• 대한한방소아과학회지
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• 제22권3호
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• pp.105-137
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• 2008

Objectives : The purpose of this study is to investigate the effect of Gupoongjeseuptang (GPJST) on atopic dermatitis by in vivo experiment using NC/Nga atopic dermatitis mouse, which has histological and clinical similarities to the atopic dermatitis of human. Methods : To investigate the effect of GPJST on atopic dermatifis, we evaluated atopic dermatitis-like skin lesions by clinical skin index and analyzed immunological parameters in peripheral blood mononuclear cells (PBMCs), splenocytes, draining lymph node (DLN) and performed skin histology in ears and dorsal skin of atopic dermatitis-like skin NC/Nga mouse in vivo. Results : In vivo, clinical skin severity score were significantly lower in GPJST group than control group. IgE, IL-6, $TNF-{\alpha}$, IgG1, IgM, IgG2a and IgG2b levels in serum decreased remarkably in GPJST group than control group. Also, total absolute number of $CD3^+CD69^+$, and $CCR3^+$ cells recovered as normal in PBMCs and $CD3^+$, $CD3^+CD69^+$ decreased significantly compared with control group in isolated DLN from NC/Nga mouse and total absolute number of $CD11b^+Gr-1^+$, $CCR3^+CD3^+$ in dorsal skin of NC/Nga mouse decreased by GPJST. We analyzed ear and neck-back skin after biopsy and dyeing by hematoxyline/eosin (H&E) and toluidine staining (mast cells marker) and obtained results that GPJST are very effective to histological symptoms (dermal and epidermal thickening, hyperkeratosis and inflammatory cell (CD4, $CCR3^+$) infiltration). Conclusions : This study demonstrates immunological activity of GPJST on atopic dermatitis-like model mice.

• 한방재활의학과학회지
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• 제20권2호
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• pp.17-34
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• 2010

Objectives : This study was carried out to know the effects of Gwanjul8-bang(here in after reffered to GJ8) on the inhibition of arthritis induced by collagen on DBA/1J mice. Methods : For this purpose, GJ8 was orally administered to mouse with arthritis induced by collagen II. Cytotoxicity, hepatotoxicity, arthritis index, value of immunocyte in draining lymph node(DLN) and paw joint, cytokine in serum were measured in vivo. Results : 1. The arthritis index was decreased significantly. 2. In total cell counts of DLN and paw joint, the cells in DLN increased significantly while there was a significant decrease in paw joint. 3. In lymph nodes, $CD19^+$, $CD3^+$, $CD4^+/CD25^+$ cells increased significantly, and $B220^+/CD23^+$ cells decreased significantly. 4. In joints, $CD3^+$, $CD4^+$, $CD11b^+/Gr-1^+$ cells decreased significantly. 5. The levels of $TNF-{\alpha}$, IL-6, IL-17, MCP-1 and vascular endothelial growth factor(VEGF) in serum was significantly decreased compared with control. 6. Anti-collagen II in serum was significantly decreased compared with control. 7. The degree of arthritis induced damage of joint of GJ8 group is slight compared with control group in histopathologic observation(hematoxylin & eosin(H&E), Masson-Trichrome(MT) staining). Conclusions : Comparison of the results for this study showed that GJ8 had immunomodulatory effects. So we expect that GJ8 should be used as a effective drugs for not only rheumatoid arthritis but also another auto-immune disease. Therefore there seems to be many clinical research needed in the future.

• 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.

• Journal of Acupuncture Research
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• 제22권1호
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• pp.131-144
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• 2005

