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Genetic alterations in Wnt family of genes and their putative association with head and neck squamous cell carcinoma

  • Aditya, Jain;Smiline Girija, A.S.;Paramasivam, A.;Priyadharsini, J. Vijayashree
    • Genomics & Informatics
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    • v.19 no.1
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    • pp.5.1-5.11
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    • 2021
  • Head and neck squamous cell carcinoma (HNSCC) is the most frequent type of head and neck cancer that usually arises from the mucosal surfaces of several organs including nasal cavity, paranasal sinuses, oral cavity, tongue, pharynx, and larynx. The Wnt signaling pathway is a crucial mechanism for cellular maintenance and development. It regulates cell cycle progression, apoptosis, proliferation, migration, and differentiation. Dysregulation of this pathway correlates with oncogenesis in various tissues including breast, colon, pancreatic as well as head and neck cancers. The present study aims to assess the gene alterations in the Wnt family of genes so as to derive an association with HNSCC. Computational approaches have been utilized for the identification of gene alterations in the Wnt family of genes. Several databases such as cBioportal, STRING, and UALCAN were used for the purpose. The frequency of alteration was high in case of Wnt family member 11 (5%). Gene amplification, deep deletions, missense and truncating mutations were observed in HNSCC patients. There was a marked difference in the gene expression profile of WNT11 between grades as well as normal samples. The survival probability measured using the Kaplan-Meier curve also presented with a significant difference among male and female subjects experiencing a low/medium level expression. The female patients showed less survival probability when compared to the male subjects. This provides the prognostic significance of the WNT11 gene in HNSCC. Taken together, the present study provides clues on the possible association of WNT11 gene alterations with HNSCC, which has to be further validated using experimental approaches.

Biocompatibility and Bioactivity of Four Different Root Canal Sealers in Osteoblastic Cell Line MC3T3-El

  • Jun, Nu-Ri;Lee, Sun-Kyung;Lee, Sang-Im
    • Journal of dental hygiene science
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    • v.21 no.4
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    • pp.243-250
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    • 2021
  • Background: Endodontic sealers or their toxic components may become inflamed and lead to delayed wound healing when in direct contact with periapical tissues over an extended period. Moreover, an overfilled sealer can directly interact with adjacent tissues and may cause immediate necrosis or further resorption. Therefore, the treatment outcome conceivably depends on the endodontic sealer's biocompatibility and osteogenic potential. This study aimed to evaluate the cell viability and osteogenic effects of four different sealers in osteoblastic cells. Methods: AH Plus (resin-based sealer), Pulp Canal Sealer EWT (zinc oxide-eugenol sealer), BioRoot RCS (calcium silicate-based sealer), and Well-Root ST (MTA-based calcium silicate sealer) were mixed strictly according to the manufacturer's instructions, and dilutions of sealer extracts (1/2, 1/5 and 1/10) were determined. Cell viability was measured using the water-soluble tetrazolium-8 (WST-8) assay. Differentiation was assessed by alkaline phosphatase (ALP) activity and mineralized nodule formation by Alizarin Red S staining. Results: The cell viability of the extracts derived from the sealers excluding Well-Root ST was concentration dependent, with sealer extracts having the least viability at a 1/2 dilution. At sealer extract dilution of 1/10, the test groups showed the same survival rate as that control group, with the exception of BioRoot RCS. Among all experimental groups, BioRoot RCS showed the highest cell viability after 48 hours. The ALP activity was significantly higher in a concentration-dependent manner. Furthemore, all four materials promoted ALP activity and mineralized nodule formation compared to the control at 1/10 dilutions. Conclusion: This is the first study to highlight the differences in biological activity of these four materials. These results suggest that the composition of root canal sealers appears to alter the form of biocompatibility and osteoblastic differentiation.

