• Title/Summary/Keyword: Negative staining

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Relationships Between Clinical Behavior of Laryngeal Squamous Cell Carcinomas and Expression of VEGF, MMP-9 and E-Cadherin

  • Akdeniz, Onder;Akduman, Davut;Haksever, Mehmet;Ozkarakas, Haluk;Muezzinoglu, Bahar
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.9
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    • pp.5301-5310
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    • 2013
  • The biological mechanisms of cancer and associations with behavior of tumours need to be studied to understand progression and determine appropriate treatments. Here we investigated expression of VEGF, MMP-9 and E-cadherin in laryngeal SCCs and their relations with clinical behavior. This prospective study was based on 38 surgical specimens from patients with primary laryngeal SCC and data recorded in their cards. Expression of the three factors in tumor tissue was examined using immunohistochemistry and correlations with clinical parameters of primary tumors, regional lymph node metastases, stage of disease, histopathologic differentiation, and vascular/cartilage invasion were investigated. Regarding the cases with positive MMP-9 expression, the difference between well and moderately/poorly differentiated tumors was statistically significant. However, differences between early stage (stage I and II) and late-stage (stage III and IV) tumours, and between positive and negative for pLN metastasis were not. No significant relationship between positive VEGF and tumor differentiation or stage was apparent, but E-cadherin levels significantly differed between well and moderately/poorly differentiated tumours and with the presence of pLN metastasis. E-cadherin staining did not vary between MMP-9 positive and negative cases. In conclusion, MMP-9 may be a negative predictor of differentiation in laryngeal SCC, while E-cadherin is a predictor of differentiation and nodal metastases. Even if the difference between VEGF expression and tumor stage was not statistically significant, it seems that there exists some relationship, which might be clarified with a greater number of cases.

BRCA1 Protein Was Not Expressed in a Normal Human Breast Epithelial Cell Type With Stem Cell and Luminal Characteristics

  • Kang, Kyung-Sun;Maki Saitoh;Angelar Cruz;Chan, Chia-Cheng;Cho, Jae-Jin
    • Toxicological Research
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    • v.14 no.2
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    • pp.123-127
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    • 1998
  • BRCA1 is a tumor suppresser gene in familial cases of breast cancer. It has been controversial whether the subcellular localization of BRCA1 is located in nuclei or cytoplasm in normal human breast cells. We found that a p220 protein was expressed in Type II Normal human breast epithelial cells (NHBEC) but not in Type I NHBEC in Western blot analysis using the 17F8 (3A2) antibody. Immunostaining using the same antibody revealed positive staining in nuclei, cytoplasm and perinuclei of Type II cells and negative staining in Type I NHBEC. The p220 protein, however, was expressed in SV40 immortalized Type I NHBEC and tumorigenic cells derived from them after x-ray and neu oncogene treatment. The subcelluar localization was mostly cytoplasmic and punctate in the nuclei. The breast carcinoma cell lines, MCF-7 and T47D, also expressed the p220 protein. Using RT-PCR, we observed the expression of BRCA1 mRNA in both Type I and Type II NHBEC. This result indicated that there might be mechanisms involved in post-translational or translational regulation of BRCA1 gene. It is speculated that the absence of BRCA1 protein expression in Type I NHBEC might playa role in their susceptibility to neoplastic transformation.

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CORRELATION BETWEEN P53, PCNA AND KI-67 EXPRESSION IN HEAD NECK SQUAMOUS CELL CARCINOMA (두경부 편평상피세포암의 p53단백과 PCNA 및 Ki-67의 발현양상)

  • Lee, Eun-Jin;Lee, Sang-Han;Sohn, Yoon-Kyung
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.27 no.2
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    • pp.142-149
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    • 2001
  • To investigate the correlation between the clinical features and the expression of p53, PCNA, and Ki-67 of the head neck squamous cell carcinoma, immunohistochemicalstaining of p53, PCNA, and Ki-67 on the paraffin embedded tissue blocks of 116 surgically removed specimens were done. The staining intensity was divided as grade 1 to grade 3 and the results were statistically analysed. 1. The positive reation rates of cell proliferation markers (PCNA and Ki-67) were higher than that of p53. There was significant correlations of the PCNA and Ki-67 expression but there was no significant correlations between p53 and PCNA or p53 and Ki-67. 2. There were no significant correlation between the expression of p53, PCNA and Ki-67 and tumor site or tumor size. 3. There was no significant differences in the positive response according to the nodal status. The node metastasis groups revealed that higher proportion of grade 3 staining of PCNA and Ki-67 than node negative group. From the above results it is concluded that p53 and cell proliferation markers PCNA and Ki-67 might have their unique mechanism involving in the growing and progression of tumor. Overexpression of p53 does not appear to represent an independent prognostic marker in head neck squamous cell carcinoma.

