• Title/Summary/Keyword: 면역화학염색법

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Expression of TGF-β1 and EGFR in Irritation Fibroma and Oral Leukoplakia (면역조직화학염색법을 이용한 자극성 섬유종과 구강 백반증에서의 TGF-β1과 EGFR 발현 비교 연구)

  • Ryu, Mi-Heon
    • Journal of dental hygiene science
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    • v.5 no.3
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    • pp.97-103
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    • 2005
  • Irritation fibroma (IF) is the most common tumor-like oral lesion that is evolved by proliferation of collagen in response to chronic irritation. Oral leukoplakia (OL) is considered as precancerous lesion characterized by proliferation of epithelial cells due to chronic irritation, smoking and drinking. TGF-${\beta}1$ and EGFR are important factors that play an essential role in extracellular matrix remodeling during normal wound healing process. The epithelial reaction by chronic irritation may be connected with pathogenesis of IF and OL. In the present study, we examined the expression of TGF-${\beta}1$ and EGFR in the IF and OL using immunohistochemistry. We used 88 cases of IF, 44 cases of OL and 9 cases of normal oral mucosa as normal control. TGF-${\beta}1$ was decreased in the epithelium of IF and OL. As for EGFR, the epithelial cells revealed the increased positive expression in IF and OL. In case of OL, the Spearman correlation coefficient of TGF-${\beta}1$ and EGFR was -0.10 (p< 0.05), which showed weak correlation. In the fibrous tissue, TGF-${\beta}1$ was increased only in IF. The expression difference of TGF-${\beta}1$ and EGFR may be involved in the pathogenesis of IF and OL.

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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.

Immunohistochemical Localization of 36 and 29 kDa proteins in sparganum (면역조직염색법으로 관찰한 스파르가눔 층체에서의 36, 29 kDa 항원 단백질의 생성위치)

  • Kim, Lee-Su;Kong, Yoon;Kang, Shin-Yong;Cho, Seung-Yull
    • Parasites, Hosts and Diseases
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    • v.30 no.1
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    • pp.25-32
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    • 1992
  • Antigenic proteins of 36 and 29 kDa were localized in Spirometra mansoni plerocercoid (sparganum) immunohistochemically by avidin biotin complex (ABC) staining. When polyclonal antibodies such as BALB/c mouse serum immunized with crude saline extract of sparganum or confirmed sparganosis sera were reacted as pri-mary antibodies, the positive chromogen (3-amino, 9-ethylcarbazole) reactions were recognized at syncytial tegument, tegumental cells, muscle and parenchymal cells and lining cells of excretory canals. A monoclonal antibody(MAb) which was reacting to 36 and 29 kDa proteins in the extract of the worm was localized at the syncytial tegument and tegumental cells. The present results suggested that the potent antigenic proteins of 36 and 29 kDa in sparganum were produced at the tegumental cells and transported to the syncytial tegument.

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Expression of VEGF, p53, Apaf-1 and Caspase-9 in Head and Neck Squamous Cell Carcinoma (두경부 편평세포암에서 VEGF, p53, Apaf-1 and Caspase-9의 발현)

