• Title/Summary/Keyword: 조직염색법

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Tissue origin of soluble component proteins in salin extract of adult Paragonimus westermani (폐흡충 성충 수용성 단백질의 성분별 생성위치)

  • Yoon Kong;Chi- Yong Park;Shin-Yong Kang;Seung-Yull Cho
    • Parasites, Hosts and Diseases
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    • v.30 no.2
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    • pp.91-100
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    • 1992
  • Tissue origin of individual component proteins in crude extract of adult Paragonimus westermani was investigated. Major soluble component proteins were separated by disc-PAGE in 8% slab gel. By predefined Rf values, strips of gel containing each band protein was cut out. Each band protein was eluted by electrophoresis. Monospecific antibodies were prepared by immunizing rabbits with each band protein. When peroxidase-antiperoxidase (PAP) staining was done, antiserum to Band 1 reacted to content of eggs both in the worm and in the infected lung tissue. Antiserum to Band 2 reacted to parenchymal tissue of the worm. Antiserum to Band 4 showed the positive reaction at intestinal content while that to Band 5 reacted to the intestinal epithelial border. Antiserum to combined proteins of Bands 617 and that to Band 8 reacted to parenchymal tissue of the worm respectively. From the results, the origin of individual proteins in crude extract of adult P. westermani could be differentiated.

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The Neurophysiological Approaches in Animal Experiments (신경생리학적(神經生理學的) 동물실험(動物實驗))

  • Cheon, Jin-Sook
    • Korean Journal of Biological Psychiatry
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    • v.5 no.1
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    • pp.3-16
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    • 1998
  • The neurophysiological study has been widely used in search of the relationship between brain and behavior. The basic techniques for the animal experiments of this kind such as stereotaxic techniques, lesioning methods, the methods of electrical stimulation and recording, and confirmation of histological location were briefly reviewed. Nevertheless, the importance of complementary neurochemical, neuroanatomical and behavioral studies can not be neglected.

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Evaluation of Brain Damage Induced by Transient Occlusion of Middle Cerebral Artery in Rats

  • Insook Jung;Lee, Juseon;Melissa D. Oh;Kim, Myungsoo;Changbae Jin
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1997.04a
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    • pp.103-103
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    • 1997
  • 일시적인 국소 뇌허혈 rat model에서 중대뇌동맥의 폐쇄시간에 따른 뇌조직의 손상정도를 비교 조사하였다. 웅성 Wistar rats를 isoflurane 홉입마취하에서 우측 내경동맥내로 17 mm의 silicone-coated 4-0 nylon monofilament를 삽입하여, 중대뇌동맥의 기저부를 30분, 60분 또는 90분동안 폐쇄한 후 이 monofilament를 다시 밖으로 뽑아내므로써 23 시간동안 recirculation 시키는 일시적 국소 뇌허혈 rat model을 사용한 결과, 수술 후 거의 모든 rats에서 left hemiparesis 또는 좌측으로의 circling과 같은 신경학적 결손이 나타나므로써 높은 성공률을 가지고서 비교적 용이하게 뇌허혈을 유발시킬 수 있었으며, 2 mm 두께의 fresh brain coronal slices에 대하여 2,3,5-triphenyltetrazolium chloride (TTC) 염색법을 시행하여 slice surface의 사진을 찍고 computerized 영상분석법을 이용하여 필요한 면적을 측정하므로써, coronal slice의 infarction size (%)는 총 면적에 대한 infarction 면적의 %로서, 부종율 (%)은 대조측 대뇌반구 면적의 2배에 대한 동측 대뇌반구와 대조측 대뇌반구 면적 차이의 %로서 산정되어졌다. 30분 허혈군에서는 본 염색법으로는 infarction이 거의 확인되지 않았으나 60분 허혈군 (n=13)에서는 우측 somatosensory frontoparietal cortex와 striatum 양자 모두 또는 일부 rats에서는 striatum에만 국한된 ulfarction이 확인되어졌으며 90분 허혈군 (n=12)에서는 거의 대부분의 rats에서 위 두 지역 모두에서 infarction이 확인되어졌다. Infarction size (%)는 60분과 90분 허혈군 각각에서, frontal pole로부터 5 mm되는 slice에서는 11.9$\pm$1.2 (평균치$\pm$표준오차), 13.7$\pm$1.9이었으며 7 mm되는 slice에서는 19.1$\pm$1.8, 21.9$\pm$2.1이었으며 9 mm되는 slice에서는 14.7$\pm$2.4, 19.7$\pm$2.2이었다. 또한 부종율 (%)은 60분과 90분 허혈군 각각에서, frontal pole로부터 5 mm되는 slice에서는 3.1$\pm$0.6, 3.8$\pm$0.7이었으며 7 mm되는 slice에서는 3.5$\pm$0.3, 5.7$\pm$1.0이었으며 9 $\pm$되는 slice에서는 3.4$\pm$0.5, 5.7$\pm$0.9이었다. 한편 90분 동안의 중대뇌동맥 폐쇄에 따른 뇌조직 손상을 cresyl violet 염색법, NADPH-diaphorase histochemistry, GFAP immunohistochemistry를 사용하여 일부 측정한 결과, 위의 손상지역에서는 뇌신경세포체들의 shrinkage 내지는 소실됨을 확인할 수 있었으며, NADPH-diaphorase-positive neuron들도 대부분 dendrite와 axon같은 cell process들의 fragmentation, shrinkage 내지는 소실되므로써 intensity의 감소현상이 나타났으며, reactive gliosis로 말미암아 GFAP immunoreactive intensity의 증가현상이 나타났다.