Circulating Tumor Cell Detection in Lung Cancer Animal Model

  • Chong, Yooyoung;Jung, Yong Chae;Hwang, Euidoo;Cho, Hyun Jin;Kang, Min-Woong;Na, Myung Hoon
    • Journal of Chest Surgery
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    • v.54 no.6
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    • pp.460-465
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    • 2021
  • Background: Metastasis and recurrence of primary cancer are the main causes of cancer mortality. Disseminated tumor cells refer to cancer cells that cause metastasis from primary cancer to other organs. Several recent studies have suggested that circulating tumor cells (CTCs) are associated with the clinical stage, cancer recurrence, cancer metastasis, and prognosis. There are several methods of isolating CTCs from whole blood; in particular, using a membrane filtration system is advantageous due to its cost-effectiveness and availability in clinical settings. In this study, an animal model of lung cancer was established in nude mice using the human large cell lung cancer cell line H460. Methods: Six-week-old nude mice were used. The H460 lung cancer cell line was injected subcutaneously into the nude mice. Blood samples were obtained from the orbital area before cell line injection, 2 weeks after injection, and 2 weeks after tumor excision. Blood samples were filtered using a polycarbonate 12-well Transwell membrane (Corning Inc., Corning, NY, USA). An indirect immunofluorescence assay was performed with the epithelial cell adhesion molecule antibody. The number of stained cells was counted using fluorescence microscopy. Results: The average size of the tumor masses was 35.83 mm. The stained cells were counted before inoculation, 2 weeks after inoculation, and 2 weeks after tumor excision. Cancer cells generally increased after inoculation and decreased after tumor resection. Conclusion: The CTC detection method using the commercial polycarbonate 12-well Transwell (Corning Inc.) membrane is advantageous in terms of cost-effectiveness and convenience.

The influence of p53 mutation status on the anti-cancer effect of cisplatin in oral squamous cell carcinoma cell lines

  • Jo, Deuk-Won;Kim, Young-Kyun;Yun, Pil-Young
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.42 no.6
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    • pp.337-344
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    • 2016
  • Objectives: The purpose of this study was to evaluate the anti-cancer activity of cisplatin by studying its effects on cell viability and identifying the mechanisms underlying the induction of cell cycle arrest and apoptosis on oral squamous cell carcinoma (OSCC) cell lines with varying p53 mutation status. Materials and Methods: Three OSCC cell lines, YD-8 (p53 point mutation), YD-9 (p53 wild type), and YD-38 (p53 deletion) were used. To determine the cytotoxic effect of cisplatin, MTS assay was performed. The cell cycle alteration and apoptosis were analyzed using flow cytometry. Western blot analysis was used to detect the expression of cell cycle alteration- or apoptosis-related proteins as well as p53. Results: Cisplatin showed a time- and dose-dependent anti-proliferative effect in all cell lines. Cisplatin induced G2/M cell accumulation in the three cell lines after treatment with 0.5 and $1.0{\mu}g/mL$ of cisplatin for 48 hours. The proportion of annexin V-FITC-stained cells increased following treatment with cisplatin. The apoptotic proportion was lower in the YD-38 cell line than in the YD-9 or YD-8 cell lines. Also, immunoblotting analysis indicated that p53 and p21 were detected only in YD-8 and YD-9 cell lines after cisplatin treatment. Conclusion: In this study, cisplatin showed anti-cancer effects via G2/M phase arrest and apoptosis, with some difference among OSCC cell lines. The mutation status of p53 might have influenced the difference observed among cell lines. Further studies on p53 mutation status are needed to understand the biological behavior and characteristics of OSCCs and to establish appropriate treatment.

