• 한국수정란이식학회지
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• 제29권4호
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• pp.351-359
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• 2014

Phasianus colchicus is not only a beautiful bird but also a great value in science and under the threat of endanger. Hence, the aim of this study was to isolate the pheasant male germ cells (mGCs) and then induce them into elongated sperm-like cells in vitro. The mGCs were purified and enriched by a two-step plating method based on the different adherence velocities of mGCs and somatic cells. The percentage of the c-kit positive cells and c-kit negative cells examined by flow cytometry analysis (FCA) was 92.87% and 2.57%, respectively. Subsequently, the mGCs were induced for 48h in DMEM/F12 medium supplemented factors such as retinol acid, testosterone and bovine FSH, followed by 5 weeks in culture. We found that some elongated sperm-like cells appeared initially in vitro under inducement of stimulated factors. The elongated sperm-like cells showed in the expression of changed morphology and post-transcriptional marker such as spermatid associated (SPERT), spermatid perinuclear RNA binding protein (STRBP), round spermatid basic protein 1 (RSBN1) and SPER1L. Moreover, in DNA content identified assay, induced cells showed that the 1C DNA population markedly increased in differentiated group but it was not change in undifferentiated group. Successful in vitro differentiation of pheasant testicular germline cells into spermatids appears to offer extremely attractive potential for the conservation of endangered birds and treatment of male infertility.

• Macromolecular Research
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• 제16권6호
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• pp.510-516
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• 2008

New antitumor active polymers, poly(methacryloyl-2-oxy-1,2,3-propanetricarboxylic acid-co-exo-3,6-epoxy-l,2,3,6-tetrahydrophthalic acid) [poly(MTCA-co-ETAc)], poly(methacryloyl-2-oxy-l,2,3-propanetricarboxylic acid-co-hydrogen ethyl-exo-3,6-epoxy-l,2,3,6-tetrahydrophthalate) [poly(MTCA-co-HEET)], and poly(methacryloyl-2-oxy-l,2,3-propanetricarboxylic acid-co-a-ethoxy-exo-3,6-epoxy-1,2,3,6-tetrahydrophthaloyl-5-fluorouracil) [poly(MTCA-co-EETFU)] were synthesized and characterized. Their antitumor activity, inhibition of DNA replication and antiangiogenesis were examined. The structures of the polymers were identified by FT-IR, $^1H$ and $^{13}C$-NMR spectroscopy. The number average molecular weights of the fractionated polymers determined by GPC ranged from 9,400 to 14,900, and polydispersity indices were less than 1.7. The in vitro cytotoxicity of these polymers was determined and their antitumor activity was evaluated. The $IC_{50}$ values (the drug concentration at inhibition of 50% tumor growth) indicated that the synthesized polymers were much better inhibitors of cancer cells and showed lower cytotoxicity than the free 5-FU. The in vivo antitumor activity of the conjugates was examined using mice bearing the sarcoma 180 tumor cell line. The life spans (TIC) of the mice treated with the conjugates were higher than those treated with the free 5-FU. In addition, the synthesized conjugates showed excellent antiangiogenic activity based on an embryo chorioallantoic membrane assay.

• 대한한방부인과학회지
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• 제25권2호
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• pp.12-22
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• 2012

Objectives: The aim of this study is to investigate immune enhancing effect of Houttuyniae Herba water extract(HW) on RAW 264.7 cell of mouse macrophages. Methods: Effects of HW on productions of nitric oxide(NO) and hydrogen peroxide($H_2O_2$) in RAW 264.7 mouse macrophages were measured. Effect of HW on production of cytokines such as interleukin(IL)-$1{\beta}$, IL-6, and tumor necrosis factor(TNF)-${\alpha}$ in RAW 264.7 cells was accessed by a multiplex bead array assay based on xMAP technology. All of results were represented P<0.05 compared to the normal. Results: 1. After 24 hr incubation, HW increased significantly NO production in RAW 264.7 cells at the concentrations of 25, 50, 100 and 200 ${\mu}g$/mL. 2. After 24 hr incubation, HW increased significantly hydrogen peroxide production in RAW 264.7 cells at the concentrations of 25, 50, 100 and 200 ${\mu}g$/mL. 3. After 24 hr incubation, HW increased significantly IL-$1{\beta}$ production in RAW 264.7 cells at the concentrations of 100 and 200 ${\mu}g$/mL. 4. After 24 hr incubation, HW increased significantly IL-6 production in RAW 264.7 cells at the concentrations of 100 and 200 ${\mu}g$/mL. 5. After 24 hr incubation, HW increased significantly TNF-${\alpha}$ production in RAW 264.7 cells at the concentrations of 50, 100, and 200 ${\mu}g$/mL. Conclusions: These results suggest that HW has immune enhancing activity related with its increasement of NO, hydrogen peroxide, IL-$1{\beta}$, IL-6, and TNF-${\alpha}$ in macrophages.