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Expression of Matrix Metalloproteinase-2 and Tissue Inhibitor of Metalloproteinase-2 in Radiation Exposed Small Intestinal Mucosa of the Rat (방사선조사를 받은 흰쥐 소장 점막의 손상과 재생과정 중 금속단백효소 및 억제자의 발현)

  • Kwag, Hyon-Joo;Lee, Kyoung-Ja;Rhee, Chung-Sik
    • Radiation Oncology Journal
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    • v.21 no.1
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    • pp.66-74
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    • 2003
  • Purpose : The matrix metalloprotelnases (MMPs) are a family of enzymes whose main function is the degradation of the extracellular matrix. Several studies have revealed that MMPs and TIMPS are related to the wound heating process and in photoaging caused by ultraviolet Irradiation. However, the expressions of MMP and TIMP after irradiation have not, to the best of our knowledge, been studied. This study investigates the expressions of MMP-2 and TIMP-2 in rat Intestinal mucosa following irradiation. Materials and Methods : The entire abdomen of Sprague-Dawley rats was irradiated using a single dose method. The rats were sacrificed on day 1, 2, 3, 5, 7 and 14 following irradiation. Histopathological observations were made using hematoxilin & eosin staining. The expressions of MMP-2 and TIMP-2 were examined using immunohistochemistry, Irnrnunoblotting and ELISA. Results : Radiation induced damage associated with atrophic villi, and infiltration of inflammatory cell was observed from the first postirradiation day, and severe tissue damage was observed on the second and the third postirradiation days. An increase in mitosis and the number of regenerating crypts, as evidence of regeneration, were most noticeable on the fifth postirradiation day. From the immunohistochemlstry, the MMP-2 expression was observed from the first postirradiation day, but was most conspicuous on the third and the fifth postirradiation days. The TIMP-2 expression was most conspicuous on the fifth postirradiation day. From the irnrnunoblotting, the MMP-2 expression was strongly positive on the third postirradlatlon day, and that of TIMP-2 showed a strong positive response on the fifth postirradiation day. In ELISA tests, the expressions of MMP-2 and TIMP-2 were increased in the postirradiation groups compared to those of the normal controls, and showed a maximum increase on the fifth postirradiatlon day. These results were statistically significant. Conclusion : The expressions of MMP-2 and TIMP-2 were increased in the intestinal mucosa of the rats following irradiation, and these results correlated with the histopathological findings, such as tissue damage and regeneration. Therefore, this study suggests that MMP-2 and TIMP-2 play roles in the mechanisms of radiation-induced damage and regeneration of intestinal mucosa of rats.

