• 대한한방소아과학회지
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• 제23권2호
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• pp.51-85
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• 2009

Objectives : The purpose of this study is to investigate the effect of concurrent administration of KKSDU and AJ on atopic dermatitis in an in-vivo experiment using an NC/Nga atopic dermatitis mouse, which has histological and clinical similarities to the condition in humans. Methods : We evaluated clinical skin score, hematology, serum total IgE and IgG1 of NC/Nga atopic dermatitis mouse and analyzed the cytoline level, total cell number, immunohistochemical staining, histological features of axillary lymph node(ALN), draining lymph node(DLN), peripheral blood mononuclear cells(PBMCs) and dorsal skin tissue in NC/Nga mouse. Results : Orally administration of KKSDU and concurrent administration of KKSDU and AJ decreased the clinical skin score, total cell number of WBC, platelet, neutrophils, eosinophils in blood, serum total IgE & IgG1, IL-5, IL-13. Also, total cell number of ALN and dorsal skin tissue, absolute cell number of CD3e+&CD19+, CD4+&CD8+, CD3+/CCR3+, CCR3+, CD3+/CD69+, CD3+/CXCR5+ in ALN, PBMCs, absolute cell number of CCR3+, CD3+/CD69+, CD11b+/Gr-1+ in dorsal skin tissue, Eotaxin2 mRNA, CCR3 mRNA in dorsal skin tissue and gene expression of IL-5 mRNA, IL-13 mRNA in ALN are significantly decreased. Furthermore, thickness of epidermis, infiltrated inflammatory immune cell & mast cell in dermis, histologic infiltration of mast cell, the size of inflammatory lymphocytes cells & plasma cells in ALN and histologic infiltration of CD4+ & CCR3+ in ALN and dorsal skin tissue are significantly decreased. However, total cell number of DLN, absolute cell number of CD3+&CD19+, CD4+&CD8+, B220+/CD23+, CD3+/CD69+ in DLN and CD4+CD25+foxp3+Treg cell, foxp3 mRNA in dersal skin tissue are increased significantly. Conclusions : Concurrent administration of KKSDU and AJ on atopic dermatitis in an in-vivo experiment using an NC/Nga atopic dermatitis mouse was very effective to the atopic detmatitis treatment.

• 대한한방소아과학회지
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• 제23권1호
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• pp.37-72
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• 2009

Objectives The purpose of this study is to investigate the suppressive effects of Atopy cream-combined with Jawoongo ointment (A-J), on the development of atopic dermatitis-like skinlesions in NC/Nga mouse. Methods We evaluated clinical skin score, hematology and Serum total IgE and IgG1 of NC/Nga atopic dermatitis mouse, analyzed the cytokine level, total cell number, Immunohistochemical staining and Histological features of axillary lymph node(ALN), draining lymph node(DLN), peripheral blood mononuclear cells(PBMCs) and dorsal skin tissue in NC/Nga mouse. Results A-J decreased the clinical skin score, total cell number of WBC, platelet, neutrophils, eosinophils in blood, Serum total IgE & IgG1, IL-5, IL-13. Also, total cell number of ALN and dorsal skin tissue, Absolute cell number of $CD3e^+$&$CD19^+$, $CD4^+$&$CD8^+$, $CD3^+/CCR3^+$, $CCR3^+$, $CD3^+/CD69^+$, $CD3^+/CXCR5^+$ in ALN, PBMCs, Absolute cell number of $CCR3^+$, $CD3^+/CD69^+$, $CD11b^+/Gr-1^+$ in dorsalskin tissue, Eotaxin2 mRNA, CCR3 mRNA in dorsal skin tissue and gene expression of IL-5 mRNA, IL-13 mRNA in ALN decreased significantly. Furthermore, thickness of epidermis, infiltrated inflammatory immune cell and mast cell in dermis, histologic infiltration of mast cell, the size of inflammatory lymphocytes cells and plasma cells in ALN and histologic infiltration of CD4+ & CCR3+ in ALN and dorsal skin tissue decreased significantly. However, total cell number of DLN, absolute cell number of $CD3e^+$&$CD19^+$, $CD4^+$&$CD8^+$, $B220^+/CD23^+$, $CD3^+/CD69^+$ increased significantly. Conclusions A-J was the successful treatment of atopic dermatitis in a NC/Nga mouse model.

