Objective To investigate the role of intracellular protein tyrosine kinase (PTK) activation in vascular smooth muscle cells (VSMCs) migration mediated by angiotensin Ⅱ (AngⅡ). Methods Intracellular PTK activities were investigated by means of ?- 32 P incorperation in cultured rat VSMCs stimulated by AngⅡ. The migrated VSMCs were enumerated in a modified Boyden Chamber. Results Without any specific stimulus, only a few migrated cells were detected. However, the number of migrated cells were increased, under the condition of AngⅡ at the concentration of 1?10 -10 , 1?10 -9 , 1?10 -8 , 1?10 -7 and 1?10 -6 mol/L, by 3.7, 4.6, 5.9, 6.6 and 6.3 times respectively, compared with the control group. Intracellular PTK activities were also increased by 1.0,1.7,2.4,3.2 and 2.4 times compared with the control group, under the condition of AngⅡ at the concentration of 1?10 -10 , 1?10 -9 , 1?10 -8 , 1?10 -7 and 1?10 -6 mol/L, respectively. Statistical analysis showed that there was a significant positive correlation between AngⅡ-induced VSMCs migration and intracellular PTK activation. Correspondingly, PTK inhibitor Genistein significantly reduced AngⅡ-induced VSMCs movement ( P

Objective To investigate the effect of antisense oligodeoxynueleotide targeting sur-vivin (survivin ASODN) on hilar cholangioearcinoma cell line FRH-0201 depressing the expression of survivin. Methods Survivin ASODN was transfected into hilar eholangiocarcinoma cell line FRH-0201by liposome. Morphologieal changes were observed by inverted phase contrast microscope. RT-PCR and Western blot methods were performed to detect the expressions of survivin mRNA and protein re-spectively. The changes in cell apoptosis were detected by flow cytometry. Results The expression of survivin mRNA and protein was significantly decreased in survivin ASODN group than that in the con-trol group(mRNA: 0.51-t-0. 03 vs 0. 82-t-0.02,P%0. 05～protein: 1.82-t-0.16 vs 3. 08--t-_0. 27, P-Q 0.05). The morphologieal apoptotic changes were observed and the apoptosis rate was increased (11.50+1.49% vs 0.39-+-0.08～, P%0.05). Conclusion Survivin AS()DN can induce hilar cholan-gioeareinoma cell line FRH-0201 into apoptosis by decreasing the expression of survivin.

Adult ; Arrhythmogenic Right Ventricular Dysplasia ; physiopathology ; Electrocardiography ; Humans ; Male ; Middle Aged

Objective To study the low dose hyper-radiosensitivity in human lung cancer cell line A549,and its possible mechanisms.Methods Exponentially growing A549 cells were irradiated with 60Co γ-rays at doses of 0-2 Gy.Together with flow cytometry for precise cell sorting,cell survival fraction was measured by mean of conventional colony-formation assay.ATM1981 Ser-P protein expression was examined by Western blot.Apoptosis was identified by Hoechst 33258 fluorescent staining,and Annexin V-FITC and propidium iodide staining flow cytometry.Cell cycle distribution was observed by flow cytometry.Results There was an excessive cell killing per unit dose when the doses were below about 0.3 Gy,and the cells exhibited more resistant response at the doses between 0.3 and 0.5 Gy,the cell survival fraction was decreased as the doses over 0.5 Gy.The expression of ATM1981Ser-P protein was first observed at 0.2 Gy,followed by an increase over 0.2 Gy,and reached the peak at 0.5 Gy(compared with 0.2 Gy group,t=7.96,P＜0.05),with no further increase as the doses at 1.0 and 2.0 Gy(t=0.69,0.55,P＞0.05).24 hours after irradiation,part cells presented the characteristic morpholos4cal change of apoptosis,and the apoptosis curve was coincident with the dose-survival curve.Compared with the control group,the cell cycle had no change post-irradiation to 0.1 and 0.2 Gy.G2/M phase arrest was manifested at 6 and 12 hours post-irradiation to 0.3,0.4 and 0.5 Gy(t=2.87,2.88,4.92 and 3.70,3.12,8.11,P＜0.05),and the ratio of G2/M phase was decreased at 24 hours post-irradiation(t=3.87,4.77,3.01,P＜0.05).Conclusions A549 cells displays the phenomenon of hyper-radiosensitivity(HRS)/induced radioresistance(IRR).The model of cell death induced by low dose irradiation is mainly apoptosis.The activity of ATM and cell cycle change might play an important role in HRS/IRR.

