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Embryogenesis and plant regeneration of Panax ginseng Meyer via anther culture and ploidy assessment using flow cytometry (인삼 약 배양을 통한 배 발생과 식물 재분화 및 유세포 분석기를 이용한 배수성 검정)

  • Jung-Woo Lee;Kyong-Hwan Bang;Dong-Hwi Kim;Jang-Uk Kim;Young-Chang Kim;Ick-Hyun Jo
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.19-26
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    • 2023
  • Korean ginseng (Panax ginseng Meyer) is an economically important plant because of it is rich in saponins. It is mainly cultivated in Asia, including Korea and China. Since ginseng requires a long breeding period due to juvenility, homozygote production techniques, such as anther culture, must be urgently established. In the present study, callus induction and embryogenesis through anther culture were observed in P. ginseng. Murashige and Skoog medium was used as the basal medium suitable for callus induction. When the medium was supplemented with 3% sucrose, the callus induction rate was high and the callus size was large. Cold pretreatment did not significantly affect callus induction and embryogenesis. Embryogenesis was the most efficient when the embryo-formation medium was supplemented with 1.0 or 3.0 mg/L 2,4-dichlorophenoxyacetic acid. Cultivar significantly affected anther culture efficiency. Specifically, 'Cheongseon' showed the highest embryo-formation efficiency, whereas no embryogenesis occurred in 'Sunun'. Ploidy assessment revealed the haploid status of the induced calli. Embryos derived from anther culture formed shoots upon transfer to germination medium, although no difference in ploidy was noted between the induced callus and control. Overall, the anther culture conditions established in the present study may contribute to the production of homozygous P. ginseng plants in the future.

Effects of Cardiotonic Pills® on Cerebrovascular CO2 Reactivity and Erythrocyte Deformability in Normal Subjects: A Pilot Study

  • Sang-Kwan Moon;Han-Gyul Lee;Seungwon Kwon;Seung-Yeon Cho;Seong-Uk Park;Woo-Sang Jung;Jung-Mi Park;Chang-Nam Ko;Ki-Ho Cho
    • The Journal of Korean Medicine
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    • v.44 no.4
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    • pp.87-103
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    • 2023
  • Backgrounds and objectives: Cardiotonic Pills® (CP) are used for vascular diseases such as coronary diseases, atherosclerosis, and cerebral infarction. This study aimed to determine the transient effects of CP on cerebrovascular CO2 reactivity (CVR) and erythrocyte deformability in normal subjects. Methods: This study had a crossover design and included 10 participants who were randomly allocated to 2 groups. The experimental group was given CP with water, while the control group was given only water. CVR was measured by hyperventilation-induced CVR of the middle cerebral artery (MCA) using transcranial Doppler (TCD). Erythrocyte deformability was measured using a Rheoscan-D microfluidic ektacytometer. All measurements were performed prior to and 1, 2, and 3 hours after CP or water administration. Blood pressure and heart rate were also measured before and after administration. Results: CP significantly improved CVR 3 hours after administration in the experimental group compared to the control group (p = 0.042). The corrected blood flow velocity at partial pressure of end-tidal carbon dioxide (PETCO2) = 40mmHg (CV40) was also significantly improved 2 and 3 hours after administration in the CP group compared to the control group (p = 0.036 and p = 0.021, respectively). CP significantly improved erythrocyte deformability 3 hours after administration in the experimental group compared to the control group (p = 0.027). Mean heart rate and mean blood pressure showed no change. Conclusions: This study demonstrated that CP increases CVR and erythrocyte deformability. These results suggested that CP improves cerebral microcirculation which provide evidence for the future use of CP for prevention of ischemic stroke and neurodegenerative diseases.

