• Title/Summary/Keyword: Immunohistochemical Staining

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E-cadherin and $\beta-catenin$ Expression and Mutation in Gastric Carcinomas (위암에서 E-cadherin과 $\beta-catenin$ 발현과 유전자 돌연변이에 관한 연구)

  • Kim Kwang Il;Park Sung-Hye;Han Sun-Ae;Chae Yang-Seok;Kim Insun
    • Journal of Gastric Cancer
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    • v.1 no.4
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    • pp.202-209
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    • 2001
  • Purpose: When cancer cels invade the stroma, they should be dissociated from the adjacent cells at first. E-cadherin and $\beta-catenin$ constitute an important protein complex associated with cellular adhesion, development, and differentiation, especially in epithelial cells. The role of E-cadherin and $\beta-catenin$ in gastric carcinogenesis were studied. Materials and Methods: The expression of E-cadherin and $\beta-catenin$ in gastric adenocarcinomas by using immunohistochemical staining and the mutation by using polymerase chain reaction- single stranded conformation polymorphism (PCR-SSCP) and sequencing were performed in 40 adenocarcinomas and 5 dysplasia of stomach. Thirteen cases, which had lymph node metastasis, were also included for immunohistochemical staining. Results: Inappropriate cytoplasmic and/or nuclear expression of a E-cadherin-$\beta-catenin$ complex was more frequent in poorly differentiated, diffuse type signet ring cell carcinomas than in well-differentiated, intestinal type adenocarcinomas (P<0.05). However, the expression was not related with clinical stage or lymph node metastasis. Mutation of E-cadherin was detected in 4 cases by using PCR-SSCP, whereas mutation of $\beta-catenin$ was detected in 2 cases. Conclusion: E-cadherin and $\beta-catenin$ seem to be important in gastric carcinogenesis, especially in poorly differentiated diffuse type.

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Demyelination of neurofilament protein 200 immune positive never fibers in human pulp (사람 치수 내에서 neurofilament protein 200 면역양성반응을 나타내는 신경섬유의 탈말이집 현상에 대한 연구)

  • Jang, Jung-Woo;Choi, So-Young;Kwon, Dae-Geon;Bae, Yong-Chul;Kim, Chin-Soo;Lee, Sang-Han
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.5
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    • pp.360-365
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    • 2010
  • Introduction: Mammalian tooth pulp is densely innervated by sensory nerves that are mostly C fibers and A delta fibers. However, there is evidence suggesting that many unmyelinated axons in the pulp are in fact parent meylinated axons. Immunohistochemical staining for neurofilament protein 200 kDa (NFP200) was performed to identify the demyelinated but parent myelinated axons. Materials and Methods: The pulp was removed from healthy premolars and 3rd molars extracted from juveniles and adults undergoing orthodontic treatment, and immunohistochemical staining were applied with NPF200 antibodies, which specifically dye myelinated axons. The specimens underwent an electron microscopy examination with diaminobenzidine (DAB) immunostaining after observation and analysis by fluorescence and confocal laser scanning microscopy. Results: The NPF200 immuno-positive axons in the radicular pulp areas were observed as bundles of many nerve fibers. Many small bundles were formed with fewer axons when firing to the coronal pulp areas and then reachrd a different direction. In the radicular pulp, unmyelinated axons and myelinated axons were present together. However, in the coronal pulp, unmyelinated axons were most common and NFP200 immuno-positive unmyelinated axons with a larger diameter than those in the radicular pulp were observed more frequently. On the other hand, most of the immuno-positive unmyelinated fibers were similar in size to that of typically well-known unmyelinated fibers. Conclusion: Myelinated fibers innervated to the dental pulp maintain their myelins in the radicular portion, but these fibers lost myelins in the coronal portion. After the loss of myelin, the size of the axoplasm also decreased.

ROLE OF DCC(DELETED IN COLORECTAL CANCER) GENE IN ORAL SQUAMOUS CELL CARCINOMA (구강편평상피암종에서 DCC 유전자의 역할)

