Anemoside B4 (AB4), as an important ingredient of Chinese herb Pulsatilla Chinensis, has been used in Chinese ancient medicine for thousands of years with exact therapeutic effects of anti-inflammation and anti-tumor. Due to its high potency and low toxicity, AB4 has caused more and more attention of researchers. In this paper, we will first review the clinical usage and fundamental research of AB4 against damaging inflammation of liver, lung, kidney, intestine and joints. Then, we will overview its application and molecular mechanisms in a range of cancers, such as liver cancer, lung cancer, colorectal cancer and cervical cancer. In addition, we pay attention to its pharmacological actions and molecular mechanisms against other diseases, for example, retinal injury, allergic conjunctivitis, pain, vulvovaginal inflammatory disease, acute infectious disease and oral ulcer. Finally, we will discuss the future perspectives of AB4, so as to improve its wide application in clinic.