Objective To analyze the clinical features and prognosis of patients with drug-induced liver injury (DILI)，so as to provide basis for its prevention and treatment. Methods The clinical materials of 416 patients with DILI in the First Affiliated Hospital of Nanjing Medical University between January 2017 to December 2020 were reviewed. The demographic data, suspicious drugs, clinical characteristics, laboratory examination results and prognosis were retrospectively analyzed. Results Among 416 DILI patients, there were 161 males (38.70%) and 255 females (61.30%), and their ages mainly distributed between 40~59 years old (46.39%), followed by ≥60 years old (31.50%). The main clinical manifestations included fatigue, darker urine, yellow skin and sclera, and gastrointestinal symptoms. The main type of injuries was hepatocellular injury (52.64%), followed by mixed type (23.80%) and cholestatic type (23.56%). The most common drug causing DILI was Chinese herbal and patent medicine (56.25%). Patients with hepatocellular type had the best overall prognosis, with cure and improvement rates of 10.05% and 88.58% respectively, and a failure rate of 1.37%. Conclusion A number of drugs may cause DILI, and the clinical manifestations were nonspecific. There were statistical differences in liver function tests of different genders, ages and clinical classification. Most patients have a good prognosis after active treatment, but some patients still progress or die, which needs more clinical attention.