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This paper describes a new formulation of magnetic nanoparticles coated by a novel polymer matrix-O-Carboxylmethylated Chitosan (O-CMC) as a drug/gene carrier. The O-CMC magnetic nanoparticles were derivatized with a peptide sequence from the HIV-tat protein and transferrin to improve the translocationai property and cellar uptake of the nanoparticles. To evaluate the O-MNPsTat-Tf as a drug carrier, Methotrexate (MTX) was incorporated as a model drug and MTX-Ioaded O-MNPs-Tat-Tf with an average diameter of 75 nm were prepared and characterized by TEM, AFM and VSM. The cytotoxicity of MTX-Ioaded OMNPs-Tat-Tf was investigated with C6 cells. The results showed that the MTXloaded O-MNPs-Tat-Tf retained significant antitumor toxicity.