線維化におけるＴＧＦ－β情報伝達の関与についての分子基盤の解明

2013 Fiscal Year Research-status Report

• Project/Area Number: 24591658
• Research Institution: Kumamoto University
• Principal Investigator: 尹 浩信 熊本大学, 生命科学研究部, 教授 (20282634)
• Keywords: 遺伝子 / 細胞・組織 / シグナル伝達

Research Abstract

全身性強皮症、肺線維症の組織および病変部の培養線維芽細胞においてDNA microarry法にて過剰発現する分子、発現低下する分子を検討し、またrealtime PCRを用いたmicro RNA arrayによって過剰発現するmicro RNA、発現低下するmicro RNAを同定した。
正常皮膚線維芽細胞を用いてintegrin αVβ5 stable transfectant作成し、integrin αVβ5の過剰発現を確認した。
COL1A2 enhancer、promoterをベクターに組み込んで作成したtransgeneを用いて線維芽細胞特異的integrin αVβ5 transgenic mouseを作成し、現在phenotypeを解析している。
線維芽細胞特異的integrinαV transgenic mouseのphenotypeを解析中であり、予想に反して皮膚の線維化は認められず、正常マウスと比較して皮膚厚の減少を認めた。
I型コラーゲン、III型コラーゲンの蛋白mRNA発現量がintegrinαV transgenic mouseで低下し、またMMP-1　ｍRNA蛋白発現量がintegrinαV transgenic mouseで増加していることが明らかとなった。これらの点がintegrinαV transgenic mouseのphenotypeに関与すると考えられ、さらに解析中である。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

線維化病変に過剰発現ないし発現低下する分子を同定することができた。さらに過剰発現するmicro RNA、発現低下するmicro RNAをも同定することができた。integrin αVβ5 stable transfectantも作成でき、線維芽細胞特異的integrin αVβ5 transgenic mouseも完成した。
さらに線維芽細胞特異的integrin αV transgenic mouse作成にも成功し、phenotypeを解析中であり、その機序も解析中である。
以上より研究は順調で十分達成できたと考える。

Strategy for Future Research Activity

integrin αVβ5 stable transfectantを用いてDNA microarry法、realtime PCRを用いたmicro RNA array法、情報伝達protein arrayを行い、線維化phenotypeを形成しているか、すなわちTGF-β受容体が過剰発現しているか、TGF-β情報伝達経路（Smad2、Smad 3、Smad4、p300/CBP、p38 MAPK、PI3 kinase）が過剰リン酸化しているか、I型コラーゲンをはじめとする細胞外マトリックス蛋白および遺伝子が過剰発現しているか、MMP-1などのproteaseの発現が低下しているか、TIMP-1などのprotease inhibitorの発現が亢進しているか詳細に検討する。
線維芽細胞特異的integrin αV transgenic mouseのphenotypeを解析を進め、コラーゲン遺伝子、MMP-1遺伝子、TIMP-1遺伝子発現をマウス皮膚でin vivoの研究を行うとともに、線維芽細胞を培養し、培養線維芽細胞を用いてin vitroで検討を行う。

Research Products

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• [Journal Article] Case of disseminated cutaneous Mycobacterium chelonae infection mimicking cutaneous vasculitis. (2014)
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• [Journal Article] Clinical evaluation of moisturizers with physiological analysis of stratum corneum TARC and TSLP. (2014)
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• [Journal Article] Phase II study of i.v. interferon- gamma in Japanese patient s with mycosis fungoides. (2014)
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• [Journal Article] Dots/ globules on dermoscopy in nail-apparatus melanoma. (2014)
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• [Journal Article] Successful treatment of skin fistulas in systemic sclerosis patients with the combination of topical negative pressure therapy and split-thickness skin grafting. (2014)
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• [Journal Article] Angiopoietin-like protein 2 accelerates carcinogenesis by activating chronic inflammation and oxidative stress. (2014)
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  • Journal Title: Mol Cancer Res, Volume: 12 Pages: 239-249
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• [Journal Article] MIRSNP rs2910164 of miR-146a is associated with the muscle involvement in polymyositis/ dermatomyositis. (2014)
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• [Journal Article] Case of Fusarium paronychia successfully treated with occulisive dressing of antifungal cream. (2014)
  • Author(s): Ikeda I, Ohno T, Ohno H, Miyazaki Y, Nishimoto K, Makino T, Ihn H.