Detection of Helicobacter pylori by Pre-embedding Immunoelectron Microscopy - Comparison with Immunoblotting Method (Pre-embedding Immunoelectron Microscopy를 이용한 Helicobacter pylori의 발견 - Immunoblotting법과 비교 -)

  • Jung, Hae-Seoung;Kim, Eun-Jung;Kim, Eun-A;Park, Ji-Hoe;Jun, Jin-Su;Seo, Ji-Hyoun;Lim, Jae-Young;Choi, Myoung-Bum;Woo, Hyang-Ok;Youn, Hee-Shang;Ko, Gyung-Hyuck;Baik, Seung-Chul;Lee, Woo-Kon;Cho, Myung-Je;Rhee, Kwang-Ho
    • Clinical and Experimental Pediatrics
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    • v.45 no.7
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    • pp.862-874
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    • 2002
  • Purpose : We tried to evaluate whether the detection rate of Helicobacter pylori in gastric biopsy specimens could be improved by using pre-embedding immunoelectron microscopy. Methods : A total of 119 children who complained of upper gastrointestinal symptoms were endoscoped at the Gyeongsang National University Hospital from July, 1996 to July, 1999. Five biopsy specimens(three for urease test, one for hematoxylin-eosin(H & E) staining, and one for preembedding immunoelectron microscopy) were obtained from each antrum and body. Immunoblotting analysis were also performed. Results : Among the 119 patients, H. pylori were found in 116 patients(97.5%) by the immunoelectron microscopy. Among three patients who were found H. pylori negative in immunoelectron microscopy, two patients showed H. pylori in H & E stained slides and one patient was urease test positive(color change within six hours). Urease tests were positive in 107 patients(89.9 %). The positive rate of immunoblotting tests was 81.5%. However, only 13 patients(10.9%) showed H. pylori on the H & E stained antrum or body tissue. Conclusion : In this study, we found H. pylori histopathologically in most of the pediatric patients who complained of upper gastrointestinal symptoms. This study showed that pre-embedding immunoelectron microscopic examinations can be used as a gold standard in the diagnosis of childhood H. pylori infection. However, this method also has limited capacity to detect widely scattered H. pylori compared to the other histopathologic diagnostic methods.

Effect of Intestinal Function Enhancer (KTG075) on Mucin 2 Secretion (장기능개선제-신소재(KTG075)의 대장관 내 mucin 2 분비에 미치는 영향)

  • Lee, You-Hui;Baik, Soon-Ok;Kim, Hyun-Kyung;Ryu, Myung-Hyun
    • Korean Journal of Food Science and Technology
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    • v.36 no.6
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    • pp.991-994
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    • 2004
  • Effects of formulation KTG075 from edible plants on intestinal function, particularly on Mucin 2 secretion, were examined by loperamide-induced constipation method using Sprague Dawley rats (SD rats, male). Crypt epithelial cells containing more mucus and mucus layer stained with alcian blue were significantly thicker in KTG075 group than control group. When Biogenex AM358 of antibody against Mucin 2 was used, crypt epithelial cells secreted more Mucin 2 in KTG075 group than control group. The Mucus layer at fecal surface was thinner and less mucus was recovered from mucosal surface in constipated rats than in KTG075 group. Mucus production of crypt epithelial cells and mucus contents at fecal and mucosal surfaces were reduced by loperamide-induced constipation. These results indicate formula KTG075 accelerates evacuation and activates intestines.

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

  • Lee, Na-Young;Park, Do-Youn;Park, Jae-Hong
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.12 no.2
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    • pp.177-182
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    • 2009
  • Purpose: Colitis is a condition associated with a spectrum of altered morphologic changes and cellular adhesion. E-cadherin plays a key role in the establishment and maintenance of epithelial tissue structure and cell-cell adhesion. The purpose of this study is to evaluate E-cadherin expression in colonic epithelium of various colitis in children. Methods: The expressions of E-cadherin were examined in 39 cases of colonic mucosal biopsy specimen using immunohistochemical staining. When more than 50 percent of cells exhibited uniformly the same intensity and pattern of immunostaining as the adjacent normal mucosa, the antigen expression was considered normal. Abnormal expression was defined when less than 50 percent of cells stained, when cells showed a heterogeneously weak or altered distribution, or when complete absence of staining was observed. Results: Fifteen cases with non-specific colitis (38.5%), 7 cases of with Crohn's disease (17.9%), 5 cases of infectious colitis and milk protein sensitive proctocolitis (12.8%), 3 cases of ulcerative colitis (7.7%), 2 cases of Henoch-Schonlein purpura colitis (5.1%), one case of Behcet's disease and ischemic colitis (2.6%) were included in this study. E-cadherin expression was decreased in all kinds of colitis. Reduced expression of E-cadherin was observed in 77 percent of cases. E-cadherin was weaker or no expression in reparative epithelium and "ulcer associated cell lineage". Conclusion: Altered expression of E-cadherin occurs during mucosal inflammation in any kinds of colitis. These changes may be involved in promoting cell migration during epithelial restitution of the gastrointestinal mucosa.

