Objective To study the effects of neferine on the expression of glutathione S-transferase-pi in vitro and to explore the multi-drug resistance reversing mechanism of neferine.Methods 50% inhibition concentration(IC_(50)) of ADM on K562/A02 was determined by MTT method.The transcription of GST-? gene was detected by semi-quantitative RT-PCR and the expression level of GST-? was determined by Western blot after neferine treatment.Results Neferine remarkably enhanced chemosensitivity to ADM of K562/A02 cells.After neferine treatment in day 1,day 3 and day 5,the relative efficiency of K562/A02 to ADM was 9.6%,41.4% and 10.7%,respectively.Semi-quantitative RT-PCR showed that mRNA transcription of GST-? gene was significantly reduced(P

Objective To investigate behavior,knowledge and attitude of cigarette smoking,and their relationship among different occupational population in Jiangsu Province.Methods Convenient sampling was used to interview 516 staff from medical institution,schools,government offices,public transportations and waiting rooms,restaurants and internet bars.The content of interview included current smoking,second-hand smoking,and related knowledge and attitude.Results Proportion of current smoking in male(51.2%)was higher than that in female(0.3%,P90%),while it's relatively lower in "second-hand smoking and exact related diseases"(

Purpose To study the exposure extent of internal carotid artery siphon (ICAS) before and after removing anterior clinoid process (ACP) using multislice spiral CT (MSCT) simulation, and to improve the tumor resection rate and ensure the operation effect. Materials and Methods MSCT three-dimensional images reconstruction simulating supraorbital keyhole approach of 100 patients (200 sides) were observed, the distance between the crotch of anterior cerebral artery and middle cerebral artery and ICAS before and after removing ACP (exposure extent) was measured. Results In 100 patients (200 sides ACP), the exposure extent before and after removing ACP were (14.3±3.9) mm and (30.5±4.2) mm, respectively on the left side with statistical difference (t=45.278, P<0.001), and were (15.9±3.8) mm and (31.8±3.9) mm, respectively on the right side with statistical difference (t=40.513, P<0.001). The exposure extent increased (16.3±3.6) mm and (15.8±3.9) mm, respectively on the left and right side with no statistical difference (t=0.251, P>0.05). Conclusion MSCT simulating supraorbital keyhole approach in removing ACP can effectively increase the exposure length of ICA, and enlarge the exposure extent of sella region, thus provide reliable imaging information for removing tumor and selecting surgical project in this region.

Objective To observe the anatomy status of the anterior clinoid process (ACP)and the anterior clinoid segment of in-ternal carotid artery (ICA)respectively by multisliced computed tomography (MSCT),and to provide useful imaging information for ACP removal surgery.Methods A total of 100 patients (200 sides)had volume rendering reconstruction of skull.Cranium was removed along cranio-orbital bone in simulation.Then the anatomical structures of the ACP and its surrounding were observed in cephalad direction.The total length,medium length,basic width,medium width of the ACP and the sagittal view curve length of anterior clinoid segment of the ICA from both sides were measured.Results Total length of left ACP was (9.82±2.48)mm,basal width was (9.47±1.88)mm,medium length was (5.03±1.55)mm,medium width was (6.1 9 ±1.75)mm;for right side total length was (10.41±2.1 6)mm,basal width was (9.66 ±2.21)mm,medium length was (5.86 ±2.48)mm,medium width was (6.66±1.5 1)mm.Left anterior clinoid segment of ICA curve length was (6.74±2.25)mm;right was (8.54±3.00)mm.Paired sample t test showed no significant difference in total length,basal width and medium width of ACP in both sides (P >0.05);while the difference in medium length and curve length of the anterior clinoid segment of ICA were statistically significant respectively (P <0.05).Conclusion MSCT can clearly display the vivisection and variation status of the ACP and the anterior clinoid segment of the ICA and can provide useful imaging information for removal of ACP in operation.

Objective To investigate the characteristic and clinical significance of airway inflammation in children with acute asth-ma. Methods Underwent sputum induction and sputum induction in children (n=34) with acute asthma was repented in recovered children ( n = 24).Induced sputum were also taken from 15 healthy children as controls.Total and differential cell counts were per-formed. Interleukin(IL)-8、IL-6 were measured,The relationship between inflammatory cells and IL-8、 IL-6、peak expiratory (PEF) were analyzed.Results The inflammatory cell infiltrate was mixed including eosmophilic granulocytes, neutrophils, and macrophages. They decreased significantly, but eosinophilic granulocyte remained a higher percentage compared with healthy subjects. There was low-er percentage of lymphocytes at acute exacerhation.Eosinophilic granulocytes were correlated with the degree of airflow obstruction. Levels of IL-8、IL-6 were elevated during the acute exacerbation and decreased at resolution.IL-8 was correlated significantly with neutrophils at acute exacerhation and resolution.IL-6 was correlated signifficantly with eosinophilic granulocytes at acute exacerbation. Conclusions Airway inflammation is chataeterized by infiltration of eosinophils, neutrophils and macrophages.IL-8、IL-6 possibly is the important cytokines of airway inflammation in children with acute asthma. Increased eosinophils in induced sputum correlates with asthma severity. Therapy to the cytokines may have potential values.J Appl Clin pediatr,2004,19(12): 1023-1025

