Objective: At present, one of the bottlenecks in gene therapy is the low gene transfection efficiency.Studies have found that, under the effect of ultrasonic, ultrasound micro-bubble generated a cavitation effect on the cell membrane and the formation of spillover hole which led to the membrane permeability increasing and the micro-bubble destruction.This facilitates the gene to release and migrate into the vessel wall or tissue space so as to promote gene trans-membrane delivery.Thus, this experiment prepared the AAV-loaded lipid ultrasound micro-bubble, to test whether the lipid ultrasound micro-bubbles exposured by ultrasound could enhance the gene transduction mediated by adeno-associated virus (AAV) in the cells with tight junction.Methods: To prepare the AAV-loaded lipid ultrasound micro-bubble, a mixture of amount of a certain proportion of AAV, phosphatidylcholine, phosphatidylethanolamine, cholesterol derivative of cholesteryl chemisuccinate and glycerol were dissolve in PBS, incubated at room temperature for 10 min, and then the perfluoropropane gas were punched into the solution, and further proceed in bath-ultrasound for 20s (100W) with interval of 10s and a total of 10 circular at room temperature.Further, the particle size and particle distribution were measured by Malvern Sizer Nano ZS90.The human bronchial epithelial cells of 16HBE14o-were selected as the model cells, which were cultured in the trans-well insert to form the tight junctions between cells of 16HBE14o-.After administration, the transduced cells were observed for positive cells with expression of green fluorescence protein.Moreover, the transduction efficiency was further estimated by the flow cytometric analysis.Results: The AAV-loaded lipid ultrasound micro-bubble prepared in this experiment was spherical with a uniform distribution (Fig.1) and with an average particle size of 2.46±1.2 μm.And the result showed that these lipid ultrasound micro-bubbles kept stable at least within 48h, but it was not appropriate for long-term storage.It could significantly enhance the gene transduction of 16HBE14o-cells after ultrasound exposure (Fig.2).Conclusion: This study presented a variety of advantages of the full use of AAV combined with micro-bubbles, established an efficient, controllable, easy and practical virus-non viral integration of gene delivery system.It was found that ultrasound with lipid micro-bubbles produced cavitation effect and could increase the permeability, reduce the cell membrane resistance.This gene delivery system may be a novel strategy for AAV in the gene therapy application, which played the foundation for AAV to be used in vivo study.