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Immunohistochemical Detection of Lymph Nodes Micrometastases in Patients of Pathologic Stage I Non-small-cell Lung Cancer (병리적 병기 1기의 비소세포폐암 환자에서 면역조직화학염색에 의한 림프절 미세전이 관찰)

  • Ryu, Jeong-Seon;Han, Hye-Seung;Kim, Min-Ji;Kwak, Seung-Min;Cho, Jae-Hwa;Yoon, Yong-Han;Lee, Hong-Lyeol;Chu, Young-Chae;Kim, Kwang-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.57 no.4
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    • pp.345-350
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    • 2004
  • Background : To evaluate the frequency and clinical significance of lymph node micrometastasis in patients of non-small-cell lung cancer pathologically staged to be T1-2,N0. Method : From consecutive 29 patients of non-small-cell lung cancer who received curative operation and routine systemic nodal dissection, we immunohistochemically examined 806 lymph nodes from mediastinal, hilar and peribronchial lesion. All slides were stained with hematoxylin and eosin staining for one section and with cytokeratin AE1/AE3 antibody for another consecutive section of same lymph node to find out micrometastasis. Results : In 806 lymph nodes examined, no tumor cell was seen on hematoxylin and eosin staining and micrometastic foci were shown to be on 0.37%(3) of 806 lymph nodes, in which were upper paratracheal, interlobar and peribronchial lymph node. These three positive stains constitute 10.3%(3) of the 29 patients with non-small-cell lung cancer. Nine patients died from disease progression(4), postoperative complication(3) and concomitant diseases(2). The four patients with disease progression did not show evidence of micrometastasis on their lymph node examination. Conclusion : The frequency of lymph node micrometastasis was in 0.37% of 806 lymph nodes examined. The study results might suggested that routine analysis of micrometastasis on the lymph node didn't give any clinical implication on patients with non-small-cell lung cancer.

The Fixation Effects of Biological specimen Using Microwave Oven Equipped With Infrared-Temperature Sensor (적외선 온도감응기를 장착한 마이크로파 오븐의 생체물질 고정효과)

  • 신길상
    • The Korean Journal of Zoology
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    • v.37 no.2
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    • pp.144-155
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    • 1994
  • 마이크로파 에너지로서 현미경 관찰을 위한 생체물질을 고정할 때, 고정효과는 화학 고정액을 사용할 때와 비교하여 보다 우수한 또는 적어도 동일한 고정효과를 볼 수 있다는 것은 알려진 사실이다. 그러나 기존의 마이크로파 오븐을 사용하여 생체물질을 고정하면 고정 정도에 일정성이 없는데, 이는 마이크로파가 균일하게 조사되지 않고 고정에 적절하도록 마이크로팍의 강도를 조절하기 어려우며, 이로 인하여 정확한 고정온도의 측정이 불가능하기 때문으로 생각된다. 이 연구에서는 마이크로파가 조사되는 동안 시료의 온도를 측정할 수 없는 기존의 마이크연.파 오븐의 단점을 보완하기 위하여 비접촉식 적외선 온도 감응기를 장착한 마이크로파 오븐을 개발하였으며 고정효과를 관찰하기 위하여는 포유류의 기준 조직으로 알려진 생쥐의 신장을 고정하였다 고정효과는 전체적인 구조와 세부 구조의 보존이 모두 우수하였고 최적의 고정온도는 28$\pm$1$^{\circ}C$(완충용액 20 ml, 10초) 이었다. 이와같이 고정된 재료는 고정효과 뿐만 아니라 염색성이 좋아서 조직화학 반응이 우수하고, 냉동절편법과 연계했을 때 전체 절편제작 과정이 2~4시간 내에 완료될 수 있었다 이때 절편의 질(질)은 화학 고정액이나 냉동절편 법에 의하여 제작된 것 보다 우수하였다.

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