• Title/Summary/Keyword: Mouse skin

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Immunotherapeutic Effects of CTLA4Ig Fusion Protein on Murine EAE and GVHD (마우스 EAE, GVHD 질환에서 CTLA4Ig 융합단백의 면역치료 효과)

  • Jang, Seong-Ok;Hong, Soo-Jong;Cho, Hoon-Sik;Chung, Yong-Hoon
    • IMMUNE NETWORK
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    • v.3 no.4
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    • pp.302-309
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    • 2003
  • Background: CTLA4 (CD152), which is expressed on the surface of T cells following activation, has a much higher affinity for B7 molecules comparing to CD28, and is a negative regulator of T cell activation. In contrast to stimulating and agonistic capabilities of monoclonal antibodies specific to CTLA-4, CTLA4Ig fusion protein appears to act as CD28 antagonist and inhibits in vitro and in vivo T cell priming in variety of immunological conditions. We've set out to confirm whether inhibition of the CD28-B7 costimulatory response using a soluble form of human CTLA4Ig fusion protein would lead to persistent inhibition of alloreactive T cell activation. Methods: We have used CHO-$dhfr^-$ cell-line to produce CTLA4Ig fusion protein. After serum free culture of transfected cell line we purified this recombinant molecule by using protein A column. To confirm characterization of fusion protein, we carried out a series of Western blot, SDS-PAGE and silver staining analyses. We have also investigated the efficacy of CTLA4Ig in vitro such as mixed lymphocyte reaction (MLR) & cytotoxic T lymphocyte (CTL) response and in vivo such as experimental autoimmune encephalomyelitis (EAE), graft versus host disease (GVHD) and skin-graft whether this fusion protein could inhibit alloreactive T cell activation and lead to immunosuppression of activated T cell. Results: In vitro assay, CTLA4Ig fusion protein inhibited immune response in T cell-specific manner: 1) Human CTLA4Ig inhibited allogeneic stimulation in murine MLR; 2) CTLA4Ig prevented the specific killing activity of CTL. In vivo assay, human CTLA4Ig revealed the capacities to induce alloantigen-specific hyporesponsiveness in mouse model: 1) GVHD was efficiently blocked by dose-dependent manner; 2) Clinical score of EAE was significantly decreased compared to nomal control; 3) The time of skin-graft rejection was not different between CTLA4Ig treated and control group. Conclusion: Human CTLA4Ig suppress the T cell-mediated immune response and efficiently inhibit the EAE, GVHD in mouse model. The mechanism of T cell suppression by human CTLA4Ig fusion protein may be originated from the suppression of activity of cytotoxic T cell. Human CTLA4Ig could not suppress the rejection in mouse skin-graft, this finding suggests that other mechanism except the suppression of cytotoxic T cell may exist on the suppression of graft rejection.

Development and application of non-invasive drug delivery systems utilizing pulse power, and its application to mouse models (펄스파워를 적용한 비침습 약물 전달기 개발 및 마우스 모델로의 적용)

  • Hwi-Chan Ham;Kyu-Sik Kim;Ji-Hwan Lee;Hyung-Jin Choi;Do-Nyun Kim;Jai-Ick Yoh
    • Journal of IKEEE
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    • v.28 no.1
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    • pp.97-103
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    • 2024
  • Some drugs can offer far better medical effectiveness as it is injected through the intradermal layer of the skin, known as a needle-free injection. However, conventional needle-free devices might deliver a relatively large amount of drug in a just single spot of skin, splitting open the tissue layer structure, which might cause bruising and bleeding. By injecting the small volume with a fast repetition rate in a large surface area of skin, the patient may get much fewer injuries and pain. To achieve that specification, the driving force must be instantaneous and short-pulsed. Such a form of an injection device has been developed but the efficacy of those devices has been rarely examined. Therefore, this study developed the laser-induced microjet device that ejects microjet whose speed is ~310 m/s, during the 400~800 ㎲ of pulse time. The device can eject ~1 µL of the drug at the rate at which each shot repeated 10 shots per second. Using this specification, we evaluated the efficacy of drug injection onto mouse models. After injecting the insulin solution into the mouse model, the blood insulin level is detected, resulting in 20 % of blood insulin level with the ordinary needle syringe injection method.

