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.

ObjectiveTo investigate portal hemodynamic features and its changes following devascularization in cirrhotics. MethodsHemodynamics of portal trunk (PT), right anterior branch (RAB) and splenic vein (SV), including maximum cross-sectional velocity(CS-Vmax), flow volume and congestion index(CI), were assessed in 69 cirrhotics and 46 normal volunteers by using a recently-developed color Doppler velocity profile(CDVP). Of 28 patients undergone devascularization procedure, portal hemodynamics were studied and compared before and after operation. ResultsCSVmax of PT and RAB was significantly lower in cirrhotic group than normal group;PT and SV flow volume and the ratio of SV to PT flow volume(SV/PT)were significantly greater in cirrhotic group compared with those of normal group;CI of PT, RAB and SV was significantly higher in cirrhotic group than normal group. Postoperative PT flow volume was significantly reduced( P ＜0.01),and the reduction was closely related to preoperative SV flow volume( r = 0.65, P ＜0.001). CS-Vmax and flow volume in RAB were decreased significantly following operation( P ＜0.01), and the reduction of RAB flow volume was highly related to preoperative RAB flow volume( r =0.74, P ＜0.001). After operation, free portal pressure(FPP)was declined by (6 ± 5)cmH2O[ (0.59 ± 0.49)kPa] ( P ＜ 0.001 ). There were no significant changes in CS-Vmax, CI in PT and CI in RAB following operation. ConclusionIn cirrhotics with portal hypertension, portal venous system coexists the elevated vascular resistance and hyperdynamics, but with different redorainance at different portion. SV hyperdynamics is the main source of increased portal blood flow. Devascularization procedure could markedly relieve portal hyperdynamics by elimination of SV inflow, which is one of the main mechanisms in obmining therapeutic goal. But the operation has no favorable effects on the increased portal resistance,and portal perfusion to the liver would be further declined after relieving portal hyperdynamics, which is unfavorable to maintenance of liver function.

Objective To conclude the characterization of renal cellular carcinoma(RCC)with contrast enhanced ultrasound.Methods Seventy patients(seventy-two nodules)with RCC,which were confirmed by operation and biopsy underwent conventional ultrasound and contrast-enhanced ultrasound(CEUS).Microbubble agents SonoVue and contrast pulse sequence(CPS)were used.The conventional uItrasonographic characterization and the enhancement patterns of lesions were analyzed.Results On baseline sonography,the numbers of lesions that showed hypoechogenicity,isoechogenicity,and hyperechogenicity were 44.4%(32/72),25.0%(18/72)and 30.6%(22/72),respectively.Only 28 lesions(38.9%)showed flow signals on color Doppler sonography,the mean maximum velocity of which WSS(43.7±16.8)cm/s(range,24.8-95 cm/s),and the mean resistance index was 0.635±0.11(range.0.52-0.83).Sixty-three(87.5%)lesions were hyper-vascular in cortical phase.Among them forty-eight(76.2%)lesions were hypo-enhanced,and fifteen(23.8%)lesions were still hyper-vascular in late phase.The remaining nine hypervascular nodules in cortical phase were still hyper-enhancing in late phase.Fifty-four(75.0%)lesions were inhomogeneous enhancement.and pseudocapsule was observed in sixty-three(87.5%)RCC lesions.Conclusions The enhancement patterns of RCC are characteristic,and CEUS may be helpful in differential diagnosis of focal renal lesions.

