After the cells were treated as mentioned above (with or without US Irradiation) and then incubated for another 16 h. Then the cells were successively stained with 200 µL annexin Calcein-AM (2 µM) and PI (4.5 µM) for 20 min. Finally, the cells were washed three times with cold 1×Assay Buffer. All the above operations were performed in the dark. Confocal fluorescence imaging was performed to observe the live and dead cells distribution through the fluorescence distribution of Calcein-AM and PI. ) and ( ), the tumors were exposed to US irradiation (1.0 W cm -2 , 1.0 MHz, 50% duty free) for 3 min after the injection for 6 h and 9 h. Mice were treated three times and observed for 10 days after treatment. The body weight and tumor volume of each mouse were monitored every two days, and after 14 days treatment, the tumors were dissected and weighed to evaluate the therapeutic efficacy. All mice were kept and treated according to protocols approved by the Institutional Animal Care and Use Committee at the Ethics Committee of Animal Experiment Center, Beijing Institute of Technology. Approval number: 2016-0006-M-2020013.