BACKGROUND : To determine the optimal timing of duodenal transection in patients undergoing laparoscopic-assisted total gastrectomy (LATG) in combination with laparoscopic spleen-preserving splenic hilar lymphadenectomy (LSPL) for advanced proximal gastric cancer (APGC). METHODS : One hundred twenty-seven patients with APGC who received LATG with duodenal transection as well as LSPL between January 2017 and July 2018 were retrospectively recruited in this study. According to the different transection timing, the patients were allocated into two groups: a conventional group (CG) who received the duodenal transection prior to the LSPL and an experimental group (EG) who were given LSPL before the duodenum was transected. Clinical short-term outcomes were compared in the two groups. RESULTS : Analysis of the demographical and clinical characteristics showed that the two groups were comparable with no significant differences between CG and EG in the study patients regardless of their body mass indices (BMI). The intraoperative and postoperative indicators for clinical short-term outcomes were compared between the CG and EC, and results indicated that the EG had significant shorter mean time of LSPL and total operation time than those in the CG (P < 0.05). Of note, the numbers of patients with intraoperative injury and the volume of blood loss during the LSPL procedure were significantly reduced in the EG versus CG (P < 0.05). For the obese APGC patients, administration of LSPL prior to duodenal transection significantly increased the number of dissected No.10 lymph nodes (LNs) (P < 0.05). The other intraoperative and postoperative indicators did not show any differences between the two comparison groups. CONCLUSIONS : Our findings demonstrated that duodenal transection timing was significantly associated with clinical short-term outcomes of APGC patients. The duodenal transection prior to the LSPL is superior overall to the conventional transection timing in the treatment of APGC patients with LATG and LSPL in combination.