족삼리(足三里) 사과락약침(絲瓜絡藥鍼)이 DBA/lJ mouse의 collagen-induced arthritis 미치는 영향(影響)을 관찰(觀察) 한 결과(結果), 다음과 같은 결론(結論)을 얻었다. 1. 세포독성(細胞毒性) 측정결과(測定結果), 사과락약침액(絲瓜絡藥鍼液) 1%의 농도(濃度)에서 가장 높은 세포생존률(細胞生存率)이 관찰(觀察)되었다. 2. In vitro screening 결과(結果), 사과락약광액(絲瓜絡藥鑛液) 1%의 농도(濃度)에서 CIA mouse 관절활막세포(關節滑膜細胞)의 TNF-${\alpha}$발현(發現)이 유의(有意)하게 감소(減少)하였다. 3. 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)에 의하여 관절염(關節炎) 발병률(發病率)이 감소(減少)하였다. 4. 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)에 의하여 CIA mouse의 각장(脚臟)무게가 감소(減少)하였다. 5. 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)에 의하여 CIA mouse의 혈청(血淸) IL-6, INF-${\gamma}$, TNF-${\alpha}$, IgG, IgM 및 Anti-collagen Ⅱ가 유의(有意)하게 감소(減少)하였다. 6. 조직학적(組織學的) 검사(檢査) 결과(結果), 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)에 의하여 CIA mouse 관절(關節)의 연골파괴(軟骨破壞)와 활막증식(滑膜增殖)이 감소(減少)되고 교원질섬유(膠原質纖維)가 정상군(正常群)과 유사(類似)하게 유지(維持)되었다. 7. 임파절내(淋巴節內) CD3e+와 CD19+세포(細胞) 비율(比率), CD4+와 CD8+세포(細胞) 비율(比率)은 사과락약광치료군(絲瓜絡藥鑛治療群)에서 정상군(正常群)과 유사(類似)하게 유지(維持)되었다. 8. 임파절내(淋巴節內) CD69+/CD3e+세포(細胞) 및 CD11a+/CD19+세포(細胞)와 관절내(關節內) CD11b+/Gr-1+세포(細胞)는 사과락약침치료군(絲瓜絡藥鍼治療群)에서 대조군(對照群)에 비하여 유의(有意) 하게 감소(減少)하였다.

• 대한한방소아과학회지
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• 제21권2호
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• pp.13-34
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• 2007

Objectives The purpose of this study is to examine of the effect of SPAR medicines on the atopy eruption control Methods This experiment is about the expression of IgE, IL-4, IL-6, IL_13, IgM, IgG2a, IgG2b, IgG1 level in serum, and $IFN-{\gamma}$ production by SPAR medicines. We assayed for $CD3e^+/CD69^+$, $CD044^+/CD19^+$ positive cells by flow cytometry in splenocytes and observed the revelation of $CD3e^+/CD69^+$, $CD4^+/CD8^+$, $CD44^+/CD19^+$ marker in PBMC, spleen and DLN. We also observed the outturn of IL-4, IL-5, CCR3, $IFN-{\gamma}$ in skin of a NC/Nga mice. We also analyzed NC/Nga mice's ear and neck-back skin after biopsy and dye by H&E staining method, measured about epidermis and dermis part in comparison with control group. Results SPAR medicines as treatment result to a NC/Nga mice, clinical skin severity score decreased remarkably than the ontrol group. Specially, experiment was results by measuring IgE and IL-6 content in serum 8 weeks, 10 weeks, 12 weeks, 16 weeks, 20 weeks respectively, and it was decreased remarkably than the control group. After experiment ended, the result that observed the revelation CD3e, CD4, CD8, CD19, CD69, CD11a marker in lymph node establishment were observed and that B/T rate becomes recover as normal with political background. In addition to that, the control group was decreased in the measured value of IL-4, IL-5, IL-13, IgM, IgG2a, IgG1's level in serum, and $IFN-{\gamma}$' production secreted in Th1 cell displayed increase by SPAR medicines. IL-4, IL-5, CCR3, and $IFN-{\gamma}$'s gene revelation amount displayed marked decrease than the control group in result that observe effect that get in skin of a NC/Nga dermatitis mouse. Moreover in culture supernatant which cultivate for 14 days after separate skin cell, IL-13 and IL-6 production, and $CD69^+/CD3e^+$, $CD44^+/CD19^+$ expression cell number was decreased than the control group's number. Course inflammation immunocyte permeated of result that effect that SPAR medicines get to NC/Nga mice's skin establishment analyzes ear and neck-back skin after biopsy, and dye by H&E method decreased about epidermis and inflammation of dermis part remarkably than the control group. Conclusions Th1 cell and Th2 cell observe to be shifted by secretion amount of IL-4 and $IFN-{\gamma}$ by SPAR medicines could know that SPAR medicines can be use for treatung allergy autoimmune disease.

• Clinical and Experimental Pediatrics
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• 제53권11호
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• pp.957-964
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• 2010