Effect of Trichostatin A on Anti HepG2 Liver Carcinoma Cells: Inhibition of HDAC Activity and Activation of Wnt/β-Catenin Signaling

  • Shi, Qing-Qiang;Zuo, Guo-Wei;Feng, Zi-Qiang;Zhao, Lv-Cui;Luo, Lian;You, Zhi-Mei;Li, Dang-Yang;Xia, Jing;Li, Jing;Chen, Di-Long
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.18
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    • pp.7849-7855
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    • 2014
  • Purpose: To investigate the effect of deacetylase inhibitory trichostatin A (TSA) on anti HepG2 liver carcinoma cells and explore the underlying mechanisms. Materials and Methods: HepG2 cells exposed to different concentrations of TSA for 24, 48, or 72h were examined for cell growth inhibition using CCK8, changes in cell cycle distribution with flow cytometry, cell apoptosis with annexin V-FTIC/PI double staining, and cell morphology changes under an inverted microscope. Expression of ${\beta}$-catenin, HDAC1, HDAC3, H3K9, CyclinD1 and Bax proteins was tested by Western blotting. Gene expression for ${\beta}$-catenin, HDAC1and HDAC3 was tested by q-PCR. ${\beta}$-catenin and H3K9 proteins were also tested by immunofluorescence. Activity of Renilla luciferase (pTCF/LEF-luc) was assessed using the Luciferase Reporter Assay system reagent. The activity of total HDACs was detected with a HDACs colorimetric kit. Results: Exposure to TSA caused significant dose-and time-dependent inhibition of HepG2 cell proliferation (p<0.05) and resulted in increased cell percentages in G0/G1 and G2/M phases and decrease in the S phase. The apoptotic index in the control group was $6.22{\pm}0.25%$, which increased to $7.17{\pm}0.20%$ and $18.1{\pm}0.42%$ in the treatment group. Exposure to 250 and 500nmol/L TSA also caused cell morphology changes with numerous floating cells. Expression of ${\beta}$-catenin, H3K9and Bax proteins was significantly increased, expression levels of CyclinD1, HDAC1, HDAC3 were decreased. Expression of ${\beta}$-catenin at the genetic level was significantly increased, with no significant difference in HDAC1and HDAC3 genes. In the cytoplasm, expression of ${\beta}$-catenin fluorescence protein was not obvious changed and in the nucleus, small amounts of green fluorescence were observed. H3K9 fluorescence protein were increased. Expression levels of the transcription factor TCF werealso increased in HepG2 cells following induction by TSA, whikle the activity of total HDACs was decreased. Conclusions: TSA inhibits HDAC activity, promotes histone acetylation, and activates Wnt/${\beta}$-catenin signaling to inhibit proliferation of HepG2 cell, arrest cell cycling and induce apoptosis.

The Clinical Experiences of Patch Angioplasty in Isolated Critical Left Main Coronary Artery Stenosis (첨포를 이용한 좌주관상동맥 협착증의 치료)

  • 윤치순;유경종;이교준;김대준;강면식
    • Journal of Chest Surgery
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    • v.31 no.7
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    • pp.674-678
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    • 1998
  • The conventional surgical treatment of isolated critical stenosis of the left main coronary artery restores a less physiologic perfusion of the myocardium, leads to occlusion of the left coronary ostium, and consumes an appreciable length of bypass material. From June 1994 to February 1996, eleven patients, three male and eight female, underwent patch angioplasty and additional bypass graft to left anterior descending artery (10 internal mammary artery, 1 saphenous vein) in isolated critical left main coronary artery stenosis. Their ages ranged from 34 to 62 years, mean 44 years. All had 60% to 90% stenosis of the left main coronary artery and Class III angina. The angiogram showed nine osteal lesion and three main stem stenosis. The operation was performed with conventional cardiopulmonary bypass and cold blood cardioplegia. We approached anteriorly and used bovine pericardium as onlay patch in all patients. There were one leg wound dehiscence, but no operative deaths and infarctions. All patients are free of symptoms after a mean follow-up of 15.5 months. Angiographic restudy at an average 14.4 months was obtained in five patients and showed widely patent left main coronary artery with excellent runoff. But additional graft to left anterior descending coronary artery were stenosed in two patients and showed diminutive flow in others. Our preliminary results suggest that angioplasty of the left main coronary artery can be carried out with low operative risks. But additional bypass graft to left anterior descending coronary artery may be unnecessary. The technique appears to be a promising alternative to conventional coronary artery bypass grafting in isolated left main coronary artery stenosis.