OBJECTIVE: To determine the effect of neferine (Nef) combined with mdr-1shRNA on the expression of mdr/P-gp in K562/A02 cell line. METHODS: MTT assay was used to observe the cell proliferation. The expression level of P-gp was determined by Western blot and the transcription of mdr-1 gene was detected by semi-quantitative RT-PCR. RESULTS: After K562/A02 cells were treated by Nef or mdr-1shRNA alone or both for 24 h, the proliferation of K562/A02 cells was significantly higher in the Nef combined with mdr-1shRNA treatment group than that of Nef or mdr-1shRNA alone group (P<0.01).The expression of mdr-1/P-gp in the Nef with mdr-1 shRNA group was significantly lower than that of Nef or mdr-1shRNA alone group. CONCLUSION Nef enhances the inhibition of mdr-1shRNA expression vector on K562/A02 cell proliferation and on P-gp protein to effectively reverse multidrug resistance induced by mdr-1 gene encoding P-gp.
ATP Binding Cassette Transporter, Subfamily B ; ATP Binding Cassette Transporter, Subfamily B, Member 1 ; genetics ; metabolism ; Benzylisoquinolines ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Humans ; K562 Cells ; RNA, Small Interfering ; genetics ; pharmacology

OBJECTIVETo construct a short hairpin RNA (shRNA) eukaryotic expression vector specific to MDR1 gene in multidrug resistance (MDR) human leukemia cell line K562/A02 to observe its silencing effect on MDR1 and P-glycprotein (P-gp) expression.

METHODSThe shRNA expression vector was constructed by gene recombination, then transfected into the cultured K562/A02 cells. The transcription of MDR1 gene was detected by semi-quantitative RT-PCR and the expression level of P-gp was determined by Western blot. 50% inhibition concentration (IC50) of ADM in K562/A02 cells was determined by MTT method. The intracellular doxorubicin (ADM) concentration was determined by HPLC.

RESULTSThe introduction of pEGFP-C1/U6/MDR1-A or pEGFP-C1/U6/MDR1-B expression vector was shown to efficiently and specifically inhibit the expression of P-gp according to results of Western blot, with an inhibitory rate of 50.67%. Semi-quantitative RT-PCR showed that mRNA transcription of MDR1 gene was reduced by (48.2 +/- 2.5)%. On the contrast, the control plasmid did not exhibit inhibitory effect on the protein expression and mRNA transcription of MDR1. The relative efficiency of K562/A02 to ADM was 40.8% or 62.4%, respectively, and the intracellular accumulation of ADM increased after shRNA treatment.

CONCLUSIONThe shRNA expression vector targeting MDR1 gene showed dramatic inhibition on RNA transcription and protein expression. It could effectively restore the sensitivity of K562/A02 cells to conventional chemotherapeutic agents.

ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Antibiotics, Antineoplastic ; metabolism ; pharmacology ; Blotting, Western ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Doxorubicin ; metabolism ; pharmacology ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Gene Silencing ; Genetic Vectors ; genetics ; Humans ; K562 Cells ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

Actins ; metabolism ; Adult ; Antigens, CD34 ; metabolism ; Carcinoma, Renal Cell ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Glomus Tumor ; metabolism ; pathology ; Hemangiopericytoma ; metabolism ; pathology ; Humans ; Hypertension ; etiology ; Juxtaglomerular Apparatus ; metabolism ; pathology ; surgery ; ultrastructure ; Kidney Neoplasms ; complications ; metabolism ; pathology ; surgery ; ultrastructure ; Nephrectomy ; Wilms Tumor ; metabolism ; pathology

Glucose ; Humans ; Hypothalamo-Hypophyseal System ; Lycium ; Pituitary-Adrenal System ; Resistance Training

Antioxidant activity of bronchial epithelial cells (BECs) plays an essential role in preventing the airway epithelium integrity from damage in structure and function. Integrin expressed by BECs is the receptor of extracellular matrix such as fibronectin (Fn), and it is involved in modulation of proliferation, differentiation and metabolism of the cells. In order to test the hypothesis that integrin-ligand binding reaction supports the ability of cells to withstand oxidant attack, the present study evaluated the antioxidant activity of primary cultured rabbit BECs treated with fibronectin or its sequence Arg-Gly-Asp (RGD peptide), by determining changes in the activity of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT) and in the level of glutathione (GSH). The results are as follows: (1) Fn (10 μg/ml) increased significantly the activity unit of GSH-Px (P<0.05, n=5), which was inhibited by calmodulin-inhibitor W7 　(10－5 mol/L) (P<0.05). Both Fn (5～20 μg/ml) and RGD (15～60 μg/ml) showed a dose-dependent upregulatory effect (respectively r=0.93 and r=0.73). (2) Treatment with Fn increased SOD activity (P<0.01,　n=7), which was abolished by W7 　(P<0.01). (3) Catalase activity was also stimulated by Fn (P<0.05, n=6) and reversed by W7 　(P<0.01). (4) A dose-dependent increase of GSH level was observed in both Fn (r=0.82) and RGD treatment (r=0.84). The data suggest that the binding of integrin with extracellular matrix can upregulate activity of antioxidant enzymes, and increase the content of GSH and improve the ability of BECs to resist oxidant injury.