• Title/Summary/Keyword: Negative staining

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Modified Toluidine Blue: an Alternative Stain for Helicobacter pylori Detection in Routine Diagnostic Use and Post-eradication Confirmation for Gastric Cancer Prevention

  • Sakonlaya, Dussadee;Apisarnthanarak, Anucha;Yamada, Nobutaka;Tomtitchong, Prakitpunthu
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.16
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    • pp.6983-6987
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    • 2014
  • Background: Modified toluidine blue staining (MTBs) is a simple, inexpensive and time saving method to detect H. pylori in gastric biopsy specimens. As a metachromatic stain, it simultaneously highlights intestinal metaplasia, a gastric cancer precancerous lesion. The aim of this study was to assess the reliability of MTBs compared with hematoxylin-eosin (H&E) for H. pylori detection using immunoperoxidase staining as the gold standard. This technique would be beneficial for a routine diagnosis and confirmation of H. pylori eradication in developing countries where endoscopic-based approaches are dominant. Materials and Methods: Esophagogastroduodenoscopy with triple site gastric biopsies was undertaken in 207 dyspeptic patients at Thammasat University Hospital, Thailand between 1997 and 1999. H&E, MTBs and immunoperoxidase staining were applied to each specimen. The presence or absence of H. pylori with each stain was interpreted separately and the sensitivity, specificity, positive and negative predictive values of H&E and MTBs were calculated. Results: A total of 282 specimens from 207 patients were evaluated. Using immunoperoxidase staining, organisms were positive in 117 specimens (41%). MTBs proved almost equally sensitive as immunoperoxidase (99%) and significantly more sensitive than H&E (85%). It has comparable specificity (96% vs 96%), PPV (95% vs 94%), and NPV (99% vs 90%) to H&E, using immunoperoxidase staining as gold standard. MTBs compared with immunoperoxidase staining, is cheaper (2 USD vs 12 USD) and faster (20 min vs 16 hrs) compared to immunoperoxidase staining. Conclusions: MTBs is effective, economical and easy to use in daily practice for the detection of H. pylori in gastric biopsy specimens. In addition to saving time in evaluating H. pylori associated gastritis, with a high sensitivity and ability to demonstrate intestinal metaplasia, the technique may have a role in confirmation of H. pylori eradication for gastric cancer prevention in a developing country setting.

C-Kit-Negative Gastrointestinal Stromal Tumor in the Stomach

  • Seo, Ho Seok;Hyeon, Ji Yeon;Shin, Ok-Ran;Lee, Han Hong
    • Journal of Gastric Cancer
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    • v.15 no.4
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    • pp.290-294
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    • 2015
  • C-kit-negative gastrointestinal stromal tumors (GISTs) are uncommon, and there have been few reports about the diagnosis and treatment of c-kit-negative GISTs in the stomach. We report the case of a patient who was diagnosed with a huge and atypical GIST in the stomach. The GIST was completely resected and finally diagnosed as c-kit-negative GIST based on immunohistochemical staining of tumor cells, which were negative for CD117 and CD34 and positive for Discovered on GIST-1 (DOG1). C-kit-negative GISTs could be treated by complete resection and/or imatinib, which is the same treatment for c-kit-positive GISTs.

Androgen Receptor Expression and its Correlation with Other Risk Factors in Triple Negative Breast Cancers: a Report from Western Iran

  • Payandeh, Mehrdad;Shazad, Babak;Madani, Seyed-Hamid;Ramezani, Mazaher;Sadeghi, Masoud
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3321-3324
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    • 2016
  • Background: Androgen receptors (ARs) are expressed in more than 70% of breast cancers (BCs) and have been implicated in BC pathogenesis. Some triple negative (TN)BC tumors express AR and may benefit from AR-targeted therapies. The aim of this study was to evaluate survival and the prevalence of AR expression and its correlation with other risk factors in triple negative BCs in women from Western Iran. Materials and Methods: In a retrospective study between 2009-2015, 41 patients with TNBC were referred to the Private Clinic of Oncology, Kermanshah city, Iran. ER, PR and AR-positive expression was defined as ${\geq}10%$ nuclear staining and also HER2 (2+), FISH was performed. Nuclear staining was considered representative for Ki67 and P53. The mean follow-up for the patients was 25 months. In this time, 5 patients died and 4 lost to follow-up were censored from survival analysis. Results: The mean age at diagnosis was 46.9 years (range, 24-71 years) and all patients were female. The OS rates for AR-positive and AR-negative patients were 90% and 85.1%, respectively, and the mean OS was 26.3 and 23.2 months. Therefore, there was no significant difference between the two groups (Hazard ratio: 0.580, 95% CI: 0.086-3.893, P=0.575). Conclusions: In TNBC patients, evaluation of AR status may provide additional information on prognosis and treatment. The results of studies showed that the prevalence AR expression may differ in the world and probably ethnicity can be an influencing factor.

