In response to a highly competitive environment, companies should explore new opportunities and at the same time exploit existing businesses to maintain profitability in the short run and achieve growth in the long run. Manager is the key person to deal well with the contradictory tasks. Researchers have discussed the constructs of ambidextrous leadership and ambidextrous activities. This research seeks to examine the relationship between ambidextrous leadership of top managers and ambidextrous activities of middle managers. Three hundred and thirty five questionnaires were distributed to 95 public-listed companies, in which the heads of R&D, manufacturing and marketing departments were respondents. We received 152 effective responses. The analysis of data showed that ambidextrous leadership has a positive influence on exploitative activities of middle managers, but not exploratory activities. Theoretical and practical implications of our findings are discussed.