  • Lee, Sung Hak;Kang, Yeo Ju;Jo, Ui Ju;Lee, Youn Soo;Kang, Chang Suk
    • Korean Journal of Head & Neck Oncology
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    • v.28 no.2
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    • pp.107-113
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    • 2012
  • 배 경 편평상피암은 두경부의 악성종양 중 가장 흔하며, 임상적인 경과가 불량하다. 따라서 나쁜 예후를 가지는 환자군을 조기에 선별하여 더 적극적인 치료의 시행을 결정짓는 표지자의 필요성이 대두된다. 우리는 일련의 두경부 편평세포암 검체에서 몇몇 분자 표지자의 예후적 유용성을 평가하고자 하였다. 방 법 23예의 두경부 편평세포암 검체를 대상으로 VEGF, p53, Apaf-1, caspase-9의 발현과 몇몇 임상병리학적 지표들간의 연관성을 면역조직화학염색을 통해 조사하였다. 결 과 환자군은 남성이 더 많았으며 평균연령은 63.7세였다. 1기가 5예, 2기가 2예, 3기가 8예, 4기가 8예였다. 평균생존기간은 37.3개월이었다. VEGF단백 발현은 종양의 크기와 통계적으로 유의한 연관성을 보였다. 이와 더불어 VEGF단백 발현은 병기, 그리고 림프관 침습과 연관적인 경향성을 보였다. 그러나 VEGF단백 발현과 생존기간과는 관련성이 없었다. 또한, Apaf-1과 caspase-9의 단백발현은 다른 임상지표, 환자의 생존기간과는 관련이 없었다. 결 론 VEGF단백 발현은 두경부 편평세포암 환자에서 나쁜 임상경과를 예측할 수 있게 하는 표지자로서의 역할을 할 수 있다. 또한 본 연구는 두경부 편평세포암에서 별로 연구되지 않은 Apaf-1과 caspase-9의 발현상태를 밝힌 점에서 의의가 있다.

Effects of histochemical staining in microwave-irradiated tissues (마이크로파 처리 고정 조직의 조직염색 효과)

  • Lee, Yoon-Jin;Lee, Sang-Han
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.8
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    • pp.417-424
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    • 2019
  • Despite its superior ability to show distinct cellular morphology and for long-term storage, conventional tissue fixation by formalin has many drawback, including slower fixation, the exposure to harmful chemicals and extensive protein modification. Herein, we assessed the effects of rapid microwave-assisted tissue fixation on histological examination and on protein integrity by comparing these microwave irradiation fixated tissues with the formalin-fixed tissues. One of the paired mouse tissues (liver and kidney) was fixed in formalin and the other was fixed by using microwave irradiation in phosphate buffered saline. Each slide from the paraffin-embedded tissues was examined by H & E staining for the adequacy of fixation and by immunohistochemical staining for antigenicity in a blinded fashion. Evaluation of protein recovery and the protein quality from the fixed tissues were analyzed by the BCA method and Western blotting, respectively. The results from H & E staining and immunohistochemical staining showed that the sections obtained from microwave-fixed tissues under our experimental conditions were comparable to those of the formalin-fixed tissues except for the integrity of RBCs. Furthermore, proteins were effectively extracted from the microwave-fixed tissues with acceptable preservation of the proteins' quality. Taken together, this microwave-assisted tissue processing yields a quick fixation and better protein recovery in higher amounts, as well as the adequacy of fixation and the antigenicity being comparable to formalin-fixed tissues, and this all suggests that this new fixation technique can be applied in an environment where rapid tissue fixation is required.

Characterization of Bovine Brucellosis in Korean Native Cattle by Means of Immunohistochemistry and Proteomics (면역조직 화학법 및 단백질체 변화 분석을 통한 한우에서 발생한 브루셀라증의 특성)

  • Jang, Seong-Jun;Do, Sun-Hee;Ki, Mi-Ran;Hong, Il-Hwa;Park, Jin-Kyu;Cho, Yu-Jeong;Park, Sang-Joon;Kim, Tae-Hwan;Kwak, Dong-Mi;Jeong, Kyu-Shik
    • Journal of Life Science
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    • v.20 no.2
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    • pp.153-160
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    • 2010
  • This study was conducted to examine the utilization of immunohistochemistry using the bovine anti-brucella immunoglobulin G (IgG) antibody in the diagnosis of brucellosis and to develop a functional biomarker relation for the progress of the disease. Anti-brucella IgG antibody was purified from the affected bovine serum using an affinity chromatography. We performed our investigation on 17 cases of brucellosis and 19 control cases with negative Rose-Bengal test results. Our purified anti-brucella IgG antibody showed a positive immunoreactivity in cytoplasmic hepatocytes of the centrilobular region, and glomeruli and tubular epithelium of the kidney. The protein pattern of the affected liver versus control was analyzed by two-dimensional electrophoresis, showing a different expression pattern of proteins between the two. Five protein spots were up-regulated and another were five down-regulated in the brucellosis liver. Significant upregulaton of catalase and 3-hydroxyacyl-CoA dehydrogenase might be due to a compensatory reaction in response to the endotoxic shock of brucella. In conclusion, the anti-brucella IgG antibody may be a good tool for discriminative diagnosis of the affected tissues and proteomics data suggest new target proteins underlying a possible pathogenic mechanism of brucellosis.