  • Journal Title: J Dermatol, Volume: 41 Pages: 340-342
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• [Journal Article] Analysis of expression pattern of serum microRNA levels in each patient with psoriasis. (2014)
  • Author(s): Koga Y, Jinnin M, Ichihara A, Fujisawa A, Moriya C, Sakai K, Fukushima S, Inoue Y, Ihn H.
  • Journal Title: J Dermatol Sci, Volume: in press Pages: in press
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• [Journal Article] Serum levels of leptin receptor in patients with systemic sclerosis. (2014)
  • Author(s): Ohyoshi Y, Makino T, Nakayama W, Fukushima S, Inoue Y, Jinnin M, Ihn H.
  • Journal Title: Rheumatol Int, Volume: in press Pages: in press
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• [Journal Article] Up-regulation of miR-18a contributes to the epidermal necrolysis in severe drug eruptions. (2014)
  • Author(s): Ichihara A, Wang Z, Jinnin M, Izono Y, Shimozono N, Yamane K, Fujisawa A, Moriya C, Fukushima S, Inoue Y, Ihn H.
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• [Journal Article] Sensitive detection of melanoma metastasis utilizing circulating microRNA expression profiles. (2014)
  • Author(s): Shiiyama R, Fukushima S, Jinnin M, Yamashita J, Miyashita A, Nakahara S, Kogi A, Aoi J, Masuguchi S, Inoue Y, Ihn H.
  • Journal Title: Melanoma Res, Volume: in press Pages: in press
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• [Journal Article] Proportion of lymphocytic inflammation with CD123 positive cells in lupus erythematosus profundus predict a clinical response to treatment. (2014)
  • Author(s): Miyashita A, Fukushima S, Makino T, Yoshino Y, Yamashita J, Honda N, Aoi J, ,Ichihara A, Jinnin M, Ihn H
  • Journal Title: Acta Derm Venereol, Volume: in press Pages: in press
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• [Journal Article] miR-205 down-regulation promotes proliferation of dermatofibrosarcoma protuberans tumor cells by regulating LRP-1 and ERK phosphorylation. (2014)
  • Author(s): Kajihara I, Jinnin M, Harada M, Makino K, Honda N, Makino T, Igata T, Masuguchi S, Fukushima S, Ihn H.
  • Journal Title: Arch Dermatol Res, Volume: in press Pages: in press
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• [Journal Article] Decreased IL-20 expression in scleroderma skin contributes to cutaneous fibrosis. (2014)
  • Author(s): Kudo H, Jinnin M, Asano Y, Trojanowska M, Nakayama W, Inoue K, Honda N, Kajihara I, Makino K, Fukushima S, Ihn H.
  • Journal Title: Arthritis Rheumatol, Volume: in press Pages: in press
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• [Journal Article] Effects of the immunosuppressant rapamycin on the expression of human α2(I) collagen and matrix metalloproteinase 1 genes in scleroderma dermal fibroblasts. (2014)
  • Author(s): Tamaki Z, Asano Y, Kubo M, Ihn H, Tada Y, Sugaya M, Kadono T, Sato S.
  • Journal Title: J Dermatol Sci, Volume: in press Pages: in press
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• [Journal Article] miR-424 levels in hair shaft are increased in psoriasis patients. (2014)
  • Author(s): Tsuru Y, Jinnin M, Ichihara A, Fujisawa A, Moriya C, Sakai K, Fukushima S, Ihn H.
  • Journal Title: J Dermatol, Volume: in press Pages: in press
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• [Journal Article] Clinical significance of serum VE-cadherin concentrations in inflammatory skin diseases. (2014)
  • Author(s): Kuriyama H, Jinnin M, Kanemaru H, Ichihara A, Fujisawa A, Moriya C, Fukushima S, Ihn H.
