Objective To explore the optimal dose-effect relationship of gene transfer according to the expression of pEGFP in tumor cells under different parameters mediated by microbubble contrast agent and ultrasound in vivo.Methods C57BL/6J mice were inoculated subcutaneously in the middle of the right flank with hepal-6 tumor cells.The mixture of SonoVue and enhanced green fluorescent protein(EGFP)plasmid was injected into the tail vein of each mouse, groups were randomly established according to different output intensity( 1 W/cm2, 2 W/cm2, 3 W/cm2 ), exposure time ( 1 min, 5 min, 10 min) and volume of SonoVue(30 μl,60 μl,90 μl), the expression of EGFP in tumor cells under different parameter was examined by flow cytometry and fluorescence microscopy.Results The expression of pEGFP in tumor cells was significantly higher with the increase of exposure time,output intensity and volume of SonoVue (1 min vs 5 min, P＜0.05; 1 W/cm2 vs 2 W/cm2, P＜0.05 ; 30/μl vs 60 μl, P＜0.05).However, the transfection efficiency of EGFP can not be enhanced continuously with the increasing exposure time,output intensity and volume of SonoVue (5 min vs 10 min, P＞0.05; 2 W/cm2 vs 3 W/cm2, P＞0.05; 60 μl vs 90 μl, P＞0.05).The highest transfection effieieney was achieved under intensities was 2 W/cm2 [(21.02±1.45)%]with 5 min[(23.22±1.91)%] exposed to ultrasound, and 60 /μl SonoVue [ (21.02±1.45) % ].Conclusions Gene transfeetion efficiency changded with the difference parameters.2 W/cm2,5 min and 60 μl SonoVue led to the optimum dose-effect relationship of gene transfer in vivo .

Objective To investigate the usefulness of parametric imaging of contrast-enhanced ultrasound(CEUS) in evaluating hepatocellular carcinoma ( HCC) and focal nodular hyperplasia(FNH). Methods Thirty clinically or pathologically proven HCCs and 30 pathologically proven FNHs that had undergone CEUS were randomly included. SonoLiver CAP software were used to quantitatively anlalyze the CEUS and reconstruct DVP parametric images. Results The rise time, time to peak and mean transit time in HCC and FNH were (16. 7 ± 11. 1)s and (21.9±9. 0)s (P = 0. 052) ,(29. 9 ± 14. l)s and (33. 2 ±11.1)s ( P =0. 322), (115. 0±90. 9)s and (271. 5 ± 147. 6)s ( P = 0. 000),respectively. The perfusion index was 90. 4 + 102. 5 in HCC and 42. 6 + 37. 1 in FNH( P = 0. 022). DVP curve and DVP parametric image could both be divided into three types: washout, non-washout and cystic type. In DVP curve, the percentages of which were 76. 7% (23/30),20. 0% (6/30),3. 3% (1/30) in HCC, and 43. 3% (13/30) , 53. 3% (16/30) , 3. 3%(l/30) in FNH,respectively ( P = 0. 023). In DVP parametric image, the percentages of which were 66. 7%(20/30) ,30. 0%(9/30) ,3. 3%(l/30) in HCC,and 33. 3%(10/30) ,60. 0%(18/30) ,6. 7%(2/30) in FNH,respectively ( P = 0. 033). 30. 0% (9/30) of FNH had a clear spoke-wheel pattern in DVP parametric image. Conclusions In parametric imaging of CEUS, the mean transit time of HCC was shorter than that of FNH,and the perfusion index of HCC was higher than that of FNH. The DVP parametric image can display the rapid change and detail of the enhancement clearly.

Objective To evaluate the clinical value of percutaneous ultrasound (US)-guided thermal ablation of intrahepatic cholangiocarcinoma. Methods 14 patients (19 nodules) with histolog-ically proven intrahepatic cholangiocarcinoma (ICC) were treated with percutaneous US-guided thermal ablation from Oct. 1998 to Aug. 2009 in our hospital. The local treatment response, complication and survival were retrospectively analyzed. Results 18 (18/19, 94.7%) nodules had complete necrosis, and 1 (1/19, 5.3%) nodule had residual tumor after ablation. There was no mortality associated with thermal ablation. Only 1 (1/14, 7.1%) patient developed portal vein thrombosis. The follow-up ranged from 1.3 to 72.9 months (mean, 19.0±15.1 months; median, 7.25 months), 10 (10/14,71.4%) patients survived less than 1 year, 2 (2/14,14.3%) survived 1-5 years, and 2 (2/14,14.3%) survived more than 5 years. Of the 14 patients, 9 (9/14, 64.3%) had died and 5 (5/14,35.7%) were alive. The causes of death included tumor progression (8/9, 88.9%) and liver failure (1/9, 11.1%). 9 (9/14,64.3%) patients developed recurrence on follow-up. The disease-free survival ranged from 1.1 to 72. 9 months (mean 16.6±26. 3 months; median 4.1 months). Conclusion Percutaneous US-guided thermal ablation therapy using microwave ablation (MWA) or radiofrequency ablation (RFA) is a safe and effective therapy for intrahepatic cholangiocarcinoma.