Immunocytochemical Staining for p16 of Atypical Squamous Cells in Cervicovaginal Smear (자궁목질 바른표본에서 비정형 편평세포의 p16에 대한 면역세포화학염색)

  • Kim, Hwal-Woong;Lee, Jong-Sil;Lee, Jeong-Hee;Ko, Gyung-Hyuck
    • The Korean Journal of Cytopathology
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    • v.15 no.1
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    • pp.28-32
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    • 2004
  • It was reported that the main cause of intraepithelial neoplasm and squamous cell carcinoma of the uterine cervix is human papilloma virus infection, and that the expression of p16 is increased in cells infected by human papilloma virus. We performed an immunocytochemical staining for protein p16 in 17 cases of cervocovaginal smears initially diagnosed as atypical squamous cells of undetermined significance, to know whether the staining could help the differentiation of neoplastic cells from reactive atypical cells. Of 17 smears, 6 were diagnosed finally as high grade intraepithelial neoplasm or invasive squamous cell carcinoma by follow-up biopsy and smear, and 5 of the 6 were positive for p16. Three were diagnosed as koilocytosis, and one of them was weakly positive for p16. Eight were diagnosed as reactive atypical cells, and all of them were negative for p16. We thought that immunocytochemical staining of p16 in cervocovaginal smears could help the differentiation of neoplastic cells from reactive atypical cells.

Opisthorchis viverrini Infection Activates the PI3K/AKT/PTEN and Wnt/β-catenin Signaling Pathways in a Cholangiocarcinogenesis Model

  • Yothaisong, Supak;Thanee, Malinee;Namwat, Nisana;Yongvanit, Puangrat;Boonmars, Thidarut;Puapairoj, Anucha;Loilome, Watcharin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10463-10468
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    • 2015
  • Opisthorchis viverrini (Ov) infection is the major etiological factor for cholangiocarcinoma (CCA), especially in northeast Thailand. We have previously reported significant involvement of PI3K/AKT/PTEN and $Wnt/{\beta}$-catenin in human CCA tissues. The present study, therefore, examined the expression and activation of PI3K/AKT/PTEN and $Wnt/{\beta}$-catenin signaling components during Ov-induced cholangiocarcinogenesis in a hamster animal model. Hamsters were divided into two groups; non-treated and Ov plus NDMA treated. The results of immunohistochemical staining showed an upregulation of PI3K/AKT signaling as determined by elevated expression of the $p85{\alpha}$-regulatory and $p110{\alpha}$-catalytic subunits of PI3K as well as increased expression and activation of AKT during cholangiocarcinogenesis. Interestingly, the staining intensity of activated AKT (p-AKT) increased in the apical regions of the bile ducts and strong staining was detected where the liver fluke resides. Moreover, PTEN, a negative regulator of PI3K/AKT, was suppressed by decreased expression and increased phosphorylation during cholangiocarcinogenesis. We also detected upregulation of $Wnt/{\beta}$-catenin signaling as determined by increased positive staining of Wnt3, Wnt3a, Wnt5a, Wnt7b and ${\beta}$-catenin, corresponded with the period of cholangiocarcinogenesis. Furthermore, nuclear staining of ${\beta}$-catenin was observed in CCA tissues. Our results suggest the liver fluke infection causes chronic inflammatory conditions which lead to upregulation of the PI3K/AKT and $Wnt/{\beta}$-catenin signaling pathways which may drive CCA carcinogenesis. These results provide useful information for drug development, prevention and treatment of CCA.

