Objective To develop an albumin aptamer that may potentially serve as a selective ligand for albumin removal from experimental samples.Methods A single-stranded 59nt DNA library that contains 21 random oligo nucleotides was synthesized in vitro.An albumin aptamer A6 was developed by SELEX technique using bovine serum albumin (BSA) as target.The enrichment of aptamer and evaluation of its binding properties were monitored by flow cytometry.The secondary structure of A6 was predicted by MFord software.Results The aptamer A6 strongly bound to BSA with a Kd of 77.4 nmol/L,and had minimal cross reactivity with control proteins including ovalbu min,IgG,and trypsin.Conclusions Aptamer A6 may be a potential tool in albumin removal.

Objective To explore whether quercetin-loaded PEG-PE micelles(M-Q) can synergize the growth-in-hibitory activity of adriamycin prepared ( ADR) by reversing the drug resistance of MCF-7 ADRr breast cancer cells in vitro.Methods M-Q was prepared by adding saline to lipid film containing quercetin and PEG-PE.The size of M-Q was characterized by dynamic light scattering ( DLS) .The inhibition of MCF-7 ADRr cells was evaluated by MTS assay after incubation with M-Q and ADR.Results The incorporation efficiency of quercetin by the micelles was above 74%.The average size of M-Q was 11.11 nm.Compared with the quercetin dissolved in ethanol , M-Q more effectively reversed the drug resistance of MCF-7 ADRr cells in vitro.Conclusions PEG-PE micelles may potentially deliver quercetin to cancer cells for reversal of drug resistance .

Objective To investigate the dynamic changes of osteopontin (OPN) in the guinea pig model of cholesterol gallstone disease.Methods Forty-four guinea pigs were randomly assigned to cholesterol gallstone group and control group.The animals were sequentially killed on Day 7,14,28,42,56 and 70 after operation.The expressions of OPN mRNA in gallbladder and liver tissues were detected by real-time PCR.The changes of OPN,mucin,α1-acid glycoprotein (AAP) and apolipoprotein A1 (APOA1) in bile and blood plasma were detected by ELISA kits.Results The expression of OPN mRNA in gallbladder and liver tissues increased gradually and reached the peak in the 6th week,and then decreased.The increased expression of OPN in bile in the gallstone group started from the 1st week and reached the peak in the 6th week (P ＜ 0.05),which gradually decreased to the baseline in the 10th week (P ＞ 0.05).The expressions of OPN in bile and blood demonstrated similar trends,while the peak time in blood samples was much earlier (4th week).The changes of APOA1 in bile and blood were similar to OPN,although there was no advanced peak value in blood.The levels of mucin and AAP in bile and blood increased after operation,and were kept at high level throughout the study.Conclusions OPN is involved in the whole process of cholesterol gallstone formation,which may be associated with other nucleation-active factors.

Animals ; Antigens, Neoplasm ; chemistry ; immunology ; Cancer Vaccines ; chemistry ; therapeutic use ; Cell Line, Tumor ; CpG Islands ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; metabolism ; Humans ; In Vitro Techniques ; Lactic Acid ; chemistry ; Leukemia ; immunology ; therapy ; Lymphocytes ; cytology ; immunology ; Nanoparticles ; chemistry ; Peptides ; chemistry ; immunology ; therapeutic use ; Polyglycolic Acid ; chemistry ; Polylactic Acid-Polyglycolic Acid Copolymer ; WT1 Proteins ; chemistry ; immunology

Objective To investigate the role of osteopontin (OPN) in the pathogenesis ot cholesterol gallstone formation in bile.Methods The nucleation time of OPN in model bile and human gallbladder bile was studied by the nucleation time assay,the effect of OPN on cholesterol crystal growth in model bile was examined by the cholesterol crystal growth assay.The effect of OPN on vesicle was detected by the transmission electron microscopy in model bile and gallbladder bile; then the content of OPN and calcium were detected via the commercial kits in human bile.Results Osteopontin prolonged nucleation time in a dose dependent manner in model bile and human bile,and this effect was correlated with calcium.Compared with control group,the nucleation times were prolonged by 1.50and 1.93 times in lithogenic bile at the concentration of osteopontin 50 μg/ml and 100 μg/ml (P＜0.01),respectively. Nucleation time were prolonged by 1.17 and 1.33 times in normal bile (P＜0.01) and by 1.29 and 1.48 times in model bile (P＜0.01),respectively.The rate of cholesterol crystals growth was not influenced by calcium ions,but inhibited by osteopontin in a dose dependent manner in the model bile.Furthermore,the formation,aggregation and fusion of vesicles were delayed by osteopontin in bile samples as indicated by the transmission electron microscopy.The concentration of osteopontin [(0.53± 0.08) mg/ml vs. (0.65 ± 0.14) mg/ml,P＜0.05] and the calcium ions [ (0.71 ± 0.17) mmol/L vs. ( 0.84 ± 0.08 ) mmol/L,P ＜ 0.05 ] were lower in lithogenic bile than in control.Conclusions Osteopontin can inhibit the cholesterol gallstone formation in model and human gallbladder bile as the anti nucleating factor.

