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The purpose of this study is to seek an effective teaching device in the education of Geometric figures in the middle school mathematics through the transformation. The analysis between of the state book of the third curriculum, of the S.M.S,G. and of the contents of current textbooks on the similarity conditions of triangle, together with the comparative study of internal and external data has been made in the area of the problem solving ability and mathematical thinking ability. The results of thinking ability. 1. The teaching device of the state book of the third curriculum and the S.M.S.G have been proved to be more effective in helping students improve their problem solving ability and in teaching students the basic concepts of the Geometric figures than other. Hence, the reconstruction of the contents of text books on the teaching of Geometric figures adding the transformation helps students develop their mathematical thinking ability. 2. The remarkable growth of mathematical thinking ability and problem solving ability has been made by the students who have mid-level scores and high-level intelligence quotient. A large number of students have improved their mathematical thinking ability by the transformation learning of Geometric figures. Therefore, the device helps students to develop their mathematical thinking ability and problem solving ability through the transformation learning of Geometric figures.
Objective : Hedyotis diffusa has been used as an arnicancer agent for several decades in oriental medicine. We test whether the methanol extract of the herb affects transcriptional activation factors including NF-κB and AP-l. Methods : 1. HL-60 cells were treated with various concentrations(from 200 to 50ug/ml) of methanol extract and H_2O extract(200ug/ml)of hedyotis diffusa, After 48h later, the cells were tested for viability by MTT assay. 2. The HL-60 cells were treated with 200ug/ml of methanol extract for the indicated periods. First. Nuclear extracts were isolated and incubated with oligonucleotide probe of NF-κB and AP-l. Second. Nuclear extracts were isolated and reacted with p50, p65. c-rel par-Jun, c-Jun, JunB. JunD antibody on ice for 30min. Finally The cell lnates were prepared and analyzed by westem blotting using anti-Fas, anti-FasL and anti-p53 antibdy. Results : l. The methanol extract decreases the viability of human lymphoid origin leukemia HL-60 cells in a dose-depemdemt manner. 2. NF-κB is rapidly activated by the addition of the methanol extract, reaches a peak at 30min and gradually returns to resting level. We confirm tha NF-κB is a heterodimer mainly composed of p65 subunit with c-Rel. 3. Transcriptional activation of AP-1 is detected at 30min and reaches a maximum at lhr after stimulation of the cells with the methanol extract. AP-l is mainly composed with Jur-D and partially Jug-B proteins. 4. the methanol extract of Hedyotis diffusa induces the expression of Fas, Fas ligand and p53 proteins of HL-60 cells in a time dependent fashion. Conclusions : These results suggest that the methanol extract of Hedyotis diffusa exerts anticancer effects to induce the death of human leukomic HL-60 cells via activation of trascriptional factors such a NF-κB and AP-1, increase in expression of Fas mediated signaling proteins, and induction of tumor suppressor gene. p53
Objectives: This experimental study was carried out to evaluate the effects of aqueous and methanol extracts of Hedyotis diffusa which has long been used for cancer treatment in oriental medicines on the induction of apoptotic cell death in human lymphoid leukemia cell line, HL-60. Methods: Cells were treated with various concentrations (200 to 0.4㎍) and periods (6 to 30 hr) of H_2O and methanol extracts of Hedyotis diffusa. Then, cells were tested for viability by MTT assay. Cells wrere treated with 200㎍/ml of methanol extract fork various periods. Genomic DNA was isolated, separated, on 1.5% agarose gels, stained with ethidium bromide and visualized under UV light. Cells were treated with 200㎍/ml of each extract for 16hr.Then, cells were treated with various doses of each extract for 12 hr and 100㎍/ml of methanol extract for various periods. Lysate from the cells used to measure the activity of caspase-1 and-3 