Ultrasonic tissue characterization with integrated backscatter (IBS) offers a promising method for the noninvasive assessment of myocardial fibrosis and contractile performance. The aim of this study was to investigate the effect of thrice-weekly in-center nocturnal hemodialysis (INHD) and conventional hemodialysis (CHD) on myocardial fibrosis and left ventricular function in end-stage renal disease patients. Thirty-two INHD and 58 matched CHD patients were enrolled; baseline and 12-month measures of blood pressure (BP), serum calcium and phosphorus, echocardiographic left ventricular mass index (LVMI) and left ventricular function, the myocardial calibrated IBS (C-IBS), and systodiastolic cyclical variations in IBS (CV-IBS) were collected. The baseline characteristics were similar between groups, except that serum phosphorus and calcium × phosphorus were higher in the INHD group. At 12-month follow-up, there was a significant decrease in the mean C-IBS (-20.2 ± 3.7 to -28.1 ± 4.0 dB, P<0.01) and a significant increase in CV-IBS (5.0 ± 1.5 to 7.1 ± 1.6 dB, P<0.01) in INHD patients. Multivariate analysis showed that the mean C-IBS was positively related to SBP, DBP, LVMI, serum phosphorus, and left atrial volume index and inversely related to midwall fractional shortening, transmitral E/A ratio, and E(m) . The mean CV-IBS was positively correlated with left ventricular midwall fractional shortening, E/A ratio, E(m) and inversely correlated with SBP, DBP, LVMI, serum phosphorus, E/E(m) , and left atrial volume index. There was no significant change in the mean C-IBS, mean CV-IBS, and LVMI in the CHD group. Compared with CHD, INHD improves myocardial fibrosis and left ventricular function, and control of serum phosphorus is associated with the improvement of myocardial fibrosis. Improvement of myocardial fibrosis contributes to the reduction of left ventricle hypertrophy.