AIM: To investigate the effect of sodium ferulate on phosphorylation of heat shock protein 27(HSP27) in vascular smooth muscle cells(VSMCs) induced by angiotensin Ⅱ(AngⅡ) and platelet derived growth factor-BB(PDGF-BB).METHODS: Cultured VSMCs derived from rat thoracic aorta were used.The activity of HSP27 was evaluated by Western blotting with specific phospho-HSP27 antibody.RESULTS: The phosphorylation of HSP27 in response to AngⅡ and PDGF-BB was suppressed by sodium ferulate in a dose-dependent manner,with maximal inhibition rates of 39.0%(P

[Objective] To study the effect on the growth of mouse Lewis lung carcinoma and on the expression of VEGF in tumor tissue.[Methods] C57BL/6 mice were inoculated the Lewis lung carcinoma cell at right armpit and divided into groups.Different groups were administered by MAALC(low,normal and high dose),CTX or saline.After 15 days,all mice were put to death to detach the tumors then weighed them.After that,we treated tumor tissue with VEGF immunohistochemistry stain,detected the integral optical density of VEGF expression with image analytical technique.[Results] Compared with model group,the average weight and VEGF expression level of the tumors of the high dose group were less and statistically significant(P

AIM:The effective abortifacient drug,mifepristone,was incorporated in the solid lipid nanoparticles (SLNs). The aim of this paper was to study the influence of ratios of drug to lipid on the characteristics of SLNs.METHOD:The physicochemical properties of the fine dispersed systems,such as the size distribution,Zeta potential had been analyzed by laser diffractometry(LD).The measurements of entrapment efficiency(EE)and thermal analysis of DSC were performed as well.RESULT:It was showed if the weight of lipid remained unchanged,the mean particle size of SLNs increased with the increase of drug amount.Drug entrapment efficiency was the highest while about 50 mg drug was loaded on weight 1 g lipid.Simultaneously,the charge of Zeta potential agreed well with the amount of free drug in the colloidal system.DSC analysis results showed that the melting peak of mifepristone at approx.190 ℃ disappeared.CONCLUSION:It was evidence that the drug incorporation would affect the average particle size and Zeta potential of the colloidal systems and the physical state of crystalline drug mifepristone in SLNs was present in the amorphous form or molecularly dispersed at even so high adding amount of 250 mg/g(lipid),which there was no report about the physical state of the drug and SLN with so high drug-loading.It was suggested that the model drug mifepristone could influence the property of nanodispersion system and the crystalline character of the drug was altered by the nanometer carrier system vice versa.

AIM:Surfactants and their blend play important roles in the preparation of solid lipid nanoparticles (SLNs).In this study,four types of surfactant were employed to investigate the influence of the surfactants on properties of SLNs in the absence of model drugs thereby avoiding the interaction between the surfactant and the drug.METHODS:The physicochemical properties of the colloidal systems,such as mean particle size,distribution range and Zeta potential,were investigated by laser diffractometry and the DSC analysis was performed as well.RESULTS:It was found that ionic surfactants,such as sodium deoxycholate, increased the Zeta potential of nanoparticles leading to improve the physical stability of the system.But it showed obviously relative low emulsification efficiency in the preparation.Non-ionic emulsifier,especially Pluronic F-68, offered additional steric stabilization effect avoiding aggregation of the fine particles in the colloidal system.CONCLUSION:The formulation in the study for the first time combined four types of additives including ionic surfactant (sodium deoxycholate),non-ionic emulsifier (Pluronic F-68 and Tween-80),and lecithin to obtain favorably stable nanosuspension,which could stabilize for more than six months without creaming.

0.05). However, monocrotaline significantly increased WT% and WA% of pulmonary arterioles (WT:39.1%?2.8% vs 50.8%?3.1%, WA:51.2%?3.0% vs 74.5%?2.9%, P

AIM: To investigate the effect and mechanism of fluvastatin on the migration induced by platelet derived growth factor-BB (PDGF-BB) and endothelin-1 (ET-1) in cultured vascular smooth muscle cells (VSMCs). METHODS: Cultured VSMCs derived from spontaneously hypertensive rats (SHR) were used. Cell migration was determined by modified Boyden chamber assays. Intracellular free calcium ([Ca 2+ ]i) was measured with fluorescent Ca 2+ indicator Fura-2/AM. RESULTS: PDGF-BB and ET-1 significantly induced VSMCs migration, which was inhibited by pretreatment of VSMCs with fluvastatin (10 -9 -10 -5 mol/L) in a dose-dependent manner, and the peak inhibition rate of migration induced by PDGF-BB and ET-1 was over 86.67%. Fluvastatin also attenuated the increase in [Ca 2+ ]i induced by PDGF-BB and ET-1, with a peak inhibition rate of 86.76% and 65.32%, respectively. CONCLUSION: PDGF-BB and ET-1 promote migration of VSMCs from SHR.Fluvastatin may have direct inhibitory effects on cell migration induced by PDGF-BB and ET-1. The increase in [Ca 2+ ]i may acts as intracellular signaling in the migration in response to PDGF-BB and ET-1 in VSMCs.