  • Ko, Seong-Kyu;Han, Se-Jin;Kim, Kyung-Wook
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.34 no.5
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    • pp.518-524
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    • 2008
  • Chromosome 18q alteration plays a key role in colorectal tumorigenesis, and loss of heterozygosity at 18q is associated with a poor prognosis in colon cancer. DCC(Deleted in Colorectal Cancer) is a putative tumor- suppressor gene at 18q21 that encodes a transmembrane protein with structural similarity to neural cell adhesion molecule that is involved in both epithelial and neuronal cell differentiation. DCC is implicated in regulation of cell growth, survival and proliferation. Thus, tumor progression in squamous cell carcinoma, stomach cancer, colorectal cancer correlates with downregulation of DCC expression. The mechanism for DCC suppression is associated with hypermethylation of the DCC gene promoter region. Hence, the goal of this study is to identify the promoter methylation responsible for the down-regulation of DCC expression in oral squamous cell carcinoma. 12 of tissue specimens for the study are excised and gathered from 12 patients who are diagnosed as SCC in department of OMS, dental hospital, dankook university. To find expression of DCC in each tissue samples, immunohistochemical staining, RT-PCR gene analysis and methylation specific PCR are processed. The results are as follows. 1. In the DCC gene RT-PCR analysis, 5(41.6%) of 12 specimens of oral squamous cell carcinoma did not expressed DCC gene. 2. In the promoter methylation specific PCR analysis, 5(41.6%) of 12 specimens showed promoter methylation of DCC gene. 3. In the immunohistochemical staining of poor differentiated and invasive oral squamous cell carcinoma, loss of DCC expression was observed. These findings suggest that methylation of the DCC gene may play a role in loss of gene expression in invasive oral squamous cell carcinoma.

Differential Expression of EGFR Protein by Immunohistochemical Staining Methods and the Relationship Between the Degree of EGFR Protein Expression and EGFR Gene Mutation (면역조직화학적 염색 방법에 따른 상피세포 성장 수용체 단백(EGFR)의 발현정도의 차이 및 EGFR의 발현정도와 EGFR 유전자의 돌연변이와의 상관관계에 대한 고찰)

  • Yoon, In-Sook;Kim, Keuk-Jun;Lee, Eun-Hwa;Seok, Sang-Hee;Kim, Sang-Hee;Kim, Hyun-Yong;Song, Ho-Jung;Lee, Tae-Jong
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.3
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    • pp.217-222
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    • 2007
  • In the last 5 years the Epidermal Growth Factor Receptor (EGFR) has emerged as one of the most important targets for drug development in oncology. Monoclonal antibodies targeting the external domain of EGFR have been shown to have clinical benefits in colorectal and head and neck cancer when combined with chemotherapy and/or radiation. Also the targeting of the epithelial growth factor receptor (EGFR) kinase domain using the closely related inhibitors gefitinib and erlotinib has generally been ineffective against solid tumors, many of which over express the receptor. We found that there were some differential expressions according to primary antibodies of the EGFR protein which being used as one of the histological tumor markers for non-small cell lung cancer (NSCLC). We also found that there are some differential expressions according to antibodies, the pH of the antigen retrieval (AR) buffer solutions and kinds of enzymes. There were some differential expressions according to the secondary antibodies and the detection systems. We analyzed the correlations between the immunohistochemical expressions of the EGFR protein and the gene mutations of the EGFR. The differences between automatic stainers and manual staining methods were also evaluated.

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Differential Expression of miR-34c and Its Predicted Target Genes in Testicular Tissue at Different Development Stages of Swine

  • Zhang, Xiaojun;Zhao, Wei;Li, Chuanmin;Yu, Haibin;Qiao, YanYan;Li, Aonan;Lu, Chunyan;Zhao, Zhihui;Sun, Boxing
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.11
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    • pp.1532-1536
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    • 2015
  • To verified the target genes of miR-34c, bioinformatics software was used to predict the targets of miR-34c. Three possible target genes of miR-34c related to spermatogenesis and male reproductive development: zinc finger protein 148 (ZNF148), kruppel-like factor 4 (KLF4), and platelet-derived growth factor receptor alpha (PDGFRA) were predicted. Then, the expression of miR-34c and its target genes were detected in swine testicular tissue at different developmental stages by quantitative polymerase chain reaction. The results suggested that the expression of PDGFRA has the highest negative correlation with miR-34c. Then immunohistochemical staining was done to observe the morphology of swine testicular tissue at 2-days and 3, 4, 5-months of age, which indicated that PDGFRA was mainly expressed in the support cells near the basement membrane during the early development stages of testicular tissue, but that the expression of PDGFRA was gradually reduced in later stages. Therefore, western blot analyzed that the highest expression of PDGFRA was generated in 2-days old testicular tissues and the expression levels reduced at 3 and 4-months old, which correlated with the results of immunohistochemical staining. In conclusion, PDGFRA is a target gene of miR-34c.

