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The purpose of the present study was to prepare psoralen-doxorubicin nanostructured lipid carriers (PSO-DOX-NLC) with better physicochemical properties for further study on its multidrug resistance reversal activity on leukemia cells. PSO-DOX-NLC was synthesized by emulsion evaporation and solidification at a low temperature. PSO-DOX-NLC morphology and particle size were examined using transmission electron microscopy, and Zeta potential was determined using photon correlation spectroscopy; encapsulation efficiency was also investigated. The results showed that PSO-DOX-NLC possessed a rounded structure, with a mean (± SD) particle size of 128.7±1.8 nm, a polydispersity index of 0.22±0.01, a Zeta potential of 20.17±0.31 mV, a psoralen encapsulation rate of 76.56% and a doxorubicin encapsulation rate of 89.48%. Thus, PSO-DOX-NLC prepared by emulsion evaporation and solidification at a low temperature had good physicochemical properties.