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.

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.

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 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.

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 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 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 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.