Objective To investigate the antitumor effect of tumor necrosis factor-related apoptosis-inducing ligand(TRAIL) gene transfection mediated by adenovirus into human pancreatic carcinoma cell line Panc-1,and the mechanisms involved in this effect.Methods TRAIL gene was transfected into pancreatic cancer cell line Panc-1 by an adenovirus vector(Ad-TRAIL).Level of TRAIL mRNA expression was determined using RT-PCR,and TRAIL protein synthesis was evaluated with Western blot.Cell-growth activities were determined by MTT assay.The bystander effect was observed by co-culturing the Panc-1 cells with and without the transfected TRAIL gene at different ratios.Apoptosis in pancreatic cancer cells was detected by flow cytometry.Proaspase-8 and procaspase-3 proteins were determined by Western blot.Results The stable overexpression of TRAIL was detected in Panc-1 cells transfected by Ad-TRAIL.Ad-TRAIL significantly inhibited cell viability of Panc-1 cells.Furthermore,co-culture of cancer cells transfected with TRAIL resulted in the nontransfected cell inhibition by bystander effect.Moreover,the percentage of apoptotic cells was significantly higher in the Ad-TRAIL-treatment group compared to the control groups(P

A double targets of high throughput screening model for xanthine oxidase inhibitors and superoxide anion scavengers was established. In the reaction system of xanthine oxidase, WST-1 works as the probe for the ultra oxygen anion generation, and product uric acid works as xanthine oxidase activity indicator. By using SpectraMax M5 continuous spectrum enzyme sign reflectoscope reflector, the changes of these indicators' concentration were observed and the influence factors of this reaction system to establish the high throughput screening model were studied. And the model is confirmed by positive drugs. In the reaction system, the final volume of reaction system is 50 μL and the concentrations of xanthine oxidase is 4 mU x mL(-1), xanthine 250 μmol x L(-1) and WST-1 100 μmol x L(-1), separately. The Z'-factor of model for xanthine oxidase inhibitors is 0.537 4, S/N is 47.519 9; the Z'-factor of model for superoxide anion scavengers is 0.507 4, S/N is 5.388 9. This model for xanthine oxidase inhibitors and superoxide anion scavengers has more common characteristics of the good stability, the fewer reagent types and quantity, the good repeatability, and so on. And it can be widely applied in high-throughput screening research.

A double targets of high throughput screening model for xanthine oxidase inhibitors and superoxide anion scavengers was established. In the reaction system of xanthine oxidase, WST-1 works as the probe for the ultra oxygen anion generation, and product uric acid works as xanthine oxidase activity indicator. By using SpectraMax M5 continuous spectrum enzyme sign reflectoscope reflector, the changes of these indicators' concentration were observed and the influence factors of this reaction system to establish the high throughput screening model were studied. And the model is confirmed by positive drugs. In the reaction system, the final volume of reaction system is 50 μL and the concentrations of xanthine oxidase is 4 mU x mL(-1), xanthine 250 μmol x L(-1) and WST-1 100 μmol x L(-1), separately. The Z'-factor of model for xanthine oxidase inhibitors is 0.537 4, S/N is 47.519 9; the Z'-factor of model for superoxide anion scavengers is 0.507 4, S/N is 5.388 9. This model for xanthine oxidase inhibitors and superoxide anion scavengers has more common characteristics of the good stability, the fewer reagent types and quantity, the good repeatability, and so on. And it can be widely applied in high-throughput screening research.
Enzyme Inhibitors ; pharmacology ; Free Radical Scavengers ; pharmacology ; High-Throughput Screening Assays ; Superoxides ; Uric Acid ; Xanthine ; Xanthine Oxidase ; antagonists & inhibitors

Objective To study the difference of expression of proteins in pancreatic cancer and adjacent fissue.Methods Proteomes of eight pairs of pancreatic ductal adenocarcinoma tissue samples and adjacent tissue samples were obtained by high resolution two-dimensional gel electrophoresis(2-DE).Comprehensive analyses of proteins were focused on total protein spots exhibiting statistical alternations between the two groups.Protein identification was done by peptide mass fingerprinting with tandem mass spectrometry(MS/MS).In addition,Western blotting and immunohistochemistry were performed to verify the expression of certain candidate protein.Results A total of 28 protein spot-features were found to be significantly increased and 17 significantly decreased in tumor tissues.Thirty of these protein spots were identified,which included enzymes,antioxidant proteins,signal transduction proteins,calcium-binding protein,structural proteins,chaperones and others.Western blotting and IHC further validated up-regulated expressions of one candidate protein(annexin II)in tumor tissues.Conclusions The analysis of proteomics with 2-DE on human tissue is a useful method for discovering valuable cancer marker candidates.These differential expressed proteins may serve as biomarkers for early detection and therapeutic targets to pancreatic cancer.

