Cells, Cultured ; Endothelial Cells ; drug effects ; metabolism ; Endothelium, Vascular ; cytology ; metabolism ; Gene Expression Profiling ; Humans ; Ouabain ; pharmacology ; Umbilical Veins ; cytology

Cells, Cultured ; Drug Antagonism ; Drug Resistance, Multiple ; drug effects ; HSP70 Heat-Shock Proteins ; pharmacology ; Humans ; K562 Cells ; drug effects ; pathology ; Quercetin ; pharmacology

AIM:To develop the method for analyzing bulleyacinitine A in plasm from mice through the treatment of subcutaneous injection of bulleyacinitine A by liquid chromatography tandem mass spectrometry(LC-MS/MS).METHODS:50 ?L mouse plasma sample added zolpidem as internal standard and pH 9.2 buffer solution was extracted with ethylether,followed by liquid chromatography and mass spectrometric detection.The mobile phase was consisted of methanol-buffer solution(85∶15)(buffer solution consisted of 20 mmol/L ammonium acetate and 0.1% formic solution).The flow rate was 300 ?L per minute.The separation column was C_ 18 column.The electrospray ionization tandem mass spectrometry and the multiple reaction monitoring mode were applied to detecting the bulleyacinitine A in plasma.RESULTS:The assay was linear from 0.1 to 1 000 ng/mL.The recovery rate was more than 80%.CONCLUSION:The method could be used to detect bulleyacinitine A level in mouse plasma,which offers advantages of sensitiveness,specificity and simpleness.

Objective To construct a luciferase reporter plasmid containing interferon regulatory factor 3(IRF-3)human gene promoter and to evaluate promoter activity in human embryonic kidney(HEK)-293 cells.Methods The 1 000 bp fragment was amplified by PCR with human genomic DNA as a template and was directionally cloned into pGL3-basic multiple cloning sites to construct the luciferase repor-ter plasmid pGL3-pIRF-3.Transfection of HEK-293 cells with the promoter-driven lucife-rase construct was performed to induce lucife-rase gene expression and calculate the relative luciferase activity unit(RLU).Promoter sequence of 1 000 bp upstream of transcription initiation site of IRF-3 was analyzed by using Promoter 2.0 Prediction software.Results DNA sequencing and restriction endonuclease analysis verified the successful construction of the plasmid pGL3-pIRF-3.This IRF-3 promoter exhibited a strong promoter activity with an increase of 42.2-fold of RLU in HEK-293 cells when compared with pGL-3 basic vector.The transfection experiment confirmed that the levels of its activation were significantly higher than that in controls in HEK-293 cells.Function analysis of IRF-3 promoter disclosed seve-ral GATA-1 and specific protein 1(Sp1) sites and E2F in minimal promoter region.Conclusion The plasmid pGL3-pIRF-3 promoter is successfully constructed and has a strong basal promoter activity in HEK-293 cells.

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Breast ; pathology ; Breast Neoplasms ; drug therapy ; pathology ; Cyclophosphamide ; therapeutic use ; Diagnosis, Differential ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Lymphoma, B-Cell ; drug therapy ; pathology ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; pathology ; Male ; Middle Aged ; Prednisone ; therapeutic use ; Sarcoma ; pathology ; Spleen ; pathology ; Splenic Neoplasms ; drug therapy ; pathology ; Vincristine ; therapeutic use

Objective To investigate the expression of HSP70 mRNA or MDR1 mRNA by quercetin in leukemia.Methods Methyl thiazoly tetrazolium(MTT) asssy was used to detect the growth suppression of Quercetin on K562 cells.The expression of HSP70 mRNA and MDR1 mRNA were detected in ADM plus Quercetin,Quercetin plus ADM group.The 2 of them were detected in K562/ADM cells in Quercetin plus ADM group.Results 1.Quercetin could suppress the growth of K562 cells.2.The expression of HSP70 mRNA and MDR1 mRNA detected by reverse transcriptase-polymerase chain reaction(RT-PCR) decreased in K562 and K562/ADM cells in Quercetin plus ADM group compared with ADM group and control(P