Expression of Nitric Oxide Synthase and Endothelin-1 in Human Uterine Artery from Full-Term Pregnancies

  • Choi, Ook-Hwan;Lee, Sun-Hee;Kim, Eun-Jin;Kim, Koan-Hoi;Rhim, Byung-Yong
    • The Korean Journal of Physiology and Pharmacology
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    • v.9 no.3
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    • pp.165-172
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    • 2005
  • The aim of this study was to determine the roles of ET-1 and NO on uterine blood flow in pregnancy. Uterine arteries were isolated from 17 nonpregnant and 12 pregnant women. Nonpregnant group included patients with median age of $48.6{\pm}2.3$ years who underwent hysterectomy, because of myoma. Pregnant group included patients with median age of $31.3{\pm}1.4$ years undergoing cesarean delivery. ET-1 and ET-2 induced concentration-dependent contraction in isolated nonpregnant and pregnant uterine arteries. The contractile response and maximal contraction were increased in pregnant uterine arteries. In nonpregnant uterine arteries, there was no contraction in response to ET-3, whereas pregnancy induced concentration-dependent contraction by ET-3. Tissue nitrite/nitrate level and immunohistochemical staining of eNOS and iNOS were increased in pregnant uterine arteries, compared with nonpregnant uterine arteries. In addition, the expressions of eNOS and iNOS mRNA were significantly increased in pregnancy. Moreover, contractions by ET isopeptides, including ET-1, were enhanced, and immunohistochemical staining of ET-1 and ET-1 mRNA expression was increased in pregnant uterine arteries. These results suggest that NO production by increased NOS activity, especially eNOS activity, is related to placental and uterine blood flow. Furthermore, ET-1 appears to play a pathophysiological role in pregnant complications such as hypertension.

Histologic effects of intentional-socket-assisted orthodontic movement in rabbits

  • Yu, Ji-Yeon;Lee, Won;Park, Jae Hyun;Bayome, Mohamed;Kim, Yong;Kook, Yoon-Ah
    • The korean journal of orthodontics
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    • v.42 no.4
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    • pp.207-217
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    • 2012
  • Objective: This study aimed to evaluate the effect of an intentionally created socket on bone remodeling with orthodontic tooth movement in rabbits. Methods: Eighteen male rabbits weighing 3.8 - 4.25 kg were used. An 8-mm deep and 2-mm wide socket was drilled in the bone 1 mm mesial to the right mandibular first premolar. The left first premolar was extracted to serve as an extraction socket. A traction force of 100 cN was applied to the right first premolar and left second premolar. Sections were obtained at the middle third of the moving tooth for both the drilled and extraction sockets and evaluated with hematoxylin and eosin staining and immunohistochemical analyses. The amount of tooth movement and tartrate-resistant acid phosphatase (TRAP)-positive cell count were compared between the 2 groups using the Mann-Whitney U test. Results: At week 2, the distance of tooth movement was significantly higher in the intentional socket group (p < 0.05) than in the extraction socket group. The number of TRAP-positive cells decreased in week 2 but increased in week 3 (p < 0.05). However, there were no significant differences between the groups. Furthermore, results of transforming growth factor (TGF)-${\beta}$ staining revealed no significant differences. Conclusions: The intentional socket group showed greater distance of tooth movement than did the extraction socket group at week 2. Osteoclast counts and results of immunohistochemical analyses suggested elevated bone remodeling in both the groups. Thus, osteotomy may be an effective modality for enhancing tooth movement in orthodontic treatment.

Expression of Neural Cell Adhesion Molecule (NCAM) and Glial Cell Line-Derived Neurotrophic Factor (GDNF) in Aganglionic Bowel of Hirschsprung's Disease (허쉬슈프렁병 환아의 무신경절 장관에서 Neural Cell Adhesion Molecule (NCAM) 과 Glial Cell Line-Derived Neurotrophic Factor (GDNF)의 발현)