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The Study on the Increased Causes of Chloride ($Cl^{-}$) Concentration of the Samyang 3rd Pumping Station in Cheju Island (제주도 삼양 3수원지의 염소이온농도 상승 원인에 관한 연구)

  • 이성복;김구영;한소라;한정상
    • Journal of the Korean Society of Groundwater Environment
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    • v.4 no.2
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    • pp.85-94
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    • 1997
  • The study is aimed to find out the causes of rapidly increasing chloride (Cl$^{-}$) concentration of the Samyang 3rd pumping station originated from coastal springs of Cheju since January 1996. The study results show that it was caused by following complicated natural and anthropogenic effects. Due to severe draught in 1996 with total rainfall of only 41.7% of annual mean of the last 36 years (1991 to 1995), it creates firstly), significant decrease of the spring discharges as well decline of the groundwater level at the site . Sea water level was in general 4.4 cm to 12.4 cm higher than the groundwater level of the site during 2 to 3.8 hours at each high tide. Those higher potential head of sea water motivates the sea water intrusion into the fresh water lens through the permeable clinkers and fracture zones situated beneath the existing grouted zone which was installed to a maximum 10 m below the ground water surface, The repeated expansion and contraction of the fresh water lens occurred by periodic changes of the sea water level at high and low tide accelerates secondly the enlargement of the transition zone between the fresh and sea water at the site. The decrease of recharge amount by rainfall shortage creates thirdly the reverse flow at the interface of sea water and groundwater. The repeated groundwater extraction of 2790${\pm}$450 $m^3$d$^{-1}$ at the time of low tide, when the fresh water lens of the sire is under the contraction stare, makes additional drawdown of the ground water level and induces the upconing of salt water into the fresh water lens. The duration of spring discharge whose Cl concentration is less than 150 mg/1 at the low tide measured at the nearby springs was about two hours with discharge rate of 532 $m^3$d$^{-1}$ and after that Cl$^{-}$ concentration is increased up to more than 1900 mg/ι.eased up to more than 1900 mg/L.

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Effects of Water Temperature and Salinity on the Oxygen Consumption Rate of Juvenile Spotted Sea Bass, Lateolabrax maculatus (점농어, Lateolabrax maculatus 치어의 산소 소비율에 미치는 수온과 염분의 영향)

  • Oh, Sung-Yong;Shin, Chang Hoon;Jo, Jae-Yoon;Noh, Choong Hwan;Myoung, Jung-Goo;Kim, Jong-Man
    • Korean Journal of Ichthyology
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    • v.18 no.3
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    • pp.202-208
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    • 2006
  • An experiment was conducted to investigate the effects of three water temperatures (15, 20 and $25^{\circ}C$) in combination with three salinities (0, 15 and 30 psu) on the oxygen consumption rate of juvenile spotted sea bass, Lateolabrax maculatus (mean body weight $5.5{\pm}0.3g$). The oxygen consumption rates of L. maculatus were measured in triplicate for 24 hours using a continuous flow-through respirometer. Water temperature resulted in significant differences in the mean oxygen consumption rate of L. maculatus (p<0.001), but salinity and combinations of salinity and water temperature did not have (p>0.05). The oxygen consumption increased with increasing water temperatures in all experimental salinity regimes (p<001). Mean oxygen consumption rates at 15, 20 and $25^{\circ}C$ ranged 328.8~342.3, 433.9~441.0 and 651.5~659.9 mg $O_2\;kg^{-1}\;h^{-1}$, respectively. $Q_{10}$ values did not vary with salinity, bud varied with water temperature. $Q_{10}$ values ranged 1.63~1.75 between 15 and $20^{\circ}C$, 2.24~2.26 between 20 and $25^{\circ}C$, and 1.92~1.98 over the full temperature range. The energy loss by metabolic cost increased with increasing water temperatures in all experimental salinity regimes (p<0.001) Mean energy loss rates at 15, 20 and $25^{\circ}C$ ranged 224.6~233.8, 296.3~301.2 and $444.9{\sim}450.7kJ\;kg^{-1}\;d^{-1}$, respectively. These data suggest that the culture of juvenile spotted sea bass is possible without energy loss by salinity difference in freshwater as well as seawater after salinity acclimation. Thus, this result has an application for culture management and bioenergetic model for growth of this species.