• 한국어병학회지
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• 제17권1호
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• pp.1-10
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• 2004

해수 환경 중의 VHSV 분포 조사를 위하여, 2003년 2월에서 5월까지 동해 및 남해안 인근 해역에 서식하는 자연산 해수 어류를 채집하였다. 바이러스 분리용 시료는 9종의 해수 어류로서 42개의 시료를 분석하였다. 각 시료의 조직여과액을 Epithelioma papulosum cyprini (EPC) cells에 접종한 결과, 고등어 Scomber japonicus와 숭어 Mugil cephalus에서 바이러스가 분리되었으며 이들 바이러스는 RT-PCR법을 이용하여 VHSV로 동정할 수 있었다. 고등어에서 VHSV가 검출되었다는 기보고는 있지만, 숭어에 대해서는 본 연구에서 처음으로 VHSV를 분리하였다. 자연산 어류에서 유래된 세 개의 VHSV isolates에 대한 glycoprotein gene을 분석한 결과, 이들 바이러스는 양식 넙치에서 질병을 유발하는 VHSV isolates와 유사하며 모두 genogroup I (American type)에 속하는 것으로 밝혀졌다.

• 방사선산업학회지
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• 제5권1호
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• pp.15-20
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• 2011

Protein kinases are the most important drug targets for the treatment of numerous diseases. The involvement of protein kinase C (PKC) in many biological processes such as development, memory, cell differentiation, and proliferation has been demonstrated. PKC is recognized as an important player in carcinogenesis. Thus, a variety of PKC inhibitors have been investigated. Among them, flavonoids have been demonstrated to affect the activity of many mammalian in vitro enzyme systems. The recent investigation was performed to evaluate the inhibitory effects of hesperidin, which is a flavonoid, on the proliferation and carcinogenesis of many cancers. In this study, an efficient kinase assay based on a biochip using radio-phosphorylation was established and performed for an examination of the inhibitory effects of hesperidin on PKC activity at different concentrations of 50, 200, 500 nM. It was found that hesperidin shows inhibitory potency on PKC, and that the biochip can be used to rapidly screen kinase inhibitors resulting in the therapeutic agents.

• Biomolecules & Therapeutics
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• 제27권2호
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• pp.193-200
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• 2019

Ceramide metabolism is known to be an essential etiology for various diseases, such as atopic dermatitis and Gaucher disease. Glucosylceramide synthase (GCS) is a key enzyme for the synthesis of glucosylceramide (GlcCer), which is a main ceramide metabolism pathway in mammalian cells. In this article, we developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to determine GCS activity using synthetic non-natural sphingolipid C8-ceramide as a substrate. The reaction products, C8-GlcCer for GCS, could be separated on a C18 column by reverse-phase high-performance liquid chromatography (HPLC). Quantification was conducted using the multiple reaction monitoring (MRM) mode to monitor the precursor-to-product ion transitions of m/z $588.6{\rightarrow}264.4$ for C8-GlcCer at positive ionization mode. The calibration curve was established over the range of 0.625-160 ng/mL, and the correlation coefficient was larger than 0.999. This method was successfully applied to detect GCS in the human hepatocellular carcinoma cell line (HepG2 cells) and mouse peripheral blood mononuclear cells. We also evaluated the inhibition degree of a known GCS inhibitor 1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) on GCS enzymatic activity and proved that this method could be successfully applied to GCS inhibitor screening of preventive and therapeutic drugs for ceramide metabolism diseases, such as atopic dermatitis and Gaucher disease.