Correlation Between the Expression of Epidermal Growth Factor Receptor and MR Features in Glioma (신경교종에서 표피성장인자수용체의 발현도와 자기공명영상 소견의 상관관계)

  • 김범수;신경섭
    • Investigative Magnetic Resonance Imaging
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    • v.1 no.1
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    • pp.125-129
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    • 1997
  • Purpose: The aim of this study was to find correlation between the expression of epidermal growth factor receptor (EGFR) and MR findings in the brain glioma. Materials and Methods: MR features including edema, margin, necrosis, heterogeneity, hemorrhage and contrast enhancement were retrospectively analyzed with preoperative MR images in 41 patients with proven brain gliomas (8 low grade astrocytomas, 12 anaplastic astrocytomas, 21 glioblastoma multiformes). Immunohistochemical study of EGFR was done and their expressions were graded by both stained distribution and intensity. Correlation analysis between the MR features and EGFR expressions was done. Results: Peritumoral edema was correlated with both distribution (r=0.71, p=0.00) and stain intensity (r=0.69, p=0.00) of EGFR expression. Other MR features showed no statistical correlation with EGFR expression. Conclusion: MRI is useful in evaluation of brain glioma, and peritumoral edema is useful finding that suggests EGFR expression as well as malignant histopathologic grade of the tumor.

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Clinical significance of loss of p16 protein by immunohistochemical staining in acute lymphoblastic leukemia (급성림프구성백혈병에서 면역조직화학염색에 의한 p16 단백질 소실의 의의)

  • Jin, Hye Young;Kang, Kyoung In;Kim, Sun Young;Youn, You Sook;Kang, Joon Won;Jo, Deog Yeon;Kwon, Kye Chul;Park, Kyung Duk
    • Clinical and Experimental Pediatrics
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    • v.51 no.1
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    • pp.73-77
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    • 2008
  • Purpose : p16 gene, mapped to the 9p21 chromosomal region, has emerged as a candidate tumor suppressor gene in human neoplasm. It is an inhibitor of cyclin-dependent kinase and inhibits Rb phosphorylation. In a variety of tumors including childhood acute lymphoblastic leukemia (ALL), deletion and/or mutation of the p16 gene has been found. Despite their high frequency, the prognostic importance of p16 alterations is still controversial in ALL and has been reported to be either unfavorable or similar to that of other patients. We studied the correlation between loss of p16 protein confirmed by immunohistochemical staining and clinical outcomes of patients diagnosed as ALL. Methods : We performed an immunohistochemical staining for p16 protein in 74 cases of bone marrow biopsy slide initially diagnosed as ALL between January 1998 and December 2006. We reviewed the clinical manifestations, laboratory findings, treatment outcomes retrospectively. Results : Of 74 slides, 12 were negative for p16 protein. Seven were males and 5 were females with a median age at diagnosis was 5.8 (1.3-18.8) years. Initial WBC were 17,225 $(500-403,300)/{\mu}L$. By immunologic surface marker analysis, 7 patients were early pre-B CALLA (+) and 5 patients were T-cell ALL. Two patients of intermediate risk group had relapsed and died. Three patients had family history of breast cancer. Four patients died and overall survival rates were $53.5{\pm}18.7%$. Conclusion : Loss of p16 protein is supposed to be an independent risk factor of childhood ALL associated with poor outcomes. In clinical setting, the clinician must take into account p16 status, not only at the genomic but also at the protein level. Further clinical experience on thoroughly investigated cases will help a better understanding between p16 status and clinical outcomes.

