Lee, Soojin;Shim, Ji Hwan;Gim, Huijin;Park, Hyun Soo;Kim, Byung Joo
Journal of Pharmacopuncture
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v.19
no.1
/
pp.51-58
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2016
Objectives: Oldenlandia diffusa is traditionally used to relieve the symptoms of and to treat various diseases, but its anti-cancer activity has not been well studied. In the present study, the authors investigated the anti-cancer effects of an ethanol extract of Oldenlandia diffusa (EOD) on HT-29 human adenocarcinoma cells. Methods: Cells were treated with different concentrations of an EOD, and cell death was assessed by using a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. Analyses of the sub G1 peak, the caspase-3 and -9 activities, and the mitochondrial membrane depolarizations were conducted to confirm cell death by apoptosis. Also, intracellular reactive oxygen species (ROS) generation was determined using carboxy-H2DCFDA (5-(and-6)-carboxy-20,70-dichlorodihydrofluorescein diacetate). Results: EOD inhibited the proliferation of HT-29 cells for 24 hours by $78.6%{\pm}8.1%$ at $50{\mu}g/mL$, $74.4%{\pm}4.6%$ at $100{\mu}g/mL$, $65.9%{\pm}5.2%$ at $200{\mu}g/mL$, $51.4%{\pm}6.2%$ at $300{\mu}g/mL$, and by $41.7%{\pm}8.9%$ at $400{\mu}g/mL$, and treatment for 72 hours reduced the proliferation at the corresponding concentrations by $43.3%{\pm}8.8%$, $24.3{\pm}5.1mV$, $13.5{\pm}3.2mV$, $6.5{\pm}2.3mV$, and by $2.6{\pm}2.3mV$. EOD increased the number of cells in the sub-G1 peak in a dose-dependent manner. The mitochondrial membrane depolarization was elevated by EOD. Also, caspase activities were dose-dependently elevated in the presence of EOD, and these activities were repressed by a pan-caspase inhibitor (zVAD-fmk). The ROS generation was significantly increased by EOD and N-acetyl-L-cysteine (NAC; a ROS scavenger) remarkably abolished EOD-induced cell death. In addition, a combination of sub-optimal doses of EOD and chemotherapeutic agents noticeably suppressed the growth of HT-29 cancer cells. Conclusion: These results indicate that EOD might be an effective chemotherapeutic for the treatment of human colorectal cancer.
The combined effects of primisulfuron and imazethapyr treated with piperonyl butoxide(PBO) on growth inhibition and acetolactate synthase(ALS) activity in corn were studied to identify the tolerance mechanism among corn cultivars. Pioneer 3751 IR showed resistance to primisulfuron and imazethapyr, but Pioneer 3751 and Chalok 2 were susceptible to them. Pioneer 3751 IR was tolerant to primisulfuron regardless of PBO treatment, but Pioneer 3751, Suwon 118 and Chalok 2 were more greatly inhibited by combined treatment of primisulfuron with PBO. Synergistic effect on growth inhibition in Pioneer 3751 IR, Pioneer 3751, Suwon 118 and Chalok 2 was not occurred when imazethapyr was treated with PBO, but Pioneer 3751, Suwon 118 and Chalok 2 tended to inhibit by combined treatment of imazethapyr with PBO. Pioneer 3751 IR showed higher ALS activity than Pioneer 3751 when primisulfuron or imazethapyr was treated. ALS activity was inhibited by primisulfuron at $0.01{\mu}M$ or above and imazethapyr at $1{\mu}M$ or above, and inhibitory effect on ALS activity was observed by primisufuron and imazethapyr treatments with PBO.
