• Journal of Ginseng Research
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• 제46권6호
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• pp.809-818
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• 2022

Background: The non-saponin fraction (NSF) of Korean Red Ginseng is a powder in which saponin is eliminated from red ginseng concentrate by fractionation. In this study, we examined the effect of NSF on age-associated sarcopenia in old mice. Methods: NSF (50 or 200 mg/kg/day) was administered orally daily to young (3-6-month-old) and old (20-24-month-old) C57BL/6 J mice for 6 weeks. Body weight and grip strength were assessed once a week during the oral administration period. The gastrocnemius and quadriceps muscle were excised, and the muscle fiber size was compared through hematoxylin and eosin staining. In addition, the effect of NSF on sarcopenia and inflammation/oxidative stress-related factors in hindlimb muscles was investigated by western blotting. Flow cytometry analysis was conducted to investigate the effect of NSF on immune homeostasis. Blood samples were collected by cardiac puncture, and the serum levels of insulin-like growth factor 1, pro-inflammatory cytokines, and glutathione were evaluated. Results: NSF significantly alleviated muscle strength, mass, and also fiber size in old mice. Age-associated impairment of immune homeostasis was recovered by NSF through retaining CD11b⁺F4/80⁺ macrophages and regulating inflammatory biomarkers. NSF also decreased the age-induced expression of oxidative stress factors. Taken together, NSF showed the effect of improving sarcopenia by inhibiting low-grade chronic inflammatory/oxidative stress factors. Conclusion: NSF exhibited anti-sarcopenia effects by regulating chronic inflammation and oxidative stress in old mice. Thus, we suggest that NSF is a promising restorative agent that can be used to improve sarcopenia in the elderly as well as maintain immune homeostasis.

• Biomolecules & Therapeutics
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• 제12권4호
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• pp.221-227
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• 2004

Nonsteroidal anti-inflammatory drugs (NSAIDs), particularly the highly selective cyclooxygenase (COX)-2 inhibitors have been shown to decrease the growth of tumor, in part, by inhibition of neovascularization. Recently, besides mature endothelial cells, endothelial progenitor cells (EPCs) have been shown to contribute neovascularization in angiogenic tissues. In this study, we addressed a question whether nimesulide, a selective COX-2 inhibitor, could affect differentiation of EPCs into adhesive endothelial cells in vitro. Total mononuclear cells were isolated from cord blood by Ficoll density gradient centrifugation, and then the cells were incubated with nimesulide or vehicle control for 7 days. The number of adherent and spindle-shaped cells decreased by nimesulide treatment in a concentration-dependent fashion at a concentration range of 5 - 200 ${\mu}M$. Moreover, the adherent cells double positive for DiI-ac-LDL uptake and lectin binding significantly decreased upon nimesulide treatment. There was no change of expression of CD31 between treatment and control groups, whereas slight reduction was detected upon treatment in expression of VE-cadherin, ICAM-1, vWF, ${\alpha}v$, and ${\alpha}5$. Nimesulide also reduced cell viability during first 3 days' culture and induced apoptosis in adherent EPCs, resulting in increased annexin-V-positive and propidium iodide-negative cells. Taken together, these results suggest that nimesulide could be applied for the inhibition of new vessel formation, in part, by inhibiting differentiation and survival of EPCs.

• Journal of Acupuncture Research
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• 제29권1호
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• pp.115-125
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• 2012

