• 한방재활의학과학회지
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• 제18권4호
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• pp.39-61
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• 2008

목적 : 좌골신경 압좌손상으로 유발된 쥐의 모델을 이용하여 손상된 말초신경의 재생효과에 관한 침자극 효과를 세포분자학적, 조직학적 관점에서 연구하였다. 아울러 손상 좌골신경을 지배하는 척수신경근과 가까운 부위 경혈자극과 좌골신경이 지배하는 말초부위 경혈자극과의 침자극 효과를 비교 연구하였다. 방법 : 한쪽 좌골신경에 압좌손상을 유발 한 실험쥐들을 1주, 2주로 나눈 뒤 각각에 대해 격일로 1주군은 3회, 2주군은 6회의 침자극을 시행하였다. 손상 좌골신경의 재생효과를 비교실험하기 위해 정상군, 압좌손상만을 유발한 실험군, 침자극 군으로 나누었다. 침자극 군 중 한 군은 손상신경을 지배하는 척수신경근에 가장 가까운 2개의 화타협척혈(EX B2)에 자침(근위부 자극군)하였고, 다른 한 군은 말초부위에 위치한 족삼리혈(ST 36)과 위중혈(BL 40) 2곳에 자침(원위부 자극군)하였다. 실험 후 각각의 조직을 분리하여 Western blotting 혹은 Hoechst staining으로 Gap-43, Cdc2, Cdk2, Erk1/2 단백질을 분석 및 좌골신경의 각 세포수를 측정하였다. Retrograde tracing을 통해 L5의 DRG와 척수에서 말초신경 재생 효과를 관찰하였고, Immunofluorescence staining을 통해 신경돌기 가지의 신장 정도를 파악하였다. 결과 : 좌골신경 손상 7일된 실험쥐의 근위부와 원위부 침자극군에서 GAP-43와 Cdc2 단백질수준이 향상된 것으로 나타났다. Cdk2 단백질수준은 압좌손상 실험군에서 강하게 증가하였지만 침자극군과 비교해서 별다른 차이는 보이지 않았다. Phospho-Erk1/2 단백질수준은 침자극군에서 향상된 것으로 나타났다. 손상 7일과 14일 된 실험쥐의 손상 원위부에서 슈반세포 수가 증가하였으며 특히 근위부 침자극군에서 더욱 증가한 것으로 나타났다. Retrograde tracing을 이용한 검사 결과 침자극군에서 L5의 DRG와 척수의 염색 세포 수가 증가된 것으로 나타나 침자극이 축삭재생에 효과적인 것으로 나타났다. L5의 DRG 감각신경의 신경돌기 가지 신장정도 및 GAP-43 단백질의 발현 정도를 측정한 결과 근위부 침자극군에서 효과적으로 GAP-43 단백질의 발현 및 신경돌기 가지가 신장된 것으로 나타났다. 결론 : 본 실험결과 침치료가 손상 좌골신경의 재생에 효과적인 것으로 보여지며, 특히 손상된 좌골신경을 지배하는 척수신경근 주위 화타협척혈에 대한 침자극이 말초부위의 침자극에 비해 신경재생에 더욱 효과적인 것으로 나타났다.

• Applied Microscopy
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• 제39권4호
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• pp.291-299
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• 2009

