Objective To study the changes of left ventricular(LV) function before and after rAAV2-GFP targeted to myocardium by the ultrasound mediated microbubbles. Methods Twenty rats were divided into 2 groups. Ten rats underwent ultrasound mediated microbubbles destruction without rAAV2-GFP as control. Another ten rats underwent ultrasound mediated microbubbles destruction containing rAAV2-GFP as experimental group. The mean value of LV rotation of rats was obtained in apical and base plane using speckle tracking imaging(STI) software. LV torsion(LVtor) was defined as apical rotation relative to the base. LV function including circumferential strain,circumferential strain rate of systolic and early diastolic and late diastolic, torsion, time to peak, ejection fraction, fractional shortening, end-diastolic volume, end-systolic volume of LV,heart rate and so on were also measured before and after 14 days. Other rats were killed after 14 days and hearts were harvested. GFP protein expression which showed rAAV2 transfer was observed under fluorescence microscope in frozen section. Results There was no significant difference of LV function of rats before and after rAAV2-GFP myocardium transfection by the ultrasound mediated microhubbles. Conclusions LV myocardial function has no significant difference before and after delivering rAAV2 to the heart in vivo by ultrasound-mediated microbubbles destruction.

Objective To explore the express of von willebrand factor(vWF) and tissue factor(TF) in ultrasound combined with urokinase and RGDS-targeted microbubbles for recanalization of occluded rabbit femoral artery.Methods A total of 42 rabbits with platelet-rich thrombi in the femoral artery were randomized into 7 treatment groups (n =6 in each group):①ultrasound alone (US) group;②ultrasound,non-targeted microbubbles (US + M) group;③urokinase alone (UK) group;④ ultrasound,non-targeted microbubble and urokinase (US + M + UK) group;⑤ ultrasound,platelet-targeted microbubble (US + R) group;⑥ platelet-targeted microbubble,urokinase (R + UK) group;⑦ ultrasound,platelet-targeted microbubble and urokinase (US + R + UK) group.A total of 6 ml of infusion liquor of urokinase,RGDS and microbubbles (SonoVue) were mixed by the direct conjugation method,infusion via vein within 20 min.Ultrasound was conducted to lyse the clot for 30 min.The recanalization and the velocity tracing change of blood flow in thrombolytic process and the express of vWF and TF were evaluated at 120 min post treatment.Results For US,UK,US + M,US + R and US + M + UK groups,recanalization were failed.The R + UK and US + R + UK groups were partly recanalizated or completed recanalized (P ＜0.001).vWF and TF were positive in US,US + M,US + R and R + UK group.vWF and TF were negative in US + M + UK,UK and US + R + UK groups.Conclusions The combined effects of low frequency ultrasound and targeted microbubble combined with urokinase can inhibit the expression of vWF and TF,and then promote the thrombolysis,however it can affect the expression of vWF and TF after reperfusion.

BACKGROUND:Stromal cel-derived factor-1 (SDF-1) is one of the most powerful chemokines in myocardial infarction region and plays a particularly pivotal role in the homing of stem cels to an injured myocardium and promoting angiogenesis. On the other hand the microbubble and acoustics active substances carrying targetable ligands can be prepared into targeted ultrasound contrast agents that can be combined with living cels used for molecular imaging. The key of ultrasonic molecular imaging is to find imaging targets, and to successfuly prepare targeted ultrasound contrast agent which can be combined with the imaging target specificaly and efficiently. OBJECTIVE:To prepare and evaluate targeted microbubble contrast agents with SDF-1 monoclonal antibody. METHODS:Targeted microbubble contrast agent with SDF-1 monoclonal antibody was prepared using the biotin-streptavidin method. The physiochemical properties of targeted microbubble contrast agent were evaluated by appearance, pH, particle diameter, optical and fluorescence microscope and flow cytometry test. Four minipigs underwent ligation of the left anterior descending coronary artery to complete the establishment of acute myocardial infarction model, and another two minipigs were subject to thoracotomy but no ligation of the coronary artery. Then, al animals were injected with microbubble contrast agents. The stability of microbubbles was assessed by immunofluorescence testin vivo. RESULTS AND CONCLUSION:SDF-1 and microbubbles were combined by biotin-streptavidin method.In vitro appearance of the contrast agent was translucent yelow or green, and stratified after standing. pH vaule was 7.02±0.12 for non-targeted contrast agent and 6.10±0.19 for targeted microbubble contrast agent. Under the fluorescence microscope, the distribution and size of targeted microbubbles were uniform, and the microbubbles were surrounded by bright and ring shaped green fluorescence that had no changes after highly shaking. The diameter of microbubbles was (2 422.62±238.82) nm after carrying the SDF-1 antibody. Flow cytometry results showed that the carrying rate of targeted contrast agents was stable in different periods.In vivo test showed that targeted microbubbles gathered in vascular endothelial cel surface after acute myocardial infarction. These findings indicate that the targeted microbubble contrast agent carrying SDF-1 monoclonal antibody prepared by biotin-streptavidin method can be combined with vascular endothelial cels, and the binding rate is high and stablein vitro.