IMMUNOHISTOCHEMICAL STUDY ON EXPRESSION PATTERNS OF TUMOR GROWTH RELATED FACTORS IN SALIVARY GLAND TUMORS (타액선 종양에서 종양증식 관련인자 발현에 관한 면역조직화학적 연구)

  • Kim, Han-Seok;Kim, Soung-Min;Park, Young-Wook
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.29 no.5
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    • pp.405-416
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    • 2007
  • Objective : Lots of papers have revealed that tumor growth related factors such as EGF, EGFR, c-erbB-2 play an important role in tumorigenesis and proliferation. These factors are found in most tumors of ectodermal origin. But, documentations of tumor growth related factors on salivary gland tumors were rare. Therefore, we determined expressions of tumor growth related factors; PCNA, p53, EGF, EGFR, cerbB2(HER-2), Maspin, DMBT-1, N-Ras in representative salivary gland tumors. Materials and methods : A few types of salivary tumors were examined by immunohistochemical assays. Each antibody was applied to specimens of tumors. Specimens were composed of 5 pleomorphic adenomas (PA), 3 mucoepidermoid carcinomas (MEC), 2 adenoid cystic carcinomas (ACC) and 2 squamous cell carcinomas (SCC) from 12 patients. One specimen was selected randomly as negative control. For evaluation of staining intensity, each stained sample was divided into 5 grade; no staining, obscure, weak staining, moderate staining, strong staining. Results : Strong expressions of PCNA were found in all tumors except of PA. EGF was expressed strongly in SCC, ACC sequently. But in both PA and MEC, EGF expression was weak. EGFR and c-erbB-2 expression showed similar patterns in all salivary gland tumor tissues. P53 showed weak expression generally in all salivary gland tumors. DMBT-1 was expressed in SCC rather than in ACC or in MEC. N-Ras showed weak expressions in all salivary gland tumors except of squamous cell carcinoma. Conclusion : Taken together, tumor growth related factors were expressed in salivay tumors as well as mucosal squamous cell carcinoma. Especially EGFR and c-erbB-2 could be candidates as diagnostic markers for estimating clinical grade of salivary gland tumors. But further studies with reliable methods will be needed to confirm the results of this study.

Correlation Between p53 and p21 Proteins Expression and Prognostic Factors Related with Colon Cancer

  • Kim, Tai-Jeon;Kim, Tae-Geun
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.2
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    • pp.128-135
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    • 2007
  • This study was designed to investigate the correlation between the expression rate of p53 and p21 proteins by immunohistochemical staining and tumor prognostic factors including the tumor size, histological differentiation and Dukes' stage of tumor prognostic factors in colon cancer, and to acquire necessary data for the presumption of diagnosis, treatment and prognosis of colon cancer patients. From January 2000 to January 2003 at Hanyang University Guri Hospital, the paraffin blocks of 35 patients diagnosed with colon cancer whose pathologic reports were possible to review were selected. Harris hematoxylin & eosin (H&E) staining and immunohistochemical staining by ABC (Avidin Biotin Conjugate) method were performed. The histological differentiation grade and stage were classified according to the classification of the World Health Organization (WHO) and modified Dukes's stage from H&E staining. The expression rate of p53 and p21 proteins were analyzed by immunohistochemical staining. The results was analyzed statistically by SPSS (Windows version 8.0). As a result, the expression rate of p53 protein was 11.4% (4 cases) in clear differentiation, 48.6% (17 cases) in moderate differentiation, and 17.1% (6 cases) in poor differentiation. In other words, the poorer the differentiation, the higher the expression rate of p53 protein (p<0.05). The expression rate of p21 was 17.1% (6 cases) in clear differentiation, 40.0%(14 cases) in moderate differentiation, and 8.6% (3 cases) in poor differentiation, According to the progression of histological malignant degeneration, the expression rate of p21 protein decreased distinctively (p<0.05). However, the correlation between the two above mentioned proteins and the tumor-size and Dukes' stage was not of statistical significance. In the comparison of the expression rate of p53 protein with that of p21 protein, in 10 cases, p53 protein expression was positive while p21 protein expression was negative, and in 6 cases, p53 protein expression was negative whereas p21 protein expression was positive. Consequently a statistically significant inverse correlation between the expression rate of p53 protein and that of p21 protein was observed (p<0.05). In conclusion, we found a significant correlation between histological differentiation and the expression rate of p53 and p21 proteins (p<0.05), and a significant inverse correlation between the expression rate of p53 protein and that of p21 protein (p<0.05). Also, it could be confirmed that the over expression of p53 and p21 proteins is closely associated with the occurrence of colon cancer and its progress. Therefore, it is thought that this study may be greatly beneficial to the presumption of diagnosis, treatment and prognosis of colon cancer patients.