  • Journal Title: Ann Dermatol, Volume: in press Pages: in press
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• [Journal Article] miR-150 down-regulation contributes to the constitutive type I collagen overexpression in scleroderma dermal fibroblasts via the induction of integrin β3. (2013)
  • Author(s): Honda N, Jinnin M, Kira-Etoh T, Makino K, Kajihara I, Makino T, Fukushima S, Inoue Y, Okamoto Y, Hasegawa M, Fujimoto M, Ihn H.
  • Journal Title: Am J Pathol, Volume: 182 Pages: 206-216
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• [Journal Article] The rs2910164 G>C polymorphism in microRNA-146a is associated with the incidence of malignant melanoma. (2013)
  • Author(s): Yamashita J, Iwakiri T, Fukushima S, Jinnin M, Miyashita A, Hamasaki T, Makino T, Aoi J, Masuguchi S, Inoue Y, Ihn H.
  • Journal Title: Melanoma Res, Volume: 23 Pages: 13-20
  • Peer Reviewed
• [Journal Article] The miR-146a rs2910164G/C polymorphism is associated with telangiectasia in systemic sclerosis. (2013)
  • Author(s): Sakoguchi A, Jinnin M, Makino T, Kajihara I, Makino K, Honda N, Nakayama W, Inoue K, Fukushima S, Ihn H.
  • Journal Title: Clin Exp Dermatol, Volume: 38 Pages: 99-100
  • Peer Reviewed
• [Journal Article] microRNA-7 down-regulation mediates excess collagen expression in localized scleroderma. (2013)
  • Author(s): Etoh M, Jinnin M, Makino K, Yamane K, Nakayama W, Aoi J, Honda N, Kajihara I, Makino T, Fukushima S, Ihn H.
  • Journal Title: Arch Dermatol Res, Volume: 305 Pages: 9-15
  • Peer Reviewed
• [Journal Article] Down-regulation of miR-124/ -214 in cutaneous squamous cell carcinoma mediates abnormal cell proliferation via the induction of ERK. (2013)
  • Author(s): Yamane K, Jinnin M, Etoh T, Kobayashi Y, Shimozono N, Fukushima S, Masuguchi S, Maruo K, Inoue Y, Ishihara T, Aoi J, Oike Y, Ihn H.
  • Journal Title: J Mol Med, Volume: 91 Pages: 69-81
  • Peer Reviewed
• [Journal Article] Intravenous immunoglobulin treatment recovers the down-regulatied levels of Th1 cytokines in the sera and skin of scleroderma patients. (2013)
  • Author(s): Kudo H, Jinnin M, Yamane K, Makino T, Kajihara I, Makino K, Honda N, Nakayama W, Fukushima S, Ihn H.
  • Journal Title: J Dermatol Sci, Volume: 69 Pages: 77-80
  • Peer Reviewed
• [Journal Article] Diccoidin domain receptor 2-micro RNA 196a -mediated negative feedback against excess type I collagen expression is impaired in scleroderma dermal fibroblasts. (2013)
  • Author(s): Makino K, Jinnin M, Aoi J, Hirano A, Kajihara I, Makino T, Sakai K, Fukushima S, Inoue Y, Ihn H.
  • Journal Title: J Invest Dermatol, Volume: 133 Pages: 110-119
  • Peer Reviewed
• [Journal Article] Serum epidermal growth factor receptor levels in patients with malignant melanoma. (2013)
  • Author(s): Kanemura H, Fukushima S, Yamashita J, Jinnin M, Sakai K, Masuguchi M, Aoi J, Makino T, Inoue Y, Ihn H.
  • Journal Title: Clin Exp Dermatol, Volume: 38 Pages: 172-177
  • Peer Reviewed
• [Journal Article] Serum miR-21 levels in patients with dermatomyositis. (2013)
  • Author(s): Shimada S, Jinnin M, Ogata A, Makino T, Kajihara I, Makino K, Honda N, Nakayama W, Inoue K, Fukushima S, Ihn H.
