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 assess the effects ofberberine on autophagy in C3H10T1/2 adipocytes.Methods C3H10T1/2 cells,a pluripotent stem-cell line of mesodermal origin,were induced to differentiate into mature adipocytes,and then were treated with berberine,The expression of autophagy marker protein LC3 Ⅱ/Ⅰ and autophagy substrate P62 was determined by Western blot ; after treating C3 H 10T1/2 cells with berberine and lysosomal inhibitor,chloroquine,autophagy flux was assessed by Western blot.Autophagosome was observed by transmission electron microscopy after berberine treatment.Results (1) After berberine treatment,the expression of LC3 Ⅱ / Ⅰin C3H10T1/2 adipocytes was reduced and P62 was increased in a time-and dose-dependent manner(P＜0.05 or P＜0.01) ; (2) Following treatment with chloroquine and berberine,the protein level of LC3 Ⅱ/Ⅰ was decreased (P＜0.05); (3) The number of autophagosome was decreased apparently after berberine treatment.Conclusions Berberine inhibits autophagy in C3H10T1/2 adipocytes.

Background and purpose:KIT mutation plays an important role during the pathogenesis of melanoma. This study was designed to investigate the mutation status ofKIT in different subtypes of melanoma. Methods:A total number of 144 cases of melanoma were analyzed forKIT mutation (exon 9, 11, 13 and 17) by DNA sequencing using paraffin-embedded tissues.Results:The total incidence ofKIT mutation in melanoma was 9.0% (13/144).KIT mutations in acral melanoma, mucosal melanoma, melanoma on skin with chronic sun-induced damage (CSD) and melanoma on skin without chronic sun-induced damage (non-CSD) was 7.7% (4/52), 20% (7/35), 14.3% (1/7) and 2.8% (1/36), respectively. Among 13 cases withKIT mutation, 1 mutation lay in exon 9, 9 lay in exon 11 and 3 in exon 13. L576P in exon 11 was the most common type of mutation.Conclusion:The most prevalent type ofKIT mutation in patients lies in exon 11.KIT mutation could be the potential drug target in melanoma therapy.

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.

Four crystalline forms of clopidogrel bisulfate were characterized by analytical techniques. Aiming to research the absorption characteristics of clopidogrel bisulfate polymorphs after taken orally by rat, and to estimate the influence of crystal form to pharmacodynamic action, four crystalline forms of clopidogrel bisulfate were administered intragastrically to rats, and high performance liquid chromatography (HPLC) was used to measure the contents of clopidogrel bisulfate and its metabolite in rat plasma. The metabolite of clopidogrel bisulfate was detected in rat plasma. There were significant deviations among four crystalline forms in the areas under curve of the metabolite of clopidogrel bisulfate. We concluded that the different crystal forms of clopidogrel bisulfate showed different pharmacokinetic characteristics, which might affect pharmacodynamic action.

Objective To identify microbubbles targeted (MBt) to alpha(v)beta(3) (αvβ3) via biotin-avidin bridge and evaluate the adhesion to human umbilical vein endothelial cells (HUVECs) in vitro.Methods MBt produced via biotin-avidin bridge were validated using fluorescence in vitro.Adhesion of αvβ3-integrin targeted MBt (MBαvβ3) to HUVECs was tested using the parallel plate flow chamber (PPFC) test.Results Bright green fluorescence was observed on the biotinylated microbubbles(MBB) incubated with fluorescein isothiocyanate labeled streptavidin (FITC-SA) and on MBB-SA incubated with FITC labeled biotin.There was no fluorescence seen on non-targeted control microbubbles,MBB incubated with FITC labeled protein A and MBB-SA incubated with FITC labeled protein A. The adherent rate of MBαvβ3 was significantly higher than MBt with non-specific antibody (MBN) in PPFC test,with 9.9±3.1 of MBαvβ3 and 0.8±0.3 of MBN adhered to HUVECs,respectively(P<0.05).Conclusions Avβ3 targeted microbubbles using biotin-avidin bridging method is highly efficient and reliable for HUVECs.

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.

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 evaluate the therapeutic efficacy and safety of radiofrequency ablation (RFA) combined with ethanol injection with a multipronged needle under ultrasound guidance for the treatment of hepatocellular carcinoma (HCC) larger than 3 cm in diameter.Methods 65 patients with 67HCC nodules ranging from 3.1 to 7.0 cm in diameter were treated percutaneously under ultrasound guidance.Tumor response and complications after treatment were observed.Results Complete ablation was achieved in 94.0％ (63/67) of HCC nodules.4 residual tumor nodules received complete ablation after additional treatment.Ablation-related major complications was occurred in 3 patients,including liver abscess in 1 case,abdominal bleeding in 1 case and massive ascites in 1 case,all were cured by conservative therapy.After a mean follow-up period of (20.0 ± 7.6) months (6.7-32.6 months),local-tumor progression was observed in 10 (14.9％) of 67 HCC nodules,and distant recurrence was observed in 32 (49.2％) of 65patients.The 1-year and 2-year survival rate were 93.1 ％ and 88.1％,respectively.Conclusions RFA combined with ethanol injetion with a multipronged needle is a safe and effective technique for the treatment of hepatocellular carcinoma larger than 3 cm,especially 3-5 cm in diameter.