AIM: To study the methods of preparing the magnetic nano-microspheres of Fe2O3 and As2O3/Fe2O3 complexes and their therapeutic effects with magnetic fluid hyperthermia (MFH). METHODS: Nanospheres were prepared by chemical co-precipitation and their shape and diameter were observed. Hemolysis, micronucleus, cell viability, and LD50 along with other in vivo tests were performed to evaluate the Fe2O3 microsphere biocompatibility. The inhibition ratio of tumors after Fe2O3 and As2O3/Fe2O3 injections combined with induced hyperthermia in xenograft human hepatocarcinoma was calculated. RESULTS: Fe2O3 and As2O3/Fe2O3 particles were round with an average diameter of 20 nm and 100 nm as observed under transmission electron microscope. Upon exposure to an altating magnetic field (AMF), the temperature of the suspension of magnetic particles increased to 41-51℃, depending on different particle concentrations, and remained stable thereafter. Nanosized Fe2O3 microspheres are a new kind of biomaterial without cytotoxic effects. The LD50 of both Fe2O3 and As2O3/Fe2O3 in mice was higher than 5 g/kg. One to four weeks after Fe2O3 and As2O3/Fe2O3 complex injections into healthy pig livers, no significant differences were found in serum AST, ALT, BUN and Cr levels among the pigs of all groups (P > 0.05), and no obvious pathological alterations were observed. After exposure to altating magnetic fields, the inhibition ratio of the tumors was significantly different from controls in the Fe2O3 and As2O3/Fe2O3 groups (68.74% and 82.79%, respectively; P < 0.01). Tumors of mice in treatment groups showed obvious necrosis, while normal tissues adjoining the tumor and intal organs did not.CONCLUSION: Fe2O3 and As2O3/Fe2O3 complexes exerted radiofrequency-induced hyperthermia and drug toxicity on tumors without any liver or kidney damage. Therefore, nanospheres are ideal carriers for tumortargeted therapy.