Objective To evaluate the clinical efficacy of compound matrine injection combined with fentanyl transdermal patch in the treatment of elderly patients with advanced gastric cancer.Methods A total of 100 elderly patients with advanced gastric cancer were enrolled in this study.All patients were randomly divided into the observation group (n =50) given compound matrine injection plus Fentanyl transdermal patch and the control group given fentanyl transdermal patch alone(n=50).The analgesic efficacy,quality of life and adverse reaction were evaluated and compared between the two groups.Results The analgesic efficacy was significantly higher in the observation group[76％(38/50)]than in the control group[(44％),(x2=4.46,P＜0.05)].Before treatment,the life quality scores were similar between two groups.After treatment,the life quality score of the two groups was significantly improved as compared with pre-treatment.In addition,life quality score was significantly higher in the observation group than in control group(t=8.61,P＜0.05).After treatment,only mild adverse events were observed in two groups,with no significant difference in adverse events between the two groups(x2 =0.00,P＞0.05).Conclusions Compound matrine injection plus Fentanyl transdermal patch are safe and effective in the treatment of carcinoma pain in elderly patients with advanced gastric cancer.

The low dose hyper-radiosensitivity (HRS) in human lung cancer cell line A549 was investigated, the changes of ATM kinase, cell cycle and apoptosis of cells at different doses of radiation were observed, and the possible mechanisms were discussed. A549 cells in logarithmic growth phase were irradiated with (60)Co gamma-rays at doses of 0-2 Gy. Together with flow cytometry for precise cell sorting, cell survival fraction was measured by means of conventional colony-formation assay. The expression of ATM1981Ser-P protein was examined by Western blot 1 h after radiation. Apoptosis was detected by Hoechst 33258 fluorescent staining, and Annexin V-FITC/PI staining flow cytometry 24 h after radiation. Cell cycle distribution was observed by flow cytometry 6, 12 and 24 h after radiation. The results showed that the expression of ATM1981Ser-P protein was observed at 0.2 Gy, followed by an increase at >0.2 Gy, and reached the peak at 0.5 Gy, with little further increase as the dose exceeded 0.5 Gy. Twenty-four h after radiation, partial cells presented the characteristic morphological changes of apoptosis, and the cell apoptosis curve was coincident with the survival curve. As compared with control group, the cell cycle almost had no changes after exposure to 0.1 and 0.2 Gy radiation (P>0.05). After exposure to 0.3, 0.4 and 0.5 Gy radiation, G(2)/M phase arrest occurred 6 and 12 h after radiation (P<0.05), and the ratio of G(2)/M phase cells was decreased 24 h after radiation (P<0.05). It was concluded that A549 cells displayed the phenomenon of HRS/IRR. The mode of cell death was mainly apoptosis. The activity of ATM and cell cycle change may take an important role in HRS/IRR.
Cell Cycle Proteins/genetics ; Cell Cycle Proteins/metabolism ; Cell Cycle Proteins/physiology ; Cell Line, Tumor ; DNA-Binding Proteins/antagonists & inhibitors ; DNA-Binding Proteins/metabolism ; DNA-Binding Proteins/*physiology ; Dose-Response Relationship, Radiation ; Lung Neoplasms/*pathology ; Protein-Serine-Threonine Kinases/*metabolism ; Radiation Dosage ; Radiation Tolerance/*physiology ; Tumor Suppressor Proteins/metabolism