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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.

Osteocalcin Expression and Mineralization in Developing Tooth of Xenopus laevis

  • Park, Jung Hoe;Kwon, Ki-Tak;Park, Byung Keon;Lee, Young-Hoon
    • International Journal of Oral Biology
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    • v.40 no.1
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    • pp.1-9
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    • 2015
  • Osteocalcin (OC) is the most abundant noncollagenous protein of extracellular matrix in the bone. In an OC deficient mouse, bone formation rates are increased in cancellous and cortical bones. OC is known as a negative regulator of mineral apposition. OC is also expressed in the tooth of the rat, bovine, and human. However, little is known about OC during tooth development in Xenopus. The purpose of this study is to compare the expression of OC with mineralization in the developing tooth of Xenopus, by using von Kossa staining and in situ hybridization. At stage 56, the developmental stage of tooth germ corresponds to the cap stage, and an acellular zone was apparent between the dental papilla and the enamel organ. From stage 57, calcium deposition was revealed by von Kossa staining prior to OC expression, and the differentiated odontoblasts forming predentin were located at adjoining predentin. At stage 58, OC transcripts were detected in the differentiated odontoblasts. At stage 66, OC mRNA was expressed in the odontoblasts, which was aligned in a single layer at the periphery of the pulp. These findings suggest that OC may play a role in mineralization and odontogenesis of tooth development in Xenopus.

Association of Human Epidermal Growth Factor Receptor-2 Expression and Clinicopathological Findings in Patients with Colorectal Cancer

  • Karaca, Halit;Deniz, Kemal;Berk, Veli;Inanc, Mevlude;Ozkan, Metin
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6221-6225
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    • 2012
  • Background: To determine the frequency of HER-2 overexpression in colorectal cancer (CRC) patients, and to explore the relationship between clinicopathological prognostic factors and their effects on survival, based on immunohistochemistry (IHC) and fluorescent in situ hybridization (FISH) analysis. Materials and Methods: The study included 80 patients with a histologically proven diagnosis of CRC that received adjuvant FOLFOX-4 chemotherapy at our department between March 2006 and September 2010. Patient data were analyzed retrospectively. Results: The median follow-up period and age of the patients were 24 months and 59 years, respectively. In immunohistochemical staining, 3+ staining was found in 2 patients (2.5%) while 2+ was in 13 (16%). FISH for HER-2 was performed for all of these 15 patients; samples which were 3+ showed positivity but the ones with 2+ were negative. There was no significant correlation between HER-2 expression and age, gender, tumor localization, histological subtype, grade, lymphovascular and perineural invasion, or pTN stage (P>0.05), even when the patients with HER-2 overexpression were analyzed separately. There was also no significant relationship between progression-free survival (PFS) and overall survival (OS), and HER-2 expression, gender, tumor localization, obstruction-perforation, bleeding, histological type, grade, lymphovascular and perineural invasion, or pT staging (P>0.05); however, there was a significant relationship between lymph node involvement, and PFS and OS (P<0.05). Conclusions: Evaluation of HER-2 overexpression in a more comprehensive, multi-center, prospective trial with standardized methods will be an appropriate approach.

Comparative Analysis between Multilevel Sectioning with Conventional Haematoxylin and Eosin Staining and Immunohistochemistry for Detecting Nodal Micrometastases with Stage I and II Colorectal Cancers

  • Wong, Yin-Ping;Shah, Shamsul Azhar;Shaari, Noorsajida;Mohamad Esa, Mohd Shafbari;Sagap, Ismail;Isa, Nurismah Md
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.4
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    • pp.1725-1730
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    • 2014
  • Management of patients with stage II colorectal carcinomas remains challenging as 20 - 30% of them will develop recurrence. It is postulated that these patients may harbour nodal micrometastases which are imperceptible by routine histopathological evaluation. The aims of our study were to evaluate (1) the feasibility of multilevel sectioning method utilizing haematoxylin and eosin stain and immunohistochemistry technique with cytokeratin AE1/AE3, in detecting micrometastases in histologically-negative lymph nodes, and (2) correlation between nodal micrometastases with clinicopathological parameters. Sixty two stage I and II cases with a total of 635 lymph nodes were reviewed. Five-level haematoxylin and eosin staining and one-level cytokeratin AE1/AE3 immunostaining were performed on all lymph nodes retrieved. The findings were correlated with clinicopathological parameters. Two (3.2%) lymph nodes in two patients (one in each) were found to harbour micrometastases detected by both methods. With cytokeratin AE1/AE3, we successfully identified four (6.5%) patients with isolated tumour cells, but none through the multilevel sectioning method. Nodal micrometastases detected by both multilevel sectioning and immunohistochemistry methods were not associated with larger tumour size, higher depth of invasion, poorer tumour grade, disease recurrence or distant metastasis. We conclude that there is no difference between the two methods in detecting nodal micrometastases. Therefore it is opined that multilevel sectioning is a feasible and yet inexpensive method that may be incorporated into routine practice to detect nodal micrometastases in centres with limited resources.