본 연구에서는 햄스터 망막의 추상과 간상 광수용체의 양적 분석을 하였다. 추상 광수용체를 과산화 효소로 표지된 피넛 렉틴(peroxidase-labeled peanut lectin)을 사용하여 염색한 뒤 광학 미분간섭 현미경(DIC, differential interference contrast optics)을 사용하여 관찰하였다. 또한 면역세포화학적 방법으로 추상 광수용체에 위치한 M/L-옵신과 S-옵신을 염색한 뒤 공초점 형광 현미경(confocal microscope)을 사용하여 관찰하였다. 추상세포의 평균 밀도는 9,307 $cells/mm^2$이며 전체 추상세포는 293,060개였다. 추상세포의 최대값(12,857 $cells/mm^2$)은 맹점(시신경 유두, optic disk)로부터 코 쪽 망막(nasal retina)으로 0.3 mm 떨어진 곳에 위치하였다. 간상세포의 평균은 300,082 $cells/mm^2$으로 전체 간상세포는 9,448,150개였다. 간상세포의 최대값(340,000 $cells/mm^2$)은 맹점으로부터 등쪽 망막(dorsal retina) 방향으로 0.3 mm 떨어진 곳에서 발견되었다. 평균적으로 전체 광수용체 중, 간상세포의 전체 분포는 96.99%이고 추상세포는 3.01%를 차지하였다. 또한 간상과 추상의 평균 비는 32.24 : 1이었다. 햄스터의 추상세포는 M/L-와 S-옵신을 가진다. 본 연구 결과는 햄스터가 일부 명소시를 가지는 강한 간상 우세 망막임을 제시한다.

• Asian-Australasian Journal of Animal Sciences
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• 제29권9호
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• pp.1239-1246
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• 2016

The breeding of yaks is highly seasonal, there are many crucial proteins involved in the reproduction control program, especially in follicular development. In order to isolate differential proteins between mature and immature follicular fluid (FF) of yak, the FF from yak follicles with different sizes were sampled respectively, and two-dimensional gel electrophoresis (2-DE) of the proteins was carried out. After silver staining, the Image Master 2D platinum software was used for protein analysis and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) was performed for differential protein identification. The expression level of transferrin and enolase superfamily member 1 (ENOSF1) was determined by Western blotting for verification analysis. The results showed that 2-DE obtained an electrophoresis map of proteins from mature and immature yak FF with high resolution and repeatability. A comparison of protein profiles identified 12 differently expressed proteins, out of which 10 of them were upregulated while 2 were downregulated. Western blotting showed that the expression of transferrin and ENOSF1 was enhanced with follicular development. Both the obtained protein profiles and the differently expressed proteins identified in this study provided experimental data related to follicular development during yak breeding seasons. This study also laid the foundation for understanding the microenvironment during oocyte development.

• BMB Reports
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• 제40권5호
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• pp.765-772
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• 2007

Eosinophils act as effectors in the inflammatory reactions of allergic diseases including atopic dermatitis. Atopic dermatitis patients and others with allergic disorders suffer from eosinophilia, an accumulation of eosinophils due to increased survival or decreased apoptosis of eosinophils. In this study, a differential phosphoproteome analysis of AML14.3D10 eosinophil cell line after treatment with IL-5 or dexamethasone was conducted in an effort to identify the phosphoproteins involved in the proliferation or apoptosis of eosinophils. Proteins were separated by 2-DE and alterations in phosphoproteins were then detected by Pro-Q Diamond staining. The significant quantitative changes were shown in nineteen phosphoproteins including retinoblastoma binding protein 7, MTHSP75, and lymphocyte cytosolic protein 1. In addition, seven phosphoproteins including galactokinase I, and proapolipoprotein, were appeared after treatment with IL-5 or dexamethasone. Especially, the phospho-APOE protein was down-regulated in IL-5 treated AML14.3D10, while the more heavily phosphorylated APOE form was induced after dexamethasone treatment. These phosphoproteome data for the AML14.3D10 cell line may provide clues to understand the mechanism of eosinophilia as well as allergic disorders including atopic dermatitis.

• 동의생리병리학회지
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• 제16권1호
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• pp.117-123
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• 2002

