• Title/Summary/Keyword: 조직염색법

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Laminin-1 Expression in Bone Marrow Stromal Cells of Cyclophosphamide-treated Rat (Cyclophosphamide가 흰쥐 골수의 기질세포에서 Laminin-1의 발현에 미치는 영향)

  • Lee, Chang-Hun;Chung, Ho-Sam;Paik, Doo-Jin;Hwang, Se-Jin;Kim, Won-Kyu;Youn, Jee-Hee;Kim, Chong-Kwan
    • Applied Microscopy
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    • v.32 no.4
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    • pp.385-398
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    • 2002
  • The purpose of the present study is to investigate whether stromal cells supporting specific microenvironment for hematopoiesis of bone marrow are affected by toxicants and therapeutic drugs such as antibiotics and anticancer drugs and whether laminin-1 is associated with such effects. SD rats were intraperitoneally injected with 75 mg/kg of cyclophosphamide which is widely used to treat infant's solid tumor, leukemia and myeloma and sacrificed after 3 days, 1 week, 3 weeks or 5 weeks of injection. The bone marrow extracted and paraffin-sectioned was analyzed using immunohistochemical staining. A part of tissues was subjected to electron microscopy following reaction with rabbit anti-laminin antibody, biotinylated goat anti-rabbit IgG conjugated with 12 nm gold particles, and staining with uranyl acetate. 1. The bone marrow tissue at day 3 post injection with cyclophosphamide displayed dilated venous sinus, partial necrotic death, and decreased number of hematopoietic cells. Laminin-1 was intensively stained in the reticular and adipose tissues. 2. Up to 5 weeks post injection, laminin-1 was stained at a low level in the stromal tissue of bone marrow and the number of hematopoietic cell was increased. 3. Deposition of the gold particle which represents laminin-1 expression was observed at the highest level in the stromal cells of bone marrow obtained 3 days after injection, and decreased after 1 to 5 weeks. These results suggest that stromal cells which play a role in supporting microenvironment for bone marrow hematopoiesis augment induction of laminin-1 expression and activation upon administration of cyclophosphamide.

The Signal Transduction Mechanisms on the Intestinal Mucosa of Rat Following Irradiation (방사선조사후 백서소장점막에서 발생하는 신호전달체계에 관한 연구)

  • Yoo Jeong Hyun;Kim Sung Sook;Lee Kyung Ja;Rhee Chung Sik
    • Radiation Oncology Journal
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    • v.15 no.2
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    • pp.79-95
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    • 1997
  • Purpose : Phospholipase C(PLC) isozymes play significant roles in signal transduction mechanism. $PLC-\gamma$ 1 is one of the key regulatory enzymes in signal transduction for cellular proliferation and differentiation. Ras oncoprotein, EGFR, and PKC are also known to be involved in cell growth. The exact mechanisms of these signal transduction following irradiation, however, were not clearly documented Thus, this study was Planned to determine the biological significance of PLC, ras oncoprotein, EGFR, and PKC in damage and regeneration of rat intestinal mucosa following irradiation. Material and Method : Sixty Sprague-Dawley rats were irradiated to entire body with a single dose of 8Gy. The rats were divided into S groups according to the sacrifice days after irradiation. The expression of PLC, ras oncoprotein, EGFR and PKC in each group were examined by the immunoblotting and immunohistochemistry. The histopathologic findings were observed using H&I stain, and the mitoses for the evidence of regeneration were counted using the light microscopy & PCNA kit. The Phosphoinositide(PI) hydrolyzing activity assay was also done for the indirect evaluation of $PLC-\gamma$ 1 activity. Results: In the immunohistochemistry , the expression of $PLC-{\beta}$ was negative for all grøups. The expression of $PLC-{\gamma}1$ was highest in the group III followed by group II in the proliferative zone of mucosa. The expression of $PKC-{\delta}1$ was strongly positive in group 1 followed by group II in the damaged surface epithelium. The above findings were also confirttled in the immunoblotting study. In the immunoblotting study, the expressions of $PLC-{\beta}$, $PLC-{\gamma}1$, and $PKC-{\delta}1$ were the same as the results of immunohis-tochemistry. The expression of ras oncoprctein was weakly positive in groups II, III and IV. The of EGFR was the highest in the group II, III, follwed by group IV and the expression of PKC was weakly positive in the group II and III. Conclusion: $PLC-{\gamma}1$ mediated signal transduction including ras oncoprotein, EGFR, and PKC play a significant role in mucosal regeneration after irradiation. $PLC-{\delta}1$ mediated signal transduction might have an important role in mucosal damage after irradiation. Further studies will be necessary to confirm the signal transduction mediating the $PKC-{\delta}1$.