Objectives : The purpose of this study is to observe the inhibitory effects of $Chrthami$ semen oil pharmacopuncture(CSOP) on CIA (collagen-induced arthritis) mice. Materials and Methods : Two types of experiments were conducted: $in$ $vitro$ assay, inhibition of MIF mRNA and TNF-${\alpha}$ mRNA expressions in synovial membranes was observed, and $in$ $vivo$ assay, $1{\mu}{\ell}/kg$ CSOP was injected every day to the left $Weizhong$ ($BL_{40}$) from day 3 to 21 after induction of CIA, and changes in paw edema, apical surface morphology, neovascularization in synovial membranes, fibrosis, pro-inflammatory cytokines production, Th-1 differentiation, and anti-inflammatory effect were investigated. Results : 1. In synoviocytes of the CIA mice treated with CSOP, MIF mRNA and TNF-${\alpha}$ mRNA expressions were down-regulated in a dose-dependent manner. 2. Paw edema of the CIA mice treated with CSOP was diminished. 3. Tissue injury in the synovial membranes, capillary distribution and fibrosis were reduced in CSOP-treated mice. 4. MIF, TNF-${\alpha}$, IL-6, MMP-9 expressions were repressed in CSOP-treated mice during the experiment to observe the inhibitory effect on cytokines production in early stage RA. 5. IL-12 and CD28 were reduced in CSOP-treated mice during the observation of inhibitory effect on Th 1 differentiation. 6. PPAR-${\gamma}$ was increased during the experiment to observe the anti-inflammatory effect of CSOP. Conclusions : The results may suggest that administration treatment using $Chrthami$ semen oil pharmacopuncture decreases the inflammatory response on an Animal Model with CIA.

• Asian Pacific Journal of Cancer Prevention
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• 제14권11호
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• pp.6949-6953
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• 2013

Background: Nowadays, the encapsulation of cytotoxic chemotherapeutic agents is attracting interest as a method for drug delivery. We hypothesized that the efficiency of helenalin might be maximized by encapsulation in ${\beta}$-cyclodextrin nanoparticles. Helenalin, with a hydrophobic structure obtained from flowers of Arnica chamissonis and Arnica Montana, has anti-cancer and anti-inflammatory activity but low water solubility and bioavailability. ${\beta}$-Cyclodextrin (${\beta}$-CD) is a cyclic oligosaccharide comprising seven D-glucopyranoside units, linked through 1,4-glycosidic bonds. Materials and Methods: To test our hypothesis, we prepared ${\beta}$-cyclodextrin-helenalin complexes to determine their inhibitory effects on telomerase gene expression by real-time polymerase chain reaction (q-PCR) and cytotoxic effects by colorimetric cell viability (MTT) assay. Results: MTT assay showed that not only ${\beta}$-cyclodextrin has no cytotoxic effect on its own but also it demonstrated that ${\beta}$-cyclodextrin-helenalin complexes inhibited the growth of the T47D breast cancer cell line in a time and dose-dependent manner. Our q-PCR results showed that the expression of telomerase gene was effectively reduced as the concentration of ${\beta}$-cyclodextrin-helenalin complexes increased. Conclusions: ${\beta}$-Cyclodextrin-helenalin complexes exerted cytotoxic effects on T47D cells through down-regulation of telomerase expression and by enhancing Helenalin uptake by cells. Therefore, ${\beta}$-cyclodextrin could be superior carrier for this kind of hydrophobic agent.

• Journal of Acupuncture Research
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• 제22권2호
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• pp.71-81
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• 2005

Objective : Dear antler (Cervus korean TEMMINCK var. mantchuricus Swinhoe) used for traditional immunosuppressive and immuno-activating action. The effect of deer antler herbal-acupuncture(DAH) solution, prepared by water extract method, on cathepsin activities in bone tissues (cartilage and synovial) cells from mouse rheumatoid arthritis (RA) model was studied. The cysteine endoprotease cathepsin mediates degradation of the MHC class II invariant chain (Ii) in human and mouse antigen-presenting cells. The studies described here examine the functional significance of cathepsin inhibition on autoantigen presentation and organ-specific autoimmune diseases in a murine model for RA. Methods : An animal model for RA in BALB/c mice thymectomized 3 days after birth (3d-Tx) was constructed All 3d-Tx BALB/c mice developed autoimmune lesions in the bone tissue cells, starting at 3 weeks of age, and the disease mediated by CD4+ T cells was chronic and progressive. Significant inhibitory effects of DAH solution on cathepsin S and L were observed in each organ in a dose-dependent manner. Moreover, we confirmed that cathepsin S and L activity in each organ were clearly inhibited by DAB solution. When we examined the inhibitory effects of DAH solution against autoantigen-specific T cell responses in vitro, in regional lymph node cells, but not in spleens, from model mice, a significant inhibitory effect of DAB solution was observed in a dose-dependent manner. DAH solution do not block T cell proliferation to Con A, indicated that the dose of DAB solution 10 to $20\;{\mu}g/m{\ell}$ was sufficient to inactivate the autoantigen-specific T cell responses in vitro. In vivo therapeutic effects of DAB solution were examined in a murine model for RA, autoantigen-specific (C-II-specific) T cell response were significantly inhibited in LNCs from DAH solution-treated mice. Results : Iinhibition of cathepsin S and L in vivo alters autoantigen presentation and development of organ-specific autoimmunity in RA model. Conclusion : These data identify selective inhibition of cysteine protease cathepsin S and L as a potential therapeutic strategy for autoimmune disease process such RA. Thus, DAH solution will served as a potent anti-inflammatory and anti-arthritic agents for treatment of human RA.

