• Title/Summary/Keyword: TdT염색

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A STUDY ABOUT THE APOPTOSIS OF EPITHELIAL CELLS IN THE FUSING FETAL RAT PALATE (흰쥐 태아 구개융합부위 상피세포의 아포프토시스에 관한 연구)

  • Lee, Hong-Joo;Cha, Kyung-Suk;Lee, Jin-Woo
    • The korean journal of orthodontics
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    • v.31 no.1 s.84
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    • pp.71-83
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    • 2001
  • The purpose of this study was to Prove that the medial edge epithelial cells covering the secondary palatal shelves were removed by apoptosis during palatal fusion. 12 mature female rats (Suprague-Dawley) were mated overnight with male rats and sacrificed on days 15.0, 16.5, 16.75, 17.0 of pregnancy. The embryos were removed from the uterus and the heads were embedded in paraffin. The paraffin blocks were sectioned and the sections were undergone H-E staining for general histologic feature and TdT staining for detection of apoptotic cells. The obtained results were as follows. k. In the section of 16.0 and 16.5 day embryos, the palatal shelves were prior to contact and no apoptotic cells wereobserved in the medial edge epithelium. At the initial contact of Palatal shelves, there was a few apoptotic cells in the fusing epithelium. 2. In the 16.75 day embryos, the samples that epithelial seams did not lost there continuity, apoptotic cells were rarely seen at the midline epithelial seam. In contrast, a lot of apoptotic cells were observed at epithelial triangles and the junction between palatal shelves and nasal septum. 3. In the 16,75 day embryos, the samples that epithelial seams lost their continuity and disrupted to epithelial islands, large number, of apoptotic cells were observed at epithelial islands and epithelial triangles. Some apoptotic cells were also observed at the oral, nasal epithelium near the midline. 4. In the 17.0 day embryos, most of epithelial islands were disappeared and mesenchymal confluence was achieved. Apoptotic cells were rarely observed in the mesenchymal tissue which replaced epithelial islands, but there were some apoptotic cells at the epithelial triangles, oral and nasal epithelium. From the results of the study, it was revealed that medial edge epithelial cells of fusing palate were removed by apoptosis. Apoptotic cells were found mainly in the disappearing midline epithelial seam and the oral and nasal epithelial triangles at some late stages of palatal fusion.

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The Induction of Human Corneal Epithelial Apoptosis by Serum-free Medium (무 혈청배지에 의한 각막상피 세포의 세포고사)

  • Ra, Myung Suk;Kim, Jai-Min
    • Journal of Korean Ophthalmic Optics Society
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    • v.4 no.1
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    • pp.1-6
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    • 1999
  • The corneal epithelium is constantly shed and apoptosis may play an important role in this turn-over. We sought to define that serum-free medium was able to induce apoptosis of corneal epithelial cells. SV-40 transfected human corneal epithelial(HCE) cells were grown to 70% confluency in culture. Serum-free medium was added to cells and the cells incubated for 1, 2, 3, or 6 days. Apoptosis of cells at different times was assessed by staining cells with Giemsa or Hoechst 33342 and measuring DNA fragmentation using the TUNEL assay. HCE cells exposed to serum-free medium demonstrated a high incidence of apoptosis, which increased over time to $50{\pm}4%$ after 3 days. They also stained positively with TUNEL assay. Serum-free medium caused time dependent apoptosis of HCE cells. Thus, serum-like nutrient might be important in corneal epithelial cell homeostasis.

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Surface Marker Analysis in Acute Leukemias (백혈병의 면역학적 표지자검사의 결과분석)