The Morphologic Changes of Parvalbumin- Immunoreactive Interneurons of the Dentate Gyrus in Kainate-Treated Mouse Hippocampal Slice Culture Epilepsy Model (Kainic Acid로 처리한 해마박편배양 마우스 간질모델에서 치아이랑 Parvalbumin 면역 반응성 사이신경세포의 형태학적 변화)

  • Chung, Hee Sun;Shin, Mi-Young;Kim, Young-Hoon;Lee, In-Goo;Whang, Kyung-Tai;Kim, Myung-Suk
    • Clinical and Experimental Pediatrics
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    • v.45 no.12
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    • pp.1551-1558
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    • 2002
  • Purpose : Loss of hippocampal interneurons in dentate gyrus has been reported in patients with severe temporal lobe epilepsy and in animals treated with kainic acid(KA). Interneurons contain $Ca^{2+}$- binding protein parvalbumin(PV). The effects of kainic acid on parvalbumin-immunoreactive (PV-IR) interneurons in dentate gyrus were investigated in organotypic hippocampal slice cultures. Methods : Cultured hippocampal slices from postnatal day nine C57/BL6 mice were exposed to $10{\mu}M$ KA, and were observed at 0, 8, 24, 48, 72 hours after a one hour KA exposure. Neuronal injury was determined by morphologic changes of PV-IR interneuron in dentate gyrus. Results : Transient(1 hour) exposure of hippocampal explant cultures to KA produced marked varicosities in dendrites of PV-IR interneuron in dentate gyrus and the shaft of interbeaded dendrite is often much thinner than those in control. The presence of varicosities in dendrites was reversible with KA washout. The dendrites of KA treated explants were no longer beaded at 8, 24, 48 and 72 hours after KA exposure. The number of cells in PV-IR interneurons in dentate gyrus was decreased at 0, 8 hours after exposure. But there was no significant difference in 24, 48 and 72 hours recovery group compared with control group. Conclusion : The results suggested that loss of PV-IR interneurons in dentate gyrus is transient, and is not accompanied by PV-IR interneuronal cell death.

Expression of Stromal Derived Factor-1 is Upregulated In Macrophages during Thymic Regeneration in Adult Rat (흰쥐 가슴샘 재생과정 동안 대식세포에서 SDF-1의 발현증가)

  • Park, Hyun-Joo;Kim, Jong-Gab;Yoon, Sik;Bae, Moon-Kyoung;Bae, Soo-Kyung
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1067-1072
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    • 2009
  • Stromal derived factor-1 (SDF-1 or CXCL12), one of the CXC chemokines, is widely expressed in many tissues, including the thymus. The thymus can regenerate to its normal mass within 14 days after acute involution induced by cyclophosphamide (CY) in adult rats. Despite the established role of SDF-1 signaling in the development of T and B lymphocytes in the thymus, it has not yet been associated with the regeneration of the adult thymus. The purpose of this study was to investigate whether SDF-1, which is expressed in thymic stromal cells, is modulated during thymic regeneration in adult rats. Here, we showed that SDF-1 mRNAs were expressed in high levels in the thymocyte and thymic stromal cells at day 7 of the CY model. SDF-1 protein was shown to be present at the cortex-medulla junction, paraseptum and within the thymic medulla. Double-immunofluorescence for SDF-1 and ED-1 showed that strong SDF-1 expression was detected in the macrophages of the medulla region displaying immunoreactivity for ED-1 during thymus regeneration. Taken together, our results demonstrated that SDF-1 expression increased in regenerating thymic macrophages, suggesting the roles of SDF-1 as a chemo-attractant for damaged cells produced in the process of thymic regeneration after acute involution induced by CY.