• 제목/요약/키워드: immunohistochemical methods

검색결과 765건 처리시간 0.023초

rmIL-5로 유도된 호산구의 활성화 및 성장에서 중루의 천식반응 억제효과 (Inhibitory effects of Paridis Rhizoma in the activation and proliferation of eosinophils: implications on its regulatory roles for asthma)

  • 신미경;길기정;이영철;김진숙;서영배;노성수
    • 대한본초학회지
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    • 제20권2호
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    • pp.159-169
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    • 2005
  • Objectives : This study was carried out for the purpose of knowing the effect from anti-arthma action of the abstraction from a extract of Paridis Rhizoma(EPR). In order to know what the effect of controlling an abstraction from Paridis Rhizoma. and about the expression of B cells and Ig E cells, mast cells it was necessary for it to be activated by ovalbumin. Methods : In order to know what the effect was on the organization of cytokine gene expression from The increase and divorce of the B cells and allergic acting by EPR, we found it necessary to examine the BALF. At the same time, as we examined the histamine release by ELISA method, we also examined the effect of EPR. Results : EPR at $100\;{\mu}g/ml$, the highest concentration examined did not have any cytotoxic effects on mLFCs. In FACS analysis, number of granulocyte/lymphocyte, $CD3e^+/CCR3^+,\;CD4^+\;and\;CD23^+/B220^+$ in asthma-induced lung cells were significantly decreased by EPR treatment compared to the control group. In RT-PCR analysis, mRNA expression for CCR3, eotaxin and histamine in asthma-induced lung cells, which was induced by rIL-3 plus rmIL-5 treatments, was significantly decreased by EPR treatment. In ELISA analysis, production levels of IL-4, IL-13 and histamine in asthma-induced lung cells, which were induced by rIL-3 plus rmIL-5 co-treatment, were significantly decreased by EPR treatment. EPR treatments significantly inhibited the proliferation of eosinohils prepared from asthma-induced mouse lung tissues compared to the non-EPR treated control cells. Immunohistochemical analysis revealed that EPR treatment significantly decreased the levels of eosipnphil activation compared to non-treated cells. Conclusion : The present data suggested that Paridis Rhizoma may have an effects on the inhibition of parameters associated with asthma responses in eosinpophils, and thus implicate the possibility for the clinical application of Paridis Rhizoma.

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In vivo Tracking of Transplanted Bone Marrow-Derived Mesenchymal Stem Cells in a Murine Model of Stroke by Bioluminescence Imaging

  • Jang, Kyung-Sool;Lee, Kwan-Sung;Yang, Seung-Ho;Jeun, Sin-Soo
    • Journal of Korean Neurosurgical Society
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    • 제48권5호
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    • pp.391-398
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    • 2010
  • Objective : This study was designed to validate the cell trafficking efficiency of the in vivo bioluminescence image (BLI) study in the setting of transplantation of the luciferase expressing bone marrow-derived mesenchymal stem cells (BMSC), which were delivered at each different time after transient middle cerebral artery occlusion (MCAO) in a mouse model. Methods : Transplanting donor BMSC were prepared by primary cell culture from transgenic mouse expressing luciferase (LUC). Transient focal infarcts were induced in 4-6-week-old male nude mice. The experiment mice were divided into five groups by the time of MSC transplantation : 1) sham-operation group, 2) 2-h group, 3) 1-day group, 4) 3-day group, and 5) 1-week group. BLI for detection of spatial distribution of transplanted MSC was performed by detecting emitted photons. Migration of the transplanted cells to the infarcted area was confirmed by histological examinations. Differences between groups were evaluated by paired t-test. Results : A focal spot of bioluminescence was observed at the injection site on the next day after transplantation by Signal intensity of bioluminescence. After 4 weeks, the mean signal intensities of 2-h, 1-day, 3-day, and 1-week group were $2.6{\times}10^7{\pm}7.4{\times}10^6$. $6.1{\times}10^6{\pm}1.2{\times}10^6$, $1.7{\times}10^6{\pm}4.4{\times}10^5$, and $8.9{\times}10^6{\pm}9.5{\times}10^5$, respectively. The 2-h group showed significantly higher signal intensity (p<0.01). The engrafted BMSC showed around the infarct border zones on immunohistochemical examination. The counts of LUC-positive cells revealed the highest number in the 2-h group, in agreement with the results of BLI experiments (p<0.01). Conclusion : In this study, the results suggested that the transplanted BMSC migrated to the infarct border zone in BLI study and the higher signal intensity of LUC-positive cells seen in 2 hrs after MSC transplantation in MCAO mouse model. In addition, noninvasive imaging in real time is an ideal method for tracking stem cell transplantation. This method can be widely applied to various research fields of cell transplantation therapy.