• 대한한방소아과학회지
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• 제28권4호
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• pp.1-29
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• 2014

Objectives Dictamni Radicis Cortex extracts (DRC) has been known to suppress allergic reaction, however the cellular target of DRC and its mode of action remain unclear. The purpose of this study is to investigate the effects of Dictamni Radicis Cortex extracts on DNCB induced atopic dermatitis-like skin lesions of NC/Nga mouse. Methods This study was designed to investigate the effects of DRC extract in the DNP-IgE-induced activation of MC/9 murine mast cell lines in vitro and in the DNCB-induced activation of NC/Nga mouse in vivo. For this investigation, We examined IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF mRNA expression by Real-time PCR, IL-13, MIP-$1{\alpha}$ production by ELISA analysis and manifestations of NFAT1, NFAT2, AP-1 and NF-${\kappa}B$ p65 transcription factors by western blotting in vitro. Then, we examined WBC, eosinophil and neutrophil in NC/Nga mouse, IL-5, IL-13 in serum, IFN-${\gamma}$, IL-4 in the spleenocyte culture supernatant, the absolute cell numbers of $CD4^+$, $CD8^+$, $^+Gr-1^+CD11b$, $B220^+CD23^+$ in the ALN, PBMCs and dorsal skin, IL-5, IL-13 in the dorsal skin by Real-time PCR and the distribution of mast cells by H&E and toluidine blue. Results In vitro the mRNA expression of IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$, GM-CSF and IL-13, MIP-$1{\alpha}$ production by ELISA analysis were completely abolished by DRC and the western blot analysis decreased the expression of mast cell-specific transcription factors including NFAT-1, NF-${\kappa}B$ p65. In vivo DRC oral adminstration also decreased the counts of WBC, eosinophils and inflammatory cytokines such as IL-13 and IgE in the serum. DRC oral adminstration elevated IL-4 level in the spleenocyte culture supernatant. DRC oral adminstration decreased total ALN cells, total skin cells, cell numbers of $CD4^+$, $B220^+CD23^+$ in the ALN, $^+Gr-1^+CD11b$ in the PBMCs and $CD4^+$, $CD8^+$ in the dorsal skin. The mRNA expression of IL-5, IL-13, thickness of epidermis, inflammation immune cells and mast cells were abolished by DRC in the dorsal skin. Conclusions Histological examination showed that infiltration levels of immune cells in the skin of AD-induced NC/Nga mouse were much improved by DRC oral adminstration. These results, therefore, suggest that DRC can regulate molecular mediators and immune cells that are functionally associated with atopic dermatitis induced in NC/Nga mouse, and may play an important role in recovering AD symptoms.

• 대한한방소아과학회지
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• 제30권3호
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• pp.1-30
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• 2016

Objectives Previous studies have found out that Forsythiae Fructus (FF) extracts have anti-atopic activities by in vitro experiment. In order to understand more about FF extracts' benefit, we subdivided FF extracts depending on systematic fractionation method by using Methylene chloride (MC), Ethyl acetate (EtOAc), n-BuOH and n-hexane (n-Hx). This study is designed to examine the effect of FF fractions on the PMA- ionomycin-induced activation of RBL-2H3 mast cell lines in vitro and on the DNCB-induced activation of NC/Nga mice in vivo. Methods For this study, we examined IL-4, IL-13 production by ELISA analysis, IL-4, IL-13, IL-31, IL-31RA and TNF-${\alpha}$ mRNA expression by real-time PCR and manifestations of AP-1 and MAPKs transcription factors by western blotting in vitro. Through in vitro experiment, we selected FF n-BuOH fraction that seems the best effective in atopic dermatitis then induced it on NC/Nga mice by DNCB. We measured mice's WBC, eosinophil and neutrophil in heart blood, IL-4, IL-5, IFN-${\gamma}$ in the spleenocyte culture supernatant, the absolute cell numbers of CD4+, CD8+, B220+CD23+, CD3+CD69+ and Gr-1+CD11b+ in the PBMCs, ALN and dorsal skin, IL-5, IL-13, IL-31, IL-31RA in the dorsal skin by real-time PCR and the distribution of immune cells by H&E on dorsal skin and ANL and toluidine blue staining on dorsal skin. Results FF n-BuOH fraction suppressed IL-4, IL-13 production and mRNA expression of IL-4, IL-13, IL-31, IL-31RA and TNF-${\alpha}$. Results from the western blot analysis showed that FF n-BuOH fraction reduced the activation of the mast cell specific transduction factors involved in AP-1 by suppressing JNK and ERK phosphorylation. In the gross, atopic dermatitis induced by DNCB in NC/Nga mice were improved by oral administration of FF n-BuOH fraction. Oral FF n-BuOH fraction also decreased the level of IgE in mice's serum and the level of IL-4 and IL-5 in the spleenocyte culture supernatant, cell numbers of CD8+, B220+CD23+ in the PBMCs, CD4+ in the ALN and CD4+, Gr-1+CD11b+ in the dorsal skin and suppressed mRNA expression of IL-5, IL-13, IL-31, IL-31RA in the dorsal skin. Histological examination showed that infiltration levels of immune cells in atopic dermatitis induced NC/Nga mice were improved by FF n-BuOH fraction. Conclusions FF n-BuOH fraction can reduce pruritus by suppressing IL-31, IL-31RA secretion and modulate molecular mediators and immune cells associated with atopic dermatitis induced in NC/Nga mice which may have played a significant role in recovering atopic dermatitis symptoms.

