The anti-cancer activities of Rhus verniciflua Stokes (GC), Ulmus davidiana var. japonica Nakai (UGP) and arsenium sublimatum (SS) extracts, which have been used Oriental medicine therapy for various diseases, were investigated. The treatment of GC, UGP and SS alone, and combined treatment with GC, UGP and SS did not affect the cell viability in the mouse normal cell lines (RAW 264.7 macrophages and C2C12 myoblasts). However, co-treatment with GC, UGP and SS markedly induces apoptosis in human gastric cancer AGS cells, but not in other various cancer cell lines (human lung cancer A549, colon cancer HCT116, liver cancer Hep3B and bladder T24 cells) as evidenced by formation of apoptotic bodies, chromatin condensation, and accumulation of annexin-V positive cells. Co-treatment with GC, UGP and SS effectively induced the expression levels of Fas and Fas ligand, and inhibited the levels IAP family proteins such as XIAP, cIAP-1 and survivin, and anti-apoptotic Bcl-xL proteins compared with treatment with either agent alone. Combined treatment also significantly induced the loss of mitochondrial membrane potential, which was associated with the activation of caspases (-3, -8, and -9) and degradation of poly (ADP-ribose) polymerase. However, the cytotoxic effects induced by co-treatment with GC, UGP and SS were significantly attenuated by pan-caspases inhibitor, z-VAD-fmk, indicating an important role for caspases. These results indicated that the caspases were key regulators of apoptosis in response to co-treatment of GC, UGP and SS in human gastric cancer AGS cells and further studies will be needed to identify the active compounds.
Kim, Jae-Do;Chung, So-Hak;Hong, Young-Gi;Choi, Jang-Seok
The Journal of the Korean bone and joint tumor society
/
v.5
no.1
/
pp.1-8
/
1999
A single fraction of 50 Gy extracorporeal irradiation, as a modality of limb-sparing operation, has been used to achieve tumor necrosis in osteosarcoma. Although this modality of radiation therapy preserving the mobility of a joint is commonly practiced, the precise knowledge on the radiobiological response of osteosarcoma cell has remained to be elucidated. We therefore observed whether a single high dose irradiation caused apoptosis in osteosarcoma cells and whether the commitment to apoptosis was associated with cell kinetics. We also investigated radiation dose response along the time course for development of apoptosis following single high dose irradiation. The morphologic change in apoptosis was observed by fluorescence with Hoechst 33258 and the degree and the fraction of cells by flow cytometry. Irradiation of osteosarcoma cells with 10, 30 and 50 Gy resulted in chromatin condensation and apoptotic body formation. The degree of apoptosis in osteosarcoma cells was $29.5{\pm}3.56%$, $39.9{\pm}4.83%$ at 24 and 48 hours after 10 Gy irradiation ; $41.1{\pm}3.93%$, $66.9{\pm}5.21%$ at 24 and 48 hours after 30 Gy irradiation ; and $48.0{\pm}3.69%$, $75.6{\pm}4.65%$ at 24 and 48 hours after 50 Gy irradiation. The fraction of cells in cell-cycle kinetic was $39.2{\pm}4.3%$ in G2/M, $22.1{\pm}4.65%$ in G1 at 24 hours after 10 Gy irradiation ; $51.0{\pm}4.3%$ in G2/M, $20.4{\pm}4.7%$ in G1 at 48 hours after 10 Gy irradiation ; $40.3{\pm}3.9%$ in G2/M, $26.1{\pm}4.7%$ in G1 at 24 hours after 30 Gy irradiation ; $59.2{\pm}3.9%$ in G2/M, $5.9{\pm}5.1%$ in G1 at 48 hours after 30 Gy irradiation ; and $44.3{\pm}4.2%$ in G2/M, $21.1{\pm}3.5%$ in G1 at 24 hours after 50 Gy irradiation. The fraction of cells at 48 hours after 50 Gy irradiation could not be observed because of irradiation induced cell death of most of cells. All values for irradiated cells showed accumulation in G2/M phase and reduction in G1 phase, irrespective of irradiation dose. The results suggest that a single fraction of high dose irradiation with 50 Gy results in accumulation of cells at G2/M phase, leading to apoptosis.
