Objective:To explore the expression levels of serum triple domain 35 (TRIM35) and tumor necrosis factor receptor associated factor 3 (TRAF3) in patients with acute pancreatitis (AP) and their correlation with the severity and prognosis of the disease.Methods:Using a prospective research method, 93 patients with AP (observation group) were selected from July 2020 to September 2022 in Qinghai Red Cross Hospital, including 40 cases of mild acute pancreatitis (MAP), 29 cases of moderate to severe acute pancreatitis (MSAP) and 24 cases of severe acute pancreatitis (SAP). During the same period, 40 healthy individuals who underwent physical examinations were selected as healthy control group. The serum TRIM35 and TRAF3 levels were detected by real time fluorescence quantitative polymerase chain reaction (RT-qPCR). The survival status after 28 d of admission was followed up. The correlation was analyzed by Pearson method. Multivariate Logistic regression analysis was used to analyze the relationship between serum TRIM35 and TRAF3 levels and the prognosis in patients with AP. The efficacy of serum TRIM35 and TRAF3 in predicting the prognosis in patients with AP was evaluated by the receiver operating characteristics (ROC) curve.Results:The serum TRIM35 and TRAF3 levels in observation group were significantly higher than those in healthy control group (3.76 ± 1.36 vs. 1.02 ± 0.19 and 5.37 ± 2.18 vs. 1.04 ± 0.16), and there were statistical differences ( P<0.01). The serum TRIM35 and TRAF3 levels in patients with MSAP and SAP were significantly higher than those in patients with MAP (4.11 ± 1.73 and 4.96 ± 1.47 vs. 2.79 ± 1.04, 5.43 ± 2.15 and 7.01 ± 2.85 vs. 4.35 ± 1.79), the indexes in patients with SAP were significantly higher than those in patients with MSAP, and there were statistical differences ( P<0.05). The follow-up results showed that 11 cases died and 82 cases survived. The serum TRIM35 and TRAF3 levels in death patients were significantly higher than those in surviving patients (4.94 ± 1.01 vs. 3.60 ± 1.67 and 7.08 ± 1.43 vs. 5.14 ± 2.57), and there were statistical differences ( P<0.05). Pearson correlation analysis result showed that serum TRIM35 level was positive correlation with serum TRAF3 level in patients with AP ( r = 0.483, P<0.01). Multivariate Logistic regression analysis result showed that serum TRIM35 and TRAF3 levels were the independent risk factors of prognosis in patients with AP ( OR = 1.86 and 1.37, 95% CI 1.12 to 3.09 and 1.02 to 1.82, P<0.05). ROC curve analysis result showed that the area under the curve of serum TRIM35 combined with TRAF3 levels for evaluating the prognosis in patients with AP was significantly larger than serum TRIM35 and TRAF3 alone (0.85 vs. 0.81 and 0.81, Z = 0.03 and 0.04, P<0.05). The optimal cutoff values of serum TRIM35 and TRAF3 levels were 4.90 and 6.11. Conclusions:The serum TRIM35 and TRAF3 levels in patients with AP are significantly elevated, and are related to the severity of the condition. The serum TRIM35 and TRAF3 levels are independent risk of prognosis in patients with AP, and their combined detection is more valuable in evaluating the prognosis in patients with AP.