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Objection: To study the relationship between different doses and biological effect of 32P-glass microspheres (32P-GMS) by percutaneous intra-tumor injection at different times and provide proofs of theory for clinical therapy. Methods: 36 Zealand rabbits and Vx-2 were used to establish the animal model of liver tumor. Six groups were randomly designed. The suspension of different radiative doses of 32P-GMS combined with lipiodol-ultrafluid (0.1 mL) was respectively injected by percutaneous intra-tumor. The tumor tissues were examined by light microscope. MRI examination of liver tumors were performed before and after the injection. Results: C and D groups were observed that the tumor volume was decreased and the rate of restrained tumor was gradually increased after injection of 32P-GMS. The living tumor tissues of E group completely disappeared after the injection for two weeks. MRI examination showed that the tumor signal of E group was equal as T2 as the signal of normal liver parenchyma. The living tumor tissues were not found in F group after the injection for three weeks.Conclusion: 111 MBq was the best radiative dose of 32P-GMS for treatment of 1 cm liver cancer by percutaneous intra-tumor injection. MRI examination was very valuable to evaluate the result and follow up after the injection to treat liver cancer.