Objective To investigate the value of ultrasound guided autobiopsy technology in interventional diagnosis of pulmo-nary peripheral disease .Methods 117 cases of pulmonary peripheral disease proved by X-ray or CT ,confirmed by ultrasound ,but could not be identified the nature by the examination above ,were biopsied under the guidance of ultrasound .Results All the 117 ca-ses were biopsed successfully .115 cases of them had definite pathological results ,the pathological positivity rate was 98 .3% ,in which contained 20 cases of chronic inflammation ,14 cases of tuberculosis ,30 cases of squamous cell carcinoma ,45 cases of adeno-carcinoma ,6 cases small cell undifferentiated carcinoma .No complication ,such as pneumatothorax ,hemoptysis ,bleeding occurred during the operation .Conclusion The ultrasound guided autobiopsy technology in interventional diagnosis of pulmonary peripheral disease is a safe and effective technique ,the tissues obtained from ultrasound guided autobiopsy were adequate to pathological slide which can provide the basis of the clinical diagnosis .

ObjectiveTo investigate characteristics in vitro and contrast-enhanced effect in vivo with self-made nano-scale ultrasoud-enhanced contrast agent.MethodsNano-lipid microbubbles were prepared by machine vibration and low-speed centrifugation.The surface potential and the microbubbles size were assessed by Zetasizer nano ZS90.The bubbles dispersity and micromorphous were observed under light microscopy and electron microscope.The characteristics of microbubbles were studied at various times at 4℃ environment.The enhancement duration time and intensity of nano-lipid microbubbles in nude mice livers,kidneys and subcutaneous implanted tumors were observed,and compared with micro-scale microbubbles.ResnltsThe mean diameter of nano-lipid microbubbles was (580.6 ± 36.3) nm.The contrast imaging study in vivo showed the nano-lipid microbubbles could significantly enhance ultrasonic imaging in nude mice livers,kidneys and subcutaneous implanted tumors.ConclusionsSelf-made nano-scale ultrasoudenhanced contrast agent showed good stability and contrast-enhancement effect in vitro,and provide a experimental basis for ultrasound imaging and therapeutics of malignant tumors.

Objective To explore the value of diagnostic ultrasound mediated microbubble destruction in improving the myocardial perfusion and left ventricular systolic function when cooperated with the mecsenchymal stem cells(MSCs) transplantation in rabbit myocardial ischemia. Methods One week after myocardial ischemia (MI) modeling,36 rabbits were divided into 3 groups,the control group(group Ⅰ) ,intravenous injection of MSCs group(group Ⅱ) and ultrasound + microbubble + MSCs group (group Ⅲ). Myocardial contrast enhancement (MCE) was performed and quantification analysis of anterior wall was assessed with Photoshop. Left ventrieular systolic function was assessed with M-mode echocardiography and bi-plane Simpson's method. CD34 expression in heart was detected with immunohistochemisty(IHC). Western blotting was applied to detect the level of VEGF in three groups. Results The differences of gray scale analyzed with histogram of Photoshop in anterior wall of ischemia myocardium between the group Ⅰ and group Ⅱ or group Ⅲ were significant,and P value was 0. 032 and 0. 000 , respectively. There were significant differences of FS between group Ⅲ (30. 43±4.09)% and group Ⅱ (26.29±2.93)%, P<0.01, and similar to group Ⅰ (19.28 ± 2.84)%. The difference of EF(%) between group Ⅲ and group Ⅱ was significant [(61.5±5.8 vs 53.6±4. 71), P<0. 05] ,or markedly significant between group Ⅲ and group Ⅰ [(61.5±5.8 vs 42.6± 5.0), P <0.01]. EF(%) assessed with bi-plane Simpson's method was significantly increased from (34.64 ± 4.59) in group Ⅰ to (41.78 ± 4.21) in group Ⅱ and (48.6±3.96) in group Ⅲ. The expression of CD34 assessed with immunohistochemistry was the highest in group Ⅲ. The level of VEGF with western blotting in group Ⅲ was significantly higher than other two groups. Conclusions It is an efficacious transplantation means of MSCs infusion under the ultrasound mediated microbubles destruction in improving the myocardial perfusion and cardiac systolic function.