• Journal of Dental Anesthesia and Pain Medicine
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• 제22권5호
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• pp.369-376
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• 2022

Background: Nonobstetric surgery is sometimes required during pregnancy, and neck abscess or facial bone fracture surgery cannot be postponed in pregnant women. However, dental surgery can be stressful and can cause inflammation, and the inflammatory response is a well-known major cause of preterm labor. Propofol is an intravenous anesthetic commonly used for general anesthesia and sedation. Studies investigating the effect of propofol on human amnion are rare. The current study investigated the effects of propofol on lipopolysaccharide (LPS)-induced inflammatory responses in human amnion-derived WISH cells. Methods: WISH cells were exposed to LPS for 24 h and co-treated with various concentrations of propofol (0.01-1 ㎍/ml). Cell viability was measured using the MTT assay. Nitric oxide (NO) production was analyzed using a microassay based on the Griess reaction. The protein expression of cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE 2), p38, and phospho-p38 was analyzed using western blotting. Results: Propofol did not affect the viability and NO production of WISH cells. Co-treatment with LPS and propofol reduced COX-2 and PGE₂ protein expression and inhibited p38 phosphorylation in WISH cells. Conclusion: Propofol does not affect the viability of WISH cells and inhibits LPS-induced expression of inflammatory factors. The inhibitory effect of propofol on inflammatory factor expression is likely mediated by the inhibition of p38 activation.

• 대한본초학회지
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• 제28권5호
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• pp.113-119
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• 2013

Objectives : The purpose of this study was to investigate the effects of Angelicae Gigantis Radix Water Extract(AG) on the production of proinflammatory mediators in RAW 264.7 cells stimulated with lipopolysaccharide(LPS). Method : RAW 264.7 cells were cotreated with AG(50 and 100 ug/mL) and lipopolysaccharide(LPS; 1 ug/mL) for 24 hours. After 24 hour treatment, using Bead-based multiplex cytokine assay, concentrations of various cytokines such as interleukin(IL)-6, IL-$1{\beta}$, IL-10, tumor necrosis factor-alpha(TNF-${\alpha}$), granulocyte colony-stimulating factor(G-CSF), granulocyte macrophage colony-stimulating factor(GM-CSF), interferon inducible protein-10(IP-10), leukemia inhibitory factor(LIF), lipopolysaccharide-induced chemokine(LIX), monocyte chemoattractant protein-1(MCP-1), macrophage colony-stimulating factor(M-CSF), macrophage inflammatory protein(MIP)-$1{\alpha}$, MIP-$1{\beta}$, MIP-2, Regulated on Activation, Normal T cell Expressed and Secreted(RANTES) and vascular endothelial growth factor(VEGF) were measured. Result : AG significantly inhibited LPS-induced production of TNF-${\alpha}$, MIP-$1{\alpha}$, G-CSF, RANTES, IL-10, and M-CSF from LPS-stimulated RAW 264.7 cells at the concentrations of 50 and 100 ug/mL. AG significantly inhibited LPS-induced production of MIP-$1{\beta}$, MIP-2, GM-CSF, and IL-6 from LPS-stimulated RAW 264.7 cells at the concentrations of 50 ug/mL. AG significantly inhibited LPS-induced production of VEGF from LPS-stimulated RAW 264.7 cells at the concentrations of 100 ug/mL. But AG did not show any significant effect on the production of MCP-1, LIF, LIX, IP-10 and IL-$1{\beta}$ from LPS-induced RAW 264.7 cells. Conclusion : These results suggest that AG has anti-inflammatory effect related with its inhibition of proinflammatory mediators such as TNF-${\alpha}$, MIP-$1{\alpha}$, G-CSF, RANTES, IL-10, MIP-$1{\beta}$, MIP-2, GM-CSF, IL-6, VEGF and M-CSF in LPS-induced macrophages.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2023년도 임시총회 및 춘계학술대회
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• pp.40-40
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• 2023