Objective To investigate the expression of Bcl-2,survivin and pancreatic cancer stem cells markers Oct-4 and ABCG2 in pancreatic cancer cells resistance to chemoradiotherapy.and explore its mechanism.Methods Concurrent ehemoradiotherapy was used to obtain pancreatic cancer cells resistant to chemoradiotherapy,the pancreatic cancer cells without chemoradiotherapy treatment were used as control.Western-blot was applied to detect the expression of Bcl-2,survivin,Oct-4,ABCG2.Results The expression of Bcl-2 was 0.7955±0.0326,0.5718±0.0212,0.6137±0.0382 and 0.8733±0.0461,respectively;the expression of survivin protein was 0.8207±0.0490,0.6973±0.0211,0.7967±0.0346 and 0.8013±0.0398,respectively;the expression of Oct-4 protein was 0.8728±0.0177,0.7861±0.0139,0.4794±0.0932 and 0.4216±0.1043,respectively;the expression of ABCG2 protein was 0.7810±0.1370,0.4957±0.1126,0.6102±0.1358 and 0.4670±0.1274,respectively.in resistant pancreatic cancer cells of SW1990,BxPC3,pc3,jf305 cell line.The corresponding values in the control group were 0.4723±0.018,0.2954±0.0103.0.3587±0.0201 and 0.2718±0.0136;0.4717±0.0274,0.3587±0.0113,0.3891±0.0147 and 0.3326±0.0124;0.6053±0.0142,0.4236±0.0086.0.2385±0.0671 and 0.1985±0.0582;0.3156±0.0582.0.2360±0.0423,0.2813±0.0512 and 0.1808±0.a0370.The expression of all the four proteins significantly increased after ehemoradiotherapy(P＜0.05).Conclusions Pancreatic cancer cells resistant to chemoradiotherapy may contain cancer stem cells.

Objective To establish an ideal focal cerebral ischemia reperfusion model in monkeys.Methods Adult healthy rhesus monkeys(Macaca mulatta) 12 cases(male 6 and female 6),aged 4-7 years and weighted 4.8-7.5kg.were used in this study.The middle cerebral artery occlusion/reperfusion(MCAO/R) model was established by inserting a standard micro-balloon catheter intraluminally from the carotid common artery or femoral artery into the proximal segment of the middle cerebral artery(MCA).The regional cerebral blood flow of MCA was occluded by expanding the micro-balloon to cause ischemia,and withdrawing the micro-ballon catheter to reperfuse the MCA.The MCAO/R model was evaluated by angiography,magnetic resonance angiography(MRA),magnetic resonance imaging(MRI),tetrazolium chloride(TTC) staining and neurological behavoral function scores.Results By inserting a micro-balloon catheter intraluminally from the carotid common artery or femoral artery into the MCA,the micro-balloon catheter could be inserted into the MCA to occlude blood flow,and no image of MCA shown on TV screen.In MCA blood flow supplied area,magnetic resonance T1,T2 and DWI showed high signals,TTC staining showed cerebral ischemic infarction,and correspondly the monkeys showed neurological function disorders.This method used a simple operatire procedure had a high successful rate,and could be repeated.Conclusion We showed ideal method to establish the MCAO/R model in monkeys by inserting intraluminally a micro-balloon catheter into the MCA.

This study was conducted to investigate the paclitaxel loaded by hydrazone bonds in poly(ethylene glycol)-poly(caprolactone) micelles (mPEG-PCL-PTX) on proliferation and apoptosis of human lung cancer A549 cells and its possible mechanisms of anti-tumor activity. The cell proliferation was measured with MTT assay. Flow cytometry were used to analyze the cell cycle. The cell apoptosis was analyzed using Hoechst/P staining. The expression levels of apoptotic genes expression in the mitochondrial apoptosis pathway were detected by RT-PCR and Western blotting, respectively. The mPEG-PCL-PTX could inhibit the proliferation of A549 cells and promote the apoptosis. The Bax, caspase-3 protein expression were increased while Bcl-2 protein expression was decreased in A549 cells. Results showed that the polymer containing hydrazone bond is non-toxic in vitro, the mPEG-PCL-PTX micelles can inhibit the proliferation and induce the apoptosis of A549 cells. Key words: paclitaxel; micelle; A549 cell; proliferation; cell cycle; apoptosis
Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Micelles ; Paclitaxel ; pharmacology ; Polyesters ; Polyethylene Glycols ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo explore the protective effect and the mechanism of Puerarin Injection (PI) on myocardial ischemia reperfusion in patients with coronary heart disease (CHD) and angina pectoris (AP).

