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 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 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 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 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.

Objective To compare contrast-enhanced ultrasound(CEUS)and contrast-enhaneed computed tomography(CECT)on the diagnosis of renal cell carcinoma(RCC). Methods CEUS and CECT were performed on 117 patients(87 men and 30 women)with 124 renal lesions(single nodule in 110 and two nodules in 7)from 2004 to 2008.Among them,there were 63 patients with 65 lesions diagnosed as RCC confirmed by pathology.The tumor enhancement pattern,extent,and dynamic change of CEUS and CECT were compared.The diagnostic efficacy of CEUS and the agreement of CECT and CEUS in diagnosing RCC were analyzed. Results The rate of displaying hypervascular performance on cortical phase,and pseudocapsule enhancement of the RCC lesions by CEUS and CECT were 87.7%(57/65)and 63.1%(41/65),89.2%(58/65)and 69.2%(45/65)(P<0.05).The rate of displaying heterogeneous enhancement were 72.3%(47/65)and 56.9%(37/65)(P>0.05)Using the diagnosis of CECT as reference diagnostic criteria,the sensitivity,specificity,positive predietive value,negative predictive value,and accuracy of CEUS in diagnosing RCC were 89.1%(57/64),91.7%(55/60),91.9%(57/62),88.7%(55/62)and 90.3%(112/124).The agreement of CECT and CEUS in diagnosing RCC was high(κ=0.806). Conclusions CEUS and CECT have the coordinate efficacy in diagnosing RCC.

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 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.