To investigate the antiinflammatory and analgesic effects of Sophorae radix extracts administered to the arthritic rat model, immunohistochemical stains for CGRP in the L4, L5 and L6 spinal cord and ganglia were done, and paw swelling thickness were measured. Complete Freund,s Adjuvant(CFA) were injected to subcutaneous tissue of left foot paw of rats to induce arthritis. Sophorae radix extracts was administered immediately after CFA injection for 10 days. The spinal cord and ganglia were frozen sectioned(30㎛). These sections were stained by CGRP immunohistochemical staining method, and observed with light microscope. The results were as follows : 1. The change of paw swelling thickness of experimental group decreased from 4 day to 10day after CFA injection compared to control group. 2. The change of differential leukocytes counts of experimental group increased the ratio of lymphocytes. and decreased the ratio of neutrophils compared to control group. 3. The change of CGRP immunoreactive nerve fiber of dorsal horn of experimental group was dense stained compared to control group. 4. The number of CGRP immunoreactive neurons of L4 and L5 spinal cord of experimental group was less than in those control group. These results suggested that Sophorae radix extracts reduces the number of CGRP immunoreactive neurons and nerve fibers of spinal cord and ganglia, and decrease paw swelling thickness in arthritic rat model, which may be closely related to analgesic and antiinflammatory effects of Sophorae radix.

• 대한세포병리학회지
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• 제9권2호
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• pp.213-219
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• 1998

Glycogen-rich clear ceil carcinoma of the breast is an unusual variant of carcinoma with a recorded incidence of $1.4{\sim}3%$ of breast carcinomas. The cytologic characteristics have not been well described. We report two cases of glycogen-rich clear cell carcinoma with corresponding fine needle aspiration(FNA) cytologic findings and compare them to infiltrating ductal carcinoma and other clear ceil malignancies with a review of literature. One was a 62-year-old woman exhibiting a palpable mass of the right breast. The smears showed atypical tight cell clusters and individually scattered single cells containing leanly or clear abundant cytoplasm with well defined cytoplasmic margins. Mild to moderate nuclear pleomorphism and a prominent nucleolus were present. The other was a 42-year-old woman who was admitted with a right breast mass. The smears showed moderately cellular, tightly cohesive tumor cells. The cytoplasmic outline was generally well demarcated. The tumor cells Contained foamy to clear abundant cytoplasm with large and small vacuoles. The nuclear pleomorphism was marked. Both tumors resected by modified radical mastectomy, were diagnosed as glycogen-rich clear cell carcinoma. Histologically, the clear cell nature of tumor cells were not characteristic enough to predict this type of the tumor. Some cytologic features can be distinguished other clear cell breast cancer from glycogen-rich carcinoma. Recognition of these unusual patterns in a breast FNAC should raise the suspicion of a clear cell carcinoma including glycogen-rich subtype. Cytological localization of glycogen using PAS and D-PAS staining may permit the correct Identification and differential diagnosis of this tumor.

• 대한약침학회지
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• 제8권3호
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• pp.5-9
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• 2005

미분간섭현미경법과 아크리딘-오렌지(acridine-orange) 형광 염색을 이용하여 쥐 혈관 내의 실 모양 구조물을 관찰하였다. 공초점현미경법과 헤마톡실린-에오신(hematoxylin-eosin) 염색을 통해 피브린, 모세혈관, 소정맥, 소동맥 혹은 림프관의 핵 분포와 뚜렷이 구별되는 혈관 내 실 모양 구조물의 핵 분포 패턴을 얻어낼 수 있었다. 이 실 모양 구조물의 생리적 기능을 침술과 연관하여 논의하였다. 특히, 이 실 모양 관을 통한 약물의 흐름이 약침의 기전에 해당할 것이라는 가설을 제기하였다.

• 생약학회지
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• 제35권4호통권139호
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• pp.330-337
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• 2004

