A new steroid, vernoanthelsterone A (1), and five known steroids were isolated from the aerial parts of Vernonia anthelmintica Willd. Compound 1 possesses a Δ(8(14))-15-one moiety. To our best knowledge, few steroids with this moiety have been reported before. Compounds 1-6 were tested for their antibacterial activities and their effects on estrogen biosynthesis in human ovarian granulosa-like cells (KGN cells). Compound 2 showed the ability to promote estrogen biosynthesis with EC(50) of 56.95 μg/mL and also exhibited the antibacterial activities against Bacillus cereus, Staphyloccocus aureus, Bacillus subtilis and Escherichia coli with MICs ranging from 3.15 to 15.5 μg/mL. The structures of 1-6 were determined on the basis of IR, MS, 1D and 2D NMR.