  • Journal Title: Clin Exp Rheumatol, Volume: 31 Pages: 161-162
  • Peer Reviewed
• [Journal Article] Serum levels of interleukin-1α in patients with systemic sclerosis. (2013)
  • Author(s): Maekawa T, Jinnin M, Ohtsuki M, Ihn H.
  • Journal Title: J Dermatol, Volume: 40 Pages: 98-101
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• [Journal Article] Suppresion of UV-induced wrinkle formation by induction of HSP70 expression in mice. (2013)
  • Author(s): Matsuda M, Hoshino T, Yamakawa N, Tahara K, Adachi H, Sobue G, Maji D, Ihn H, Mizushima T.
  • Journal Title: J Invest Dermatol, Volume: 133 Pages: 919-928
  • Peer Reviewed
• [Journal Article] Hidradenocarcinoma showing prominent mucinous and squamous differentiation and associated with pgetoid cells. (2013)
  • Author(s): Honda Y, Tanigawa H, Harada M, Fukushima S, Masuguchi S, Ishihara T, Ihn H, Iyama K.
  • Journal Title: J Cutan Pathol, Volume: 40 Pages: 503-506
  • Peer Reviewed
• [Journal Article] Case of paraneoplastic pemphigus with follicular lymphoma treated with rituximab. (2013)
  • Author(s): Aoi J, Makino K, Sakai K, Masuguchi S, Fukushima S, Jinnin M, Inoue Y, Koga H, Hashimoto T, Ihn H
  • Journal Title: J Dermatol, Volume: 40 Pages: 285-256
  • Peer Reviewed
• [Journal Article] The down-regulation of microRNA let-7a contributes to the excessive expression of type I collagen in systemic and localized scleroderma. (2013)
  • Author(s): Makino K, Jinnin M, Hirano A, Yamane K, Eto M, Kusano T, Honda N, Kajihara K, Makino T, Sakai K, Masuguchi S, Fukushima S, Ihn H.
  • Journal Title: J Immunol, Volume: 190 Pages: 3905-3915
  • Peer Reviewed
• [Journal Article] Down-regulation of miR-223 contributes to the formation of Gottron’s eruption in dermatomyositis via the induction of PKCε (2013)
  • Author(s): Inoue K, Jinnin M, Yamane K, Makino T, Kajihara I, Makino K, Honda N, Nakayama W, Fukushima S, Ihn H.
  • Journal Title: Eur J Dermatol, Volume: 23 Pages: 160-167
  • Peer Reviewed
• [Journal Article] Pyodermatitis vegetans associated with multiple myeloma. (2013)
  • Author(s): Kajihara I, Ichihara A, Higo J, Kidou M, Ihn H.
  • Journal Title: J Dermatol, Volume: 40 Pages: 222-223
  • Peer Reviewed
• [Journal Article] Decreased miR-7 expression in the skin and sera of patients with dermatomyositis. (2013)
  • Author(s): Oshikawa Y, Jinnin M, Makino T, Kajihara I, Makino K,Honda N, Nakayama W, Inoue K, Fukushima S, Ihn H.
  • Journal Title: Acta Derm Venereol, Volume: 93 Pages: 273-276
  • Peer Reviewed
• [Journal Article] Expression analysis of multiple microRNAs in each patient with scleroderma (2013)
  • Author(s): Koba S, Jinnin M, Inoue K, Nakayama W, Honda N, Makino K, Kajihara I, Makino T, Fukushima S, Ihn H.
  • Journal Title: Exp Dermatol, Volume: 22 Pages: 489-491
  • Peer Reviewed
• [Journal Article] Scleroderma dermal fibroblasts overexpress vascular endothelial growth factor due to autocrine transforming growth factor βsignaling. (2013)
  • Author(s): Kajihara I, Jinnin M, Honda N, Makino K, Makino T, Masuguchi S, Sakai K, Fukushima S, Inoue Y, Ihn H.