이중탕(理中湯)이 Mite Antigen으로 유발된 NC/Nga 생쥐의 아토피 피부염에 미치는 영향 (Therapeutic Effects of Yijungtang on Atopic Dermatitis-like Skin Lesions of NC/Nga Mouse Induced by Mite Antigen)

  • 서희연;한재경;김윤희
    • 대한한방소아과학회지
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    • 제25권1호
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    • pp.1-27
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    • 2011
  • Objectives: The purpose of this study is to investigate the effects of Yijungtang(YJT) on atopic dermatitis in an in-vitro and in-vivo experiment using a RBL-2H3 mast cells and a NC/Nga atopic dermatitis mouse. Methods: In-vitro experiment, IL-4, IL-13 mRNA expression were evaluated by a real-time PCR, IL-4, IL-13 production by ELISA and transcription factor as GATA-1, GATA-2, NF-AT1, NF-AT2, AP-1 and NF-kB by western blotting. In-vivo experiment, clinical skin score we evaluated by, hematology and Serum total IgE and IgG1 of NC/Nga atopic dermatitis mouse, cytokine level, total number of cell, Immunohistochemical staining and Histological features of auxiliary lymph node(ALN), draining lymph node(DLN), peripheral blood mononuclear cells(PBMCs) and dorsal skin tissue in NC/Nga mouse. Results: YJT decreased IL-4, IL-13 mRNA expression, IL-4, IL-13 production and prominently decreased the expression of mast cell specific transcription factors including GATA-2, NF-AT2, c-Fos and NF-kB. YJT oral administration reduced the levels of skin severity scores. It also decreased the level of inflammatory cytokines such as IL-5, IL-13, histamine and IgE in the serum. It elevated IFN-gamma level in the spleenocyte culture supernatant but decreased. $CD3e^+$, $CD19^+$, $CD4^+$, $CD8^+$, $CD3e^+CD69^+$, $CD11b^+Gr-1^+$, $CCR3^+$ in the PBMCs, $CD4^+$, $CD8^+$, $CD3e^+CD69^+$, $B220^+CD23^+$ in the ALN, $CD4^+$, $CD3e^+CD69^+$ in the ALN and $CD4^+$, $CD11b^+Gr-1^+$ in the dorsal skin. Histological examination showed that infiltration levels of immune cells in the skin of AD-induced NC/Nga mice were much improved by YJT oral administration. Conclusions: The anti-allergic activities of YJT may be mediated by down-regulation of Th2 cytokines, such as IL-4 and IL-13, through the regulation GATA-2, NF-AT2 and NF-kB transcription factors in mast cells. YJT would be regulate molecular mediators and immune cells which are functionally associated with atopic dermatitis induced in NC/Nga mice, and may play an important role in recovering AD symptoms.

아토피양(樣) 피부염 NC/Nga 생쥐에서 소풍도적탕가미(消風導赤湯加味)와 아토피크림, 자운고(紫雲膏) 및 소풍도적탕가미(消風導赤湯加味)의 병용투여가 피부염에 미치는 영향 (Effects of SPDJTK(SoPungDoJeokTangKami) and Concurrent Administration of AJ (Atopy cream, Jawoongo) Plus SPDJTK on Atopic Dermatitis-like Skin Lesions in NC/Nga Mouse Induced by BMAC)