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 value of contrast-enhanced ultrasound (CEUS) in the diagnosis of ischemie-type biliary lesion (ITBL) after liver transplantation. Methods Twenty-five liver transplantation recipients suspected with biliary complications were examined using CEUS. Percutaneous transhepatic cholangiography or endoscopic retrograde cholangiopancreatography confirmed the diagnosis. The enhancement patterns of the thickened hilum bile duct wall were observed. Results Enhancement patterns of bile duct wall could be divided into 3 sorts:① No-enhancement, six cases, showed no-enhancement all along. ②Hypo-enhancing level, four cases, presented hypo-enhancing at arterial phase and persistent hypo- or no-enhancing until late phase. ③Hyper-enhaneing level,fifteen cases,appeared as hyper- or iso-enhancing at arterial phase and persistent iso- or hypo-enhancing until late phase. The enhancement pattern of bile duct wall showed significant difference ( P = 0.00). Thirteen ITBL patients included 10 cases ( 76.9 % ) with no- or hypo-enhaneement, 3 patients (23.1 % ) with hyper-enhancement. However, all of 12 non-ITBL cases appeared hyper-enhancing. Conclusions CEUS provides a new and effective method to estimate the microcirculation of the bile duct wall. It may be help to early diagnosis of ITBL.

Objective To investigate the value of contrast-enhanced ultrasound (CEUS) in percutaneous nephrostomy for the treatment of complex renal calculi without hydronephrosis.Methods 22 patients with complex renal calculi underwent puncture and catheterization under the conventional ultrasound guidance,and then were injected with ultrasound contrast agent (SonoVue) through the needles and tubes to confirm appropriate puncture and catheterization.The display rates of puncture needle,renal pelvis,calyces,drainage tube body and distal end under CEUS were observed,and the results were compared with those of routine ultrasonography.The dosage of contrast agent,success rate and complications were also recorded.Results The display rates of puncture needle,renal pelvis,calyces,drainage tube body and distal end under routine ultrasonography were 63.64％,36.36％,63.64％,18.18％,as compared with CEUS the display rates were 100％,100％,100％,81.18％.The differences were statistically significant between CEUS and routine ultrasonography (P ＜ 0.05).Conclusions CEUS guided percutaneous catheterization makes up for the inadequacy of conventional ultrasound in patients of complex renal calculi without hydronephrosis,and it can be worthy for clinical application.

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 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 characteristic of the gallbladder diseases and to evaluate the diagnostic efficacy in differential diagnosis of the gallbladder diseases between contrast-enhanced sonography (CEUS) and contrast-enhanced computed tomography (CECT).Methods Seventy-two patiens with gallbladder lesions were examined by CEUS and CECT before operation and all final diagnoses were confirmed by surgery and/or pathological diagnosis.Results 1)In early phase,84.6％ (33/39) and 79.5％ (31/39) of benign diseases showed hyper-enhancement on CEUS and CECT,and 97.0％ (32/33) and 87.9％ (29/33) of malignant diseases showed hyper-enhancement on CEUS and CECT respectively (P =0.250).In later phase,91.2％ (31/34) and 88.2％ (30/34) of benign diseases showing hypo-enhancement (P =1.000),and 100％ of the malignant diseases showing hypo-enhancement on CEUS and CECT respectively.2)The time of enhancement from hyper-to hypo in CEUS for benign and malignant diseases were (39.9 ± 15.7)s and (29.9 ± 5.6)s respectively (t =3.61,P =1.000).3)The inhomogeneous enhancement on CEUS and CECT were 41.0％ (16/39) and 53.8％ (21/39) in the benign diseases respectively (P =0.063),84.8 ％ (28/33) and 97.0 ％ (32/33) in the malignant diseases respectively(P =0.125).4)The destruction of the gallbladder wall on CEUS and CECT are 87.9％ (29/33) and 90.9％ (30/33) respectively(P =1.000).5)The accuracy,diagnostic sensitivity,specificity of CEUS and CECT were 91.7％(66/72) and 87.5％(63/72),97.0％(32/33) and 93.9％(31/33),87.2％(34/39) and 82.15％ (32/39) respectively (P =0.250,1.000,0.500).Conclusions The enhancement pattern of the gallbladder diseases on CEUS and CECT were much similar.CEUS has equal diagnostic efficacy in comparison with CECT,but CEUS can supply more diagnostic information than CECT.