Objective To investigate the role of osteopontin (OPN) in cholesterol gallstone formation.MethodsGallbladder bile was obtained from patients with cholelithiasis (n=36,the experimental group) and from donors of liver transplantation (n=19,the control group).OPN,calcium ion and lipid were analysed quantitively.The nucleating role of OPN in bile was evaluated using nucleating time (NT) approach.ResultsOPN inhibited cholesterol nucleation in a dose dependent manner.OPN (50 μg/ml and 100 μg/ml) prolonged NT by 48.90％ (91.51％) and 17.07％ (32.93％) in lithogenic and control bile,respectively.OPN (100 μg/ml) also inhibited the nucleating effect induced by calcium ion.Furthermore,a combination of OPN (50 μg/ml) and calcium prolonged NT by 75.78％ and 33.96％ in lithogenic and control bile,respectively.A combination of OPN (100 μg/ml) and calcium prolonged NT by 125.9％ and 62.26％ in the 2 groups.The contents of osteopontin and calcium were significantly lower in lithogenic bile than control bile (P＜0.05).On the other hand,the cholesterol saturation index and the contents of cholesterol,phospholipid and bile acid were significantly higher (P＜0.05).ConclusionsOPN inhibited cholesterol gallstone formation.It may be involved in the pathogenesis of cholelithiasis.

Objective To observe the effect of combined treatment with rhTRAIL(recombinant human TNF-related apoptosis inducing ligand) and selective Cox-2 inhibitor Celecoxib on gallbladder carcinoma in vitro and to explore the possible mechanism of the effect. Methods Western blot analysis was used to detect the expression of c-FLIP and death receptors after treatment by Celecoxib. Apoptosis of gallbladder cell line SGC-996 after the combined treatment with Celecoxib and rhTRAIL was detected in three ways: (1) phase microscopy of the cells, (2) detection of effector caspase-3 and caspase-7 activity, and (3) determination of the proportion of apoptotic cells labeled by Annexin V-PI flow cytometric analysis using CELLQUEST software. Results Celecoxib down-regulated the expression of c-FLIPs and up-regulated the expression of DR5 in a dose- and time-dependent mode on cell line SGC-996. Apoptotic levels in the combined treatment group in cell line SGC-996 were significantly higher than those in the single drug treatment group and control group. Conclusion Celecoxib markedly sensitized rhTRAIL-induced apoptosis through the down-regulation of c-FLIPs and up-regulation of DR5 in gallbladder carcinoma cell line SGC-996.

ObjectiveTo investigate the effect of α-Zearalanol(α-ZAL) on lipometabolism and hemorheology in ovariectomized(OVX) hyperlipidemia rabbits.Methods44 adult virgin female rabbits were divided into 5 groups,group A: normal control;group B: sham+CHO;group C: OVX+CHO;group D: OVX+CHO+17βE_2;group E: OVX+CHO+α-ZAL.Cholesterol(CHO) was fed to rabbits for 12 weeks.Before and after feeding CHO,the serum lipid(TC,TG,LDL-C,HDL-C) were measured;Blood viscosity,plasma viscosity,aggregation index of RBC(AIRC) and fibrinogen were also assayed respectively.ResultsThe serum levels of TC,TG and LDL-C in group B and E were significantly decreased compared with those in group C(P<0.05);the level of blood viscosity,plasma viscosity and AIRC platelet aggregation rate in group D and E were also significantly decreased compared with those in group C(P<0.05).Conclusionα-ZAL can improve vascular function through the adjustment of lipometabolism and hemorheology.

ObjectiveTo investigate the effect of Jiuqiang Naoliqing(JNQ) on the TXA2 and PGI2 level in spontaneous hypertension rat (SHR) plasma.MethodsThe plasma was separated after the SHR and Wistar rats were treated with JNQ at the dose of 0.133g/kg,0.265g/kg,0.530g/kg and 1% carboxymethyl cellulose respectively for 5 weeks. The level of TXB2 and 6 keto PGF1α ,stable metabolin of TXA2 and PGI 2,in SHR plasma was tested by radioimmunoassay.ResultsThe level of TXB2 and the ratio of TXB2/6 keto PGF1α (T/P) in SHR plasma increased significantly(P<0.01),and there was no significant difference in the concentration of 6 keto PGF1α between Wistar rats and SHR plasma(P>0.05). JNQ could increase the generation of 6 keto PGF1α and decrease the level of TXB2 and T/P in SHR plasma after treated with different dosages for 5 weeks.ConclusionJNQ may improve the balance between TXA2 and PGI2 in SHR plasma.

AIM:The objective of this study was to observe the effects of AP-1 element on endothelin-1(EDN1)gene transcription in homocysteine(Hcy)-induced human umbilical vein endothelial cells(HUVECs).METHODS:The redox level in Hcy-induced HUVECs was determined by using the fluorescent product DCF.The influences of Hcy on expressions of EDN1 mRNA and protein of c-jun/AP-1 in HUVECs were measured by the methods of RT-PCR and Western blotting,respectively.The EDN1 secretion level of HUVECs induced by Hcy was determined by enzymatic immunoassay.The transient transfection assay was performed and luciferase activity of transcriptional factor AP-1 reporter gene induced by Hcy was detected.RESULTS:The Hcy significantly increased EDN1 secretion,EDN1 mRNA expression and oxidative stress in HUVECs.Hcy remarkably induced the expression of pGL3-EDN1-AP-1(115/+135)luciferase reporter gene plasmid.However,basic expression of mutation of pGL3-EDN1-AP-1(-115/+135)luciferase reporter gene plasmid was downregulated markedly in HUVECs induced by Hcy.CONCLUSION:Redox-active and release of ROS are changed by Hcy.Activated AP-1 element plays an important role in EDN1gene transcription in HUVECs induced by Hcy.