Aim This study was designed to investigate the inhibition of matrine on U251 glioma cell line and its mechanism. Methods MTT was used to measure the levels of the proliferation of U251 cultured with matrine in different concentrations.The effects of matrine on cell cycle of U251 were observed by FCM. The expression of proto oncogenes c myc was measured by RT PCR. Results The proliferation of U251 was obviously inhibited by matrine in a dose dependent manner. The inhibitory rate was (53 7?6 0)%,when cultured with matrine at 0 10 g?L -1 . The outcome of FCM showed that the proportion of G 0/G 1 phase cells were decreased. The proportion of S phase cells were reduced obviously,when cultured with matrine at 0 10 g?L -1 in 3 days.The outcome of RT PCR showed that the expression of proto onco gene C myc was notably decreased, when the dose of matrine was increased. Conclusion Matrine can inhibit the proliferation of U251 and inhibit the expression of proto onco gene C myc.

Objective To observe the effect of consciousness-restoring obstruction-clearing needing technique combined with swallowing function training on pseudobulbar paralysis after stroke.Methods 80 stroke patients with pseudobulbar paralysis were randomly divided into the trial group and control group with 40 cases in each group.The patients of the trial group were treated with consciousness-restoring obstruction-clearing needing technique combined with swallowing function training and routine medicine,those of the control group were treated only with routine medicine.Results After treatment,the whole effective rate of the trial group was 92.5%,that of the control group was 60.0%,there was a significant difference between two groups(P<0.05).Conclusion The therapeutic effect of consciousness-restoring obstruction-clearing needing technique combined with swallowing function training and routine medicine on pscudobulbar paralysis after stroke is superior to simply routine medicine.

Objective To explore the effect of erythropoietin(EPO) on apoptosis of human renal tubular(HK-2) cells induced by postasphyxial-serum of neonates.Methods HK-2 cells were used as target cells.The experiment was divided into 4 groups,control group(n=8):HK-2 cells were maintained in standard medium;asphyxia group(n=8):HK-2 cells were treated with serum obtained from neonates with asphyxia.Each culture medium replaced with 200 mL/L suffocation DMEM/F12 serum culture medium;EPO pretreatment group(n=8):HK-2 cells were pretreated 24 h with serum contains 5?104 IU/L EPO,and then deal as asphyxia group;EPO and 5-hydroxydecanoic acid sodium salt(5-HD) pretreatment group(n=8): HK-2 cells were pretreated 24 h with serum contains 5?104 IU/L EPO and 500 ?mol/L 5-HD,and then deal as asphyxia group.All cells were cultured at 37 ℃ in humidified atmosphere with 50 mL/L CO2 for 24 h.The apoptosis rate of HK-2 cells was detected by flow cytometer.The expressions of Caspase-3 and X-linked inlnibitor of apoptosis protein(XIAP) of HK-2 cells were detected by using immunohistochemical method.Results Compared with control group,after stimulated with postasphyxial-serum,the apoptosis rate and the expression of Caspase-3 of HK-2 cells were significantly increased(Pa

Objective To investigate the role and intracellular signal transduction mechanism in the injury of renal tubular cells induced by postasphyxial-serum in neonate.Methods Human renal proximal tubular cell(HK-2 cell) was used as target cell. The experiment was designed as:control group, asphyxia group ,and pyrrolidine dithiocarbamate (PDTC)blocking group. The attacking concentration of serum was 20%, and the apoptosis rate of HK-2 cells was detected by flow cytometer.Results Compared with controls[(13.3?1.70)%],after being stimulated with postasphyxial-serum, the apoptosis rate of HK-2 cells of asphyxia group [(46.73?3.68)%] and PDTC blocking group [(31.19?2.79)%]were significantly increased(P

Objective To investigate the role of postasphyxial-serum of neonate in inducing injury of human renal proximal tubular cells(HK-2 cells).Methods HK-2 cells were used as target cell.The neonatal different concentration postasphyxial-serum of 1,3,7 days after asphyxia were used as attacking means.The experimental groups were divided into 15 groups:the 2.5%,5.0%,10.0%,(20.0%) attacking concertration groups of 1,3,7 day after asphyxia and control group of each concertration.The culture medium and concertration of the control group and the experimental groups were the same.The changes of morphology were observed under inverted microscope,the cell viability was measured by 3-(4,5-dimethyl-2-thiazoly1)-2,5-diphenyl-2H-tetrazolium bromide(MTT) method and the leakage rate of lactate dehydrogenase(LDH) was determined by biochemical methods.Results Compared with control group,the changes in morphology of HK-2 were most serious and obvious,the cell viability were obviously decreased(all P

Objective To explore the effect of postasphyxial-serum in neonate on the expressions of Bcl-2-antagonist of cell death(BAD)and Bcl-2-associated X protein(BAX)in renal tubular cells(HK-2).Methods HK-2 cells were used as target cells.The experiment were divided into control group,asphyxia group and pyrrolodine dithiocarbamate(PDTC)blocking group.Control group:DMEM culture fluid was not contained asphyxia blood serum in every group;asphxia group:DMEM culture fluid contained 20 mL/L asphyxia blood serum in every group;PDTC blocking group:DMEM culture fluid contained 20 mL/L asphyxia blood serum and 40 ?mol/L PDTC in every group.The expressions of both BAD and BAX on cytoplast were detected by immunohistochemical method.Results Calculated Points according to HSCORE,compared with controls group[(1.97?0.26)and(1.77?0.11)],after stimulated with postasphyxial-serum,the expressions of both BAD and BAX of HK-2 cells of asphyxia group[(2.73?0.20)and(2.44?0.13)] and PDTC blocking group[(2.38?0.13)and(2.17?0.08)] significantly increased[F(BAD)=28.61,F(BAX)=15.51 Pa