Shim, Hye-Young;Park, Jong-Hwa;Paik, Hyun-Dong;Nah, Seung-Yeol;Kim, Darrick S.H.L.;Han, Ye Sun
Molecules and Cells
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v.24
no.1
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pp.95-104
/
2007
The mechanism of acacetin-induced apoptosis of human breast cancer MCF-7 cells was investigated. Acacetin caused 50% growth inhibition ($IC_{50}$) of MCF-7 cells at $26.4{\pm}0.7{\mu}M$ over 24 h in the MTT assay. Apoptosis was characterized by DNA fragmentation and an increase of sub-G1 cells and involved activation of caspase-7 and PARP (poly-ADP-ribose polymerase). Maximum caspase 7 activity was observed with $100{\mu}M$ acacetin for 24 h. Caspase 8 and 9 activation cascades mediated the activation of caspase 7. Acacetin caused a reduction of Bcl-2 expression leading to an increase of the Bax:Bcl-2 ratio. It also caused a loss of mitochondrial membrane potential that induced release of cytochrome c and apoptosis inducing factor (AIF) into the cytoplasm, enhancing ROS generation and subsequently resulting in apoptosis. Pretreatment of cells with N-acetylcysteine (NAC) reduced ROS generation and cell growth inhibition, and pretreatment with NAC or a caspase 8 inhibitor (Z-IETD-FMK) inhibited the acacetin-induced loss of mitochondrial membrane potential and release of cytochrome c and AIF. Stress-activated protein kinase/c-Jun $NH_4$-terminal kinase 1/2 (SAPK/JNK1/2) and c-Jun were activated by acacetin but extracellular-regulated kinase 1/2 (Erk1/2) nor p38 mitogen-activated protein kinase (MAPK) were not. Our results show that acacetin-induced apoptosis of MCF-7 cells is mediated by caspase activation cascades, ROS generation, mitochondria-mediated cell death signaling and the SAPK/JNK1/2-c-Jun signaling pathway, activated by acacetin-induced ROS generation.
Follicular fluid(FF) of mammalian Graafian follicles contains various kinds of proteins and proteinases that are believed to play important roles during follicular growth oocyte maturation and ovulation of mature oocytes. Previous studies of human FF(hFF) demonstrated the presence of many serine/threonine proteinases and matrix metalloproteinases such as gelatinases, however, little is known about the caseinases. Present study was aimed to examine the presence and the property of caseinolytic enzyme in hFF. Using casein zymographic method, it was found that hFF, human adult serum and cord serum exhibited one intense 80 kDa and another weak 78 kDa bands having caseinolytic activity. When inhibitors were added to the zymographic substrate buffer, caseinolytic activity of both 80 kDa and 78 kDa proteins were inhibited by othylenediarnine tetraacetic acid(EDTA) or soybean trypsin inhibitor(SBTI), but not by E-64, phenylmethylsulfonyl fluoride(PMSF) or 1,10-phenanthroline. Thus both enzymes appear to belong to a family of trypsin-like enzyme. Addition of EDTA to the zymographic substrate buffer almost abolished the caseinolytic activity of both enzymes. However, further addition of a divalent metal ion such as CaC $l_2$, MgC $l_2$, MnC $l_2$ or ZnC $l_2$ to the same buffer fully restored the enzyme activity at 5 mM concentration despite the presence of EDTA. Based upon these observations, 80 kDa and 78 kDa caseinolytic enzymes are present in human follicular fluid and they appear to be trypsin-like enzymes of which caseinolytic activity needs the presence of $Ca^{++}$, aM $g^{++}$, M $n^{++}$ or Z $n^{++}$././././.
Journal of agricultural medicine and community health
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v.22
no.2
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pp.277-293
/
1997
This study was conducted to provide basic materials required to enforce and develop welfare policies, as well as the health system, for the aged, by surveying the status of health care utilized by the daily increasing old population and the importance of receiving physical therapy. Data that need in this research was gathered from over ages of 65, during the period from Jan 4, 1996 to Jan 31, using the inquiries previously made by geriatric researchers and through literatures investigator by this writer. The data were analyzed by $X^2$, Z-test, Likert scale. The findings were as follows : 1) General characteristics of subjects. People in the age group between 65 and 69 were 55.6% and the highest number, while male were 37% and female 63%. Analysis of income group disclosed 60.6% whose monthly income, including the pocket money given by children, was less than 200,000 won. 91.1% of the elderly people surveyed owned houses; only 36.4% live with spouses; while 15.6% live alone. 2) Characteristics with respect to utilization of health care institution. 56% of the total medical institutions used by the elderly people were clinics and the rates of chronic disease and musculoskeletal disease were 73.2%. 3) Characteristics with respect to approach of health care institution. 45.1% of the respondent stated it took 20 minutes to arrive at hospital, and bus accounted for 48.6% of all transportation means used to go to hospital. 4) Degree of cognition with respect to the rights of geriatric patients. (1) There is no financial support from the government for geriatric patients(71.4%). (2) Government financial support is needed for geriatric patients(95.3%). (3) Have never been regionally surveyed or called upon for interviews with respect to treatment desire and problems relating to geriatric patients(87.2%). (4) Health and medical policies for geriatric patients must be established rapidly(98.4). (5) Expansion and construction of specialized medical facilities for geriatric patients such as elderly hospital and medical center are needed(90.2%). (6) Government's welfare policies for the elderly people is insufficient(82.0%) 5) Degree of cognition on importance of physical therapy with respect to geriatric patient. (1) Physical therapy is considered most effective in treating geriatric patients(82.9%). (2) Physical therapists specializing in only elderly people must be need of separately(76.2%). (3) It is desirable for medical specialists to visit geriatric patients at home to provide physical therapy(82.9%). (4) Hospitals specializing in physical therapy for geriatric patient are required(85.6%). Based on the result for this research, the following suggestions are presented to facilitate the utilization of health care institution for the welfare of geriatric patients. Medical facilities such as elderly hospital and geriatric patient's medical center specializing in elderly people must be constructed as early as possible; and home-visiting physical therapist system must be important to treat chronic geriatric patients; our government must establish policies to provide the old ages with means for the health care and curing chronic diseases, and carry out the plans of reasonable distribution and effective untilization of medical resources.