In this research, $5\times5cm^2$ unit cells were fabricated via liquid condensation process and uniaxial pressing followed by the screen printing of electrolyte and cathode layer. The SOFC stack was assembled with unit cells, gasket-type sealant and metal interconnect. The stack was designed to have a single column with internal-manifold and cross-flow type gas-channels. The SOFC stack produced 15 W, which is $50\%$ of the maximum power being expected from the maximum power density of the unit cell. Controlling factors for the proper operation of the SOFC stack and other designing factors of stack manifold and gas channels were discussed.
This experimental study was carried out to evaluate the effect of Yipahnsan on angiogenic inhibition mechanism. This study investigates the effects of Yipahnsan on angiogenic inhibition mechanism evaluate cell adhesive inhibition effect, DNA fragmantaion analysis, nuclear condensation assay, FACScan analysis, angiogenic lumen formation assay, immunocytochemistry analysis, RT-PCR for mRNA expression, western blot analysis, confocal analysis for $Ca^{2+}influx$. The results were summarized as follows : 1. The cell adhesive inhibition ability was strongly increased from $5{\mu}g/ml$ on ECV304 cell line and ECVPAR cell line. 2. YY water extract caused $G_0/G_1$ arrest peak to existed on the ECV304 cell line. 3. YY water extract caused inhibition of proliferation and inducement of apoptosis on the collagen coated plate in ECV304 cell line. 4. YY water extract inhibited the lumen formation on the matrigel coated plate in ECV304 cell line. 5. YY water extract inhibited the expressions of LFA-1 and ELAM-1 on ECV304 cell line and ECVPAR cell line. 6. YY water extract inhibited the expressions of MMP-9 and uPA on ECV304 cell line and ECVPAR cell line. 7. YY water extract inhibited the expression of integrin ${\alpha}_v{\beta}_3$ on ECV304 cell line and ECVPAR cell line. 8. YY water extract decreased the change of $Ca^{2+}$ in intracellular on ECV304 cell line and ECVPAR cell line. According to the results, Yipahnsan showed to be a key antagonist of integrin ${\alpha}_v{\beta}_3$, and to be induction of apoptosis by p53 through flow cytometry. This report also demonstrated that expressions of MMP-9 and uPA were blocked under the angiogenesis model. Thus, we suggested that Yipahnsan blocks angiogenesis by inducing apoptosis in ECV304 and ECVPAR cell lines, and another oriental herbal medicine that treats qi-stagnation and blood-stasis type also has angiogenic inhibition effects.
This experimental study was carried out to evaluate the effects of Hwallakhyoreungdan on angiogenic inhibition mechanism. In order to investigate the effects of Hwallakhyoreungdan on angiogenic inhibition mechanism, MTT assay, cell adhesive inhibition effect, DNA fragmantaion analysis, Nuclear condensation assay, FACScan analysis, Angiogenic lumen formation assay, Immunocytochemistry analysis, RT-PCR for mRNA expression, Western blot analysis and Confocal analysis for $Ca^{2+}$ change were performed. The results were summarized as follows: 1. The cell adhesive inhibition ability was strong from $5{\mu}g/ml$. 2. The $G_0/G_1$ arrest peak was existed on ECV304 cell-line. 3. The cells on Collagen plate were inhibition of proliferation and inducement of apoptosis by HR water extract. 4. Angiogenic lumen formation was inhibited by HR water extract. 5. LFA-1 and ELAM-1's expression were inhibited by HR water extract. They are commenly participation on inflammation and tumor regeneration. 6. The expression of MMP-9 and uPA were inhibited by HR water extract. 7. The expression of integrin ${\alpha}_v{\beta}_3$ was inhibited by HR water extract. 8. The expression of intracellular molecule were successively inhibited by HR water extract therefore the proliferation of ECV304 cell line was stopped and apoptosis was induced. 9. The change of $Ca^{2+}$ was decreased by HR water extract it cause confusion of signal transduction pathway therefore it was take part in apoptosis. According to the results, Hwallakhyoreungdan showed to be a key antaonist of integrin ${\alpha}_v{\beta}_3$, and to be induction of apoptosis by p53 through flow cytometry. This report also demonstrated that expressions of MMP-9 and uPA was blocked under the angiogenesis model. Thus, we suggests that Hwallakhyoreungdan blocks angiogenesis by inducing apoptosis of ECV304 and ECVPAR cell lines and another oriental herbal medicine that treats blood-stasis type also has angiogenic inhibition effects.
