number of daily influenza cases increased steadily since 2016. High RH was related to an increased risk of influenza A (maximum RR = 1.683, 95%CI: 1.365–2.076), especially among males, females, and school-age children. Low RH was associated with an increased risk of influenza B (maximum RR = 1.252, 95%CI: 1.169–1.340). The contrasting relationships of RH with influenza A and B remained significant in cold seasons. High RH and low RH were significantly associated with the increased risk of influenza A and B, respectively. The findings of our study may provide clues for proposing new effective interventions. [ABSTRACT FROM AUTHOR]