• Archives of Plastic Surgery
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• 제37권6호
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• pp.721-725
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• 2010

Purpose: The survival of composite graft is dependent on three steps, (1) plasmatic imbibitions, (2) inosculation, and (3) neovascularization. Among the many trials to increase the survival rate of composite graft, prostaglandin E1 (PGE1) has beneficial effects on the microcirculatory level with vasodilating, antithrombotic, anti-inflammatory and neoangiogenic properties. Lipo-PGE1 which is lipid microspheres containing PGE1 had developed to compensate the systemic and local side effects of PGE1. This study was proposed to determine whether Lipo-PGE1 administration enhanced the survival of composite graft through neovascularization quantitatively in a rabbit ear model. Methods: Fourteen New Zealand White Rabbits each weighing 3~4 kg were divided in two groups: (1) intravenous Lipo-PGE1 injection group and (2) control group. A $2{\times}1\;cm$ sized, full-thickness rectangular composite graft was harvested in each auricle. Then, the graft was reaaproximated in situ using a 5-0 nylon suture. For the experimental group, $3{\mu}g$/kg/day of Lipo-PGE1 ($5{\mu}g$/mL) was administered intravenously through the marginal vein of the ear for 14 days. The control group was received no pharmacologic treatment. On the 14th postoperative day, composite graft of the ear was harvested and immunochemistry staining used Monoclonal mouse anti-CD 31 antibody was performed. Neoangiogenesis was quantified by counting the vessels that showed luminal structures surrounded by the brown color-stained epithelium and counted from 10 random high-power fields (400x) by independent blinded observer. Statistical analysis (Wilcoxon Signed Ranks test for nonparametric data) was performed using SPSS v12.0, with values of p<0.05 considered significant. Results: The mean number of the microvessels was $15.48{\pm}8.65$ in the experimental group and $9.82{\pm}7.25$ in the control group (p=0.028). Conclusion: The use of Lipo-PGE1 facilitated the neoangiogenesis, resulted in the improvement of the survival rate of graft. On the basis of this results, we could support wider application of Lipo-PGE1 for more effective therapeutic angiogenesis and successful survival in various cases of composite graft in the human.

• Tuberculosis and Respiratory Diseases
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• 제39권6호
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• pp.453-473
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• 1992