Objective To explore the effect of postasphyxial-serum in neonate on expression of serine protease Omi/HtrA2 in renal tubular cells(HK-2).Methods Human renal proximal tubular cell line HK-2 cell was used as target cell.The cultural cells in orifice were divided into control group and asphyxia-serum attacking group.Blood was cowected from asphyxia newborns by means of femoral venous puncture,then the serum was garthered,anticoagulated by liquemie,3 000 r/min centrifuged 20 min,abstracted serum,thermostatic waterbathed the serum at 56 ℃,so that to inactivate addiment,filtered germ by micropore filte,the attacking concentrtion of serum was 200 mL/L,the cells of the asphyxia-serum attacking group were attacked by asphyxia-serum,and the cells of control group were cultivated with normal nutritive medium when the cells was needed.After 24 hours,the cells were tixed,then the expression of Omi/HtrA2 in cytoplast was detected by the use of immunohistochemical method.Results Omi/HtrA2 was inaurate or yellow brown and localized to the cytoplast.The rate of the cell expressed Omi/HtrA2 was(9.0?2.5)% in control group,after stimulated with postasphyxial-serum,in asphyxia group the rate of the cell expressed Omi/HtrA2 was(25.15?3.5)%,there was significant difference between 2 groups(t=-15.322 P

Animals ; HSP70 Heat-Shock Proteins ; metabolism ; Male ; Physical Conditioning, Animal ; Physical Endurance ; physiology ; Quadriceps Muscle ; metabolism ; physiology ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; metabolism

BACKGROUND:Studies have shown that bisphosphonates inhibit osteoclast resorption, but whether cathepsin K, a key cytokine of bone resorption, plays an effect has rarely been reported.
OBJECTIVE:To study the effect of bisphosphonate on capthesin K and bone resorption function during osteoclast differentiation.
METHODS:Osteoclasts were cultured by mouse monocyte-macrophage cellline-RAW264.7. The cells were divided into two groups:control group, treated with 100μg/L receptor activator of nuclear factorκB ligand factor;alendronate group, treated with 100μg/L receptor activator of nuclear factorκB ligand factor+10-7 mol/L alendronate. Osteoclastogenesis and resorption function of osteoclasts were examined at 7 days of culture and gene expression of capthesin K was detected by immunofluorescence method at 72 hours of culture. Western blot assay was used to detect capthesin K protein expression at 72 hours of culture.
RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase positive multinuclear cells were observed and resorption lacunae formed in two groups. Control group showed the higher number of tartrate-resistant acid phosphatase positive multinuclear cells and larger size of resorption lacunae than the alendronate group (P<0.01). Immunofluorescence showed expression of capthesin K was higher in the control group than the alendronate group (P<0.01);furthermore, the protein expression of capthesin K was also lower in the alendronate group than the control group (P<0.01). These findings indicate that bisphosphonates could strongly inhibit osteoclastogenesis and its resorption function by inhibiting gene expression of capthesin K.

Objective To investigate the killing effect of polymethylmethacrylate (PMMA) on spinal metastasis of transplanted VX2 carcinoma in experimental rabbit models. Methods Spinal metastasis of transplanted VX2 carcinoma model was successfully established in 18 rabbits. The experimental rabbits were randomly and equally divided into three groups with 6 rabbits in each group. Under CT guidance , PMMA or saline was injected into the center of VX2 tumor; in group A 0.3 ml of PMMA was used, in group B 0.1 ml of PMMA was used and in group C (control group) 0.3 ml saline was used. Twenty-four hours after the injection, the animals were sacrificed. Four tissue samples were obtained from the sites at 1 mm , 5 mm, 10 mm and 15 mm away from the PMMA mass in each rabbit of group A and group B , while four tissue samples were collected from different four sites from the tumor ’s center to border in each rabbit of group C. TdT-mediated dUTP nick-end labeling (TUNEL) method was used to determine the tumor cell apoptosis rate. Results After successful establishment of rabbit model, injection of PMMA was performed in sixteen among the eighteen rabbits. Technical success rates were 83.3% in both group A and B, and the success rate was 100% in group C. The difference in technical success rate was not significant. The mean tumor cell apoptosis rates of spinal VX2 carcinoma at 1 mm, 5 mm and 10 mm away from the PMMA mass in group A were (65.75±18.81)%, (50.00±14.24)% and(14.95±8.98)% respectively. The mean apoptosis rate in the control group was (9.79 ±5.24)%; the differences between the group A and the control group were statistically significant (P<0.05). The mean tumor cell apoptosis rate of spinal VX2 carcinoma at 15 mm away from the PMMA mass in group A was (10.30 ±8.13)%, which was not significantly different with that of the control group. The mean tumor cell apoptosis rates of spinal VX2 carcinoma at 1 mm and 5 mm away from the PMMA mass in group B were (49.20±15.57)% and(17.75±9.28)% respectively, which was significantly different with that of the control group(P<0.05); the mean tumor cell apoptosis rates at 10 mm and 15 mm away from the PMMA mass in group B were not significantly different with those of the control group. Statistically significant differences in the mean tumor cell apoptosis rates determined at 1 mm, 5 mm and 10 mm away from the PMMA mass existed between group A and group B(P<0.001). Conclusion PMMA can promote the apoptosis of tumor cells, properly increasing the injected amount of PMMA can enlarge the extent of tumor cell apoptosis.