Objective To investigate the role of contrast-enhanced ultrasound(CEUS) for detection of hepatic artery stenosis(HAS) in recipients following orthotopic liver transplantation(OLT). Methods CEUS was performed in 50 OLT recipients (42 men and 8 women) with abnormal liver function test and/or abnormal findings on color Doppler ultrasound(CDUS). Digital subtraction angiography (DSA), computed tomographic angiography(CTA) or follow-up CDUS was used as the reference standard. The degree (mild,narrowing rate＜50 %; moderate, narrowing rate 50 % ～ 75 %; severe, narrowing rate＞ 75 % ), location and type (single or multiple) of HAS were evaluated. Moderate and severe stenosis were defined as substantial stenosis. Results CTA or DSA depicted substantial HAS in 39 patients, 8 patients with mild HAS or normal HA were depicted on CTA,and the remaining 3 patients were diagnosed as non-substantial HAS on clinical and CDUS follow-up. CEUS depicted substantial HAS in 38 cases. Moreover,CEUS corrected falsepositive findings on CDUS in 9 of 50 cases(18.0% ). The accuracy, sensitivity, specificity, positive predictive value and negative predictive value of CEUS in diagnosing HAS were 90.0% ,92.3% ,81.8% ,94. 7% and 75.0%,respectively. Conclusions CEUS is able to provide comprehensive information including presence,degree,location and type of HAS, which may facilitate the further interventional procedure or surgical treatment.

The DNA structures could be altered or even damaged by exogeous or endogenous factors during cell proliferation. Failure of effective and timely repair will lead to cell cycle arrest or apoptosis. By taking the advantage of the quick proliferation of cancer cells, DNA damage induction, cell cycle arrest and apoptosis promotion have become important strategies for ant-cancer chemotherapy. Previous reports showed that an array of natural compounds inhibit cancer cell proliferation by inducing DNA damage, which have therapeutic potentials for anti-cancer drug research and development.
Animals ; Biological Products ; pharmacology ; therapeutic use ; DNA Damage ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Neoplasms ; drug therapy

Objective To evaluate the prognosis influencing factors of early non-small cell luny cancer (NSCLC) after radiotherapy.Methods 81 early NSCLC patients received definitive radiotherapy and were eligible.Among these patients,60 were diagnosed as squamous cell carcinoma,16 were adenocarcinoma and 5 were diagnosed through imaging instead of pathology.45 patients received conventional radiotherapy,36 patients received three dimensional conformal radiotherapy (3D-CRT),All of them received a total dose of 50-96 Gy with a median dose of 67.8 Gy. Kaplan-Meier survival curves and Cox regression model analysis were applied to evaluate the survival and prognostic factors. Results The median survival time was 34 months.The 1-,3- and 5-year survival rates (OS) were 88.7 ％,41.9 ％,21.8 ％,respectively.Karnofsky performance status≥80,Clinical stage, diameter≤4 cm and the therapeutic effect were associated with improving overall survival.Cox hazards model showed that Karnofsky performance status≥ 80 and diameter≤4 cm were likely to be independent positive prognostic factors. Conclusion Karnofsky performance status and tumor diamater can be used to evaluate the prognosis of early NSCLC after radiotherapy.

Objective To develop a computer-assisted multiangular guiding system for interventional ultrasound.Methods A computer-assisted multiangular guiding system was developed which included a high-accuracy electromagnetic tracker and a passive arm with 5 degree-of-freedom.The electromagnetic tracker was used to track ultrasound probe and needle.The position of B-mode ultrasound image plane and needle were displayed in real-time in the reconstructed 3D ultrasound.The passive arm was used to assist needle delivering.A 10mm ball immersed in a transparent water tank and a formal acoustical prototype was punctured with random angle by using this system.The systematic precision was assessed.Results The whole progress was finished successfully,the ball was touched by the tip of the needle for all the occasions,the error of the system was less than 5mm.Conclusion The computer-assisted multiangular guiding system can get rid of the limitation of angle and make the operation flexible and accurate.