Effects of calcium elevation on intracellular oxidative stress in human lens epithelial cells SRA01/04 / 眼科新进展

Objective To investigate the influence of calcium elevation on oxida tive stress in human lens epithelial cells (HLEC) SRA01/04.Method The cells (2 x 103 cells/well) which in the period of logarithmic phase were seeded into 96-well plates with three replicates for the two groups;and in the experimental group,SRA01/04 cells were exposed to a CaCI2 concentration gradient (3 mmol · L-1,5 mmol · L-1,7 mmol · L-1,9 mmol · L-1,11 mmol · L-1,13 mmol · L-1,15 mmol · L-1,17 mmol · L-1,19 mmol · L-1) for 0 h,12 h,24 h,36 h;while the cells in the control group were cultured in complete 1640 medium.Cell counting kit-8 (CCK-8) assay was used to measure cell viability.The levels of intracellular superoxide dismutase (SOD),glutathione (GSH) content and oxidized glutathione (GSSG) / total glutathione (T-GSH) were determined by using the microplate-reader method with the commercial total/oxidized glutathione and sod quantification kit.Results At first,the survival rate of SRA01/04 cells treated with 3 mmol · L-1,5 mmol · L-1,7 mmol· L-1 CaCL2 for 24 h showed a significant decrease with the increase of CaCl2 concentration by CCK-8 assays,but gradually increased when the concentration increased to 9 mmol · L-1,and the difference approached statistical significance (P ＜ 0.05).Meanwhile,there was significant difference in the viability of the control group (0.592 + 0.055) and cells exposed to 15 mmol · L-1 CaCI2 (0.293 + 0.02) (t =7.811,P ＜0.05).Cell treatment with 15 mmol· L-1 CaC12 for 24 h was the most appropriate condition for HLEC apoptosis,followed by the appearance of nuclear fragmentation and dissolution,enhanced intracellular SOD viability (t =-6.417,P ＜ 0.05),decreased T-GSH content (t =13.816,P ＜ 0.05),and increased ratio of GSSG/T-GSH (t =-4.396,P ＜ 0.05) when compared with the control group,and the differences were statistically significant.Conclusion Intracellular calcium elevation can inhibit the cell viability and increase the levels of SOD and GSSG in HLEC to aggravate the intracellular oxidative damage.

Effects of calcium elevation on intracellular oxidative stress in human lens epithelial cells SRA01/04 / 眼科新进展

Objective To investigate the influence of calcium elevation on oxida tive stress in human lens epithelial cells (HLEC) SRA01/04.Method The cells (2 x 103 cells/well) which in the period of logarithmic phase were seeded into 96-well plates with three replicates for the two groups;and in the experimental group,SRA01/04 cells were exposed to a CaCI2 concentration gradient (3 mmol · L-1,5 mmol · L-1,7 mmol · L-1,9 mmol · L-1,11 mmol · L-1,13 mmol · L-1,15 mmol · L-1,17 mmol · L-1,19 mmol · L-1) for 0 h,12 h,24 h,36 h;while the cells in the control group were cultured in complete 1640 medium.Cell counting kit-8 (CCK-8) assay was used to measure cell viability.The levels of intracellular superoxide dismutase (SOD),glutathione (GSH) content and oxidized glutathione (GSSG) / total glutathione (T-GSH) were determined by using the microplate-reader method with the commercial total/oxidized glutathione and sod quantification kit.Results At first,the survival rate of SRA01/04 cells treated with 3 mmol · L-1,5 mmol · L-1,7 mmol· L-1 CaCL2 for 24 h showed a significant decrease with the increase of CaCl2 concentration by CCK-8 assays,but gradually increased when the concentration increased to 9 mmol · L-1,and the difference approached statistical significance (P ＜ 0.05).Meanwhile,there was significant difference in the viability of the control group (0.592 + 0.055) and cells exposed to 15 mmol · L-1 CaCI2 (0.293 + 0.02) (t =7.811,P ＜0.05).Cell treatment with 15 mmol· L-1 CaC12 for 24 h was the most appropriate condition for HLEC apoptosis,followed by the appearance of nuclear fragmentation and dissolution,enhanced intracellular SOD viability (t =-6.417,P ＜ 0.05),decreased T-GSH content (t =13.816,P ＜ 0.05),and increased ratio of GSSG/T-GSH (t =-4.396,P ＜ 0.05) when compared with the control group,and the differences were statistically significant.Conclusion Intracellular calcium elevation can inhibit the cell viability and increase the levels of SOD and GSSG in HLEC to aggravate the intracellular oxidative damage.