  • Moon, Jin-Young;Lee, Chae-Hoon;Kim, Kyung-Dong;Kim, Chung-Sook
    • Journal of Yeungnam Medical Science
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    • v.14 no.2
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    • pp.359-369
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    • 1997
  • We studied the expression of the cell surface antigen associated with myeloid and lymphoid leukemias on bone marrow or peripheral blood blast cells from 153 leukemic patients including 61 cases of acute myelogenous leukemias(AML), 46 of acute lymphocytic leukemias(ALL) and 12 of acute leukemias. They were analyzed by direct or indirect immunofluorescence method for reactivity with the monoclonal antibodies to B cells(CD10, CD19, SmIg), T cells(CD2, CD5, CD7, CD3, CD4, CD8), myeloid antigen(CD13, CD14, CD33, CD61) and a nonspecific antigen, HLA-DR. Lymphoid associated markers detected on AML is CD7 32.8%, CD10 14.8%, CD5 13.1%, CD2 6.6% and CD19 1.6%. TdT was positive in 4.9% of AMLs. Hybrid leukemias were 8 cases out 61 AML cases and were mainly composed of monocytic lineage, M4 and M5a. Myeloid markers detected in ALL were CD13 2.2% and CD33 2.2%. In this study, immunologically classified ALLs were composed of 65.2% of CALLA (+) B precursor type, 10.9% of CALLA (-) B precursor pattern, 8.7% of T cell type, 2.2% of B cell type, 4.5% of mixed lymphoid lineage(B&T), 2.2% of undifferentiated leukemia, and 6.5% of hybrid leukemia. Twelve cases of acute leukemias ware finally diagnosed to be 5 cases of hybrid leukemia, 3 cases of B lineage, 3 case of T lineage and 1 case of mixed lymphoid(B&T) leukemia. In summary, we think the best method for typing acute leukemias is by using a combination of FAB classification and immunophenotying.

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Attenuation of Reperfusion Injury with Angiotension $AT_1$ Receptor Blockade in Rat Myocardial Ischemic Model (백서 심근 허혈 모델에서 angiotension $AT_1$수용체 차단제의 재관류 손상 감소 효과)

  • Choi, Jun-Young;Choi, Dong-Ju;Ahn, Hyuk
    • Journal of Chest Surgery
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    • v.34 no.3
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    • pp.203-211
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    • 2001
  • 배경: AT$_1$수용체의 길항제가 세포 수준에서 심근을 재관류 손사으로부터 보호할수 있다는 것으로 알려져 있지만, 생체내에서의 효과나 그 기전은 아직 명확히 밝혀지지 않았다. 본 연구에서는 백서 심근 허혈 모델을 이용하여, AT$_1$ 수용체의 길항제들 중 하나인 irbesartan이 심근이 재관휴 손상에 미치는 효과를 알아보고, 재관류 손상을 매개하는 한 각지 기전으로서 세포자멸의 기여에 대하여 연구하고자 하였다. 대상 및 방법: Sprague-Dawley 백서에서 무작용 부형약(10% gum arabic: 1군, 개체수=14관) irbesartan(50mg/kg/day :II 군, 개체수=12)을 각각 3일 동안 24시간마다 경구로 투여하였다. 실험동물의 좌 관상 동맥을 45분간 결찰하였다가, 그 후 2시간 동안 재관류시킨 다음 심장을 적출 하였다. TTC(triphenyltetrazolium chloride) 염색법을 이용하여, 허혈 노출 부위에 대한 심근 경색 부위의 비율을 측정하였다. Agarose gel 전기영동상의 DNa 분절 양상과 TUNEL(TdT-mediated dUCP nick end labeling) 염색을 관찰하여 세포자멸이 일어난 정도를 평가하였다. 세포자멸을 조절하는데 관여하는 것으로 알려진 Bcl-2(B-cell lymphoma 2 gene), Bad 등의 단백과 ERK (extracellular signal-regulated kinase), p-38 등 신호전달체계에 작용하는 MAPKs(mitogen-activated protein kinases)의 발현을 측정하기 위하여 Western blot을 시행하였다. 결과: 허혈 노출부위에 대한 심근 경색부위의 비율은 II군(42$\pm$2.7%)이 I군( 64.1$\pm$4.65)에 비해 유의하게 작았다.(p< 0.05), Agarose gel 전기영동상의 DNA laddering 양상은 I군에서 보다 높게 발현되었다. Bad와 ERK2의 발현은 두 군간에 유의한 차이가 없었다. 결론: AT$_1$수용체 길항제인 irbesartan은 생체에서 심근의 재관류 손상을 줄이는 효과가 있었다. 이 효과는 적어도 부분적으로 나만 심근세포의 세포자멸이 감소한 것에 기인한 것으로 설명할 수 있으며, 이 항-세포 자멸 효과는 Bcl-2의 발현증가와 관련이 있는 것으로 추정되었다.

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