proteases by using fluorogenic peptide substrates including acetyl-YVAD-AMC and acetyl-DEVD-AMC, respectively, Cells were treated with 200㎍/ml of each extract for various periods. Cell lysates were immunoprecipated with anti-JNKl antibodies. The immune complex was reacted with 32^p-ATP and c-Jun as a substrate. The phosphotransferase activity of JNKI was measured by suing Phosphoimage analyzer (Fuji Co., Japan). Nuclear extracts were isolated and incubated with oligonucleotide probe of NF-kB was measured by using EMSA and visualized by PhosphoImage analyzer (Fuji Co, Japan). Cell lysates were prepared and analyzed by Western blotting with anti-Bc12 antibodies and anti-Bax antibodies. Cells were pretreated with various doses of methanol extract for 2 hr. Then, the extract was removed by centrifugation. Cells were resuspended with RPMI-1640 media containing 0.3% agarose, 10% FBS, overlayred onto bottom layer agarose and incubated at CO_2 incubator for 6 days. The number of colony was cunted under light microscopy (×100). Results: The death of HL_60 cells was markedly induced by the addition of methanol extract of Hedyotis diffusa in a dose and time-dependent manners. The apoptotic characteristic ladder pattern of DNA strand break was observed in death of HL_60 cells. In addition, it was shown nucleus chromatin condensation under Hoechst staining. Therefore, Hedyotis diffusa extract-induced death of HL-60 cells is mediated by apoptotic signaling processes. The activity of Caspaxe 3-like proteases remained in a basal level in HL-60 cells treated with aqueous extract of Hedyotis diffusa. However, it was markedly increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. In addition, the phosphotransferase activity of JNKl was increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. Furthermore, the activation of transcriptional activator, NF-kB was markedly induced by methanol extract of Hedyotis diffusa. Anti-apoptotic Bc12 was cleaved into 23Kda fragment by treatment of methanol extract of Hedyotis diffusa. However, expression of proapoptotic Bax protein was increased by treatment of methanol extract of Hedyotis diffusa in a time-dependent manner. Furthermore, methanol extract markedly inhibited the colony forming efficiency of HL-60 cells in semisolid agar culture. Conclusions: Above results suggest that methanol extract of Hedyotis diffusa induces the apoptotic death of human leukemic HL-60 cells via activations of Caspase-3 proteases, JNKI, transcriptional activator NF-κB, In addition, our results also suggest that methanol exthanol extract of Hedyotis diffusa reduces the malignant potential of HL-60 cells via down regulation of colony forming effciency through cleavage of Bc12 as well as induction of Bax.
Objecives: Hedyotis diffusa is used to treat cancer in traditional Korea Medicine. So this study was carried out to examine the expression of cell cycle related genes in HL-60 cells undergoing apoptosis by the methanol extract of Hedyotis diffusa. Methods: 1. HL-60 cells were treated with various concentrations (from 200 to 50㎍/ml)of metnanol extract and H20 extract (200 ㎍/ml) of hedyotis diffusa. After 48 h later, the cells were tested for viability by MTT assay. 2. The HL-60 cells were treated with 200 ㎍/ml of methanol extract for the indicated periods. The whole cell lysates were prepared and analyzed by westem blotting using anti-p53 antibody. 3. The nuclear extract were prepared and analyed by western blotting using anti-p21 antibody, anti-p27 antibody, anti-cyclen A antibody, anti-cylin E antibody and anti-CDK2 antinbody. Results: 1. The methanol extract of Hedyotis diffusa induced the death of HL-60 cells in a dose dependent manner. 2. The methanol extract of Hedyotis diffusa makedly decreased the level of p21/Cipl and cyclin A in a time dependent manner. 3. The methanol extract of Hedyotis diffusa markedly increased tje ;eve; pf p27/Kip and cyclin E in a time dependent mammer. 4.The methanol extract of Hedyotis diffusa markedly did not affect the level of CDK2. Conclusions: These results provide evidence that expression of cell cycle related genes in HL-6- cells undergoing apoptosis by the methanol extract of Hedyotis diffusa mainly results from decreased level of p21/cipl and increased level of p27/Kipl of the cell cycle related genes.