Skin Regeneration, Anti-wrinkle, Whitening and Moisturizing Effects of Cheongsangbangpung-tang Aqueous Extracts with Cytotoxicity (청상방풍탕 열수 추출물의 피부재생, 주름개선, 미백, 보습 효과 및 세포독성 평가)

  • Woo, Chang-Yoon;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.30 no.2
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    • pp.49-70
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    • 2017
  • Objectives: The present study is to observe the skin-regeneration, anti-wrinkle, whitening and skin moisturizing effects of Cheongsangbangpung-tang (CSBPT) with cytotoxicity. Methods: In the present study, cytotoxicity of CSBPT lyophilized aqueous extracts (yield=18.71%) was experimented against human normal fibroblast cells and B16F10 murine melanoma cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide (MTT) assay, and skin regeneration and anti-wrinkle effects were also showed through the assay of collagen type I synthesis by an enzyme immunoassay (EIA) kit as comparing with transforming growth factor (TGF)-${\beta}1$, hyaluronidase, collagenase and matrix metalloproteinase (MMP)-1 inhibitory assays as comparing with oleanolic acid (OA), and elastase inhibitory effects as comparing with phosphoramidon disodium salt (PP). In addition, whitening effects of CSBPT were observed by tyrosinase inhibitory assay and melanin formation test in B16/F10 melanoma cells as comparing with arbutin, and skin moisturizing effects were measured through mouse skin water contents test, respectively. Results: No CSBPT treatment related cytotoxic effects were demonstrated against human normal fibroblast cells and B16/F10 murine melanoma cells. CSBPT concentration-dependent increased collagen type I synthesis at human normal fibroblast cells. It also effectively suspreessed hyaluronidase, collagenase, elastase and MMP-1 activities, which were enzymes that related to declining of ECM and formation of wrinkle. CSBPT supressed B16/F10 melanoma cells's melanin productions with tyrosinase activity, which was an enzyme connected with melanin formation, and dose-dependent and significant increases of skin water contents were detected in CSBPT treated mouse skin as compared with vehicle control skins. Conclusions: CSBPT showed favorable and enough skin regeneration, anti-wrinkle, whitening and skin moisturizing effects at least in a condition of this experiment. However, more detail mechanism and in vivo skin protective efficacy studies should be conducted in future with the screening of the biological active compounds in individual herbs of Cheongsangbangpung-tang.

Mechanism of Action of Various Vehicles That Enhance the Permeation of Ketoprofen (케토프로펜의 피부투과도를 증진시키는 다양한 용매의 작용기전)

  • Cho, Young-Joo;Choi, Hoo-Kyun
    • Journal of Pharmaceutical Investigation
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    • v.28 no.3
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    • pp.165-169
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    • 1998
  • The effect of various vehicles on the permeation of a model drug, ketoprofen in solution formulation was evaluated using a flow-through diffusion cell system at $37^{\circ}C$. To investigate the mechanism of permeation rate enhancement, the effects of pretreatment with various vehicles on the permeation of the drug were evaluated using 5 mg/ml solution and saturated solution. The order of permeation rate of ketoprofen across hairless mouse skin after pretreatment with various vehicles was similar to the case where the vehicles and the drug were coadministered except ethanol and oleic acid. The results indicate that the mechanism of enhancement can be direct action of the vehicles on the barrier property of the skin and/or carrier mechanism.

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3,4-Dihydroxytoluene suppresses UVB-induced wrinkle formation by inhibiting Raf-1

  • Park, Sang-Hee;Kang, Nam Joo
    • Korean Journal of Food Science and Technology
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    • v.52 no.4
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    • pp.385-395
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    • 2020
  • This study examined the effect of 3,4-dihydroxytoluene (DHT) on UVB-induced photoaging and determined its molecular mechanisms, using HaCaT human keratinocytes and SKH-1 hairless mice. DHT suppressed UVB-induced matrix metalloproteinase-1 (MMP-1) expression in HaCaT cells. In vivo data from mouse skin supported that DHT decreased UVB-induced wrinkle formation, epidermal thickness, and matrix metalloproteinase-13 (MMP-13) expression. DHT appeared to exert its anti-aging effects by suppressing UVB-induced Raf-1 kinase activity and subsequent attenuation of UVB-induced phosphorylation of MEK, ERK, and p90RSK in HaCaT cells. In vitro and in vivo pull-down assays revealed that DHT bound with Raf-1 in ATP-noncompetitive manner. Overall, DHT appears to anti-photoaging effects in vitro and in vivo through the suppression of Raf-1 kinase activity and may have potential as a treatment for the prevention of skin aging.

Effect of Ginseng Saponin on the Skin (인삼(人蔘) Saponin이 피부(皮膚)에 미치는 영향(影響))

  • Kim, Nak-Doo;Woo, Lin-Keun
    • Korean Journal of Pharmacognosy
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    • v.7 no.2
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    • pp.119-121
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    • 1976
  • To investigate untoward effects which might be produced by local application of Ginseng saponin, sensitization test and eye irritation test were performed in guinea pigs and rabbits. Any reaction on the skin was not found at a dose of 0.1ml of 0.1% solution in sensitization test. Instillation of Ginseng saponin (0.1ml of 1.0% and 10% soultion) into the right eye of rabbits, did not produce any irritation. Median lethal dose of the total saponin in mouse was 695 mg/kg by i.p. and 1490mg/kg by s.c. injection.