  • 한달수;한재경;김윤희
    • 대한한방소아과학회지
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    • 제24권1호
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    • pp.9-35
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    • 2010
  • Objectives The purpose of this study is to investigate the effect of SPDJTK(SoPungDoJeokTangKami) and concurrent administration of AJ(Atopy cream, Jawoongo)+SPDJTK on atopic dermatitis-like skin lesions by using in NC/Nga atopic dermatitis mouse induced by BMAC-induced mice. Methods Clinical skin score, hematology and Serum total IgE and IgG1 of NC/Nga atopic dermatitis mice were evaluated. Moreover, 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 were used in NC/Nga mice. Results Orally administrated SPDJTK with concurrent administration of SPDJTK and AJ decreased the clinical skin score, total cell number of WBC, eosinophils in blood, serum total IgE & IgG1, IL-5, IL-13, IFN-$\gamma$. Also, total cell number of ALN and dorsal skin tissue, absolute cell number of CD4+, CD8+, CD3+CD69+, CD3+CCR3+, CCR3+, CD4+CXCR5+ in ALN, absolute cell number of CD3+CCR3+, CCR3+ in DLN, granulocytes in PBMCs, activation cell number of CD3+CD69+, CCR3+, total cell number of CD3+ T cell in dorsal skin tissue were significantly decreased. Furthermore, thickness of epidermis, infiltrated inflammatory immune cell and mast cell in dermis, amount of Eotaxin2 mRNA, CCR3 mRNA in dorsal skin tissue, gene expression of IL-5, IL-13 mRNA in ALN, CD4+ Th cell in dorsal skin tissue and CCR3+ eosinophils in ALN were all significantly decreased. However, total number of DLN, absolute number of CD3e+ T cell and CD19+ B cell, absolute number of CD4+, number of Th cell in DLN and gene expression of foxp3 mRNA were significantly increased significantly. Conclusions Concurrent administration of SPDJTK and AJ on atopic dermatitis in NC/Nga atopic dermatitis mouse was very effective treatment for atopic dermatitis.

Wound Healing Effects of Rose Placenta in a Mouse Model of Full-Thickness Wounds

  • Kim, Yang Woo;Baek, Seung Ryeol;Lee, Eun Sook;Lee, Sang Ho;Moh, Sang Hyun;Kim, Soo Yun;Moh, Ji Hong;Kondo, Chieko;Cheon, Young Woo
    • Archives of Plastic Surgery
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    • 제42권6호
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    • pp.686-694
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    • 2015
  • Background Rosa damascena, a type of herb, has been used for wound healing in Eastern folk medicine. The goal of this study was to evaluate the effectiveness of rose placenta from R. damascena in a full-thickness wound model in mice. Methods Sixty six-week-old C57BL/6N mice were used. Full-thickness wounds were made with an 8-mm diameter punch. Two wounds were made on each side of the back, and wounds were assigned randomly to the control and experimental groups. Rose placenta ($250{\mu}g$) was injected in the experimental group, and normal saline was injected in the control group. Wound sizes were measured with digital photography, and specimens were harvested. Immunohistochemical staining was performed to assess the expression of epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), transforming growth factor-${\beta}1$ (TGF-${\beta}1$), and CD31. Vessel density was measured. Quantitative analysis using an enzyme-linked immunosorbent assay (ELISA) for EGF was performed. All evaluations were performed on postoperative days 0, 2, 4, 7, and 10. Statistical analyses were performed using the paired t-test. Results On days 4, 7, and 10, the wounds treated with rose placenta were significantly smaller. On day 2, VEGF and EGF expression increased in the experimental group. On days 7 and 10, TGF-${\beta}1$ expression decreased in the experimental group. On day 10, vessel density increased in the experimental group. The increase in EGF on day 2 was confirmed with ELISA. Conclusions Rose placenta was found to be associated with improved wound healing in a mouse full-thickness wound model via increased EGF release. Rose placenta may potentially be a novel drug candidate for enhancing wound healing.

뇌허혈 유발 후 전침자극이 전뇌세포사 중 Bax와 Caspase-3 발현 변화에 미치는 영향 연구 (A Study on the Effects of the Electro-Acupuncture(EA) Stimulation after Global Ischemia(GI) on Changes in Bax and Caspase-3 Expression among Forebrain Apoptosis)

  • 최정현;민경옥;김순희;김동일;이상빈;안호정;김지성;최완석;김보경;송치원
    • 동의신경정신과학회지
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    • 제21권1호
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    • pp.59-69
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    • 2010
  • Objectives: The study aims to look into how the application of EA(Electro-acupuncture) in the initial step of ischemic brain injury affects the changes in Bax and Caspase-3 expression among forebrain apoptosis. Methods: It caused GI by using common carotid artery occlusion and conducted the test by dividing into the no applied EA group after 6 hours and 12 hours, the LI 4 EA group that applied EA to LI4, and GV 20 EA group that was applied to GV 20. After that, the following results were obtained by comparing Bax and Caspase-3 expression, which were apoptosis-related factors, with its application time through immunohistochemical staining after extracting brain of each group. Results: According to each group's Bax expression by the application time, the order of expression increase after 6 hours is shown to be the no applied EA group, LI 4 group, and GV 20 group. The stimulation after 12 hours was most lowly expressed in the GV 20 group, and the no applied EA group and LI 4 group showed similar level. Conclusions: EA stimulation in the initial step after ischemia seems to affect positively Bax and Caspase-3 expression that are index of forebrain apoptosis.