Polysaccharides with multiple biological activities are usually considered as one of the major bioactive compounds in Chinese medicines (CMs). At present, the development of drug and functional foods related to polysaccharides have attracted a great deal of attention due to their great potential effects and diverse action mechanisms. However, quality control of polysaccharides is the bottleneck and a challenge due to their complexity and chemical diversity. Actually, the bioactivities of polysaccharides are closely related to their molecular structures. In order to ensure their safety and efficacy, the development of novel approaches based on the molecular structures for the improvement of quality control of polysaccharides is significantly important. Therefore, in this article, the relationship between biological activities and chemical structures, as well as the action mechanisms of polysaccharides from CMs were summarized first. Furthermore, saccharide mapping, a novel strategy for quality control of bioactive polysaccharides from CMs, was introduced and the application and perspectives were also discussed.
Carbohydrate Sequence ; Drugs, Chinese Herbal ; chemistry ; Molecular Sequence Data ; Polysaccharides ; chemistry ; Quality Control

Objective To explore the effects of electroacupuncture at Shenting (DU24) and Baihui (DU20) on learning and memory ability in the cerebral ischemia-reperfusion rats and its possible mechanism. Methods 45 male Sprague-Dawley rats were randomly divided into sham group (n=15), ischemia group (n=15) and electroacupuncture group (n=15). The latter 2 groups were modeled as focal cerebral ischemia-reperfusion injury, and the electroacupuncture group received electroacupuncture at Shenting (DU24) and Baihui (DU20) for 7 days. They were tested with Morris Water Maze, observed with Nissl's staining. The protein expression of RhoA was detected with Western blotting. Results The learning and memory ability improved in the electroacupuncture group (P<0.05), the injury of the neurons reduced (P<0.05) and the expression of RhoA in hippocampus decreased compared with the ischemia group (P<0.05). Conclusion Electroacupuncture could ameliorate the learning and memory ability in ischemia- reperfusion rats, which may relate with the inhibition of the expression of RhoA in hippocampus.

Background and purpose:Drug-resistance is the main obstacle in terms of efficacy of chemotherapy for leukemia, RNA interference(RNAi) strategy possesses the characteristics of specilization, high-efficiency and low-toxicity, and can effectively and specifically inhibit the overexpression of given gene. This study was designed to investigate the effect of small interfering RNA (siRNA) on expression of mdr1 gene and drug-resistance in multidrug-resistant human leukemia K562/ADM cell.Methods:Human multidrug-resistant leukemia cell line K562/ADM over-expressing mdr1 gene was used as the target cells, Two siRNAs (si-mdr1-1 and si-mdr1-2) targeted mdr1 gene were chemically synthesized and transfected into K562/ADM cells. Expression of mdr1 mRNA was determined by RT-PCR, P-glycoprotein (P-gp) expression was measured using flow cytometry (FCM), and the sensitivity of K562/ADM cells to adriamycin was assessed with a MTT colorimetric assay.Results:Two siRNAs (si-mdr1-1 and si-mdr1-2) specially designed in this study could markedly down-regulate the expression of mdr1 mRNA and its product P-gp in K562/ADM cells. After cells transfected with si-mdr1-1 or si-mdr1-2 for 24h and 48h, the inhibition of mdr1 mRNA expression in the cells for si-mdr1-1 was 55.5% and 22.5%; and for si-mdr1-2, 16.0% and 57.6%, respectively. Treated with siRNA for 72h, the expression intensity of P-gp in the two transfected cell lines decreased 74% and 85%, respectively. Both si-mdr1-1 and si-mdr1-2 significantly enhanced the sensitivity of K562/ADM cells to adriamycin and reversed their drug-resistance, the reversal efficiency was 2.52-folds and 1.96-folds, respectively.Conclusions:The siRNA could effectively and specifically silence the expression of mdr1 gene and overcome the drug-resistance mediated by P-gp in K562/ADM cells.