• 대한본초학회지
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• 제24권4호
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• pp.215-224
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• 2009

Objectives : Atopic dermatitis is a chronic inflammatory disease characterized by typically distributed eczematous skin lesion with pruritus, lichenification and dry skin. In this study, we performed to assess the therapeutic effects of co-treatment of Chenilyeomgamibang (CGB) and Chenggihaedok-san (CHS, C&C) on the TNCB(trinitrochlorobenzene)-induced atopic dermatitis in NC/Nga mice, characterized by the onset of atopic dermatitis along with an increase the number of inflammatory cells and dysregulation of Th2 cytokines. Methods : Defined amount of CGB was sprayed on mice skin and CHS was simultaneously orally administrated to TNCB treated NC/Nga mice for 5 weeks. The immune cell types were caracterized by flow cytometry using each specific antibody. The amount of Th2 cytokines in serum and splenocytes culture supernatant were measured by ELISA. Results : Administration of C&C significantly reduced clinical dermatitis severity including pruritus, edema, eczematous and erythema. Histological findings indicated that the thickening of epidermis/dermis and dermal infiltration of inflammatory cells were dramatically reduced. Flow cytometry analysis showed that infiltrated immune cell numbers of CCR3+, B220+/IgE+, Gr-1+/CD11b+, and CD117+ were significantly reduced in C&C-treated dorsal skin lesion. Furthermore, T cell composition rate in PBMC was also dramatically decreased by the treatment. C&C greatly down-regulated production of Th2 cytokines including IL-4, IL-5, IL-13 in the serum. The down- regulatory effects of C&C on these Th2 cytokines production were also detected in CD3/ CD28 activated splenocytes. Conclusions : These results indicated that C&C is a plausible therapeutic agent for treatment of atopic dermatitis through regulating the Th2 skewed immune system.

• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5909-5913
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• 2012

Aims: We aimed to analyze the phenotype of tumor-infiltrating lymphocytes (TILs) and non-tumor infiltrating lymphocytes (NILs) in HCC and non-tumor tissues, and evaluate relationships between changes in these cells and the prognosis of HCC. Methods: Lymphocytes were isolated from HCC and corresponding non-tumor tissues and tested by flow cytometry. For comparison, clinical parameters were analyzed. Results: Compared with the non-tumor tissue, tumor tissue had a lower intensity of NK, NKT andCD8+T cell infiltration. TILs had higher intensity of CD4+CD25+Foxp3+regulatory T cell (Treg cells) infiltration compared with that in NILs. The prevalence of Treg cells was associated with fewer CD8 + T lymphocytes in the HCC immune microenvironment. The frequencies of NK cells and CD8+T cells in TILs of HCC patients with metastasis less than 12 months were lower than those without metastasis. However, the frequency of Treg cells was higher than those without metastasis. Conclusion: These results suggest that the frequencies of CD8+T, NK and NKT cells as well as Treg cells in the tumor tissue of HCC are significantly associated with patient survival, and could be applied as predictive indicators for HCC prognosis.

• Restorative Dentistry and Endodontics
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• 제24권1호
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• pp.212-221
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• 1999

The periapical response to injury is a complex interaction of inflammatory, immune, neural, vascular and synthetic activity. Nitric oxide(NO), synthesized by nitric oxide synthetase(NOS) from L-arginine, is becoming recognized as an important bio-regulatory molecule in a variety of tissue, but little is known about its possible role in periapical tissue. The purpose of this study was to investigate the expression of nitric oxide synthetase(NOS) in tooth follicle, periapical abscess, granuloma and cyst. The expression of NOS in periapical lesions was evaluated by immunohistochemical staining for $NOS_2$, and $NOS_3$. The immunoreactivity was evaluated by staining intensity, and inflammatory cell infiltration. Correlationship between the periapical lesion in immunoreractivity were statistically analyzed by SPSS. The degree of $NOS_2$ and $NOS_3$ expression in periapical abscess was higher than in any other periapical lesions, and stastically significant. The expression degree of $NOS_2$ and $NOS_3$ was not correlated with periapical abscess and granuloma, but expression of $NOS_2$ showed very significant in periapical cyst. The increased expression of $NOS_2$ and $NOS_3$ was correlated with inflammatory cell infiltration degree of the periapical cyst. These results suggested that NO should play an important role in progress and/or mediation of periapical lesions.

