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.

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.

BACKGROUND:Currently, human umbilical cord derived-mesenchymal stem cel s are mainly for local transplantation, which has some shortcomings, such as large trauma, bleeding, complications, that limit its widespread application in clinical practice. OBJECTIVE:To investigate the feasibility of intravenous transplantation of human umbilical cord derived-mesenchymal stem cel s for repair of spinal cord injury. METHODS:Eighty Wistar rats with spinal cord hitting were divided into five groups:blank control group with no transplantation (n=10), DMEM local transplantation group (n=15), DMEM intravenous transplantation group (n=15), cel local transplantation group (n=20), cel intravenous transplantation group (n=20). The functional recovery of spinal cord injury was observed with Basso, Beattie and Bresnahan scores at regular time as wel as hematoxylin-eosin staining and immunohistochemistry staining. RESULTS AND CONCLUSION:During 1 day to 2 weeks after transplantation, there was no significant difference in the Basso, Beattie and Bresnahan scores between the five groups;within 4-12 weeks after transplantation, the Basso, Beattie and Bresnahan scores were significantly higher in the two cel transplantation groups than the other three groups, but there was no difference between these two cel transplantation groups (P>0.05). Histological observation showed that the number of voids and glial scars was less in the cel local transplantation group and cel intravenous transplantation group compared with the other three groups, and there was also no difference between the two cel transplantation groups. These results indicate that the intravenous transplantation of human umbilical cord derived-mesenchymal stem cel s is similar to the local transplantation in the repair of acute spinal cord injury, which is simple and avoids secondary injuries and various complications. It is recommended that this method provide a new approach for cel transplantation.

Objective To investigate the effect of a self made liposome perfluocarbon ultrasound contrast agent in enhancing grey scale imaging of normal kidney.Methods The kidneys of 10 normal rabbits were imaged with second harmonic imaging before and after intravenous injection of the liposome contrast agent at a dose of 0.01 ml/kg. The subjective scores and video intensities were used to evaluate contrast effects. Results The renal cortex,medulla and sinus grey scale imaging were greatly enhanced after injection, and the notable enhancement time of renal cortex,medulla and sinus could last for 52 min,2 min 20 s,1 min 40 s,respectively. Conclusions The self made liposome perfluocarbon contrast agent can significantly long time enhance the grey scale of the kidney.

Objective To promote in vitro tissue-plasminogen activator(t-PA)thrombolysis by using microbubbles augmented low frequency ultrasound(US) insonication.Two kinds of microbubbles,albumin-coated vs.lipid-coated were compared.Methods Human blood clot weighted 200～300 mg was made from of 0.8 ml fresh blood.Twenty kHz ultrasound,lipid and albumin-coated microbubbles were applied to separate clots groups with or without t-PA administration.The clots before and after processing were weighed and then the clot dissolution ratios were calculated.Results The simple ultrasound group and the basic t-PA group dissolved((24.72)?(4.83))% and((35.66)?(3.34))% of the clots,respectively.The clot dissolution ratio rised to((42.06)?(4.20))% when coordinated US and t-PA(P(0.05)).Conclusions The in vitro US augmentation of t-PA thrombolysis can be significantly promoted by introduction of microbubbles.But there is no significant thrombolysis difference between albumin and lipid-coated microbubbles.

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 .

BACKGROUND: Olfactory ensheathing cell transplantation is a better theray for spinal cord injury, and it becomes one of the most promising treatment methods. Local transplantation is applied currently, with the disadvantages of complex operation, large trauma, repeated transplantation. Looking for a simple and effective way for cell transplantation becomes a hotspot for scholars from various countries.OBJECTIVE: To investigate the effect and possibility of transplantation of olfactory ensheathing cells for treatment of spinal cord injury.METHODS: Wistar rats with T 10 spinal cord hemisecti on were divided into 4 groups: intramedullary local transplantation group (A),vein transplantation group (B), D/F12 transplantation group (C) and control group (D). The functional recovery of rats with spinal cord injury was observed with combined behavioral score at different phases. The tissue sections of each group were made at 5 and 10 weeks postoperatively to observe the axon regeneration and the survival of olfactory ensheathing cells.RESULTS AND CONCLUSION: The experiment showed that the rats transplanted with OECs at injured site and through the vein had more improvement in functional recovery and histological changes than the other two groups. The effect between A group and B group had not significant difference. The method of treating spinal cord injury by transplanting OECs via the vein not only simplifies the operation and avoids many complications but also has good curative effect similar to local transplantation.

BACKGROUND: A proper preservation method would be of important significance for experiments and clinical application ofolfactory ensheathing cells (OECs) OBJECTIVE: To explore proper cyropreservative systems for OECs.METHODS: OECs during the logarithmic growth phase were harvested, cryopreserved for 1, 3 and 6 months and then revitalized.RESULTS AND CONCLUSION: MTT assay and tryplan blue staining showed that cells exhibited highest viability after treatmentwith 5% dimethyl sulfoxide (DMSO)-6% hydroxyethyl starch (HES), followed by 10% DMSO, and lastly the 5% DMSO. Use ofrefrigerator or cryogenic control system with different cryopreservation time did not yield obvious effects on viability of OECs.Therefore, 5% DMSO-6%HES is recommended as a cryopreservative agent for OECs.

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.