Hydro-Mechanical Modelling of Fault Slip Induced by Water Injection: DECOVALEX-2019 TASK B (Step 1) (유체 주입에 의한 단층의 수리역학적 거동 해석: 국제공동연구 DECOVALEX-2019 Task B 연구 현황(Step 1))

  • Park, Jung-Wook;Park, Eui-Seob;Kim, Taehyun;Lee, Changsoo;Lee, Jaewon
    • Tunnel and Underground Space
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    • v.28 no.5
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    • pp.400-425
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    • 2018
  • This study presents the research results and current status of the DECOVALEX-2019 project Task B. Task B named 'Fault slip modelling' is aiming at developing a numerical method to simulate the coupled hydro-mechanical behavior of fault, including slip or reactivation, induced by water injection. The first research step of Task B is a benchmark simulation which is designed for the modelling teams to familiarize themselves with the problem and to set up their own codes to reproduce the hydro-mechanical coupling between the fault hydraulic transmissivity and the mechanically-induced displacement. We reproduced the coupled hydro-mechanical process of fault slip using TOUGH-FLAC simulator. The fluid flow along a fault was modelled with solid elements and governed by Darcy's law with the cubic law in TOUGH2, whereas the mechanical behavior of a single fault was represented by creating interface elements between two separating rock blocks in FLAC3D. A methodology to formulate the hydro-mechanical coupling relations of two different hydraulic aperture models and link the solid element of TOUGH2 and the interface element of FLAC3D was suggested. In addition, we developed a coupling module to update the changes in geometric features (mesh) and hydrological properties of fault caused by water injection at every calculation step for TOUGH-FLAC simulator. Then, the transient responses of the fault, including elastic deformation, reactivation, progressive evolutions of pathway, pressure distribution and water injection rate, to stepwise pressurization were examined during the simulations. The results of the simulations suggest that the developed model can provide a reasonable prediction of the hydro-mechanical behavior related to fault reactivation. The numerical model will be enhanced by continuing collaboration and interaction with other research teams of DECOLVAEX-2019 Task B and validated using the field data from fault activation experiments in a further study.

Effect of irradiation on cell cycle, cell death and expression of its related proteins in normal human oral keratinocytes (방사선조사가 사람 정상 구강각화 세포의 세포주기, 세포사 및 수종 단백질의 발현에 미치는 영향)

  • Kang Mi-Ae;Heo Min-Suk;Lee Sam-Sun;Oh Sung-Ock;Lee Sul-Mi;Jeon In-Seong;Choi Soon-Chul;Park Tae-Won
    • Imaging Science in Dentistry
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    • v.33 no.3
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    • pp.143-149
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    • 2003
  • Purpose: To investigate the radiosensitivity of the normal human oral keratinocytes (NHOK), and the effect of irradiation on cell cycle and protein expression. Materials and Methods: To evaluate the radiosensitivity of NHOK, the number of colonies and cells were counted after irradiation and the SF2 (survival fraction at 2Gy) value, and the cell survival curve fitted on a linear-quadratic model were obtained. LDH analysis was carried out to evaluate the necrosis of NHOK at 1, 2, 3, and 4 days after 2, 10, and 20 Gy irradiation. Cell cycle arrest and the induction of apoptosis were analyzed using flow cytometry at 1, 2, 3, and 4 days after 2, 10, and 200y irradiation. Finally, proteins related cell cycle arrest and apoptosis were analysed by Western blot. Results: The number of survived cell was significantly decreased in a dose-dependent manner. The cell survival curve showed SF2, α, and β values to be 0.568, 0.209, and 0.020 respectively. At 200y irradiated cells showed higher optical density than the control group. After irradiation, apoptosis was not observed but G2 arrest was observed in the NHOK cells. 1 day after 10 Gy irradiation, the expression of p53 remained unchanged, the p2l/sup WAF1/Cipl/ increased and the mdm2 decreased. The expression of bax, bcl-2, cyclin B1, and cyclin D remained unchanged. Conclusion: These results indicate that NHOK responds to irradiation by G2 arrest, which is possibly mediated by the expression of p21/sup WAFl/Cipl/, and that cell necrosis occurs by high dose irradiation.