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Local Irritation of DA-125 (DA-125의 국소자극성)

  • 안병옥;백남기;이순복;김원배;양중익;김부영;김대병;이종권
    • Biomolecules & Therapeutics
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    • v.1 no.1
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    • pp.20-25
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    • 1993
  • DA-125, a new anthracycline derivative, shows significant anticancer activities. We conducted a study to examine the local irritating effect of DA-125 using mice and rabbits. In the skin test in mice, intradermal injection of 0.4 mg of DA-125, compared to a dosage of 0.2 mg of adriamycin, had weak irritating potentials to induce skin ulceration and erythematous induration. A dosage of 0.6 mg of DA-125 produced similar degree of lesions in perivascular irritation model to that of 0.2 mg of adriamycin, but the healing time was shorter in the case of mice treated with DA-125. In ocular irritation study in rabbit, the highest M.O.I.(mean ocular irritation index) of 0.5% DA-125 solution was 0.67, therefore DA-125 could be considered as a practically non-irritating anticancer agent. These results suggest that substitution of DA-125 for Adriamycin would reduces the possibility of outbreaks of local irritation and the severity of the lesions.

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(Dendritic cells in the skin) (피부의 수상돌기 세포)

  • 이민걸
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.25 no.2
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    • pp.35-44
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    • 1999
  • Dendritic cells(DC) are a system of highly efficient antigen-presenting cells that initiate the primary immune response. There are two kinds of dendritic cells in the skin, Langerhans cell in the epidermis and dermal dendritic cell in the dermis. The knowledge of DC, which are very important in the immune reponse of cancer, autoimmune disease, transplantation and infection, has been known through the study about Langerhans cells. In this paper, the role of Langerhans cell in the contact hypersensitivity and atopic dermatitis is discussed and culture methods of mouse Langerhans cells and human U from pheripheral blood monocytes are described.

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Anti-inflammatory effects of herbal medicines(Rubus coreanus, Rehmanniae Radix, Houttuynia cordata, Betulae cortex) EtOH extract on acute atopic dermatitis mice (급성 아토피 피부염 마우스 모델에서 한약조합약물의 염증반응 진정효과)

  • Jung, Jae-Hoon;Kim, Gyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.28 no.1
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    • pp.68-84
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    • 2015
  • Object : The object of this study was effected of composed of 4 herbal medicines(Rubus coreanus, Rehmanniae Radix, Houttuynia cordata, Betulae cortex) on acute atopic dermatitis mice. Methods : BALB/c mice were divided 4 groups ; normal group, negative control group of acute atopic dermatitis mice induced by DNCB, treated apply to the back skin with the herbal medicine group, and treated orally with herbal medicine group. Treated with herbal medicine and DNCB during 2 weeks. After treated, measured WBCs, RBCs, and platelet in the blood, and analysed mRNA expression of spleen. Result : Composed herbal medicines could reduce WBCs, lymphocytes, monocytes, neutrophils, and eosinophils in the mouse blood. And, could down TNF-${\alpha}$ levels in spleen. In the skin tissue histology results, herbal medicines reduce the cells and T cells. Conclusion : Composed herbal medicines help to another inflammatory disease because they reduce the cells and T cells in the skin tissues. And they have inhibitory effects of TNF-${\alpha}$ mRNA expression, and Th 2 immune responses. Therefore herbal medicines use for an acute atopic dermatitis treatment.

The pathophysiological role of dendritic cell subsets in psoriasis

  • Kim, Tae-Gyun;Kim, Dae Suk;Kim, Hyoung-Pyo;Lee, Min-Geol
    • BMB Reports
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    • v.47 no.2
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    • pp.60-68
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    • 2014
  • Psoriasis is a chronic inflammatory disorder characterized by an erythematous scaly plaque of the skin and is occasionally accompanied by systemic complications. In the psoriatic lesions, an increased number of cytokine-producing dendritic cells and activated T cells are observed, which indicate that psoriasis is a prototype of an immune-mediated dermatosis. During the last decade, emerging studies demonstrate novel roles for the dendritic cell subsets in the process of disease initiation and maintenance of psoriasis. In addition, recently discovered anti-psoriatic therapies, which specifically target inflammatory cytokines produced by lesional dendritic cells, bring much better clinical improvement compared to conventional treatments. These new therapies implicate the crucial importance of dendritic cells in psoriasis pathogenesis. This review will summarize and discuss the dendritic cell subsets of the human skin and their pathophysiological involvement in psoriasis based on mouse- and patient-oriented studies.