Objective To study the construction of tissue engineering fat filler and the survival of fat particles.Methods Stromal vascular fraction (SVF) was derived from inguinal subcutaneous fat of rabbits,mixed with autologus mature fat particles and with or without neuropeptide Y to construct PLGA composite tissue.The complex was subcutaneously transplantef into back sites of the rabbits.Based on the different combination five groups were divided:Group A:PLGA + mature fat particles without NPY;Group B:PLGA + mature fat particles + NPY;Group C:PLGA + complex of SVF with mature fat particles without NPY;Group D:PLGA + complex of SVF with mature fat particles + NPY;Group E:Complex of SVF with mature fat particles + small ball with NPY.Diffenrence of virous constructive ways and fat particle survival was evaluated by general observation,histological staining,fluorescence tracing at two weeks,one month and three month after operation.Results Group D was superior to groups A,B,C and E in survival volume,graft texture and vascularization at one mouth and three mouths.The fat average srvival rate was 57.5±2.5％.Fat cell grew well,and precursor cells proliferated and differentiated actively.Conclusions High quality tissue engineering materials are successfully established with SVF-mature fat particle complex,PLGA and NPY,which could obviously improve fat particle transplantation.

Objective To elucidate potential activities of phthalides and terpenoids from Angelica sinensis by theoreticaldocking calculation.Methods Eleven components of phthalides and terpenoids were selected as ligand.Thecrystalline structures of targets related to common diseases were used as the receptors for calculation.Thecalculations were conducted with Schr(o)dinger software package.The grading standard of selectivity was developedaccording to G-score between ligands and receptors.Results Selective targets of phthalides and terpenoids wererelated to nevous system diseases,cancer,pain,diabetes,cardiovascular disease,liver cirrhosis,nephrotic syndrome,inflammatory diseases,rheumatoid arthritis,dermatosis,leukemia,microbial inflections,immune diseases,andhypercholesterolemia.In addition to the medical treatments reported in the literature,our research also indicated thatthese two classes of compounds may be used for tumor,diabetes,rheumatoid arthritis,dermatosis,leukemia,livercirrhosis,and nephrotic syndrome.According to our research,the effects of phthalides and terpenoids may be not sostrong.Conclusion The effects of phthalides and terpenoids on diseases founded through virtual evaluation accord greatly with those reported in experiment and clinic.The combination of computer-aided drug evaluation technique and experiment is definitely an important and fast way to investigate the effects and mechanisms of traditional Chinese medicine.

Objective To explore the investigation method of complicated and profound traditional Chinese herbal medicine,the potential action mechanisms of flavonoids from Scutellaria baicalensis were studied by docking calculation.Methods In total, eight flavonoids (aglycones and their glicosides) from S. baicalensis were selected as ligands.The crystalline structures of targets related to common diseases were used as the receptors for calculation. The calculations were conducted with Schr(o)dinger software package. The grading standard of selectivity was developed according to G-score between ligands and receptors. Results Twenty-six pharmacologic actions have been reported.Among all effects in literature, nine of them can be deduced from the docking calculation of aglycone. From glycosides with grade ++, 25 reported effects can be estimated by calculation. Apparently, the target selectivity of aglycones and their glycosides are different form the virtual evaluation. The virtual evaluation results of glycosides were closer to the reported effects. Conclusion Our proposed virtual evaluation method seems an effective way to investigate the complicated system of traditional Chinese herbal medicine. It suggests that aglycones may be effective as the form of glucoside in vivo, and metabolism is a very important factor for virtual evaluation.

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 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 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 investigate the usefulness of parametric imaging of contrast-enhanced ultrasound(CEUS) in imaging hepatocellular carcinoma (HCC) using dynamic vascular patterns (DVP). Methods Thirty clinically or pathologically proven HCCs that had undergone CEUS were randomly included. SonoLiver CAP sofeware was used to analyze the CEUS images and reconstruct DVP parametric images. Results The rise time, time to peak and mean transit time were (16. 72±11. 07) s, (29. 92±14. 13) s,(115. 03±90. 91)s in HCC versus (26. 59±9. 60) s, (41.67±12. 59) s, (159.26±123. 74) s in the surrounding liver parenchyma (all P <0. 05). The perfusion index was (90. 41±102. 49) % in HCC versus (54. 10±24. 99)% in surrounding liver parenchyma( P = 0.044). DVP curve and DVP parametric image could both be divided into three types:washout,non-washout and cystic type. The percentages of which were 76.7% (23/30), 20.0% (6/30) and 3.3% (1/30) in DVP curves, respectively, and 66.7% (20/30), 30.0% (9/30) and 3.3% (1/30) in DVP parametric images,respectively. Conclusions Parametric image of CEUS could demonstrates the difference of flow perfusion static between HCC and surrounding liver parenchyma dynamically and directly.