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The effects of PEP-1-FK506BP on dry eye disease in a rat model

  • Kim, Dae Won;Lee, Sung Ho;Ku, Sae Kwang;Lee, Ji Eun;Cha, Hyun Ju;Youn, Jong Kyu;Kwon, Hyeok Yil;Park, Jong Hoon;Park, Eun Young;Cho, Sung-Woo;Han, Kyu Hyung;Park, Jinseu;Eum, Won Sik;Choi, Soo Young
    • BMB Reports
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    • v.48 no.3
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    • pp.153-158
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    • 2015
  • As FK506 binding proteins (FK506BPs) are known to play an important role in the regulation of a variety of biological processes related to cell survival, this study was designed to examined the protective effects of FK506 binding protein 12 (FK506BP) on low humidity air flow induced dry eye in a rat model using transduced PEP-1-FK506BP. After the topical application of PEP-1-FK506BP, tear volumes were markedly increased and significant prevention of cornea damage was observed compared with dry eye rats. Further, immunohistochemical analysis demonstrated that PEP-1-FK506BP markedly prevented damage to the cornea, the bulbar conjunctiva, and the palpebral conjunctiva epithelial lining compared with dry eye rats. In addition, caspase-3 and PARP expression levels were found to be decreased. These results demonstrated that topical application of PEP-1-FK506BP significantly ameliorates dry eye injury in an animal model. Thus, we suggest that PEP-1-FK506BP can be developed as a new ophthalmic drop to treat dry eye diseases.

Interleukin-7 Enhances the in Vivo Anti-tumor Activity of Tumor-reactive CD8+ T cells with Induction of IFN-gamma in a Murine Breast Cancer Model

  • Yuan, Chun-Hui;Yang, Xue-Qin;Zhu, Cheng-Liang;Liu, Shao-Ping;Wang, Bi-Cheng;Wang, Fu-Bing
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.265-271
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    • 2014
  • Interleukin-7 (IL-7) is a potent anti-apoptotic cytokine that enhances immune effector cell functions and is essential for lymphocyte survival. While it known to induce differentiation and proliferation in some haematological malignancies, including certain types of leukaemias and lymphomas, little is known about its role in solid tumours, including breast cancer. In the current study, we investigated whether IL-7 could enhance the in vivo antitumor activity of tumor-reactive $CD8^+$ T cells with induction of IFN-${\gamma}$ in a murine breast cancer model. Human IL-7 cDNA was constructed into the eukaryotic expression plasmid pcDNA3.1, and then the recombinational pcDNA3.1-IL-7 was intratumorally injected in the TM40D BALB/C mouse graft model. Serum and intracellular IFN-${\gamma}$ levels were measured by ELISA and flow cytometry, respectively. $CD8^+$ T cell-mediated cytotoxicity was analyzed using the MTT method. Our results showed that IL-7 administration significantly inhibited tumor growth from day 15 after direct intratumoral injection of pcDNA3.1-IL-7. The anti-tumor effect correlated with a marked increase in the level of IFN-${\gamma}$ and breast cancer cells-specific CTL cytotoxicity. In vitro cytotoxicity assays showed that IL-7-treatment could augment cytolytic activity of $CD8^+$ T cells from tumor bearing mice, while anti-IFN-${\gamma}$ blocked the function of $CD8^+$ T cells, suggesting that IFN-${\gamma}$ mediated the cytolytic activity of $CD8^+$ T cells. Furthermore, in vivo neutralization of $CD8^+$ T lymphocytes by CD8 antibodies reversed the antitumor benefit of IL-7. Thus, we demonstrated that IL-7 exerts anti-tumor activity mainly through activating $CD8^+$ T cells and stimulating them to secrete IFN-${\gamma}$ in a murine breast tumor model. Based on these results, our study points to a potential novel way to treat breast cancer and may have important implications for clinical immunotherapy.