Immunohistochemical Expression of Galectin-3, Cytokeratin 19 and HBME-1 in Papillary Microcarcinoma of the Thyroid Gland (갑상선 미세 유두암종에서 Galectin-3, Cytokeratin 19와 HBME-1의 발현)

  • Kim, Jin-Hwan;Lee, Sang-Sook
    • Korean Journal of Head & Neck Oncology
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    • v.23 no.2
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    • pp.133-137
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    • 2007
  • With the wide use of ultrasonography and fine needle aspiration of the thyroid gland, the incidence of papillary microcarcinoma of the thyroid gland is rapidly increasing nowadays. To improve the diagnostic accuracy of histopathologic findings of papillary thyroid carcinoma, various molecular markers have been used recently. We analysed the expression of galectin-3, cytokeratin 19 and HBME-1, using immunohistochemical technique in 37 cases of papillary microcarcinoma of the thyroid gland to evaluate the diagnostic value of these molecular markers. Immunohistochemically, galectin-3 expression was found in 37 cases of papillary microcarcinoma. Its localization was mostly cytoplasmic. Cytokeratin 19 expression was found in 36 cases. It was mostly localized to the cytoplasm and membrane. HBME-1 expression was found in all cases. Its localization was plasma membrane. The expression of these three molecular markers was negative in the adjacent normal thyroid tissue and accompanying benign lesions, although there are scattered foci of incomplete positive staining in cases of Hashimoto's thyroiditis. Our findings suggest that the immunohistochemical staining using antibodies for galectin-3, cytokeratin 19 and HBME-1 is an useful adjunctive method for the histopathological diagnosis of a papillary microcarcinoma of the thyroid gland.

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.

Histopathological Study on Teeth, Jawbone, and Oral Tissues in Poisoned Rabbit (독극물중독 가토의 치아 및 악골조직의 병리조직학적 변화에 관한 연구)

  • 김종열
    • Journal of Oral Medicine and Pain
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    • v.9 no.1
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    • pp.11-22
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    • 1984
  • The authors administered KCN, NaF,AS2O3 orally to rabbits and caused acute and chronic poisoning, then studied the teeth, jaw bones, and other oral tissues histopathologically. The results were as follows : 1. There was no significant difference between acute poisoned group by NaF and control group. But, vasodilatation in the connective tissues, esepcially marginal area of jaw bone, atrophy and destrution of glandualr cells was observed. 2. Chronic poisoned group by NaF showed degeneration and thicking of subcutanece fibrosis ective tissues, atrophy and degeneration of subcutaneous connective tissues, atrophy and degeneration of muscle fibers, vasodilation of subcutaneous in bone cavities(lacunae), and degeneration of odotlblasts in pulp tissue. 3. Acute poisoned group by KCN showed almost similar appearances as control group, and chronic poisoned group showed hyperplasia of baal layer in epitheilium, degeneration of subcutaneous connective tissues, vasodilation and huperemia, severe hemorrhage of marginal area of jaw bone. hyperplasia of salivary gland ducts, but normal arrangement of muscle fibers and narrow bone carity(lacunae) due to active osteoblastic action, osteodentin were observed. 4. Acute poisoned group by AS2O3 showed degeneration of basal cell, atrophy of blood vessels in palatal muscosa. Chronic poisoned group showed irregular cell arrangement and degeneration, reduction of capillaries in palatal mucosa. Osteoclasts in jaw bone were observed. 5. In Masson's Trichrome and Van Gieson Staining, chronic poisoned group by NaF showed thicking and loosening of subcutaneous connective tissues. Hyperplasia of intermuscular connective tissue was observed in chronic poisoning by KCN and NaF. In PAS staining, negative reation in outer layer of palatalmucosa, positive reaction in keratin layer and mild reaction of basal layer in palate and tongue mucosa was observed.

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