C57BL/6N 생쥐에서 천남성 추출물과 분획물의 발모효과에 대한 실험적 연구 (Experimental Studies on the Hair Growth Activity of Fractions and Extract of Arisaematis Rhizoma in C57B/6N Mice)

  • 권경숙;이문원;정일국;정한솔;송범용;송정모;이창현
    • 동의생리병리학회지
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    • 제23권3호
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    • pp.619-630
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    • 2009
  • To investigate the hair growth activity of fractions and extract of Arisaematis Rhizoma in the hair removed skin of normal and spontaneous alopecia areata model in C57B/6N mice. These experiments were performed with the macroscopic, microscopic, immunohistochemical(VEGF, c-kit, PKC-${\alpha}$, TGF and FGF) and RT-PCR(TGF-${\beta}$, IGF, prolactin and placenta lactogen) methods. The results were as follows: Macroscopic observation after topical application of vehicle, 50% EtOH as control and extract of Arisaematis Rhizoma to the hair removed skin of C57BL/6N mice on the 9th, 11th and 15th day. Extensive hair growth activity was observed in treated group with extract of Arisaematis Rhizoma on the 9th, 11th and 15th day. In Arisaematis Rhizoma extracts treated group, hair follicles of middle stage of anagen was observed and it were grown down to subcutaneous tissue of skin in all the normal mice on 15th day. But in control group, most of hair follicles of telogen phase was observed in skin. The treatment of extract of Arisaematis Rhizoma increased expression of IGF(145%) and placenta lactogen(108%) in the skin of normal C57BL/6N mice on the 11th day compared to control group(100%). But expression of TGF-${\beta}$(90%) and prolactin(91%) decreased in the skin of normal C57B/6N mice on the 11th day compared to control group(100%). After application of fractions(chloroform, ethyl acetate and water fractions) of Arisaematis Rhizoma extract for 9th day, hair growth effect was observed in whole skin area in 50% of normal mice. But in control group, hair growth effect was not observed in whole skin area of normal mice. Immunoreactive density of VEGF, c-kit, PKC-${\alpha$ and FGF in skin of fractions of Arisaematis Rhizoma extracts was strongly stained in epidermis, bulge, secondary hair germ cells, cutaneous trunci m., subcutaneous tissue, root sheath compare to control group on the 9th day. In spontaneous alopecia areata model, The hair growth activity of Arisaematis Rhizoma extrat treated group(75%) was observed to be strong compared to control group(O%) on 7th day. These experiments suggest that fractions and extracts of Arisaematis Rhizoma may stimulate the topical hair growth activity. Thus it can be useful for treatment of alopecia areata.

소아에서 발생한 대장의 염증성 질환에서 E-cadherin의 발현 (E-cadherin Expression in Colonic Epithelium of Various Colitis in Children)

  • 이나영;박도윤;박재홍
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • 제12권2호
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    • pp.177-182
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    • 2009
  • 목 적: 소아에서 다양한 원인에 의해 발생한 대장염에서 점막의 형태학적 변화와 세포 접합에 다양한 변화가 있으리라 예상되어 세포 간의 결합을 유지하는 E-cadherin의 변화를 살펴보았다. 방 법: 1998년 1월부터 2003년 8월까지 부산대학교병원 소아청소년과에서 하부 위장관 내시경술과 대장점막 조직 검사를 통해 대장염으로 진단된 39명을 대상으로 하였다. 파라핀 블록에서 면역조직화학염색법을 이용하여 E-cadherin의 세포 내 발현을 조사하였다. 주변의 정상 조직과 비교하여 E-cadherin의 발현이 동일한 강도와 양상을 가진 세포가 50% 이상인 경우를 정상으로 판정하였고, 발현이 정상인 세포가 50% 미만이거나 염색 분포의 이상이 있거나 전혀 염색되지 않은 경우를 이상으로 판정하였다. 결 과: 1) 본 연구에서 비특이성 대장염 15예(38.5%), 크론병 7예(17.9%), 감염성 대장염 5예(12.8%), 음식 단백 과민성 직결장염 5예(12.8%), 궤양성 대장염 3예(7.7%), Henoch-Schonlein purpura 대장염 2예(5.1%), 그외 베체트병, 허혈성 대장염 1예가 포함되었다. 2) 모든 종류의 대장염에서 상피세포 E-cadherin 발현 감소가 관찰되었으며, 77%의 대상 표본에서 E-cadherin 발현감소가 있었다. 3) 활동성 염증이 심한 부위에서 Ecadherin 발현 감소가 현저하였으며 병변부에서 떨어진 상피세포에서는 정상 발현을, 궤양 주위나 재생 상피가 있는 부위는 심한 발현 감소를 보였다. 결 론: 모든 종류의 염증성 대장 질환에서 E-cadherin 발현 감소가 있었다. 이러한 변화는 염증과 궤양이 있는 부위에서 상피세포 접합을 느슨하게 함으로써 상피세포의 재생을 위한 세포의 이동을 용이하게 하기 위한 작용이라고 판단된다.

