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 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.

Objective To explore the effects of vasoactive intestinal peptide (VIP)on the ratio of CD4+CD25+Treg/CD4+T cell and the expression of TGF-β1 in experimental autoimmune encephalomyelitis (EAE)rats. Methods We randomly divided 60 healthy female Wistar rats into normal control group,EAE control group,VIP low-dose group and VIP high-dose group.We used myelin basic protein (MBP)+ complete adjuvant (CFA)to establish EAE model. Since the day of model construction, the low- and high-dose VIP groups received intraperitoneal injection of 4 nmol/kg (0.2 mL)and 16 nmol/kg (0.8 mL)of VIP every other day,respectively;normal control group and EAE group received injection of saline of 0.8 mL for 10 days in a row.We recorded the peak of neurological dysfunction score (NDS)changes in the rats,observed the pathological changes and GFAP+astrocyte activation in the brain at the morbidity peak of rats with HE staining,and detected the ratio of CD4+CD25+Treg/CD4+T in the spleen with FACS and TGF-β1 cytokine level in brain tissue with ELISA.Results The peak nerve dysfunction score was decreased in each VIP dose group.In normal control group,there were decreased inflammatory cell infiltration and decreased number of active astrocytes in the brain tissue.The degree of infiltration of inflammatory cells and astrocyte activation in VIP control groups were significantly lower than those in EAE group.The CD4+CD25+Treg/CD4+T cell ratio of the spleen tissue in each dose VIP treated group rats was higher than that in EAE control group.The cytokine level of TGF-β1 in the brain tissue increased in each VIP dose group in the dose-dependent manner.Conclusion Through up-regulating the ratio of CD4+CD25+Treg/CD4+T cell in the spleen tissue,increasing TGF-β1 content in brain tissue,and inhibiting the infiltration of inflammatory cells and the astrocyte activation,VIP plays an important role in prevention and control of EAE.

Objective To evaluate the diagnostic performance of real-time contrast-enhanced ultrasound (CEUS)in characterizing complex cystic focal liver lesions(FLLs).Methods Sixty seven complex cystic FLLs in 65 patients were examined with baseline ultrasound(BUS)and CEUS.BUS and CEUS images were reviewed by a resident radiologist and a staff radiologist independently.Diagnostic performance was evaluated using receiver operating characteristic(ROC)analysis and the interobserver agreement was analyzed by weighted k statistics.Results After ROC analysis,the areas under the ROC curve(Az)were 0.917 for the staff radiologist and 0.774(P=0.044)for the resident radiologist on BUS,and were 0.935 and 0.922(P=0.42)on CEUS.A significant difference in Az between BUS and CEUS was found for the resident radiologist(0.774 versus 0.922,P=0.047),whereas not found for the staff radiologist(0.917 versus 0.935,P=0.38).Better results of specific diagnosis were obtained on CEUS[28.4%(19/67)before versus 58.2%(39/67)after review of CEUS images for resident radiologist,and 26.9%(18/67)versus 76.1%(51/67)for staff radiologist,both P＜0.001].Interobserver agreement was improved after review of CEUS images[K=0.325(95%confidence interval:0.214-0.436)on BUS versus k=0.774(95%confidence interval:0.688-0.860)on CEUS].Conclusions Real-time CEUS improves the capability of discrimination between malignant and benign lesions and specific characterization for complex cystic FLLs,as well as the interobserver agreement.

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.

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.