Atopic dermatitis is a chronic inflammatory skin diseases caused by skin barrier dysfunction. Allium victoralis var. Platyphyllum (AVP) is a perennial plant used as vegetable and herbal medicine. The purpose of this study was to suggest that AVP is a new cosmetic material by examining the effects of AVP on the skin barrier and inflammatory response. A bibliometric network analysis was performed through keyword co-occurrence analysis by extracting author keyword from 69 articles retrieved from SCOPUS. We noted the anti-inflammatory activity shown by the results of clustering and mapping from network visualization analysis using VOSviewer software tool. HPLC-UV analysis showed that AVP contains 0.12 ± 0.02 mg/g of chlorogenic acid and 0.10 ± 0.01 mg/g of gallic acid. AVP at 100 ㎍/mL was shown to increase the mRNA levels of filaggrin and involucrin related to skin barrier function by 1.50-fold and 1.43-fold, respectively. In the scratch assay, AVP at concentrations of 100 ㎍/mL and 200 ㎍/mL significantly increased the cell migration rate and narrowed the scratch area. In addition, AVP suppressed the increase of inflammation-related factors COX-2 and NO and decreased the release of β-hexosaminidase. This study suggests that AVP can be developed as a functional cosmetic material for atopy management through skin barrier protection effects, anti-inflammatory and anti-itch effects.

• 대한화장품학회지
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• 제30권1호
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• pp.63-71
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• 2004

태양광선과 산소는 피부세포에 자유라디칼(free radical)과 활성산소종(Reactive oxygen species, ROS)을 생성시켜 DNA의 손상과 세포막 지질의 과산화를 유발한다. 또한 콜라겐과 같은 세포 외 기질을 파괴하는데 관여하는 효소(matrix metallo-proteinhse, MMPs)의 발현에 영향을 주어, 결합조직을 손상시키며, 피부 염증을 유발시켜 조직을 파괴하고 노화를 가속화시킨다. 본 연구는 자외선과 SLS에 의한 피부손상에 대한 향나무추물물의 피부 보호 효과에 대한 것이다. 실험 결과 프리라디칼 및 슈퍼옥사이드 라디칼 소거 효과가 우수하게 나타났으며, 피부 콜라겐과 같은 메트릭스를 분해하는 효소인 MMP-1의 활성 및 발현 저해 효과는 섬유아세포에서 UVA 조사에 의한 실험에서 우수하게 나타났으며, 피부세포 배양액에 대해 zymography를 실시하여 활성이 감소됨을 확인하였다. 피부 각질형성세포에서 자외선에 의해 유도된 염증관련 사이토카인인 인터루킨 6의 발현량 실험에서도 무처리군에 비해 향나무추출물이 인터루킨 6를 30% 정도 저해효과가 나타났으며, 염증반응 중 cyclooxygenase (COX)에 의한 경로에서 생성되는 프로스타글란딘 E$_2$의 생성량도 감소시켰다. 사람 피부에서 SLS (0.5%)첩포로 유발된 자극성 피부염의 항염증 효과 평가 시 SLS에 의해 유발되는 자극정도가 피검자의 피부상태에 따라 다양하게 나타났으며, 향나무추출물 함유 에멀젼 제품 도포 실험에서는 피부 홍반 완화 효과와 피부장벽 회복 효과가 우수하게 나타났다. 따라서 향나무추출물은 자외선 조사 및 SLS에 의한 피부손상에 대한 보호 효과를 가진 화장품에서 우수한 소재로 적용될 수 있다. 녹두추출물을 화장품에 사용하였을 경우 우수한 항자극 효과를 보임을 확인하였다.완화시키는데 기여하였다. 이러한 결과를 종합하여 한국에서 삼림쇠퇴를 유발할 수 있는 스트레스요인들의 잠재적 상호작용을 모식화하였다. 나아가 그것을 완화할 수 있는 복원방안을 토양개량과 식생복원의 측면에서 제시하였다. mg/l)치 농도가 높을지라도 이것이 낮은 cinnamic acid와 protocatechuic acid에서 현저한 억제현상을 보였다. 서양등골나물 임상의 토양보다 서양등골나물이 분포하지 않는 토양의 중금속 함량이 전반적으로 높았다. 특히 Al, Fe 및 Mn의 함량이 높았으며 이들 중금속은 total phenolic compound 함량이 높은 잎에 대부분이 축적되었다." separating himself from the real world. Thirdly, The nature of "Gang-Ho" is 'absence of worldly desire' and 'cordiality' that one could be deserved his diligency becoming a part of the harmonious idealistic living place. Fourthly, on the character of story teller. Originally he is a incomer of "Gang-Ho" from real world. so that reason, he is showing dualism not to deny the loyalty oath to his king, while he intends to satisfy with the life in "Gang- Ho" separating himself from real world. As