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Availability of the lacZ gene as a Reporter Gene for Production of Transgenic Artemia franciscana (형질전환 아르테미아(Artemia franciscana) 생산을 위한 리포터 유전자로서 lacZ 유전자의 유용성 검토)

  • Jung, Hyo Sun;Kim, Dong Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.46 no.6
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    • pp.901-906
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    • 2013
  • We examined the availability of the lacZ gene (${\beta}$-galactosidase gene) as a reporter of foreign gene transfer in the cysts of Artemia franciscana (A. franciscana) to conduct a risk assessment of living genetically modified organisms (LMOs) in the marine ecosystem. The LacZ gene was transferred to decapsulated cysts by particle bombardment, and its insertion and expression were assessed by means of polymerase chain reaction (PCR) and X-gal staining. X-gal staining indicated lacZ expression in all A. franciscana examined (including the control group), which exhibited not only negative but also positive PCR amplification. Endogenous ${\beta}$-galactosidase is highly active in the whole body of A. franciscana during all stages of the life cycle. Thus, the lacZ gene is unsuitable as a reporter for foreign gene transfer in A. franciscana cysts, because it is difficult to discriminate between exogenous and endogenous ${\beta}$-galactosidase activity.

c-KIT Positive Schistosomal Urinary Bladder Carcinomas are Frequent but Lack KIT Gene Mutations

  • Shams, Tahany M.;Metawea, Mokhtar;Salim, Elsayed I.
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.15-20
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    • 2013
  • Urinary bladder squamous cell carcinoma (SCC), one of the most common neoplasms in Egypt, is attributed to chronic urinary infection with Schistosoma haematobium (Schistosomiasis). The proto-oncogene c-KIT, encoding a tyrosine kinase receptor and implicated in the development of a number of human malignancies, has not been studied so far in schistosomal urinary bladder SCCs. We therefore determined immunohistochemical (IHC) expression of c-KIT in paraffin sections from 120 radical cystectomies of SCCs originally obtained from the Pathology Department of Suez Canal University (Ismailia, Egypt). Each slide was evaluated for staining intensity where the staining extent of >10% of cells was considered positive. c-KIT overexpression was detected in 78.3% (94/120) of the patients, the staining extents in the tumor cells were 11-50% and >50% in 40 (42.6%) and 54 (57.4%) respectively. The positive cases had 14.9%, 63.8%, 21.3% as weak, moderate and strong intensity respectively. Patients with positive bilharzial ova had significantly higher c-KIT expression than patients without (95.2% vs. 38.9%, P=0.000). Mutation analysis of exons 9-13 was negative in thirty KIT positive cases. The high rate of positivity in SBSCC was one of the striking findings; However, CD117 may be a potential target for site specific immunotherapy to improve the outcome of this tumor.

Staining patterns of squamous cell carcinomas of the larynx by monoclonal anti-cytokeratin antibodies (Monoclonal anticytokeratin antibodies 에 의한 후두편평세포암의 발현양상)

  • 도남용;전세영;이성재;최봉남
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1993.05a
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    • pp.88-88
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    • 1993
  • Immunohistochemical staining for keratin proteins may be useful as a diagnostic parameter in head and neck neoplasm. Our study evaluates the keratin antibody staining properties of normal tissues and squamous cell carcinoma of the vocal folds from surgical procedures performed on 27 cases. In normal epithelia, low molecular weight cytokeratins were strongly positive in basal layer but apparently reduced in suprabasal layers and completely negative in superficial layer. In invasive squamous cell carcinomas, low molecular weight anti-ck Ab were positive in all carcinoma cells of poorly differentiated carcinomas. On the other hand, high molecular weight anti-ck Ab were positive in almost carcinoma cells of well differentiated carcinomas.

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