Objective To investigate the best pacing sites of right ventricular septum (RVS) by comparing QRS wave width, serum N-terminal pro B-type natriuretic peptide (NT-proBNP) level and hemodynamic indicators in the patients undergoing different electrode positions in RVS. Methods A total of 177 patients who met the indications for artificial permanent pacemaker implantation and underwent single-chamber pacemaker implantation in our hospital from July 2014 to February 2018 were recruited in this study. According to the implantation site of active fixation electrode compared with height of vertebral shadow under X-ray at the posteroanterior position, they were divided into right ventricular high-septal (n=54), median-septal (n=68), and low-septal groups (n=55). The width of QRS complex, serum NT-proBNP level and hemodynamic indicators were recorded before and after operation. Left ventricular end diastolic diameter (LVEDD), left ventricular ejection fraction (LVEF), stroke volume (SV) and left ventricular fractional shor-tening (LVFS) were measured before and 6 months and 1 year after pacemaker implantation. SPSS statistics 26.0 was used for statistical analysis. One-way analysis of variance, non-parametric test, Chi-square test or Fisher exact probability test was adopted for intergroup comparison depending on different data types. Results There was no statistical difference in preoperative QRS width, serum NT-proBNP level, LVEDD, LVEF, LVFS and SV in the three groups (P>0.05). After operation, QRS width was increased in all groups, and it was the narrowest in the median-septal group, and widest in the low-septal group (P<0.05). Post-operative NT-proBNP was significantly lower in the median-septal groups than before(P<0.05), and both the low-septal group and high-septal group increased significantly at 6 months and 1 year after operation (P<0.05).The LVEDD of high-septal group and low-septal group was greater than that of median-septal group at 6 months and 1 year after operation (P<0.05); LVEDD of median-sptal group was less than that before operation; LVEDD of low-sptal group was more than that before operation. There was no significant difference between preoperative and postoperative LVEF, LVFS and SV in the median-septal group(P>0.05). Compared with low-septal group, the SV at 6 months, and LVEF, LVFS, and SV at 1 year postoperatively of median-septal group were significantly increased; compared with high-septal group, only LVEF at 1 year after operation was significantly increased (P＜0.05). Conclusion Median-septal site for RVS pacing shows little effect on cardiac function, and is the optimal site for RVS.