Journal of the korean academy of Pediatric Dentistry
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v.30
no.2
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pp.263-271
/
2003
The purpose of the present study was to evaluate the patterns of hybrid layer according to the application time and the frequency and its effects on the shear bonding strength of All-In-One system in primary teeth. A single bonding agent(Scotchbond Multi-Purpose Plus, 3M) and an All-In-One system(Prompt L-pop, 3M ESPE) were applied on the dentin varying the application time and the frequency in primary teeth. Shear bond strength was measured and the patterns of hybrid layers were observed by SEM. The following results were obtained ; 1. The shear bonding strength of single bonding agent was significantly higher than that of All-In-One system(P<0.05). 2. The shear bonding strength of All-In-One system applied twice or 3 times were higher than that of applied once (P<0.05). And thickness of the hybrid layer was increased when applied twice or 3 times compared to once. 3. The shear bonding strength of All-In-One system when applied for 15 second and 30 second were higher than that of 7 second (P<0.05). And the hybrid layer thickness of 15 second and 30 second's application time were higher than that of 7 second. 4. Thickness of hybrid layer applied with single bonding agent was $2-4{\mu}m$ and that of All-In-One system was $1-2{\mu}m$.
An in vitro gas production technique was used in this study to elucidate the effect of two strains of active live yeast on methane ($CH_4$) production in the large intestinal content of pigs to provide an insight to whether active live yeast could suppress $CH_4$ production in the hindgut of pigs. Treatments used in this study include blank (no substrate and no live yeast cells), control (no live yeast cells) and yeast (YST) supplementation groups (supplemented with live yeast cells, YST1 or YST2). The yeast cultures contained $1.8{\times}10^{10}$ cells per g, which were added at the rates of 0.2 mg and 0.4 mg per ml of the fermented inoculum. Large intestinal contents were collected from 2 Duroc${\times}$Landrace${\times}$Yorkshire pigs, mixed with a phosphate buffer (1:2), and incubated anaerobically at $39^{\circ}C$ for 24 h using 500 mg substrate (dry matter (DM) basis). Total gas and $CH_4$ production decreased (p<0.05) with supplementation of yeast. The methane production reduction potential (MRP) was calculated by assuming net methane concentration for the control as 100%. The MRP of yeast 2 was more than 25%. Compared with the control group, in vitro DM digestibility (IVDMD) and total volatile fatty acids (VFA) concentration increased (p<0.05) in 0.4 mg/ml YST1 and 0.2 mg/ml YST2 supplementation groups. Proportion of propionate, butyrate and valerate increased (p<0.05), but that of acetate decreased (p<0.05), which led to a decreased (p<0.05) acetate: propionate (A: P) ratio in the both YST2 treatments and the 0.4 mg/ml YST 1 supplementation groups. Hydrogen recovery decreased (p<0.05) with yeast supplementation. Quantity of methanogenic archaea per milliliter of inoculum decreased (p<0.05) with yeast supplementation after 24 h of incubation. Our results suggest that live yeast cells suppressed in vitro $CH_4$ production when inoculated into the large intestinal contents of pigs and shifted the fermentation pattern to favor propionate production together with an increased population of acetogenic bacteria, both of which serve as a competitive pathway for the available H2 resulting in the reduction of methanogenic archaea.
