• Journal of Plant Biotechnology
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• 제6권3호
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• pp.145-150
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• 2004

Cabbage plants were transformed with the potato proteinase inhibitor II (PINII) gene, bar gene, and hpt gene using Agrobacterium. The expression of the PINII gene was driven by its own promoter which was wound-inducible. Ten transgenic plants were obtained from medium containing hygromycin as a selection antibiotic. The integration and expression of PINII and bar genes were confirmed by Southern and Northern hybridization. Growth and development of diamondback moths (Plutella xylostella) and tobacco cutworm (Spodoptera litura) larvae were examined on $T_1$ plants. The weight of the larvae and pupae of these two insects grown on transgenic plants was not different compared to those grown on wild type plants. However, the pupation and emergence rate of diamondback moths and tobacco cutworms fed on some transgenic plants was lower than on wild type plants. These results suggest that the PINII transgene under the control of a wound-induced promoter may be used for control of insects in transgenic cabbage through reduction of insect progeny number.

• 한방재활의학과학회지
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• 제33권2호
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• pp.1-18
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• 2023

Objectives The purpose of this study was to evaluate the antioxidant, anti-inflammatory and wound healing effects of Samul-tang-ga-dansam water extract (SD) in wound-induced mice. Methods The mice were divided into five groups (n=7): the normal group, the control group, the positive control group, the low-dose SD group and the high-dose SD group. The normal group had no wounds and the other groups were wounded on the back with a leather punch. Distilled water was administered to the control group, 200 mg/kg of vitamin E was administered to the positive control group. In the low-dose SD group and the high-dose SD group, 1.23 g/kg and 2.47 g/kg of SD were administered, respectively. Antioxidant and anti-inflammatory protein levels were evaluated using western blot analysis. Skin tissue was analyzed by H&E, Masson's trichrome staining method. Results Oral administration of the SD significantly reduced the visible skin damage and decreased the reactive oxygen species and ONOO^- activity of the serum. It significantly increased heme oxygenase-1, superoxide dismutase, catalase, GPx-1/2, Nrf2 and Keap-1 which are antioxidant-related factors in skin tissue and reduced NF-κB p65, inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor α, interleukin (IL)-1β, IL-6 which are inflammation-related factors. Also, SD significantly decreased NOX2, p22^phox and p47^phox and increased α-smooth muscle actin and COL1A1 protein expression in fibroblasts involved in connective tissue repair. According to histological examination, the thickened epithelial layer was thinned and collagen fibers were increased to accelerate wound healing. Conclusions It is suggested that Samul-tang-ga-dansam has antioxidant and anti-inflammatory effects and promotes wound tissue repair.

• 한국환경과학회지
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• 제8권6호
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• pp.669-676
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• 1999

The effects of wounding and jasmonic acid(JA) on polyphenol oxidase(PPO) in tomato(Lycopersicon esculentum Mill.) seedlings were investigated. PPO was strongly induced by wounding or JA, and the response was also shown to be systemically induced by wounding. Mechanical wounding in cotyledon or leaf produced a signal that caused the concentration of PPO to increase in the unwounded cotyledon, in the first leaves but not in the second leaves. Severity of wounding and light intensity also affected wound induced change in PPO activity, JA showed a stimulatory effect on the loss of chlorophyll and the rapid increase in PPO activity. The PPO was clearly more active in the wounded leaves than in controls. The potency and specificity of the JA indicate a close relationship between JA and wound-induced changes in PPO in tomato species. JA and abscisic acid(ABA) acted similarly on both unwounded and wounded leaves, but the amount of PPO in the wounded leaves was always more than the respective controls. The highest increase in PPO activity occurred in woundand JA-induced leaves of seedlings kept under bright lighting. Benzyladenine(BA) completely abolished JA- and ABA-induced PPO activity. The results suggest that JA-induced PPO activity is due to de novo PPO synthesis. Histochemical tests for PPO in stems of wound- and JA -treated tomato plants indicate that PPO was localized primarily, in the. outer .cortex . and xylem parenchyma. It is concluded that exogenously applied JA acts as stress agents and PPO may be a component of the inducible anti-hervivore defense response.