Purpose: Our study attempted to determine the prognostic significance of minimal residual disease (MRD) detected by a simplified flow cytometric assay during induction chemotherapy in children with B-cell acute lymphoblastic leukemia (B-ALL). Methods: A total of 98 patients were newly diagnosed with precursor B-ALL from June 2004 to December 2008 at the Asan Medical Center (Seoul, Korea). Of those, 37 were eligible for flow cytometric MRD study analysis on day 14 of their induction treatment. The flow cytometric MRD assay was based on the expression intensity of CD19/CD10/CD34 or aberrant expression of myeloid antigens by bone marrow nucleated cells. Results: Thirty-five patients (94.6%) had CD19-positive leukemic cells that also expressed CD10 and/or CD34, and 18 (48.6%) had leukemic cells with aberrant expression of myeloid antigens. Seven patients with ${\geq}1%$ leukemic cells on day 14 had a significantly lower relapse-free survival (RFS) compared to the 30 patients with lower levels (42.9 % [18.7%] vs. 92.0% [5.4%], $P$=0.004). Stratification into 3 MRD groups (${\geq}1%$, 0.1-1%, and <0.1%) also showed a statistically significant difference in RFS (42.9% [18.7%] vs. 86.9% [8.7%] vs. 100%, $P$=0.013). However, the MRD status had no significant influence on overall survival. Multivariate analysis demonstrated that the MRD level on day 14 was an independent prognostic factor with borderline significance. Conclusion: An MRD assay using simplified flow cytometry during induction chemotherapy may help to identify patients with B-ALL who have an excellent outcome and patients who are at higher risk for relapse.

• 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.

• 대한한방내과학회지
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• 제26권1호
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• pp.143-155
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• 2005

Objective : The purpose of this research is to examine the effects of Gleditsiae Spina (GS) extract on immune cells and cytokines in ovalbumin (OVA)-induced asthmatic mice. Methods : In vivo, C57BL/6 mice were sensitized and handicapped by OVA for 12 weeks. During this experiment, the one group was then treated with GS extract for the later 8 weeks (3 times per week) and analyzed by ELISA, flow cytometer and RT-PCR. Results : In vivo, there were significant decreases in eosinophils, IL-4, IL-5, IL-13, IgE in BALF (bronchoalveolar lavage fluid). However, $IFN-{\gamma}$ in BALF of GS group increased significantly, compared with that of control group. Additionally, the population of $CD3e^-/CCR3^+,\;CD69^+/CD3e^+,\;IgE^+/B220^+,\;CD11b^+/Gr-l^+$ cells in the GS group decreased. Conclusion : The results of this study support a role for GS as an effective treatment for asthma in its experimental success in significantly decreasing inflammation and asthma reactions, and in increasing $IFN-{\gamma}$, which helps prevent such reactions.

• 대한한방내과학회지
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• 제26권1호
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• pp.156-168
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• 2005

Objective : The purpose of this research is to examine the effects of Pinelliae Rhizoma (PR) extract on immune cells and cytokines in ovalbumin (OVA)-induced asthmatic mice. Methods : In vivo, C57BL/6 mice were sensitized and handicapped by OVA for 12 weeks. During this experiment, the one group was then treated with PR extract for the 8 weeks (3 times per week) and analyzed by ELISA and flow cytometer. Results : In vivo, there were significant decreases in eosinophils, IL-4, IL-5, IL-13, IgE in BALF (bronchoalveolar lavage fluid) compared with that of control group. However, $IFN-{\gamma}$ in BALF of GS group increased significantly, compared with that of control group. Additionally, the population of $CD3e^-/CCR3^+,\;CD69^+/CD3e^+,\;IgE^+/B220^+,\;CD11b^+/Gr-l^+$ cells in the PR group decreased, compared with that of control group. Conclusion : The results of this study support a role for PR as an effective treatment for asthma in its experimental success in significantly decreasing inflammation and asthma reactions, and in increasing $IFN-{\gamma}$, which helps prevent such reactions.

• 한국가축번식학회지
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• 제26권2호
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• pp.165-171
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• 2002

난포액에는 난자의 성숙을 억제하는 인자를 함유하고 있으나 화학적 성질이 정확히 알려져 있지않다. 본 연구에서는 1차 난모세포에서 난구세포의 해리와 제1극체의 형성을 억제하는 난포액 성분을 화학적으로 동정하기 위하여 수행되었다 이를 위하여 돼지 난포액을 methanol로 추출한 다음이 추출물을 Superose 12 및 Superdex column을 이용 연속적으로 분리하였으며, Superdex 분절은 PITC로 처리한 다음 아미노산 분석용 column을 이용 분석하였다. 얻어진 결과는 다음과 같다. 난포액은 1차 난모세포에서 난구세포의 해리와 제1극체의 형성을 억제하였다 난포액에서 추출 분리한 Superdex분절 RV2.11 역시 난구세포의 해리와 제1극체의 형성을 억제하였다. 아미노산 분석 결과, 분절 RV2.11은 proline으로 추정되며, proline은 난구세포의 해리와 제1극체의 형성을 억제하였다. 결론적으로 난포액에는 난구세포의 해리와 제1극체의 형성을 억제하는 성분인 proline을 함유하고 있으며, 이는 난자의 성숙을 억제하는 성분일 것으로 추정된다.