Objective To investigate the effect of microbubbles mediated ultrasound insonation on proliferation and apoptosis of vascular smooth muscle cells (VSMCs) in different phase of cell cycle. Methods Rat thoracic aortic VSMCs were cultured in vitro by the method of tissue adherence. The cells were synchronized by the methods of serum starvation and double thymidine block. The synchronization results were detected by flow eytometer. VSMCs in different phases of cell cycle were exposed to 1 MHz continuous waves ultrasound for 120 s at intensity 0.3 W/cm2 in the presence of lipid-coated microbubbles (1 ml/L). Apoptosis of VSMCs was analyzed by AnnexinV/PI staining using flow eytometry. The proliferation and the proliferating cell nuclear antigen(PCNA) protein expression of VSMCs were detected by MTT assay and immunoeytochemistry, respectively. Results The synchronized G0/G1 and S phase VSMCs were achieved, with synchronized rates to 89.53 % and 66.87 %, respectively. Ultrasound sonication for 120 s with microbubbles could significantly inhibit the proliferation and downregulate the PCNA expression of S phase VSMCs,but the proliferation and PCNA expression of G0/G1 phase VSMCs were not affected. After treatment of ultrasound with microbubbles, the apoptotic ratio were found to reach (7.05 ± 2.04)% in G0/G1 phase VSMCs and (27.01 ±3.87)% in S phase VSMCs. Conclusions Microbubbles mediated ultrasound insonation can significantly inhabit the proliferation and induce apoptosis in VSMCs at proliferation stage.

Objective To investigate the effect of ultrasound irradiation combined with liposome membrane microbubbles on the reorgnization of cytoskeleton in vascular smooth muscle cells (VSMCs). Methods Rat thoracic aortic VSMCs were cultured in vitro. VSMCs were exposed to 1 MHz continuous waves ultrasound radiation for 120 s at intensity 0.3 W/cm2in the presence of liposome membrane microbubbles (1 μl/ml) after treated with platelet derived growth factor-BB (PDGF-BB). The reorganizations of microfilaments, microtubules and intermediate filaments were examined by using immunofluorescence and fluorocytochemistry techniques. Results There was a substantial increase in the expression of F-actin and assembly of long bundles of stress fibers in the transversed cell body when treated with PDGF-BB. Neither alterations of β-tubulin nor of vimentin cytoskeletal protein organization were observed in PDGF-BB treated cells as compared to those of the contol group. After ultrasound irradiation combined with liposome membrane microbubbles, the expression of F-actin, β-tubulin and vimentin were reduced along with the simultaneous changes in microfilaments, microtubles and intermediate filaments array. Conclusions Ultrasound irradiation combined with liposome membrane microbubbles can induce significant changes in cytoskeleton structure of VSMCs cultured in vitro.

Objective To investigate characteristics and ultrasonic imaging of the nano-scale ultrasound-enhanced contrast in vivo. Methods Nano-scale microbubbles were prepared by machine vibration and low speed centrifugation. The surface morphology and average size distribution of microbuble were measured. The contrast-enhanced effect of the hahn-scale microbubbles in the normal rabbit liver was observed,and compared with micro-scale microbubbles. Results The nano-scale microbubbles had a good shape and uniform distribution by light microscope and transmission electron microscope, with average diameter of 623.4 nm and surface electric potential of 1.3 inV. The contrast imaging study in vivo showed the nano-scale microbubbles could significantly enhance ultrasonic imaging of rabbit livers, which had no obvious difference with micro-scale microbubbles. Conclusions The nano-scale ultrasound contrast agent is stable and effective for the enhancement of ultrasound imaging, which is based on development of miniaturizing targeting ultrasound contrast agent.