  • Oh, Jung-Tak;Han, Ai-Ri;Son, Suk-Woo;Choi, Seung-Hoon;Han, Seok-Joo;Hwang, Eui-Ho;Yang, Woo-Ick
    • Advances in pediatric surgery
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    • v.7 no.1
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    • pp.15-20
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    • 2001
  • The pathophysiology of Hirschsprung's disease (HD) is not fully understood, but recent studies have disclosed that neural cell adhesion molecule (NCAM) and glial cell line-derived neurotrophic factor (GDNF) play important roles in the formation of aganglionic bowel of Hirschsprung's disease. To evaluate the roles of NCAM and GDNF in HD, immunohistochemical analysis was performed using formalin-fixed and paraffin-embedded tissue sections. On the basis of the results, we tried to evaluate them as diagnostic markers. The specimens were obtained from 7 patients with HD who underwent modified Duhamel operation. The diagnosis was based on the clinical findings and the absence of ganglion cells in the nerve plexuses by routine microscopy. NCAM immunoreactivity was found in the nerve plexuses and scattered nerve fibers in the smooth muscle layers of ganglionic segments. In aganglionic segments, the number of NCAM positive nerve fibers in the smooth muscle layers was significantly reduced compared with ganglionic segments. In two cases the nerve plexuses in aganglionic segments, NCAM was negligible. The smooth muscle cells showed diffuse immunoreactivity for GDNF and the staining intensity was not different in the aganglionic and ganglionic segments. However, higher expression of GDNF in the nerve plexus of the ganglionic segments was noted comparing to aganglionic segments. These data suggest that both NCAM and GDNF may play important roles in pathogenesis of Hirschsprung's disease and immunohistochemical staining for NCAM can be used as an ancillary diagnostic tool for HD.

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Alleviation of Atopic Dermatitis through Probiotic and Mixed-probiotic Treatments in an Atopic Dermatitis Model

  • Seo, Jae-Gu;Chung, Myung-Jun;Lee, Hyun-Gi
    • Food Science of Animal Resources
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    • v.31 no.3
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    • pp.420-427
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    • 2011
  • The use of lactic-acid bacteria (LAB) is effective for preventing and curing immune disorders by activating the immune system in the digestive tract and the consequent immune response in the blood. In this study, LAB and mixed LABs were used in an atopic dermatitis (AD) mouse model. Alleviation of AD was observed based on the change in cytokine level and immunohistochemical staining. An ex vivo test showed that immunoglobulin-E and interleukin (IL)-4 levels were significantly lower in all groups treated with LAB than in the group treated with only 1-chloro-2,4-dinitrobenzene. Results of an in vivo test based on the ex vivo results showed that the scratch score decreased in all groups treated with the LAB and particularly decreased in the group treated with mixed LABs. Additionally, the T helper (Th) 1 cytokines interferon-gamma and IL-12p40 were upregulated by the LAB and mixed-LABs, whereas levels of the Th2 cytokine IL-4 were downregulated in a mouse model of AD-like skin lesions. Furthermore, hematoxylin & eosin and immunohistochemical staining of the dorsal area of the mice in each group showed that AD improved in the LAB-treated groups. These results suggest that LAB and mixed LABs inhibit the development of AD in NC/Nga mice by suppressing the Th2 cell response and increasing the Th1 cell response. Our results indicate that mixed LABs are better than LAB for treating AD-like skin lesions.

Expression of Pituitary Tumor Transforming Gene 1 is an Independent Factor of Poor Prognosis in Localized or Locally Advanced Prostate Cancer Cases Receiving Hormone Therapy

  • Cao, Xi-Liang;Gao, Jiang-Ping;Wang, Wei;Xu, Yong;Shi, Huai-Yin;Zhang, Xu
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3083-3088
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    • 2012
  • We investigated the prognostic value of pituitary tumor transforming gene 1 (PTTG1) expression according to clinicopathological features among localized or locally advanced prostate cancer cases receiving hormone therapy. A retrospective study involved 64 patients receiving combined androgen blockade treatment was performed. PTTG1 expression was determined by immunohistochemical staining using initial needle biopsy specimens for diagnosis. Associations of PTTG1 with various clinicopathological features and disease-free survival were examined via uni- and multivariate analyses. No association between PTTG1 expression and clinical T stage, Gleason score, pretreatment PSA levels, risk groups was found (p =0.682, 0.184, 0.487, 0.571, respectively). Univariate analysis revealed that increased PTTG1 expression, T3 stage and high risk group were associated with increased risk of disease progression (p =0.000, 0.042, and 0.001), and high PSA level had a tendency to predict disease progression (p =0.056). Cox hazard ratio analysis showed that PTTG1 low expression (p =0.002), PTTG1 high expression (p =0.000) and high risk group (p =0.0147) were significantly related to decreased disease-free survival. In conclusion, PTTG1 expression determined by immunohistochemical staining in needle biopsy specimens for diagnosis is a negative prognostic factor for progression in localized or locally advanced prostate cancer receiving hormone therapy.