Objective To investigate the features of low grade dysplastic nodule(LGDN)and high grade dysplastic nodule(HGDN)in cirrhotic liver on the baseline ultrasound(BUS)and contrast-enhanced ultrasound (CEUS).Methods Seventeen LGDNs and 16 HGDNs which were pathologically proved underwent baseline and CEUS examination.CEUS was performed with contrast pulse sequence(CPS)and contrast harmonic imaging (CHI)and used SonoVue as contrast agent.Results Significant differences(all P>0.05)were not shown between LGDN and HGDN for manifestations on both the BUS and CEUS.After contrast agent injected, hyperenhancement in arterial phase and hyper-or iso-enhancement in late phase,hypo-enhancement in early arterial phase and iso-enhancement through late arterial,portal and late phase,hyper-enhancement in arterial phase and hypo-enhancement in late phase,iso-enhancement in arterial,portal and late phase,iso-enhancement in arterial phase and hypo-enhancement in late phase were showed in 10(30.3%),10(30.3%),8(24.3%),4(12.1%),1(3.0%)cases of DN, respectively.Conclusions LGDN and HGDN displayed similar feature on both the BUS and CEUS.There were complicated manifestations of DN on the CEUS and the main contrast-enhanced sonographic feature of DN was,different from regenerative nodule and hepatocellular carcinoma.CEUS was helpful in diagnosing DN in cirrhotic liver.

Objective This study was carried out to compare chnical efficiency of methylprednisolone (MP) and dexamethasone (Dex) and their effects on Th cytokines in asthmatic children.Methods A total of 39 children with moderate and severe asthma at acute exacerbation were randomly divided into two groups, one (A) with MP 1 -2 mg/kg (n=21) and the other (B) with Dex 0.25 -0.75 mg/kg (n= 18),every 12 -24 h by intravenous drip.Scores of respiratory effort and peak expiratory flow rate prior to and three days after treatment in the asthmatic children aged over five years were evaluated,respectively,as well as the time of wheezing vanishing after treatment.Serum levels of interleukin-2 (IL-2) and IL-4 were determined by radioimmunoassay (RIA) prior to and three days after treatment,respectively.Results There was statistically significant difference in the time of wheezing vanishing between groups A and B (P = 0.042).Three days after treatment,PEER was significantly higher in group A than that in group B (P = 0.025).No statistically significant difference in serum IL-2 and IL-4,as well as ratio of IL-2/IL-4,was found in group B prior to and three days after treatment (P>0.05).Statistically significant difference in serum IL-2 was not observed in group B prior to and three day after treatment (P>0.05),and serum IL-4 decreased significantly (P=0.001) and ratio of serum IL-2/IL-4 (P=0.027) increased significantly three days after treatment than those prior to treatment.No significant correlation between respiratory-effort scores and ratio of serum IL-2/IL-4 prior to treatment was found (P=0.613).Conclusions Up-regulation of IL-2,or inhibition of release of IL-4,probably is not the main anti-inflammatory mechanism of dexamethasone.Methylprednisolone has little effect on serum IL-2,but can effectively reduce serum IL-4, thus increasing the ratio of serum IL-2/IL-4 and counterbalancing function of the Thl/Th2 cells.

Objective:To investigate the effect of IL-17A on the differentiation and maturation of murine bone marrow-derived dendritic cells( BMDCs ) . Methods: Murine bone marrow cells were isolated and cultured in RPMI1640 complete medium in the presence of GM-CSF(20 ng/ml) for 8 days to induce differentiation of murine bone marrow cells to DC progenitors. Then these cells were treated with LPS(1 μg/ml) for 36 h which polarized immature DCs into mature DCs. Different concentrations of rmIL-17A(10 or 100 ng/ml) was added to the culture medium at different stages of BMDC differentiation and maturation. Co-stimulatory molecules expression on BMDC were analyzed by flow cytometry,and the culture supernatants were analyzed for IL-12p40 and IL-10 level by ELISA. Results:rmIL-17 could promote co-stimulatory molecules( CD40,CD80,CD86 and MHCⅡ) expression on BMDCs in a does-dependent manner,especially,the expression of CD40 and MHCⅡhad a significant increase in high concentration of rmIL-17A group;rmIL-17A was added while LPS induced maturation of BMDCs. CD40,CD80,CD86 and MHCⅡexpression on BMDC increased sharply in LPS plus rmIL-17A stimulation group,besides,CD86,MHCⅡ showed a higher level expression on BMDC with the increase of con-centration of rmIL-17A. Furthermore,secretion of IL-12p40 and IL-10 increased significantly in the group of DCs treated with LPS plus low concentration of rmIL-17 compared with the group without rmIL-17(P<0. 001). However,high concentration of rmIL-17A group showed significantly higher levels of IL-12p40(P<0. 001),but there was no difference in IL-10. Conclusion:IL-17A promotes the phe-notypic development of BMDC progenitors propagated in GM-CSF and cooperate with LPS to induce BMDC differentiation and matura-tion.