• Journal of Periodontal and Implant Science
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• 제23권1호
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• pp.56-66
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• 1993

The Langerhans cells are dendritic nonkeratinocytes found suprabasally in most stratified squamous epithelia, such as human epidermis and the epithelium of the oral mucosa including that of gingiva. After Paul Langerhans found it in the skin in 1968, there have been sturdies of it's function and distribution . Stingle et al. reported that the Langerhans cells seem able to present antigens and to stimulate T-lymphocytes. Shelley et al. discovered that they can take up contact allergens. Accordingly it has been suggested that Langerhans cells are important elements of p Peripheral cell mediated immune system. In this study, the gingival tissue of a adult periodontitis patient was taken and freeze dried. In one specimen, we used the CD1 monoclonal antbody to staining the Langerhans cell. The other specimen, we embedded in paraffin and staining it with S-100 monoclonal antibody. The purpose of this study was to use these specimens to find out the distribution, orientation, morphology of the Langerhans cell and to discover the increase or decrease of Langerhans cell in an increased inflammatory state. The results were obtained as follows : 1. Langerhans cells were distributed between the basal cell layer and spinous cell layer against the CD1 & S-100 monoclonal antibody. 2. Langerhans cessl were plentiful in the oral eptihelium, and there was very little in the sulcular epithelium. 3. There were no Langerhans cell in the junction epithelium and pocket lining epithelium. 4. The number of Langerhans cells that responsed to the CD1 & S-100 monoclonal antibody had a statistically difference. 5. As the infiltration of the lymphocyte into the connective tissue were increased, the number of Langerhans cells in the epithelium were increased. 6. As the inflammation was increased, Langerhans cells in the spinous cell layer were more increased than those of the basal layer.

• The Korean Journal of Physiology and Pharmacology
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• 제21권3호
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• pp.335-343
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• 2017

Atopic dermatitis (AD) is an inflammatory skin condition accompanied by symptoms such as edema and hemorrhage. Kimchi is a traditional fermented Korean dish consisting of various probiotics. In this study, the therapeutic effect of Lactobacillus plantarum CJLP55 isolated from Kimchi was studied in AD-induced mice. Orally administered Lactobacillus strain, CJLP55, suppressed AD symptoms and high serum IgE levels. CJLP55 administration reduced the thickness of the epidermis, infiltration of mast cells and eosinophils into the skin lesion, enlargement of axillary lymph nodes, and increase in cell population in axillary lymph nodes. CJLP55 treatment decreased the production of type 2 cytokines, such as interleukin (IL)-4, IL-5, IL-10, IL-12, interferon (IFN)-${\gamma}$, and IL-6,which were stimulated by house dust mite extracts, in the axillary lymph node cells. Orally administered CJLP55 exhibited a therapeutic effect on house dust mite-induced AD in NC/Nga mice after onset of the disease by altering immune cell activation. The Lactobacillus strain, CJLP55, isolated from Kimchi, suppressed AD. Our results suggest its possible use as a potential candidate for management of AD.

• 생명과학회지
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• 제25권6호
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• pp.686-692
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• 2015

암조직에 대한 면역반응은 종양의 성장을 억제하거나 유도한다. Foxp3 양성 T 림프구는 종양에 대한 면역억제 반응을 유도하여 종양의 성장을 유도하는 반면 CD8 양성 T 림프구는 종양의 성장을 억제한다. 본 연구에서는 피부에서 발생한 편평세포암종, 편평세포 암종의 전암병변인 광선각화증과 보웬병 그리고 기저세포암종 조직에 면역조직화학염색을 시행하여 CD8 양성 T 림프구 및 Foxp3 양성 T 림프구의 분포 그리고 두 종류의 림프구의 비(CD8/Foxp3)를 조사하여 다음과 같은 결과를 얻었다. CD8 양성 T 림프구와 Foxp3 양성 T 림프구는 보웬병과 평평세포암종의 침습부위에서 광선각화증과 기저세포암종의 침습부위에 비하여 더 많이 침윤하였으며 CD8/Foxp3는 보웬병에서 광선각화증과 기저세포암종의 침습부위에 비하여 낮았다. CD8 양성 T 림프구와 Foxp3 양성 T 림프구의 침윤 정도 및 CD8/Foxp3는 보웬병과 편평세포암종의 침습부위에서 유의한 차이가 없었다. 기저세포암종과 편평세포암종에서 CD8 양성 T 림프구와 Foxp3 양성 T 림프구는 침습부위에서 종양의 중심부위에 비하여 더 많이 침윤하였다. 이러한 결과로 보아 편평세포종양과 기저세포암종은 서로 다른 면역반응을 보이며 종양내에서도 부위에 따라 상이한 면역반응을 보인다고 생각된다.