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It is urgent to find a technology accurately to better diagnose and treat to brain tumor. Eu-doped Gd2O3 nanorods (Eu-Gd2O3 NRs) with paramagnetic and fluorescent properties were conjugated with doxorubicin (Dox) and chlorotoxin (CTX) via PEGylation, hydrazone bond and sulfur bond (named as CTX-NRs-Dox), and these NRs could release more Dox in lower pH environment. The results of cell experiments indicated that CTX-NRs-Dox had obvious targeting and toxic effects on U251 cells, as well as good fluorescence imaging behavior. The orthotopic glioma-transplanted mice models were constructed via the intracranial injection of glioma cells (U87MG). The result of experiments after the tail-vein injection of the prepared NRs suggested that CTX-NRs-Dox could target to brain tumors via the long-time blood circulation, leading to their obvious contrast enhancement of MR imaging of the intracranial tumor and their significant inhibitory effect on the growth and metastasis of brain tumors. A mechanism of synergistic effect of CTX-NRs-Dox on targeting and inhabiting the brain tumor was proposed. Our research suggested that CTX-NRs-Dox had potential application prospect in the detection and treatment of glioma.