Objectives: This study examined the relative efficacies of a derivative of betulinic acid (dBA) and its poly (lactide-co-glycolide) (PLGA) nano-encapsulated form in A549 lung cancer cells in vivo and in co-mutagen [sodium arsenite (SA) + benzo[a]pyrene (BaP)]-induced lung cancer in mice in vivo. Methods: dBA was loaded with PLGA nanoparticles by using the standard solvent displacement method. The sizes and morphologies of nano-dBA (NdBA) were determined by using transmission electron microscopy (TEM), and their intracellular localization was verified by using confocal microscopy. The binding and interaction of NdBA with calf thymus deoxyribonucleic acid (CT-DNA) as a target were analyzed by using conventional circular dichroism (CD) and melting temperature (Tm) profile data. Apoptotic signalling cascades in vitro and in vivo were studied by using an enzyme-linked immunosorbent assay (ELISA); the ability of NdBA to cross the blood-brain barrier (BBB) was also examined. The stage of cell cycle arrest was confirmed by using a fluorescence-activated cell-sorting (FACS) data analysis. Results: The average size of the nanoparticles was ~ 110 nm. Confocal microscopy images confirmed the presence of NdBA in the cellular cytoplasm. The bio-physical properties of dBA and NdBA ascertained from the CD and the Tm profiles revealed that NdBA had greater interaction with the target DNA than dBA did. Both dBA and NdBA arrested cell proliferation at G0/G1, NdBA showing the greater effect. NdBA also induced a greater degree of cytotoxicity in A549 cells, but it had an insignificant cytotoxic effect in normal L6 cells. The results of flow cytometric, cytogenetial and histopathological studies in mice revealed that NdBA caused less nuclear condensation and DNA damage than dBA did. TEM images showed the presence of NdBA in brain samples of NdBA fed mice, indicating its ability to cross the BBB. Conclusion: Thus, compared to dBA, NdBA appears to have greater chemoprotective potential against lung cancer.
Journal of the Korean Society of Propulsion Engineers
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v.19
no.1
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pp.61-67
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2015
The cooling of hot exhaust gas is an important issue for the construction of combustor test facility. Water spray is an effective method for exhaust gas cooling due to its large latent heat in process of evaporation. In this study, 1-D analysis has been performed based on continuity, energy conservation, and saturated vapor property to understand water spray cooling of combustion gas. In the exhaust duct of combustor test facility, the injected water decreases combustion gas temperature, and evaporates in the combustion gas. However, some of the injected water is collected in the sump due to condensation. The evaporation of water helps combustion gas cooling, but causes pressure increase inside the exhaust duct due to increase of vapor pressure. These phenomena has been analyzed by 1-D modeling in this study. From 1-D analysis, the adequate mass flow rate of water spray to cool combustion gas and to avoid excessive pressure rise inside the exhaust duct has been decided.
In this study, the energy conversion equipment from the radiation energy to mechanical energy by using npentane as the operating fluid was constructed and the performance to pump the water was tested for the utilization of solar powered water pump. The equipment was designed optimally, after the theoretical analyses of the water pumping head and water quantity per cycle were done. The pentane vapour temperature in the condenser and the temperature of the outlet water from the condenser became lowered and the heat transfer rate became higher with decreasing the water inlet level to the condenser. The temperature difference between the condenser and the water tank was significant. Therefore, the distance between the water tank and condenser was recommended to be shorten and the diameter of their connecting pipe was recommended to be narrow in order to reduce the resistance of the fluid passage and improve the heat transfer rate. The amount of water pumped was 1.62.4 liters. Mass flow rate of the cooling water became lowered when the cooling water pipe was prolonged from the condenser to improve the heat transfer rate.