연구배경 : 전국 실태조사를 통하여 현재까지 확진된 전국의 유육종증 발생현황을 파악하고 이를 분석하고 외국의 보고와 비교하여 일반 개업의 및 내과 전문개업의 진료활동에 도움이 되도록 하고자 하였다. 방법 : 조사와 분석기간은 1991년 1월부터 1992년 12월까지 2년간으로 정하였으며 자료수집방법은 후향적 방법으로 입원실이 100병상 이상인 213개 종합병원의 내과, 안과, 피부과, 신경내과에 서신을 통한 경험증례를 수집 하였으며 확진예나 의심예가 있었던 경우에 유육종증 증례 기록지를 보내어 정확한 임상 및 검사정보를 수집하여 분석하였다. 결과 : 1) 제 1 차 설문엽서 발송은 입원실이 100병상 이상인 213개 종합병원의 내과, 안과, 피부과, 신경내과의 523개과 과장 및 호흡기 내과를 담당하는 전문의에게 발송하였다. 내과가 213개과로 41%, 피부과가 1087개과로 20%, 안과가 104개과로 20%, 신경과가 100개과로 19%를 차지하였다. 2) 제 1 차 설문조사에 대한 답신은 241개과에서 연락이 있었다(48% 회신율). 3) 확진예 및 의심예는 총 50개과 113예 및 10예가 있다고 보고되었으며 내과가 92예로 81%, 피부과 15예로 13%, 안과, 신경과가 각 3예로 3%를 차지하였다. 자료분석은 총 78예에서 시행되었고 내과 72예로 92%, 피부과 4예로 5%, 신경과 2예로 3%를 차지하였다. 4) 1980년 1에보고 이후 증례가 많은 수로 증가하여 1989년 13예, 19901건 17예, 1991년 18예가 보고되었다. 5) 진단 당시의 연령은 20대, 30대, 40대가 84.4%로 대부분을 차지하고 있었다. 6) 남녀비는 1 : 1.5의 비율로 여자 환자가 많았다. 7) 진단을 받게된 동기로 주소의 빈도로는 호흡기 증상이 48%로 가장 많았고, 피부의 병변, 전신증상, 시각장애, 관절통, 복통, 연하곤란의 순서이었고 무증상이 12예(16%)이었다. 8) 초증상에서 확진을 받을때까지 중앙치는 2개월이었다. 9) 증상의 반도로는 호흡기 증상, 전신증상, 피부증상, 눈 증상, 말초림프구 종창, 신경계증상, 심장증상의 순이었다. 10) 말초혈액검사 소견상 혈색소, 혈구용적치 및 혈소판수는 대부분 정상범위를 나타냈다. 58%의 증례에서 30%미만의 림프구 감소증을 나타냈으며, 임파구 아형으로 T4/T8비는 1.73{\pm}1.16$(0.43~4.62)였다. 43%에서 혈침속도가 상승하였다. 11) 혈청 생화학검사는 대부분 정상범위를 이었다. 혈청 ACE 농도의 평균치는 $66.8{\pm}58.6$(8.8~265) U/L로서 46.2%에서 증가되었다. 42.9%에서 하루 150 mg 이상의 단백뇨 소견을 나타냈고, 36.8%가 하루 300 mg이상의 단백뇨 소견을 나타냈다. 12) 면역학적 검사상 혈청 IgG는 43.5%, IgA는 45.5%, lgM은 59.1%, IgE는 46.7%에서 상승되었고, C3 및 C4는 대부분 정상범위 였으며, ANA는 11%에서 양성을 보였다. Kveim test는 시행된 3예에서 모두 양성을 나타냈다. 13) 폐기능검사중 FVC는 80% 미만인 경우가 17.3%, FEV1은 70% 미만인 경우가 11.5% FEV1/FVC는 70% 미만인 경우가 10%, TLC는 80% 미만인 경우가 15.2%, DLCO는 100% 미만인 경우가 64.7%이었다. 14) 동맥혈가스 분석소견은 PaO2가 90mmHg 미만인 경우가 48.6%, PaCO2는 평균 45mmHg보다 증가된 경우보다 5.7%이었다. 15) 기관지 폐포세척액 검사상 대식세포의 평균 백분율은 $51.4{\pm}19.2%$였고, 임파구의 평균 백분율은 $44.4{\pm}21.1$이였다. 임파구아형은 T4/T8 ratio는 $3.41{\pm}2.07$이였다. 16) 전 검사항목에 대한 남녀 성별간의 유의한 차이는 없었다. 17) 흉부방사선의 분석에서 폐문 임파절 비대를 보인례는 87.9%였고, 78.8%에서는 양측성병변을 9.1%에 일측성 병변을 보였다. 18) Siltzbach의 분류에 의한 병기 구분시 stage 0 : 5%, stage 1 : 58.3%, stage 2 : 28.3%, stage 3 : 8.3%의 분포를 보였다. 19) 흉부 CT를 시행한 54예에서 폐문임파절비대는 50예(92.6%)에서 보였고 이들 중 47예는 양측성 비대였다. 20) 고해상도 CT는 16예에서 시행하였는데, 2예에서는 병변이 없었으며, 폐실질의 병변은 결정성 병변, 소엽간 중격비후, 반점상 국소 폐음영의 순으로 나타났다. 21) 부수적으로 시행한 방사선학적 검사에서 골병변은 1예에서 수근골의 병변을 보였고, 초음파검사에서 2에에서 비장비대의 소견을 보였다. 22) 갈륨스캔은 12에에서 시행하여 폐문 및 종격동 임파절의 흡수증가 소견이 8예에서 보였고, 폐장의 흡수증가는 2예에서 보였다. 23) 조직검사상 가장 많이 시행한 진단방법은 경기관지 폐생검 47.3%이었고, 피부와 종격동 임파절 생검은 각각 34.5%, 말초임파절생검 23.6%, 개흉 폐생검 18.2%, 기관지생검이 16.2%이었다. 2가지 이상의 부위에서 조직검사를 시행한 경우는 52.7%이었다. 한 부위에서만 조직검사를 시행한 26예중 종격동 임파절 생검은 10예, 피부생검 6예, 말초임파절 생검 5예, 개흉폐생검 3예 그리고 경기관지 폐생검 2예이었다. 24) 병리소견상 비건락성 육아종은 100% 모두에서 관찰되었며, 다핵거대 세포는 47.3%, 봉입체는 10.9%, 초자질 섬유화는 34.5%, 괴사 9.1%, reticulin fibrils network 20%, lymphangitic distribution of granuloma 1.8%에서 관찰되었다. 결론 : 유육종 전국 실태조사에 따른 영상소견, 검사실소견, 방사선소견, 병리소견을 분석해 본 결과, 우리나라 유육종 환자들도 다소 차이는 있었지만 외국의 보고들과 유사한 소견들을 나타냈으며, 향후로는 이들에 대한 결과들을 추가분석하여, 유육종 진단에 도움을 줄 수 있고, 임상적 지표로 이용될 수 있는 검사항목을 찾아내는 것이 과제라 할 수 있겠다. 유육종증에 대한 자료는 대한 결핵 맺 호흡기학회에 보관이 되어 있음을 알려드립니다.