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Early Development of Olfactory Organs in White rat (백서 후각기의 초기발생)

  • 백경기;홍세표
    • The Korean Journal of Zoology
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    • v.10 no.1
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    • pp.10-16
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    • 1967
  • 본 연구는 백서후각기의 발생과정을 세포조직학적으로 규명보고하는 바이며 사용된 stage는 stage 20부터 32까지는 Christe(1964)의것을 인용하였으며 그 외에 명시된 stage 33,34, 35 는 저자가 편의상 구분하였다. 재료는 백서 5 필을 온실서 충분한 영양으로 사육후 임신시켜 요구되는 배를 stage 별로 얻어 Bouin's solution 에 고정시켜 paraffin법을 따랐으며 5-7$\mu$의 두께로 절편을 만들어 Delafield's hematoxylin, eosin, thionin 에 염색하였다. Stage 별 후각기의 발생은 state 20 에 후순, state 22에 후와 및 조비기(Jacobson's organ), stage23A에 구비강막(oro-nasal membrane), state 23 B 에 수신경섬유 와 원시내비공, stage 24 에 조비기신경섬유, stage 27에는 골갑개상피에 후세포와 호흡상피상에 섬유 또한 악골갑개상피에서 seromucinous 선들이 최초로 나타났다. 본 실험결과 백서후각기의 형성과정은 인체와 생쥐( mouse) 에서와 유사하나 조비기의 성장, 비갑개의수, 및 후역의 범위에 있어서는 극히 차가 심한 것같다.

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Detection of HHV6 and EBV in histiocytic necrotizing lymphadenitis (Histiocytic necrotizing lymphadenitis에서 HHV6과 EBV의 검출)

  • Park, Kyung Hee;Park, Sung Shik;Kim, Ji Yeon;Park, Su Eun
    • Clinical and Experimental Pediatrics
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    • v.51 no.9
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    • pp.987-991
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    • 2008
  • Purpose : Kikuchi-Fujimoto disease (KFD), also known as histiocytic necrotizing lymphadenitis (HNL), is a self-limited disease characterized by cervical lymphadenopathy and fever. The etiology of KFD remains unknown; however, the self-limiting nature of HNL suggests the cause of this disease could be viral infection. For this reason, several viruses have been evaluated as possible etiologies of HNL, including Epstein-Barr virus (EBV), human herpesvirus 6 (HHV6), human herpesvirus 8 (HHV8), and cytomegalovirus (CMV). The aim of this study was to examine the relationship of EBV and HHV6 to HNL. Methods : Data pertaining to 51 cases with biopsy-confirmed HNL were collected between January 1999 and December 2005, from the Department of Pathology, College of Medicine, Pusan National University, Busan, Korea. The clinical records-including data regarding age, gender, duration of fever, and lymph node involvementwere reviewed retrospectively. The in situ hybridization (ISH) assay was performed by EBER PNA probe (Dako, Capinteria, CA, USA), and immunohistochemistry testing was performed with anti-HHV type 6 monoclonal antibodies (Chemicon, Temecula, CA, USA). Results : The HNL patients in this study were 24 males and 27 females, ranging in age from seven to 61 years (median: 25.9). ISH for EBV was positive in 8/51 (15.7%) biopsies, and immunohistochemistry for HHV6 was positive in 15/51 (29.4%) biopsies. Serologic analysis of EBV IgM was performed in 23 cases; only one patient was positive for EBV IgM and EBV ISH. Conclusion : Our study could not provide supportive evidence of a viral pathogenesis for HNL; therefore, cases of HNL may not have a dominant viral cause. However, some rare exceptional cases may have been caused by viral infection.

Study on DNA Content and Ki-67 Antibody Expression by Means of Image Analyzer for the Benign and Malignant Lesions of the Larynx (후두 편평상피의 전암성 및 악성병변에서 화상분석기를 이용한 DNA 배수성검사와 Ki-67 항체 양성세포의 분석에 관한 연구)

  • 주형로;이선희;최종욱;김인선
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1993.05a
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    • pp.89-89
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    • 1993
  • The laryngeal epithelial cell kinetics of 26 laryngeal lesions(invasive squamous cell carcinoma 14, epithelial hyperplasia 5, laryngeal nodule 7) were studied by immunehistochemical analysis with the monoclonal antibody Ki-67, which reacts with nuclear antigen in proliferating cells using paraffin embedded tissue. For DNA analysis, touch implint with fresh biopsy specimens were stained with feulgen and analyzed by image analyzer in 22 cases. 1) The proportion of Ki-67-positive cells were 32.65$\pm$ 11.59% in invasive squamous cell ca, 20.14$\pm$3.38% in epithelial hyperplasia lesion and 11.66$\pm$3.02% in laryngeal nodule. 2) DNA aneuploidy was found in 7 cases of 10(70%) invasive squamous cell carcinomas, 2 cases of 5(40%) epithelial hyperplasia lesions and all cases of laryngeal 3) Proliferation index(S phase+G2/M phase) show 23.42$\pm$11.33% in squamous cell carcinoma, 13.09$\pm$ 10.90% in epithelial hyperplasia lesion and 4.50$\pm$1.19% in laryngeal nodule. As the results, measuring the DNA content from touch imprint method together positivity of Ki-67 antibody from the microtissue during the laryngeal microscopic surgery, cell kinetics can be assessed as an effort of deciding the prognosis and provide a key to the management of precancerous lesions.