Objective To compare two effective preparation methods for paclitaxel-loaded lipid microbubbles, and to evaluate the physiochemical properties as for acoustic activated drug delivery. Methods Paclitaxel-loaded lipid microbubbles were prepared with two methods: one was mixed with phospholipids directly (Ⅰ); the other was added in triacetin, then mixed with phospholipids (Ⅱ). Concent ration, size, pH, drug entrapment efficiency, drug-loading amounts of these two kinds of paclitaxel-loaded lipid microbubbles were studied, while drug release with ultrasound and tumor imaging enhanced on rabbit breast tumor were observed. Results There was no significant difference in tumor imaging between two kinds of microbubbles which could be ruptured by low energy ultrasound. Compared with Ⅰ, the mean diameter of Ⅱ decreased significantly ([ 1.07±0.38] μm vs [2.79± 0.41] μm, P<0.01), the surface potencial was higher ([19.10±0.32] mV vs [-5.90±0.21] mV, P<0.01), whereas entrapment efficiency and drug-loading amounts increased markedly ([ 95.00±1.22]% vs [36.10±4.74]%, P<0.01; [5.60±0.11]% vs [0.50±0.04]%, P<0.01). Conclusion The Ⅱ paclitaxel-loaded lipid microbubbles added triacetin have an important clinical value.

Objective To investigate the mechanism of the increased permeability of the blood-brain barrier during transcranial ultrasound contrast imaging. Methods Sprague-Dawhy (SD) rats were performed transcranial ultrasound contrast imaging, the lanthanum nitrate and the evans-blue were used as tracers,the distribution of the tracers were observed and the transports mechanism were also investigated. Results The opening of the tight junction and increased permeability of the cellular membrane were observed after the transcranial ultrasound contrast imaging. Conclusions The main mechanism of the increased permeability of the blood-brain barrier was the opening of the tight junction and increased permeability of the cellular membrane.

Objective To explore the targeting homing capacity of bone-marrow derived mesenchymal stem eells(MSCs) on rabbit myocardial ischemia by intravenous injection of MSCs under the mediation of diagnostic ultrasound and microbubble. Methods Density gradient centrifugation and adherent culture method were used in the isolation and cultivation of MSCs. MSCs were labeled with DAPI. Rabbit myocardial infarction(MI) models were builded by totally ligation of left anterior descending branch of coronary artery. DAPI labeled MSCs were implanted by intravenous injection with or without the mediation of diagnostic ultrasound and microbubbles. Forty-eight hours after cell transplantation, the hearts of MI rabbits were made of frozen section and observed under fluorescent microscope. The DAPI positive cells were counted in the MI and border area of rabbit heart and compared between two groups. Pathological changes of MI area were observed with HE staining under light microscope and transmission electronic microscope. Results The number of DAPI positive cells in MI and border area of rabbit in both groups were counted under fluorescent microscope. There were more DAPI positive cells in the MI area in ultrasound + microbubble + MSCs group (213.2±26.5) than that in the intravenous injection group (146.8±18.78, P<0.01). There were erythrocytes leaking out of the vessels in MI area in HE staining section under light microscope in ultrasound + microbubble + MSCs group while there were nearly none in the intravenous injection MSCs group. The intercellular space of endothelial cells of the vessels wall was increased and serum component leaked out of the vessel wall in ultrasound + microbubble + MSCs under transmission electronic microscope. Conclusions The targeted homing capacity of BM-MSCs in the MI area of rabbit heart can be enhanced under the mediation of diagnostic ultrasound and microbubbles.

Objective To access the relation between the Doppler waveform of the hepatic venous flow and the hepatic fibrosis.Methods To construct fibrosis models,20 experimental rabbits were fed with thioacetamide as the only drinking water at the concentration of 1.2 g/L,then the Doppler waveforms of the hepatic vein were detected at the time before constructing the fibrosis model,then later the 8th week,12th week and 16th week during constructing the fibrosis models,the results were compared with the hepatic biopsy ones.Results With the process of constructing the fibrosis models,the experimental rabbits had more abnormal Doppler waveforms of the hepatic venous flow.Conclusions The changes of the Doppler waveforms of the hepatic venous flow reflect the change of hepatic parenchyma.