• Preventive Nutrition and Food Science
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• 제2권4호
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• pp.333-337
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• 1997

Trans-cinnamic acid-4-hydroxylase(tC4H) is the first cytochrome P450-dependent monooxygenase of the phenylpropanoid pathway. The roots of avocado seedlings were wounded and examined to determine whether the tC4H would be activated in response to wounding and/or whether tC4H activity be modulated by the application of exogenous p-coumarate. At the specified length of times, the wounded and treated roots were either frozen in liquid nitrogen or used immediately to extract microsomal proteins. The microsomal proteins were subjected to immunoblot analysis using polyclonal antibodies against CYP73 of tC4H gene. In this study, tC4H was induced in wounded roots sealed in bags within 6 hours, and in low level({TEX}$10^{-8}${/TEX}M) of p-coumarate solution within 24 hours, whereas the olution without p-coumarate and high levels of p-coumarate solution repressed tC4H induction in wounded roots. These results indicate that tC4H is induced by wounding in the root of avocado, and is inhibited by the application of exogenous p-coumarate.

• 대한한방내과학회지
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• 제34권1호
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• pp.31-45
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• 2013

Objectives : Gokgisaeng (Korean mistletoe) is used for the treatment of inflammatory and cancer diseases in traditional Korean medicine and its major component lectins have been reported to induce nitric oxide (NO) in RAW 264.7 macrophages, and also induce apoptosis of various types of cancer cells, although its modulatory effects on cancer cell migration and macrophage activation is poorly understood. The aim of this study is to clarify molecular mechanisms of action responsible for the anti-inflammatory and antitumor migration potentials of Korean mistletoe extract (KME). Methods : We investigated the anti-inflammatory activity of KME on NO production and inducible nitric oxide synthase (iNOS) expression by lipopolysaccharide (LPS) in both RAW 264.7 macrophages and rat C6 glioma cells, and also evaluated inhibitory efficacy on glioma cell growth and migration. For assessment, XTT assay, nitrite assay, RT-PCR, scratch-wound and Boyden chamber assay, and western blot analysis were performed. Results : Previously reported, unlike the efficacy of Gokgisaeng lectin, KME inhibited NO production and iNOS expression, and suppressed pro-inflammatory mediators including IL-$1{\beta}$, IL-6, COX-2, iNOS in LPS-stimulated RAW 264.7 cells. Furthermore, KME suppressed tumor cell growth and migration, and it also inhibited LPS-induced NO release and iNOS activation by down-regulating expression of protein kinase C (PKC) and phosphorylation of ERK in C6 glioma cells. Conclusions : Our research findings provide evidence that KME can play a significant role in blocking pro-inflammatory reaction and malignant progression of tumors through the suppression of NO/iNOS by down-regulating of inflammatory signaling pathways, PKC/ERK.

• Journal of Ginseng Research
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• 제42권3호
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• pp.288-297
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• 2018

Background: The incidence of colorectal cancer (CRC) is increasing, with metastasis of newly diagnosed CRC reported in a large proportion of patients. However, the effect of Korean Red Ginseng extracts (KRGE) on epithelial to mesenchymal transition (EMT) in CRC is unknown. Therefore, we examined the mechanisms by which KRGE regulates EMT of CRC in hypoxic conditions. Methods: Human CRC cell lines HT29 and HCT116 were incubated under hypoxic (1% oxygen) and normoxic (21% oxygen) conditions. Western blot analysis and real-time PCR were used to evaluate the expression of EMT markers in the presence of KRGE. Furthermore, we performed scratched wound healing, transwell migration, and invasion assays to monitor whether KRGE affects migratory and invasive abilities of CRC cells under hypoxic conditions. Results: KRGE-treated HT29 and HCT116 cells displayed attenuated vascular endothelial growth factor (VEGF) mRNA levels and hypoxia-inducible $factor-1{\alpha}$ ($HIF-1{\alpha}$) protein expression under hypoxic conditions. KRGE repressed Snail, Slug, and Twist mRNA expression and integrin ${\alpha}V{\beta}6$ protein levels. Furthermore, hypoxia-repressed E-cadherin was restored in KRGE-treated cells; KRGE blocked the invasion and migration of colon cancer cells by repressing $NF-{\kappa}B$ and ERK1/2 pathways in hypoxia. Conclusions: KRGE inhibits hypoxia-induced EMT by repressing $NF-{\kappa}B$ and ERK1/2 pathways in colon cancer cells.