Wang, Xin;Wang, Chao;Sun, Yu-Ting;Sun, Chuan-Zhen;Zhang, Yue;Wang, Xiao-Hua;Zhao, Kai
Asian Pacific Journal of Cancer Prevention
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v.16
no.1
/
pp.125-131
/
2015
Currently, taxol is mainly extracted from the bark of yews; however, this method can not meet its increasing demand on the market because yews grow very slowly and are a rare and endangered species belonging to first-level conservation plants. Recently, increasing efforts have been made to develop alternative means of taxol production; microbe fermentation would be a very promising method to increase the production scale of taxol. To determine the activities of the taxol extracted from endophytic fungus N. sylviforme HDFS4-26 in inhibiting the growth and causing the apoptosis of cancer cells, on comparison with the taxol extracted from the bark of yew, we used cellular morphology, cell counting kit (CCK-8) assay, staining (HO33258/PI and Giemsa), DNA agarose gel electrophoresis and flow cytometry (FCM) analyses to determine the apoptosis status of breast cancer MCF-7 cells, cervical cancer HeLa cells and ovarian cancer HO8910 cells. Our results showed that the fungal taxol inhibited the growth of MCF-7, HeLa and HO8910 cells in a dose-and time-dependent manner. IC50 values of fungal taxol for HeLa, MCF-7 and HO8910 cells were $0.1-1.0{\mu}g/ml$, $0.001-0.01{\mu}g/ml$ and $0.01-0.1{\mu}g/ml$, respectively. The fungal taxol induced these tumor cells to undergo apoptosis with typical apoptotic characteristics, including morphological changes for chromatin condensation, chromatin crescent formation, nucleus fragmentation, apoptotic body formation and G2/M cell cycle arrest. The fungal taxol at the $0.01-1.0{\mu}g/ml$ had significant effects of inducing apoptosis between 24-48 h, which was the same as that of taxol extracted from yews. This study offers important information and a new resource for the production of an important anticancer drug by endofungus fermentation.
Inhibitory effects of Maclura amboinenesis Bl, one plant used traditionally for the treatment of cancers, on metastatic potential of highly metastatic B16F10 melanoma cells were investigated in vitro. Cell proliferation was assessed using the MTT colorimetric assay. Details of metastatic capabilities including invasion, migration and adhesion of B16F10 melanoma cells were examined by Boyden Chamber invasion and migration, scratch motility and cell attachment assays, respectively. The results demonstrated that n-hexane and chloroform extracts exhibited potent anti-proliferative effects (p<0.01), whereas the methanol and aqueous extracts had less pronounced effects after 24 h exposure. Bioactivity-guided chromatographic fractionation of both active n-hexane and chloroform extracts led to the isolation of two main prenylated xanthones and characterization as macluraxanthone and gerontoxanthone-I, respectively, their structures being identified by comparison with the spectral data. Interestingly, both exhibited potent effective effects. At non-toxic effective doses, n-hexane and chloroform extracts (10 and $30{\mu}g/ml$) as well as macluraxanthone and gerontoxanthone-I (3 and $10{\mu}M$) significantly inhibited B16F10 cell invasion, to a greater extent than $10{\mu}m$ doxorubicin, while reducing migration of cancer cells without cellular cytotoxicity. Moreover, exposure of B16F10 melanoma cells to high concentrations of chloroform ($30{\mu}g/ml$) and geratoxanthone-I ($20{\mu}M$) for 24 h resulted in delayed adhesion and retarded colonization. As insights into mechanisms of action, typical morphological changes of apoptotic cells e.g. membrane blebbing, chromatin condensation, nuclear fragmentation, apoptotic bodies and loss of adhesion as well as cell cycle arrest in the G1 phase with increase of sub-G1 cell proportions, detected by Hoechst 33342 staining and flow cytometry were observed, suggesting DNA damage and subsequent apoptotic cell death. Taken together, our findings indicate for the first time that active n-hexane and chloroform extracts as well as macluraxanthone and gerontoxanthone-I isolated from Maclura amboinensis Bl. roots affect multistep of cancer metastasis processes including proliferation, adhesion, invasion and migration, possibly through induction of apoptosis of highly metastatic B16F10 melanoma cells. Based on these data, M. amboinensis Bl. represents a potential candidate novel chemopreventive and/or chemotherapeutic agent. Additionally, they also support its ethno-medicinal usage for cancer prevention and/or chemotherapy.
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