Comparative Study on the Salivary Gland between Two Species (Achatina fulica and Incilaria fruhstorferi) of the Snails in Stylommatophora ( Mollusca, Gastropoda ) (병안목 달팽이류 두 종간 (Achatina fulica and Incilaria fruhstorferi)의 타액선에 관한 비교 연구)

  • 한종민;장남섭
    • The Korean Journal of Malacology
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    • v.12 no.2
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    • pp.109-121
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    • 1996
  • Histochemical experiment was carry out respectively to confirm the properties of the salis (Achatina fulica and Incilaria fruhstorferi). SDS-PAGE was carried out to compare and invertigate the distribution aspects of protein patterns between the two species. Five types(A, B, F, H and I)of gland cells with four neutral mucopolysaccharide cells and one acid mucopolysaccharide cells and one acid mucopolysaccharide cell were observed in acinous of Achatina fulica, while six types were observed in acinous of Incilaria fruhstorferi: ond acid mucopolysaccharide cell(type-A) and four neutral mucopolysaccharide cells(type-B, C, D and F) and one cell that acid mucopolysaccharide is only mimbrane that surrounded granule(type-E). The results are follows:The thpe-A fland cell is commonly observed between the two species. The type-A gland cell in Achatina fulica possesses a nucleus with a developed heterdchromatin, and the cytoplasm was filled with round granules. The granules were surrounded with an uncertain boundary mimbrane and confirmed with neutral mucopolysaccharides, but is confirmed acid mucopolysaccharide in Incilaria fruhstorferi.The type-B gland cell is obwerved in the two species, too. The type-B gland cell in Achatina fulica was round shaped, and included an evenly alrge nucleus. The uncleoplasm included granules that were confirmed in the neutral mucopolysaccharides of the two species. The type-C and D gland cells exist only in Incilaria fruhstorferi, nucleoplasm was well developed heterochromatins. The type-E gland cell appears in the acinous surrounded the salivary gland of Incilaria fruhstorferi. Thdse granules appear irregular irregular shape and size and the cytoplasm is formed in alveolar. The type-F gland cells are commonly observed in the salivary glands of the two species. They are similar with the type-B gland cell, but the granular shape is comparatively small and irregular, and possess the neutral mucos granules. The type-H gland cells are mainly seen in only Achatina, and in nucleus is a well developed heterochromatin. The cytoplasm is filled with round small granules with acid mucopolysaccharide for alcianophilia observed. The type-I cell was small cell with an irregular shape and only observed in the gland cells of Achatina fulica. The heterochromatins were developed in the nucleus and the granules are not observed in cytoplasm.Secretory ducts of saliva are composed of the interlobular duct and interlobar secretory duct. In Achatina fulica the interlobular duct consists of a simple cuboidal epithelium, while the endothelium of intralobar secretory duct of Incilaria fruhstorferi consists of a simple squamous epithelium and in the cytoplasm is filled with granules(type-G secretory cell). A SDS-PAGE was carried out to confirm that the protein band pattern consist of salivary gland. In conclusions, five more bands in Achatina fulica and three bands in Incilaria fruhstorferi were confirmed in MW<29 kDa. one main band coincides comparatively with both and is between 29-45 kDa. There are four main bands in Achatina fulica and two main bands in Incilaria fruhstorferi between 45-66.5 kDa respectively. The bands in Achatina fulica seem more complex than in incilaria fruhstorferi.

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Immunohistochemical Expression of Nuclear Retinoid Receptor and CREB(cAMP Response Element Binding Protein) in Lung Cancers (폐암종에서 Nuclear Retinoid Receptor 및 CREB의 면역조직화학적 발현 양상)