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폐 편평세포암에서 자발성 아포토시스와 원격전이 (Spontaneous Apoptosis and Metastasis in Squamous Cell Carcinoma of the Lung)

  • 오윤경;기근홍
    • Radiation Oncology Journal
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    • 제17권3호
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    • pp.203-208
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    • 1999
  • 목적 : 폐암 환자의 대부분은 진단 당시 수술을 받을 수 없는 병기로 발견되기 때문에 수술 조직이 아닌 기관지내시경 조직에서 자발성 아포토시스 정도를 평가하여 이의 임상적 의의에 대한 기초 자료로 삼고자 본 연구를 시행하였다. 대상 및 방법 : 1990년 9월부터 1994년 9월까지의 4년동안 흉부에 방사선치료를 받은 폐 편평세포암 환자 중 조직표본이 충분히 보관되어 있으며 추적이 가능하였던 19명을 대상으로 하였다. 병기는 II기가 1명, IIIa기 8명, IIIb기 5명, IV기 5명이었다. 면역조직화학적 염색법으로 자발성 아포토시스율(Al)과 p53 단백질 양성률을 관찰하였다. 결과 : 19명 중 16명은 $5\~15$ 개월 후에 사망하였으며 3명은 55, 67, 67 개월간 생존하고 있다. 중앙생존기간은 17개월, 평균 생존기간은 24 개월이었다. AI는 $0\~1\%$의 범위로 중앙값이 $0.4\%$였다. AI가 낮은 군에서 진단 당시 원격전이가 있었던 경우가 $50\%$ (5/10) 였고, 높은 근에서는 원격전이가 전혀 없었다(0/9). 생존기간에 영향을 줄 수 있는 예후인자들의 분석 결과 단변량 분석에서는 M병기가 통계학적으로 유의한 차이를 보였고, 다변량 분석에서는 AI, 화학요법, M병기, T병기, 병기가 의의가 있었다. 자발성 아포토시스와 p53 변이 사이의 관련은 관찰되지 않았다. 결론 : AI는 진단 당시 원격전이와 관련이 있으며, p53 변이와는 관련이 없었다. AI가 낮은 군에서 높은 군보다 생존기간이 짧은 경향을 보였다.

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사물탕가미방(四物湯加味方)이 원형탈모 생쥐의 발모효과에 관한 형태학적 연구 (Morphological Studies on Hair Growth Effect of Samultang-gamibang)

  • 권강주;이창현;이문원;문연자;최두호;이학재;김호민;우원홍
    • 동의생리병리학회지
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    • 제19권3호
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    • pp.696-704
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    • 2005
  • This experiment examined the effect of Samultang-gamibang (SGB) on hair growth in spontaneous alopecia areata C57BL/6N mice. We first investigated hair growth effect of SGB compare to control groups after apply to oral administration for 10 weeks and regional treatment in skin for last 4 weeks. We second investigated the number of hair follicle and mast cells after treatment of SGB in spontaneous alopecia areata C57BL/6N mice for 10 weeks. We third investigated immunoreactive density of neuropeptides in skin of spontaneous alopecia areata C57BL/6N mice by immunohistochemical methods. The results were as follows : Hair growing effect of experimental group was observed from 7 weeks after administration of SGB (87.5%). In experimental group, the number of mast cells and eosinophils was significantly decreased compare to control group. Immunoreactive density of substance P and corticotropin releasing factor (CRF) in skin of experimental group was weakly stained in epidermis and subcutaneous tissue compare to control group. Immunoreactive density of CRF-receptor (CRF-R), CRF-binding protein (CRF-BP) in skin of experimental group was increased in epidermis, sebaceous gland, inner root sheath, outer root sheath and secondary hair germ epithelium compare to control group. These results suggest that SGB may be used in treatment of alopecia areata.