METHODSSeventy-eight patients with AP planned to receive the PTCA and stenting treatment were randomly divided and single-blindedly into the conventional group and the PI group. Based on the conventional treatment and pre-operational preparation, the PI group was given 200 ml of PI by intravenous dripping once a day, beginning from one week before operation, but to the conventional group, normal saline was given for instead. The condition of AP attack in balloon dilatatory stage of PTCA was observed and change of ST segment of ECG detected by a 12-lead ECG monitor. The blood levels of von Willebrand factor (vWF:Ag), nitric oxide (NO) and endothelin-1 (ET-1) were also observed before and after treatment.

RESULTSAs compared with those in the conventional group, number of patients having AP attack and ST segment change in PTCA process was lessened in the PI group, with blood levels of vWF:Ag and ET-1 obviously lower, and NO content obviously higher than those in the conventional group,

CONCLUSIONSPI could protect the myocardium in 2-3 days after ischemia reperfusion, one of the possible reasons is that PI can simulate the late phase of ischemic preconditioning, which may be related to its effect in lowering plasma vWF:Ag and ET-1, and increasing the serum NO content.

Angina Pectoris ; therapy ; Angioplasty, Balloon, Coronary ; Antigens ; blood ; Endothelin-1 ; blood ; Female ; Humans ; Infusions, Intravenous ; Isoflavones ; therapeutic use ; Male ; Middle Aged ; Myocardial Reperfusion Injury ; prevention & control ; Nitric Oxide ; blood ; Stents ; von Willebrand Factor ; immunology

Objective To observe the sequence of fat deposit and its relationship with insulin resistance in SD rats fed by high fat diet.Methods Normal 8-week-old male SD rats were randomly divided into normal chow (NC,n=40)and high fat diet(HF,n=40)groups.Triglyceride(TG)in serum,liver and muscle were measured;glucose infusion rate(GIR)and the mRNA level of genes related to lipid metabolism in liver and muscle were determined in different stages.GIR was detected by eugiyeemic-hyperinsulinemic clamp for evaluating the insulin sensitivity.Gene expression was determined by real-time PCR.Results(1)As compared with NC group,serum TG was not increased after high fat feeding for4 and 8 weeks,it began to increase after 12 weeks [0.52(0.15-1.00) mmol/L vs O.31(0.09-0.53)retool/L, P0.05)in skeletal muscle.After 8 weeks,the expression of ACC1 in liver in HF group was increased by 20.6%,CPT-1 was decreased by 27.1%(P

Ginsenosides in the decoction of Radix Ginseng, Radix Ginseng with Flos Lonicerae, Radix Polygoni Multiflori or Radix Astragali have been investigated by high performance liquid chromatography (HPLC) and electrospray ionization mass spectrometric method (ESI-MS). Change of the content of ginsenosides was nonlinear in diverse combinative proportion of Radix Ginseng with Flos Lonicerae, while the stripping of ginsenosides was promoted by a small amount of Radix Polygoni Multiflori. In the combinative decoction of Radix Ginseng with Radix Astragali, ginsenosides contents were increased compared to single decoction of Radix Ginseng. Besides, ferric reducing antioxidant power (FRAP) method was developed for determination of the total antioxidative activity of n-butanol and water-soluble extracts from the decoction. The experimental results showed that antioxidative activity was better in the combinative decoction than that in single decoction, and the FRAP values of n-butanol extract were also greater compared with that of water extract.
Antioxidants ; analysis ; chemistry ; Astragalus Plant ; chemistry ; Caprifoliaceae ; chemistry ; Chromatography, High Pressure Liquid ; Drug Combinations ; Drugs, Chinese Herbal ; analysis ; chemistry ; isolation & purification ; Flowers ; chemistry ; Ginsenosides ; analysis ; isolation & purification ; Oxidation-Reduction ; Panax ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Polygonaceae ; chemistry ; Spectrometry, Mass, Electrospray Ionization