Effects of Sinopodophyllum hexundrum on apoptosis in K562 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of Sinopodophyllum hexundrum on apoptosis in K562 cells.</p><p><b>METHODS</b>K562 cells were treated with Sinopodophyllum hexundrum at different concentrations and for different lengths of time to determine the optimal conditions of SinoPodophyllum hexandrum treatment for K562 cells using CCK8 assay. The cell apoptotic rate was detected by flow cytometry, and the cell morphology and nuclear morphology of K562 cells were observed with Wright staining and DPAI staining, respectively. The protein expressions of BCR/ABL, p-BCR/ABL, STAT5, p-STAT5 and the apoptosis-related proteins PARP, caspase-3 and cleaved-caspase-3 were determined with Western blotting.</p><p><b>RESULTS</b>The cell proliferation was inhibited in a concentration-and time-dependent manner by 1, 2, and 3 µg/mL Sinopodophyllum hexundrum. The treatment was optimal with a Sinopodophyllum hexundrum concentration of 2 µg/mL a treatment time of 48 h, and the cell apoptotic rate increased in a time-dependent manner and significantly increased at 48 h (P<0.001). The expression of apoptosis-related proteins PARP, caspase-3 and cleaved-caspase-3 were also activated in a time-dependent manner. The cells showed typical apoptotic changes after treatment with 2 µg/mL Sinopodophyllum hexundrum for 48 h with significantly reduced expressions of BCR/ABL, p-BCR/ABL, STAT5, AND p-STAT5.</p><p><b>CONCLUSION</b>Sinopodophyllum hexundrum promotes K562 cell apoptosis possibly by inhibiting BCR/ABL-STAT5 survival signal pathways and activating the mitochondrion-associated apoptotic pathways.</p>

Application of Magnetoencephalography with Synthetic Aperture Magnetometry in Localization of Motor Cortex and Epileptogenic Focus / 中国康复理论与实践

@#Objective To investigate the value of synthetic aperture magnetometry (SAM) in localizing motor cortex and epileptogenic focus for brain lesions near the central sulcus and to clear its advantage in the localization. Methods 12 patients (6 patients with epilepsy) were enrolled in this study. Before the operation, the patients were all taken Karnofsky Performance Status Score (KPS), examined with MEG by SAM technique in the localization of motor cortex and epileptogenic focus to determine their position relationship, and guide the scheme of surgery programme. During the operation, the location of hand-motor functional area were identified with evoked potential monitoring awaking test, and epileptogenic focus with electrocorticogram (ECoG) monitoring. The accuracy of location was assessed with the hand movement and KPS score, and the epileptic attack were evaluated with Engel curative effect grading. They were followed up for 2 years. Results The motor cortex of all the patients were located near the precentral gyrus with SAM and the localization of epileptogenic focus in 6 patients by SAM was consistent with that by ECoG. All the operations were based on and guided by the SAM. After the operations, the motor function and KPS score of 8 patients improved. No extra functional lesions happened in all patients. Epilepsy was well controlled in 5 cases. Conclusion SAM can correctly localize the motor cortex and epileptogenic focus. Meanwhile position relationship between the intracranial lesions and motor functional areas and epileptic focus can be clear. It is a valuable method for surgical planning and epilepsy controlling and will decrease the occurrence of neurological deficits after operation.