Based on the traditional application of Carthamus tinctorious L. (CF) as a component of Korean medicinal decoctions, in the present study, we investigated in vitro an immunomodulatory activity of water extract of CF(WECF). Water extract of CF significantly increased the in vitro proliferative responses of spleen cells (SPC). However, addition of WECF during anti-CD3 activation resulted in a significant decrease in SPC proliferation. Flow cytometric analysis showed that WECF addition chanced T and B cell frequencies in anti-CD3-activated spleen cell populations. Using purified cells, it was revealed that WECF is mitogenic to B cells but rather inhibitory to T cell Proliferation. Upon anti-CD3 stimulation, high concentration (1 mg/ml) of WECF significantly inhibited T cell proliferation until day 2 of stimulation. At day 3, anti-CD3-activated cells exposed to WECF recovered their proliferation to the level comparable to control. Although B cell proliferation was also inhibited in proliferation at day 1, it recovered sooner and then was rather augmented by WECF at day 3. These data indicate that WECF down-regulates lymphocyte proliferation at early phase of activation but T cells are more vulnerable than B cells to WECF, However, CD4+ and CD8+ T cells did not differ in WECF-mediated immunotoxicity. Data of propidium iodide (PI) staining showed that WECF accelerates activated T cell, but not B cell, apoptosis and WECF concurrently inhibited cytokine production of activated T cells. Taken together, WECF exhibits B cell mitogenic activity and differential toxicity more pronounced to T cells, suggesting a possible in vivo application of WECF for specific control of T cells without alteration of B cell activity.

• Restorative Dentistry and Endodontics
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• 제24권1호
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• pp.200-211
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• 1999

The periapical response to injury is a complex interaction of inflammatory, immune, neural, vascular and synthetic activity. TGF-${\beta}$ is a potent modulator of proliferation and differentiation in various tissue, seems to lead to an increase in extracellular matrix. MMP are a family of proteolytic enzyme that mediate the degradation of extracellular matric macromolecules, but little is known about theirs possible role in periapical tissue. The purpose of this study is to investigate the differential expression of TGF-${\beta}$ and MMP-1 in tooth follicle, periapical abscess, granuloma and cyst. The expression of TGF-${\beta}$ and MMP-1 in Periapical tissue was evaluated by immunohistochemical staining and Western blot analysis. Correlationship among the periapical lesions were stastically analyzed. The degree of MMP-1 expression in periapical abscess was higher than in any other periapical lesion, and stastically significant. TGF-${\beta}$ expression is the prominent in granuloma than other periapical lesion, which was stastically significant. The increased expression of MMP and TGF-${\beta}$ was not co-related with inflammatory cell infiltration degree of the periapical cyst. The expression degree of MMP and TGF-${\beta}$ was not co-related with periapical abscess and cyst, but expression of MMP and TGF-${\beta}$ showed strong positive co-relationship with periapical granuloma, which was stastically significant. TGF-${\beta}$ expression by Western blot analysis was prominent in granuloma and cyst, and similar to the results by imunohistochemistry. MMP-1 expression is less than TGF-${\beta}$, but there is not extreme difference between periapical lesion. These results suggest that TGF-${\beta}$ and MMP may be involved in tissue remodeling and has an important role in progress or mediation of periapical lesions.

• Asian-Australasian Journal of Animal Sciences
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• 제24권9호
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• pp.1199-1203
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• 2011

The objective of this study was to examine the binding potential of sperm to the epithelium of the sperm storage tubules (SST) in vitro and in vivo to assess the functional maturation of fowl sperm. Sperm from the testis, epididymis, as well as the proximal, middle and distal vas deferens were incubated in vitro with either the uterovaginal junction (UVJ)- or infundibular tissue containing SST at $39^{\circ}C$ for 30 min. Aliquots of sperm were also artificially inseminated into the uteri of hens, and the UVJ and infundibulum were collected 24 h post artificial insemination (AI). After incubation and AI, tissues were washed to remove loosely adhered sperm and subjected to fluorescence staining with 4', 6-diamidino-2-phenylindole, dihydrochloride (DAPI) for counting the number of bound sperm per 0.25 mm2 of surface area. Sperm from the testis, epididymis, and the three segments of the vas deferens exhibited their differential (p<0.05) binding capacity, which increased gradually from the testicular to distal vas deferens sperm under both in vitro and in vivo conditions. Existing similar trend, sperm, regardless of their source had a lesser affinity to bind to the epithelium of the infundibular SST than to the UVJ-SST. These experimental results suggested that fowl sperm may undergo gradual changes in the process of functional maturation, whereby they gain the ability to bind to the epithelium of the SST during their passage through the male reproductive tract (MRT).