• 동의생리병리학회지
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• 제20권1호
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• pp.163-170
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• 2006

Nardostachyos Rhizoma (N. Rhizoma) belonging to the family Valerianaceae has been anti-arrhythmic effect, and sedation to the central nerve and a smooth muscle. We reported that the water extract of N. Rhizoma induced apoptotic cell death and differentiation in human promyelocytic leukemia (HL-60) cells. Cytotoxicity of N. Rhizoma was detected only in HL-60 cells (IC50 is about 200 ${\mu}g/ml$). The cytotoxic activity of N. Rhizoma in HL-60 cells was increased in a dose-dependent manner. We used several measures of apoptosis to determine whether these processes were involved in N. Rhizoma-induced apoptotic cell death. The high-dose (200 ${\mu}g/ml$) treatment of N. Rhizoma to HL-60 cells showed cell shrinkage, cell membrane blobbing, apoptotic bodies, and the fragmentation of DNA, suggesting that these cells underwent apoptosis. Treatment of HL-60 cells with N. Rhizoma time-dependently induced activation of caspase-3, caspase-8, and caspase-9 and proteolytic cleavage of poly(ADP-ribose) polymerase. Also, we investigated the effect of N. Rhizoma on cellular differentiation and proliferation in HL-60 cells. Differentiation and proliferation of HL-60 cells was determined through expression of CD11b and CD14 surface antigens using flow cytometry and nitroblue tetrazolium (NBT) assay, and through analysis of cell cycle using propidium iodide assay, respectively. N. Rhizoma induced the differentiation of HL-60 at the low-dose (100 ${\mu}g/ml$) treatment, as shown by increased expression of differentiation surface antigen CD11b, but not CDl4 and increased reducing activity of NBT. When HL-60 cells were treated with N. Rhizoma at concentration of $50{\mu}g/ml\;and\;100{\mu}g/ml$, NBT-reducing activities induced approximately 1.5-fold and 20.0-fold as compared with the control. In contrast, HL-60 cells treated with the N. Rhizoma-ATRA combination showed markedly elevated levels of 26.3-fold at $50{\mu}g/ml$ N. Rhizoma-0.1 ${\mu}M$ ATRA combination and 27.5-fold at 50 ${\mu}g/ml$ N. Rhizoma-0.2 ${\mu}M$ ATRA combination than when treated with N. Rhizoma alone or ATRA alone. It may be that N. Rhizoma plays important roles in synergy with ATRA during differentiation of HL-60 cells. DNA flow-cytometry indicated that N. Rhizoma markedly induced a G1 phase arrest of HL-60 cells. N. Rhizoma-treated HL-60 cells increased the cell population in G1 phase from 32.71% to 42.26%, whereas cell population in G2/M and S phases decreased from 23.61% to 10.33% and from 37.78% to 33.98%, respectively. We examined the change in the $p21^{WAF1/Cip1}\;and\;p27^{Kip1}$ proteins, which are the CKIs related with the G1 phase arrest. The expression of the CDK inhibitor $p27^{Kip1},\;but\;not\;p21^{WAF1/Cip1}$ were markedly increased by N. Rhizoma. Taken together, these results demonstrated that N. Rhizoma induces apoptotic cell death through activation of caspase-3, and potently inhibits the proliferation of HL-60 cells via the G1 phase cell cycle arrest in association with $p27^{Kip1}$ and granulocytic differentiation induction .