AIM: To investigate the effect of sodium ferulate (SF), one of the principal components of rhizoma ligustici wallichii, on the attachment and migration induced by fibronectin (FN) and fibrinogen (Fg) in vascular smooth muscle cells (VSMCs). METHODS: Cultured VSMCs derived from spontaneously hypertensive rats (SHR) were used. Cell attachment was conducted using a flat-bottomed 96 well polystyrene plate. Cell migration was determined by modified Boyden chamber assays. Intracellular free calcium (Ca~ 2+ i) was measured with fluorescent Ca~ 2+ indicator Fura-2/AM. RESULTS: FN and Fg significantly induced the attachment and migration of VSMCs in a dose-and time-dependent manner, which was inhibited by pre-treatment of VSMCs with SF (10~ -7 -10~ -3 mol/L) a dose-dependentl fashion. The peak inhibition rate of attachment induced by FN and Fg was 67.12% and 70.23%, respectively, and the peak inhibition rate of SF on migration induced by FN and Fg was 69.79% and 87.06% (P

Aim To construct the recombinant lentiviral RNA interference(RNAi) sequence targeting rat heat shock protein 27(HSP27)gene.Methods Three complementary DNA sequences targeting rat HSP27gene were designed and synthesized.After phosphorylation and annealing,these double strands DNA were cloned into vector pSUPER-basic respectively.Next the ds-oligo DNA which expressed short hairpin RNA(shRNA) was subcloned into lentiviral vector.The transfer plasmid(pNL-HSP27-IRES2-EGFP) was constructed and confirmed by electrophoresis and sequencing.Then the three plasmids of the lentiviral vector: pNL-HSP27-IRES2-EGFP,VSVG,and pHeler were cotransfected to 293T cells by Lipofectamine 2000 to package lentiviral particles.Forty eight hours later the supernatant was collected and the titer of virus was measured by detecting expression leve1 of enhanced green fluorescent protein(EGFP).Results The analysis using restriction endonuclease digestion and sequencing confirmed that the ds-oligo DNA was successfully inserted into the lentiviral vector.The recombinant lentivirus was harvested from 293T cells with a viral titer of 3.12 ?109 fu?L-1.Vascular smooth muscle cells(VSMCs) were infected by lentivirus,and the interfering efficiency was detected by RT-PCR and Western blot.The plasmid with the highest interfering efficiency was pNL-HSP27-IRES2-EGFP-1,and the interfering efficiency was 0.771.Conclusions The recombinant lentivirus containing RNAi targeting HSP27 gene has been successfully constructed,which lay a foundation for further study of the function and gene therapy of HSP27.

Objective To investigate the influence of Angiopoietin-1(Ang1)gene modified mesenchymal stem cells on the expression of adhension factors from endothelial cells,to clarify the feasibility of modulating inflammation reaction by gene modification after stem cells transplantation.Methods Genetic engineering rMSCs were constructed by lentivirally-tranduced Ang1 gene,the expressions of ICAM-1 and VCAM-1 at different time points after 20 ?g/L VEGF stimulating were investigated,the supernatants of gene modified rMSC were collected and then added into HUVEC culture with different concentrations of Ang1,the expressions of adhension factors after VEGF stimulated were observed at the same time.Results Genetic engineering rMSCs were successfully constructed,the expressions of ICAM-1 and VCAM-1 mRNA were significantly increased after VEGF stimulated,the maximal value was at 8 h,the ICAM-1 and VCAM-1 mRNA expressions were decreased after incubation with Ang1 at different concentrations,Ang1 suppressed the VEGF-stimulated protein levels of ICAM-1 and VCAM-1.Conclusion The supernatants of Ang1 gene modified stem cells can suppresse VEGF-stimulated inflammation reaction of HUVEC.

AIM: To investigate the effects of fibronectin (FN) on the proliferation and collagen synthesis in cultured rat cardiac fibroblasts (CFb) derived from SHR (CFb SHR ) and WKY (CFb WKY ). METHODS: CFb derived from 12-week-old spontaneously hypertensive rat (SHR) and WKY was cultured by outgrowth of tissue block. Cell proliferation of CFb was measured by cell number counting and [ 3H]-TdR incorporation using 24-well plates pre-coated with 5 ?g/cm 2 of FN. Collagen synthesis was determined by [ 3H]-proline incorporation. RESULTS: As compared with control, the cell number of fibroblasts derived from SHR and WKY were significantly increased to 163.75% and 170.42% respectively after 72 h incubation with FN in the presence of 0.4% FCS from a intial cell density of 1?10 4 cells/mL. DNA synthesis of CFb was markedly promoted by FN [CFb SHR : ( 44?734 ? 8?981 )counts/min vs ( 29?233 ? 6?746 ) counts/min, P