0.05 ). 3DUS was superior to 2DUS in displaying the expanded tips (P 0.05 ) in the ablation modality. 3DUS enhanced the confidence level significantly in judging the position relationship between applicators and lesions in both modalities (both P

Objective To assess the characterization and usefulness of contrast enhanced ultrasound (CEUS)to diagnose solid focal lesions of pancreas(s-FLPs)in comparison to contrast enhanced computed tomography(CECT).Methods Forty-nine s-FLPs with final diagnosis were studied with low mechanical index CEUS and CECT.The enhanced patterns and diagnostic capability of CEUS were analyzed,and these results were compared with those from CECT.Results ①In CEUS,the perfusion of capillary could be detected in vascular phase.②The concordance of dual-phases enhanced patterns between CEUS and CECT was 73.5%(36/49).The concordance of enhanced level between early parenchymatous perfusion phase in CEUS and pancreatic phase in CECT could achieve 87.8%(43/49).Hypo/hypo enhancement was the most common enhanced pattern of pancreatic carcinoma in CEUS and CECT.③There was no statistical significance between CEUS and CECT in the differential diagnostic.Conclusions The concordance of enhanted patterns between CEUS and CECT is satisfactory.CEUS is similar to CECT in the differential diagnosis of s-FLPs.CEUS would become an important non-invasive diagnostic method for s-FLPs.

Objective To investigate the features of hepatic angiomyolipoma(AML) on contrast enhanced ultrasound (CEUS). Methods Ten pathologically proved AML lesions in 9 patients were evaluated by using baseline ultrasound and CEUS. CEUS was performed with contrast pulse sequencing technique and the contrast agent of SonoVue. Results On baseline ultrasound, 7 lesions exhibited mixed echoic,which was obvious hyperechoic combined with partly of hypoechoie; 3 lesions exhitited obvious hypereehoic. On color Doppler flow imaging artery signals were detected in all lesions. And the lesions had plenty or slightly plenty of artery supply,with resistive index about 0.53 4±0.10 (0.48~0.62). On CEUS, in the arterial phase, 9 lesions exhibited hyper-enhanced. In the portal phase, 5 lesions exhibited iso-enhancement,1 lesion was slightly hyper-enhancement and 3 lesions was hypo-enhancement. In the late phase 6 lesions were iso-enhancement and 3 lesions were hypo-enhancement. One lesion was all hypo-enhancement in all three phases. Conclusions There are some characteristic manifestations of hepatic AML in CEUS, which are helpful for the diagnosis of hepatic AML in a way,but we still need more experiences.

Objective To evaluate the feasilibility of ultrasonic cholangiography(USC)with contrast agent SonoVue in evaluating the obstructive bile duct diseases.Methods Twenty patients with obstructive jaundice(choledocholithiasis in 4,post-operative biliary strictures in 2 and malignant tumors in 14)underwent uhrasound-guided percutaneous transhepatic cholangiodrainge drainage(PTCD).Afterward,USC with contrast agent SonoVue administrated from the PTCD tube and percutaneous transhepatic cholangiography(PTC)were carried out in each patient.By using the surgical or PTC findings as reference standard,accuracy of USC in determining the obstructive level and cause of bstructive was evaluated.Results ①The visualization of contrast agent SonoVue was successfully obtained in all 20 patients,three order of bile duct was visualized in 19 patients.The visualization of the ultrasound contrast agent persisted more than 6 min.There was no side effect occurred during the procedure and no uncomfortable complaint after the procedure.②USC determined correctly in 18 cases.Compared with surgical or PTC findings,the accuracy of USC in determining obstructive levels was 90.0%(18/20),and the accuracy in determining the obstructive cause was 85.0%(17/20).Conclusions As a new and safe technique for cholangiography,USC is comparable to PTC in displaying the intrahepatic bite duct and determining the obstructive levels in patients with obstructive jaundice.

