Scanning electron microscope (SEM, TESCAN vega3, Czech Republic) and transmission electron microscopy (TEM, FEI TECHNI, G2-USA) were used to analyze the morphology and size of calycosin-loaded nanoliposomes, respectively. Zeta Sizer (Malvern, UK) was employed to assess the particle size and charge distribution of calycosin-loaded nanoliposomes by dissolving them in water solution at room temperature. Fourier transform infrared spectroscopy (FTIR) within 4000–400 cm−1 range was used employing on PerkinElmer Spectrum FT-IR spectrometer (USA).

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