  • Shin, Jong Wook;Gi, Seung-Seok;Paik, Kwang Hyun;Choi, Won;Park, In Won;Kim, Mi Kyung
    • Tuberculosis and Respiratory Diseases
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    • v.59 no.6
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    • pp.631-637
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    • 2005
  • Background : Transcriptional factors of the CREB(cAMP Response Element Binding Protein) are involved in the regulation of gene expression in response to a variety of signaling pathways. Proteins produced by the CREB genes play key roles in many physiological processes, including memory and long-term potentiation. The retinoic acid receptor (RAR) axis mediates epithelial cell differentiation and proliferation in many tissues including the lung. Material and method : The RAR and CREB expression levels were examined in 60 adenocarcinomas and 60 squamous cell carcinomas of the lung using immunohistochemical staining. Results : 1) RAR protein expression was found in 58.3%(35/60) of adenocarcinomas and 36.7%(22/60) of squamous cell carcinomas(P<0.05). 2) RAR protein expression was found in 80%(16/20) of well differentiated adenocarcinomas, 60%(12/20) of moderately differentiated adenocarcinomas, and 35%(7/20) of poorly differentiated adenocarcinomas (P<0.01). 3) RAR protein expression was found in 45%(9/20) of well differentiated squamous cell carcinomas, 35%(7/20) of moderately differentiated squamous cell carcinomas, and 30%(6/20) of poorly differentiated squamous cell carcinomas (P>0.05). 4) CREB expression was found in 61.7%(37/60) of adenocarcinomas and 40%(24/60) of squamous cell carcinomas( P<0.05). 5) CREB expression was found in 85%(17/20) of well differentiated adenocarcinomas, 60%(12/20) of moderately differentiated adenocarcinomas, and 40%(8/20) of poorly differentiated adenocarcinomas (P<0.01). 6) CREB expression was found in 45%(9/20) of well differentiated squamous cell carcinomas, 35%(7/20) of moderately differentiated squamous cell carcinomas, and 35%(8/20) of poorly differentiated squamous cell carcinomas(P>0.05). 7) RAR and CREB expression was found in 68.5% of lung cancers, and there was a significant correlation between them(P<0.05). Conclusion : RAR and CREB expression can be used to indirectly determine the malignant potentiality of a cell.

Investigation of Carcinogenic Potential of TCDD in the Human Breast Epithelial Cell line (사람의 유방상피세포에서 TCDD에 의한 발암성 연구)

  • 김정환;나혜경;서영준
    • Environmental Mutagens and Carcinogens
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    • v.22 no.4
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    • pp.312-318
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    • 2002
  • Dioxin represents a group of halogenated aromatic hydrocarbons of which 2,3,7,8-tetrachlorod-ibenzo-p-dioxin (TCDD) is well known for its extremely toxic properties as well as ubiquitous presence in our environment and ecosystems. In order to better assess the carcinogenic mechanism of dioxin, we should utilize the reliable biomarkers that can precisely and correctly reflect multi-stage carcinogenesis. When MCF10A cells were exposed to TCDD (10 nM), expression of both CYP1A1 and CYP1B1 was induced in a time-related manner. The expression as well as activity of ornithine decarboxylase was transiently induced by TCDD treatment. In contrast, the induction of COX-2 that is implicated in carcinogenesis as well as inflammation, was not induced by TCDD. In another study, gap-junctional intercellular communication (GJIC) was attenuated by TCDD treatment as revealed by the dye-transfer assay. Based on these findings, TCDD has both tumor initiating and promoting potential in human breast epithelial cells in culture. Also, treatment of MCF10A cells with the carcinogen 7,12-dimethylbenz[a]anthracene plus TCDD resulted in malignant cell transformation as revealed by increased anchorage-independent growth of exposed cells. Additional studies may be necessary to assess the effects of TCDD on multi-stage carcinogenesis in vivo.