Objective To compare the enhancement pattern of hilar cholangiocarcinoma on contrast-enhanced sonography(CEUS) and contrast-enhanced computed tomography(CECT). Methods Thirty-two hilar cholangiocarcinoma lesions in 32 patients were evaluated by CEUS and CECT. CEUS was performed with contrast agent of SonoVue and low mechanical index contrast specific mode. CECT was performed with contrast media of Ultravist and a standard biphasic helical CT scanning protocol. Results In arterial phase of CEUS and CECT,the numbers of the lesions showing hyper-enhancement were 14 (43.8 % ) and 12 (37.5 % ), iso-enhancement were 14 (43.8%) and 9(28. 1%),hypo-enhancement were 4(12.5%) and 11(34.4%),respectively (P = 0. 162).Three lesions (9.4%) showed periphery rim-like hyper-enhancement and 29 (90.6%) showed diffuse homogeneous or heterogeneous enhanced of the whole tumor on CEUS, whereas for CECT the number were 2 (6. 3%) and 30 (93. 8%) respectively ( P = 1. 000). In portal phase, the numbers of the lesions showing hypo-enhancement on CEUS and CECT were 30 (93.8 % ) and 23 ( 71.90% ), iso-enhancement were 1 ( 3.1 % ) and 8 ( 25.0 % ), hyper-enhancement were 1(3. 1%) and 1 (3. 1%), respectively ( P = 0. 046). The portal vein invasion was correctly detected in 16 ( 84. 2 % ), 17 (89. 5 % ), 15 (78.9 % ) lesions with baseline ultrasound, CEUS and CECT, respectively. CEUS and CECT correctly diagnosed 30(93.8% ) and 25 (78. 1% ) lesions prior to operation (P = 0. 125). Conclusions The enhancement pattern of hilar cholangiocarcinoma on CEUS was similar with that on CECT in arterial phase, whereas in portal phase hilar cholangiocarcinoma was prone to show hypo-enhancement on CEUS. CEUS and CECT has similar diagnostie efficacy prior to operation.

Objective To evaluate the accuracy and utility of percutaneous ultrasonic cholangiography in Bismuth staging of hilar cholangiocarcinoma. Methods Thirty patients who underwent surgery and obtained pathologic diagnosis of hilar cholangiocarcincoma were perspectively ruled in this study. All patients with hilar obstruction underwent baseline ultrasound (BUS), percutaneous ultrasonic cholangiography (PUSC) and percutaneous transhepatic cholangiography(PTC) respectively. Taking operative findings as reference standard,the accuracy of the three imaging modalities in staging of hilar cholangiocarcinoma was compared. Results Among 30 patients, the accuracy of classification among BUS, PUSC and PTC was 23.3 % (7/30), 73.3 % ( 22/30 ), 73.3 % (22/30), respectively. There was statistically significant difference between BUS and PUSC, but the difference between PUSC and PTC was not statistically significant.Conclusions As a new technique for cholangiography, PUSC expands the application of ultrasound in evaluating hilar cholangiocarcinoma and is comparable to PTC in classifying hilar cholangiocarcinoma by Bismuth classification.

Objective To prepare targeted micorbubble with low immunogenicity. Methods The microbubbles were produced with different phospholipids and identified by the fluorescent method. Detect the level of C3a after reaction with human serum in vitro with enzyme-linked immunosorboent assay (ELISA) method and the number of microbubble binding with the streptavidin packed on the dish by using the parallel plate flow chamber. Results The level of C3a was (1.037±0.047)ng/ml in MBb group,(1. 326 ± 0. 042)ng/ml in MBe group and ( 1.004 ± 0.031 ) ng/ml in MBc group. The level of C3a in MBb group was significantly lower than that in MBe group( P ＜0.05),and there was no significantly difference between MBb group and MBc group ( P ＞ 0. 05). The parallel plate flow experiments showed that the number of MBb(15.2 ± 11.3) in each field of view binding with the streptavidin packed on the dish was significantly fewer than that of MBe ( 103.2 ± 28.3) ( P＜0.05 ), and there was no significantly difference between MBb and MBc(17.8 ± 11.9) ( P ＞0.05). Conclusions The targeted microbubble with low immunogenicity has been prepared successfully,which can be used for further experiment in vivo.