  • Journal Title: Mod Rheumatol, Volume: 23 Pages: 516-524
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• [Journal Article] Geographical flushing of the children’s face: A new clinical entity. (2013)
  • Author(s): Jinnin M, Fukushima S, Inoue Y, Ihn H.
  • Journal Title: Case Rep Dermatol Med, Volume: - Pages: 802130
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• [Journal Article] A randomized, double-blind, placebo-controlled trial: intravenous immunoglobulin treatment in patients with diffuse cutaneous systemic sclerosis. (2013)
  • Author(s): Takehara K, Ihn H, Sato S.
  • Journal Title: Clin Exp Rheum, Volume: 31 Pages: 151-156
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• [Journal Article] Serum levels of tenascin-C in collagen diseases. (2013)
  • Author(s): Inoue K, Jinnin M, Hara Y, Makino K, Kajihara I, Makino T, Sakai K, Fukushima S, Inoue Y, Ihn H.
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• [Journal Article] Case of neuro-Behcet’s disease successfully maintained a remission using infliximab for 2 years. (2013)
  • Author(s): Kanemaru H, Makino T, Jinnin M, Yonemitsu A, Makino K, Ihn H.
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• [Journal Article] Role of c-Jun N-termainal kinase isoforms in the cellular activity of melanoma cell lines. (2013)
  • Author(s): Kogushi-Nishi M, Jinnin M, Kobayashi Y, Muchemwa FC, Hirano A, Makino T, Fukushima S, Masuguchi S, Ihsihara T, Ihn H.
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• [Journal Article] Association of linear IgA bullous disease with ulcerative colitis: A case of successful treatment with infliximab. (2013)
  • Author(s): Yamada S, Makino T, Jinnin M, Sakai K, Fukushima S, Inoue Y, Ihn H.
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• [Journal Article] Detection of hair root miR-19a as a novel diagnostic marker for psoriasis. (2013)
  • Author(s): Hirao H, Jinnin M, Ichihara A, Fujisawa A, Makino K, Kajihara I, Sakai K, Fukushima S, Inoue Y, Ihn H.
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• [Journal Article] Hair miR-29a levels are decreased in patients with scleroderma. (2013)
  • Author(s): Takemoto R, Jinnin M, Wang Z, Kudo H, Inoue K, Nakayama W, Ichihara A, Igata T, Kajihara I, Fukushima S, Ihn H.
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  • Peer Reviewed
• [Journal Article] Detection of hair-microRNAs as the novel potent biomarker: Evaluation of the usefulness for the diagnosis of scleroderma. (2013)
  • Author(s): Wang Z, Jinnin M, Kudo H, Inoue K, Nakayama W, Honda N, Makino K, Kajihara I, Fukushima S, Inoue Y, Ihn H.
  • Journal Title: Detection of hair-microRNAs as the novel potent biomarker: Evaluation of the usefulness for the diagnosis of scleroderma. Volume: 72 Pages: 134-141
  • Peer Reviewed
• [Journal Article] Serum chemokine levels as prognostic markers in patients with early systemic sclerosis: a multicentre, prospective, observational study. (2013)
  • Author(s): Hasegawa M, Asano Y, Endo H, Fujimoto M, Goto D, Ihn H, Inoue K, Ishikawa O, Kawaguchi Y, Kuwana M, Ogawa F, Takahashi H, Tanaka S, Sato S, Takehara K.
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• [Journal Article] CEA (carcinoembryonic antigen) and CEACAM6 (CEA-related cell adhesion molecule 6) are expressed in psoriasis vulgaris. (2013)
  • Author(s): Fujisawa A, Egawa K, Honda Y, Kuroki M, Jinnin M, Ihn H.
  • Journal Title: The Open Dermatol J Volume: 7 Pages: 47-52
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• [Journal Article] Serum levels of leptin receptor in patients with malignant melanoma as a new tumor marker. (2013)
  • Author(s): Mizutani H, Fukushima S, Masuguchi S, Yamashita J, Miyashita A, Nakahara S, Aoi J, Inoue Y, Jinnin M, Ihn H.
