Thymosins are small peptides, originally identified from the thymus, but now known to be more widely distributed in many tissues and cells. Thymosins are divided into three main groups, alpha-, beta-, : and gamma-thymosins, based on their isoelectric points. alpha-thymosins (ProTalpha, Talphal) have nuclear localization and are involved in transcription and/or DNA replications; whereas beta-thymosins (Tbeta4, Tbeta10, Tbetal5) have cytoplasmic localization and show high affinity to G-actin for cell mobility. Furthermore, it is well known that both alpha- and beta-thymosins play important roles in modulating immune response, vascular biology, and cancer pathogenesis. More importantly, thymosins may have significant clinical applications. They may serve as molecular markers for the diagnosis and prognosis of certain diseases. In addition, they could be molecular targets of certain diseases or be used as therapeutic agents to treat certain diseases. However, the molecular mechanisms of action of thymosins are largely unknown. This review not only presents recent advances of basic science research of thymosins and their clinical applications but provides thoughtful views for future directions of investigation on thymosins.