Detection and diagnosis of neoplastic and inflammatory gastrointestinal (GI) diseases are typically based on endoscopic and pathologic examination. In GI neoplastic diseases, diagnosis can be delayed due to the expense and invasive nature of this approach. Recently, PIWI-interacting RNAs (piRNAs), a group of small non-coding RNA molecules containing 24-31 nucleotides, have been thought to serve as biomarkers in many disease processes. For example, piRNAs are differentially expressed in GI cancer but their biologic role remains unclear. Using next-generation sequencing and microarray analyses, researchers have suggested that monitoring piRNAs could facilitate diagnosis and prognosis in GI disease. Herein, we reviewed the use of piRNAs in neoplastic, inflammatory, functional, and other diseases of the digestive system, which could shed new light on cancer screening, early detection, and personalized treatment.