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  • Peer Reviewed
• [Journal Article] 創傷・熱傷ガイドライン (2013)
  • Author(s): 尹 浩信
  • Journal Title: Asahi Medical Volume: 42 Pages: 71-73
• [Journal Article] 乾癬治療における生物学的製剤の適応と使い方 (2013)
  • Author(s): 尹 浩信
  • Journal Title: 日経CME Volume: 1 Pages: 1-6
• [Journal Article] 実地医科のための強皮症早期診断とガイドラインの考え方 (2013)
  • Author(s): 尹 浩信、藤本 学、佐藤伸一
  • Journal Title: マルホ皮膚科セミナー Volume: 224 Pages: 4-12
• [Journal Article] 関節症性乾癬へのアプローチ (2013)
  • Author(s): 尹 浩信
  • Journal Title: 日経CME Volume: 2 Pages: 1-6
• [Journal Article] 褥瘡治療の最近の進歩 褥瘡ガイドラインをどう生かすか？ (2013)
  • Author(s): 尹 浩信
  • Journal Title: Geriat. Med. Volume: 51 Pages: 1273-1279
• [Presentation] Effects of sarpogrelate for Raynaud’s phenomenon secondary to systemic sclerosis. (2014)
  • Author(s): Makino T, Jinnin M, Kajihara I, Makino K, Ihn H.
  • Organizer: 3 rd Systemic Sclerosis World Congress
  • Place of Presentation: Rome
  • Year and Date: 20140206-20140208
• [Presentation] Scleroderma dermal fibroblasts overexpress vascular endothelial growth factor due to autocrine transforming growth factor-β signaling. (2014)
  • Author(s): Kajihara I, Jinnin M, Ihn H.
  • Organizer: 3 rd Systemic Sclerosis World Congress
  • Place of Presentation: Rome
  • Year and Date: 20140206-20140208
• [Presentation] 色素レーザーが有効であった毛細血管拡張症をを伴った全身性強皮症の1例 (2014)
  • Author(s): 牧野貴充、梶原一亨、神人正寿、尹 浩信
  • Organizer: 第37回皮膚脈管・膠原病研究会
  • Place of Presentation: 東京医科歯科大学M&Dタワー
  • Year and Date: 20140124-20140125
• [Presentation] The effect of IL-17 on the extracellular matrix expression in dermal fibroblasts. (2013)
  • Author(s): Jinnin M, Ihn H.
  • Organizer: The American Society for Biochemistry and Molecular Biology
  • Place of Presentation: Boston (アメリカ）
  • Year and Date: 20130810-20130811
• [Presentation] V beamが有効であった全身性強皮症に伴う毛細血管拡張症の1例 (2013)
  • Author(s): 牧野貴充、神人正寿、尹 浩信
  • Organizer: 第31回日本美容皮膚科学会
  • Place of Presentation: 神戸国際会議場（神戸）
  • Year and Date: 20130810-20130811
• [Book] 看護学大辞典 (2013)
  • Author(s): 尹 浩信
  • Total Pages: 2480
  • Publisher: メジカルフレンド社
• [Book] Monthly Book Derma No.202治りにくい皮膚病変 治療の実際 (2013)
  • Author(s): 尹 浩信
  • Total Pages: 80
  • Publisher: 全日本病院出版会
• [Book] 皮膚科臨床アセット12 新しい創傷治療のすべて 褥瘡、熱傷、皮膚潰瘍 (2013)
  • Author(s): 尹 浩信
  • Total Pages: 368
  • Publisher: 中山書店
• [Book] 創傷と痛み (2013)
  • Author(s): 尹 浩信
  • Total Pages: 168
  • Publisher: 金原出版
• [Book] 皮膚疾患最近の話題 (2013)
  • Author(s): 尹 浩信
  • Total Pages: 304
  • Publisher: 南江堂
• [Book] 一冊で分かる最新皮膚科治療 (2013)
  • Author(s): 尹 浩信
  • Total Pages: 328
  • Publisher: 文光堂