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Priming Effect of Endotoxin in Human Alveolar Macrophage (사람 폐포대식세포에서 내독소의 Priming 효과)

  • Chung, Man-Pyo;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.1
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    • pp.46-53
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    • 1996
  • Background: Endotoxin or lipopolysaccharide(LPS) can prime phagocytic cells such as polymorphonuclear leukocytes, monocytes or animal peritoneal macrophages to generate increased amounts of secretory products such as oxygen free radicals and tumor necrosis factor, which play an important role in developing adult respiratory distress syndrome in gram negative sepsis. Human alveolar macrophages(HAM) are continuously exposed to various stimuli inhaled into the alveoli, and the response to LPS might be different in HAM. Therefore, we investigated the effect of LPS pre-exposure on HAM adhered to plastic surface and A549 cell(type II human alveolar epithelial cell line) monolayer. Methods: HAM were isolated from bronchoalveolar lavage fluid from normal lung of the patients with localized lung cancer and esophageal cancer. LPS was exposed to HAM for 2hrs before or after adherence to plastic surface of 24-well Linbro plate and A549 cell monolayer. And then HAM was stimulated with PMA(phorbol myristate acetate) or fMLP(N-formyl-methionylleucyl-phenylalanine). The amount of hydrogen peroxide($H_2O_2$) production in the supernatant was measured on the principle of peroxidase-dependent oxidation of phenol red by hydrogen peroxide. Results: LPS pre-exposure could not enhance $H_2O_2$ production in neither HAM adhered to plastic surface nor one to A549 cell monolayer. But LPS even in the absence of PMA or fMLP stimulation directly increased $H_2O_2$ release in HAM if added after the adherence to A549 cell monolayer. Conclusion: Endotoxin does not prime HAM, but may directly activate HAM adhered to alveolar epithelial cells. Further investagation will be necessary.

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Performance Analysis of the UPC/NPC Algorithm for Guaranteed QoS in ATM Networks

  • Kim, Yong-Jin;Kim, Jang-Kyung;Lee, Young-Hee;Park, Chee-Hang
    • ETRI Journal
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    • v.20 no.3
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    • pp.251-271
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    • 1998
  • It is well known that if usage parameter control/network parameter control (UPC/NPC) functions are used together with a cell loss priority control scheme in ATM networks, the measurement phasing problem can occur. This makes it difficult for a network provider to define and commit the cell loss ratio as a QoS parameter. To solve the problem, we propose a new UPC/NPC algorithm. By using the proposed UPC/NPC algorithm, we can define the cell loss ratios for CLP = 0 and CLP = 0+1 cell streams without the measurement phasing problem under any conditions. We analyzed the performance of the proposed UPC/NPC algorithm. Using a discrete time model for the UPC/NPC architecture with a discrete-time semi-Markov process (DSMP) input model, we obtained the cell discarding probabilities of CLP = 0 and CLP = 0+1 cells streams and showed that more CLP = 0 cells are accepted compared to what was proposed in ITU-T.

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The Cytologic Analysis of Microinvasive Squamous Cell Carcinoma of the Uterine Cervix on Cervical Smear (자궁경부 세포도말 검사에시 미세침윤성 편평세포암종의 세포학적 분석)

  • Choi, Hyun-Joo;Park, In-Ae
    • The Korean Journal of Cytopathology
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    • v.12 no.1
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    • pp.31-37
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    • 2001
  • Whlie cytologic characteristics of squamous dysplasia, carcinoma in situ, and invasive squamous cell carcinoma of the uterine cervix are well documented, relatively few studios have dealt with the cellular features of microinvasive carcinoma. In order to describe the cellular characteristics of microinvasive squamous cell carcinoma, we retrospectively reviewed 45 cervovaginal smears(15 carcinoma in situ, 15 microinvasive cancer, 15 invasive cancer) which were confirmed by histologic examination of specimens obtained by hysterectomy at the Seoul National University Hospital during S years from 1995 to 1999. The cytologic features about tumor diathesis, inflammatory background, ceil arrangement, anisonucleosis, nuclear membrane irregularity, nuclear chromatin pattern, and nucleoli were observed. The cytologlc characteristics of microinvasive squamous cell carcinoma of the uterine cervix are syncytial pattern, mild tumor diathesis, the irregularity of nuclear membrane, irregularly distributed nuclear chromatin, and occurrence of micronucleoli. But, correlation between the depth of Invasion and the cytologic feature had limited value.

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