$(Pb,La)TiO_3$(PLT) thin films were prepared on Pt/SiO$_2$/Si substrates by the sol-gel method and investigated the crystalline and electrical properties according to La concentration and post-annealing temperatures. The PLT films annealed at above $600^{\circ}C$ were exhibited the typical perovskite structures regardless of La contents. When the $(Pb,La)TiO_3$(PT) films were doped with La concentration up to 10mol%(PLT-10), the degree of z-axis orientation was greatly decreased from 63% to 26%. From AES depth profiles for the PLT-10 samples, no remarkable inter-reaction between PLT film and lower Pt electrode was found. The remanent polarization$(2Pr,Pr_+-Pr_-)$ were increased from $4\muC\textrm{cm}^2 to 16\muC\textrm{cm}^2$ as the annealing temperature increased from $600^{\circ}C to 700^{\circ}C$. This result may be ascribed to the improvement of crystallinity by the high temperature post-annealing. The dielectric constant$({\varepsilon}r)$ and tangent loss(tan$\delta$) of the PLT-10 films annealed at $650^{\circ}C$ were about 193 and 0.02, respectively with the pyroelectric coefficient($\gamma$) of around $4.0nC/\textrm{cm}^2{\cdot}^{\circ}C at 30^{\circ}C$.
The study investrgated the properti es of lh$\xi$ dust~ from felToalloy manufacture, The chemical composition, composItron material, particle size and shapes of the bulk dust, sized dust and magnetically separated dust were llivestigated. As the result, we s suppose that the dust from HLgh Carbon Fenomauganesc Manufacturing Process is not sufficient as soource material of Mn because of the low Mn conteut (13.5%) aud complicated composition material The dust from Bag Filter of AOD Process is m mainly made up of $0.2~2\mu\textrm{m}$$Mn_3O_4$ (Hausmatmite) particle in spherical shape and the Mn content is 63.1%. The dust from Cooler of AOD Process is mainly made up of coarse $Ca(OH)_2$ Mn, $Fe_yO_2$$SiO_2$ and fine $Mn_3O_1$.
The purpose of this study was to compare microleakage and marginal hybrid layer in class V restorations using two one-bottle adhesives and one self-etching adhesive. Class V cavity preparations with occlusal margins in enamel and gingival margins in dentin were pre-pared on buccal and lingual surfaces of 30 extracted human molar teeth. Prepared teeth were randomly divided into three treatment groups (n=30) and restored with three adhesives and composites: Single Bond/Filtek Z-250 (Group 1), Prime&BondNT/Esthet.X (Group 2), UniFil Bond/UniFil F (Group 3). For microleakage, samples were stored in room temperature water for 24 hours, thermocycled stained with 2% methylene blue dye, sectioned into halves, scored and analysed using Mann-whitney test and Wilcoxon signed rank sum test. For marginal hybrid layer, samples were sectioned into halves, treated with 10% phosphoric acid for 5 seconds, stored in 5% NaOCL solution for 24 hours, dried and gold coated. Occlusal and gingival margins of each sample were inspected under SEM. The results of this study were as follows ; 1. Microleakage at the occlusal margins was not evident in group 1 and group 2, but it showed in group 3 (p<0.05). 2. Microleakage in group 1 and group 3 was significantly lower than in group 2 at gingival margins (p<0.05). 3. Microleakage at gingival margins was greater than at occlusal margins in group 1 and group 2, but microleakage at occlusal margins was greater than at gingival margins in group 3 (p<0.05). 4. In group 1 and group 2, no gaps at occlusal margins showed. But gaps showed in group 3. Occlusal margins were free from a hybrid layer in all groups 5. The thickness of the marginal hybrid layers was 2.5~5 $\mu\textrm{m}$ thick in group 5 $\mu\textrm{m}$ thick in group 2 and 1.5 $\mu\textrm{m}$ thick in group 3. 6 There was no corelation between microleakage and thickness of marginal hybrid layer. In coclusion, the effect of dentin adhesives on microleakge in class V composite restorations was excellent when one-bottle adhesives were applied on enamel margin, and it was good when a self-etching adhesive was applied on dentinal margin. There was no corelation between microleakage and thickness of marginal hybrid layer.
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