Background: This study aimed to evaluate the role of two-dimensional speckle-tracking imaging (2D-STI) and myocardial layer-specific analysis in evaluating early left ventricular (LV) myocardial function and systolic dyssynchrony in young and middle-aged uremic patients undergoing peritoneal dialysis (PD). Methods: We enrolled 31 PD patients aged ≤65 years with preserved LV ejection fraction (LVEF, ≥54%) as the PD group and 49 age-matched healthy people as the control group. Echocardiography was used to assess the left atrial diameter index (LADI, LAD/BSA), LV mass index (LVMI), LVEF, peak early diastolic velocity/late diastolic velocity (E/A) (measured by pulsed Doppler), and peak early diastolic velocity (by pulsed Doppler)/peak velocity of the early diastolic wave (by pulsed-wave tissue Doppler) (E/e′). Next, we used 2D-STI and myocardial layer-specific analysis to obtain longitudinal strains (LS) of the endocardium (LSendo), the myocardium (LSmyo), the epicardium (LSepi), and the global myocardium (GLS). Then, we measured the postsystolic index (PSI) to evaluate LV myocardial function. Time to peak LS (TTP) and peak strain dispersion (PSD) from 17 consecutive segments were assessed to quantify LV dyssynchrony. Results: Compared with the controls, PD patients had significantly increased LADI (p = 0.041), LVMI (p = 0.000), and E/e′ (p = 0.009), but reduced LVEF (p = 0.000) and E/A (p = 0.000). The average values of GLS (GLS avg) (p = 0.01) and GLS of the apical 2-chamber view (p = 0.003), including the LSendo (p = 0.024), LSmyo (p = 0.024), and LSepi (p = 0.032), were significantly decreased in patients with PD compared with controls. In PSI, segments of LS were markedly delayed in the anterior septum (p = 0.047), anterior (p = 0.000) and septum wall (p = 0.024) from basal segments, anterior wall (p = 0.001) from middle segments, and anterior (p = 0.024) and inferior (p = 0.024) wall from apical segments. Moreover, PSD was significantly increased in PD patients (p = 0.015), while TTP was evidently delayed in the anterior septum (p = 0.004), anterior (p = 0.000) and posterior (p = 0.042) wall from basal segments, and inferior wall (p = 0.048) from apical segments. Conclusions: Despite preserved LVEF, young and middle-aged PD patients developed LV dysfunction and myocardial systolic dyssynchrony earlier compared with controls.