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Laminin Expression in the Rat Lung Development (흰쥐 폐 발생시 Laminin의 발현에 대한 연구)

  • Chung, Ho-Sam;Park, Chul-Hong;Paik, Doo-Jin;Baik, Tae-Kyung;Kim, Won-Kyu;Youn, Jee-Hee;Suh, Yun-Kyung
    • Applied Microscopy
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    • v.31 no.1
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    • pp.71-83
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    • 2001
  • Laminin, a kind of multidomain glycoproteins, is mainly localized in the basement membranes of various tissues. It is known that laminin plays an important part in mammalian lung morphogenesis. The authors have undertaken this study to investigate the changes in the distribution of laminin, and to find out cells which synthesize laminin during the organogenesis and differentiation of the lung. The fetal and neoantal rats (Sprague-Dawley strain) were used as experimental animals. The immunohisto-chemical methods were employed for detection of laminin within the developing lung tissue and the immunegold cytochemical methods were performed for detection of cells which synthesize laminin according to each stage of development. The results are as follows; 1. During fetal life, strong immunoreactivity for laminin is maintained in the basement membranes of the blood vessels and the bronchioles, the extracellular matrix of the mesenchyme, and basal lamina of the alveolar septum in the fetal rat lung. 2. After birth, laminin immunoreactivity at the alveolar septum is gradually reduced. 3. During fetal life, laminin is mainly detected within the cytoplasm of the mesenchymal cells, the endothelial cells of blood vessels and the fibroblasts in fetal rat lung. 4. According to the differentiation of type I and type II pneumocyte after birth, laminin is detected within cytoplasm of the type I pneumocytes, type II pneumocytes and fibroblasts. It is consequently suggested that laminin is largely expressed in the developing lung and laminin may be also synthesized by the type II pneumonocytes at early newborn stages.

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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.

The Relationship Between Loss of Blood Group Antigen A in Cancer Tissue and Survival Time in the Antigen A Positive Non-Small Cell Lung Cancer (A 항원 양성 원발성 비소세포폐암 조직에서의 A 항원 소실과 생존기간과의 관계)

  • Yang, Sei-Hoon;Jeong, Eun-Taik
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.3
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    • pp.339-346
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    • 2000
  • Background : The moot important prognostic factor in non-small cell lung cancer is the TNM stage. Even after complete resection in early non-small cell lung cancer, the five-year survival rate is still low. However, new prognostic factors, including molecular biologic factors, have recently been found to guide the treatment of patients with non-small cell lung cancer. We evaluated the prognostic value of the loss of blood-group antigen A in tumor tissue, which has been implicated as an important prognostic factor for overall survival and the timing of the disease progression. Methods : The loss of blood-group antigen A was assessed immunohistochemically in paraffin-embedded tumor samples from 26 patients with blood types A or AB, who had undergone curative surgery. Monoclonal antibody was used to detect the blood group antigen A expression. Results : Fifteen patients (58%) expressed antigen A in their tumor tissue, whereas 11 patients (42%) did not show antigen A. The median survival time of the blood A antigen positive group was 11 months, while the median survival time of the blood A antigen negative group was 18 months. The difference in survival between the two groups was not statistically significant. Conclusion : The loss of blood-group antigen A in tumor tissue was not found to be a significant prognostic factor in patients with non-small cell lung cancer. This study needs to be extended for further evaluation.

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Studies on the Histological Observation of Removing the Skin from Squid by Various Treatments (각종처리(各種處理)에 의한 오징어의 박피(剝皮)에 관(關)한 조직학적관찰(組織學的觀察))

  • Youn, Jung-Eui
    • Korean Journal of Food Science and Technology
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    • v.8 no.4
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    • pp.207-211
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    • 1976
  • Treated with various methods, body muscle of squid was cross section, stained by Van Gieson staining method, and observed microscopically to study on the state of thin layer of skin separated from meat. Results obtained were summarized as follows : 1. By general hot spring treatment were 1st. 2nd. and 3rd. layers of skin removed from squid muscle but 4th. not. 2. By boiling treatment was tissue of muscle fibre fissured largely toward the muscle fibre. 3. Sodium acetate treatment was superior to sodium nitrate, sodium sulfate and sodium tartrate treatment in the effect that the skin was separated from meat. Especially, concentrate solution of sodium acetate had the most excellent effect in the chemical reagent treatment. 4. By proteolytic enzyme treatment were 1st. 2nd. 3rd. and 4th. layers of skin removed all from squid muscle and the boundary fibre between skin and meat swelled in particular. 5. Two kinds of skin removing method, proteolytic enzyme treatment and sodium acetate treatment, were desirable to the actual processing.

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