• 한국자원식물학회지
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• 제24권5호
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• pp.584-590
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• 2011

의성종 마늘의 유묘로부터 상처(wound)에 특이적으로 발현되는 cytochrome P450 유전자군의 하나인 P450-Esg cDNA를 탐색하였다. P450-Esg는 1,419 bp의 open reading frame(ORF)을 가지고 473개의 아미노산을 가진 polypeptide를 코딩하는 것으로 나타났다. 국내와 몽골로부터 수집한 12개의 재배종으로 부터 P450-Esg 유사 유전자의 염기서열을 비교한 결과 시작코돈(ATG)에서 472~510 bp 및 1,210~1,240 bp 부위의 염기에서 재배종간에 차이를 보이는 염기서열을 다수 확인하였다. cDNA 1,210~1,240 bp의 부위는 P450 유전자에서 공통적으로 알려진 heme binding domain으로, 각 지역에서 수집된 재배종은 염기서열뿐만 아니라 아미노산 서열에 있어서도 특이적인 변이를 보였다. cDNA 472~510 bp 부위에서 코딩하는 13개 아미노산의 서열은 12개 재배종에서 모두 동일하였으나, 13개 아미노산 중 7개에서 재배종 마다 각각 다른 DNA 염기로 코딩하는 단일 염기다형성(single nucleotide polymorphism)을 보이는 서열을 확인하였다. 이 결과는 다양하게 존재하는 국내외 마늘 재배종을 구분하는 marker로 사용될 것이며, 외국산 마늘에 대한 유전적 우선권을 확보하는 수단으로 사용될 것이다.

• Preventive Nutrition and Food Science
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• 제2권4호
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• pp.328-332
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• 1997

The ripe fruit of Hass avocado contains one of the highest elvels of cytochrome P-450 protein found in the plant kingdom. To determine whether wounded roots of avocado contain P-450 protein, the roots of avocado were wounded by slicing, and then allowed to incubate in sealed plastic bags, in 0.4M mannitol, and in the solution to make protoplast preparation containing cellulysin and macerase during the specified times. The microsomal proteins were extracted from the samples, separated by SDS-PAGE, and then subjected to Western blot analysis using polyclonal antibodies which are generated against the CYP71A1 protein. wounded roots in sealed bags produced CYP71A1 within 6 hours after cutting, and those in 0.4M mannitol did not produce CYP71A1 even after 72 hours, but those in the protoplast preparation by cellulysin and macerase induced and produced CYP71A was induced in only 24 hours. These results indicate that CYP 71A1 plays a role for wound healing for root tissue o avocado, and would-inducible P-450 protein was not detected in the mannitol solution by preventing a synthesis of ethylene in a liquid state, but the softening of tissues by cellulysin and macerase to make protoplast preparation was involved in an activation of CYP 71A1 even in the liquid state.

• The Plant Pathology Journal
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• 제15권1호
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• pp.8-13
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• 1999

Hypersensitive cell death of plants during incompatible plant-pathogen interactions is one of the efficient defense mechanisms of plants against pathogen infections. For better understanding of the molecular mechanisms involved in the plant hypersensitive response (HR), TMV-induced biotic plant cell death and CuSO4-induced abiotic plant cell death were compared in terms of expression patterns of ten different defense-related genes as molecular markers. The genes include five pathogenesis-related protein genes, two plant secondary metabolite-associated genes, two oxidative stress-related genes and one wound-inducible gene isolated from tobacco. Northern blot analyses revealed that a same set of defense-related genes was induced during both biotic and abiotic cell death but with different time and magnitude. The expression of defense-related genes in tobacco plants was temporarily coincided with the time of cell death. However, when suspension cell cultures was used to monitor the expression of defense-related genes, different patterns of the gene expression were detected. This result implies that three are common and, in addition, also different branches of signaling pathways leading to the induced expression of defense-related genes in tobacco during the pathogen- and heavy metal-induced cell death.

• Journal of Plant Biotechnology
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• 제29권2호
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• pp.99-104
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• 2002

Suppression subtractive hybridization (SSH)를 통해 저온에 의해 발현이 유도되는 cDNA들을 분리하였고, 그 중 하나인 slti182는 asparaginase 유전자들과 높은 유사성을 보였다. Slti182의 전체 cDNA인 GmASP1은 1258 bp 길이에 326개 아미노산으로 구성된 긴 Open reading frame (ORF)를 포함하고 있었다. GmASP1 단백질은 애기장대의 putative asparaginase (AB012247)와 84%의 유사성을 보였으나, 애기장대의 다른 isoform(Z34884)와는 55%의 유사성을 나타내었다. GmASP1 유전자는 저온 처리 후 3시간째부터 발현이 유도되어 6시간째에 최대발현을 나타내었고, 이후 점차 감소하여 48시간에는 저온처리전 수준으로 되돌아왔다. 또한, 식물체를 저온에서 48시간 둔 후에 다시 상온으로 옮겼을 때, 정상수준으로 떨어진 GmASP1의 발현이 다시 증가하는 양상을 보였다. 이러한 결과로 볼 때, GmASP1이 저온 스트레스에 반응 초기의 단백질 합성 촉진에 중요한 역할을 할 것으로 사료된다.