• 한국식품과학회지
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• 제28권5호
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• pp.953-958
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• 1996

발암성 진균 독소의 하나인 fumonisin을 신속, 간편하게 분석할 수 있는 효소면역측정법 (ELISA)을 개발하고자 하였다. 먼저 Fumonisin $B_1\;(FB_{1})$에 특이적인 항체를 생산하기 위하여 $FB_1$을 keyhole lympet hemocyanin (KLH)에 공유결합시키고 이를 Freund's adjuvant와 함께 토끼에 3차례 피하주사하여 면역하였다. 높은 항체가와 가장 양호한 경합반응을 보인 항혈청으로부터 항체를 정제하였으며, 이 항체의 fumonisin 유사독소에 대한 교차반응은 fumonisin $B_1,\;B_2$및 $B_3$에 대하여 각각 100%, 69% 및 166%이었다. $FB_1$의 검출을 위하여 확립한 직접법 경합ELISA (cdELISA)로 옥수수에 인위적으로 오염시킨 $FB_1$을 정제없이 ELISA로 분석하는 경우, 75% methanol로 시료의 추출 후 완충액으로 1/100 희석하였을 때 양호한 회수율을 보였다. 이 조건하에서 행한 ELISA의 FB₁분석회수율은 $1-30\;{\mu}g/g$의 오염농도범위에서 평균 67%이었으며, 농도별 회수율의 분산은 3.4%로 분석치가 매우 안정하였다. 본 연구에서 개발한 fumonisin 분석용 ELISA system은 수입 및 국내산 농산물로부터 이 독소의 정제없이 손쉽게 다량의 시료를 일차로 분석하는 데 유용할 것으로 판단된다.

• Molecules and Cells
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• 제19권3호
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• pp.402-407
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• 2005

Bone marrow mesenchymal stem cells (MSCs) have shown potential for cardiac repair following myocardial injury, but this approach is limited by their poor viability after transplantation. To reduce cell loss after transplantation, we introduced the fibroblast growth factor-2 (FGF-2) gene ex vivo before transplantation. The isolated MSCs produced colonies with a fibroblast-like morphology in 2 weeks; over 95% expressed CD71, and 28% expressed the cardiomyocyte-specific transcription factor, Nkx2.5, as well as ${\alpha}$-skeletal actin, Nkx2.5, and GATA4. In hypoxic culture, the FGF-2-transfected MSCs (FGF-2-MSCs) secreted increased levels of FGF-2 and displayed a threefold increase in viability, as well as increased expression of the anti-apoptotic gene, Bcl2, and reduced DNA laddering. They had functional adrenergic receptors, like cardiomyocytes, and exposure to norepinephrine led to phosphorylation of ERK1/2. Viable cells persisted 4 weeks after implantation of $5.0{\times}10^5$ FGF-2-MSCs into infarcted myocardia. Expression of cardiac troponin T (CTn T) and a voltage-gated $Ca^{2+}$ channel (CaV2.1) increased, and new blood vessels formed. These data suggest that genetic modification of MSCs before transplantation could be useful for treating myocardial infarction and end-stage cardiac failure.