• Title/Summary/Keyword: Paraffin-embedded tissue

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High Resolution Melting Analysis for Epidermal Growth Factor Receptor Mutations in Formalin-fixed Paraffin-embedded Tissue and Plasma Free DNA from Non-small Cell Lung Cancer Patients

  • Jing, Chang-Wen;Wang, Zhuo;Cao, Hai-Xia;Ma, Rong;Wu, Jian-Zhong
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6619-6623
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    • 2013
  • Background:The aim of the research was to explore a cost effective, fast, easy to perform, and sensitive method for epidermal growth factor receptor (EGFR) mutation testing. Methods: High resolution melting analysis (HRM) was introduced to evaluate the efficacy of the analysis for dectecting EGFR mutations in exons 18 to 21 using formalin-fixed paraffin-embedded (FFPE) tissues and plasma free DNA from 120 patients. Results: The total EGFR mutation rate was 37.5% (45/120) detected by direct sequencing. There were 48 mutations in 120 FFPE tissues assessed by HRM. For plasma free DNA, the EGFR mutation rate was 25.8% (31/120). The sensitivity of HRM assays in FFPE samples was 100% by HRM. There was a low false-positive mutation rate but a high false-negative rate in plasma free DNA detected by HRM. Conclusions: Our results show that HRM analysis has the advantage of small tumor sample need. HRM applied with plasma free DNA showed a high false-negative rate but a low false-positive rate. Further research into appropriate methods and analysis needs to be performed before HRM for plasma free DNA could be accepted as an option in diagnostic or screening settings.

Application of Terahertz Spectroscopy and Imaging in the Diagnosis of Prostate Cancer

  • Zhang, Ping;Zhong, Shuncong;Zhang, Junxi;Ding, Jian;Liu, Zhenxiang;Huang, Yi;Zhou, Ning;Nsengiyumva, Walter;Zhang, Tianfu
    • Current Optics and Photonics
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    • v.4 no.1
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    • pp.31-43
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    • 2020
  • The feasibility of the application of terahertz electromagnetic waves in the diagnosis of prostate cancer was examined. Four samples of incomplete cancerous prostatic paraffin-embedded tissues were examined using terahertz spectral imaging (TPI) system and the results obtained by comparing the absorption coefficient and refractive index of prostate tumor, normal prostate tissue and smooth muscle from one of the paraffin tissue masses examined were reported. Three hundred and sixty cases of absorption coefficients from one of the paraffin tissues examined were used as raw data to classify these three tissues using the Principal Component Analysis (PCA) and Least Squares Support Vector Machine (LS-SVM). An excellent classification with an accuracy of 92.22% in the prediction set was achieved. Using the distribution information of THz reflection signal intensity from sample surface and absorption coefficient of the sample, an attempt was made to use the TPI system to identify the boundaries of the different tissues involved (prostate tumors, normal and smooth muscles). The location of three identified regions in the terahertz images (frequency domain slice absorption coefficient imaging, 1.2 THz) were compared with those obtained from the histopathologic examination. The tissue tumor region had a distinctively visible color and could well be distinguished from other tissue regions in terahertz images. Results indicate that a THz spectroscopy imaging system can be efficiently used in conjunction with the proposed advanced computer-based mathematical analysis method to identify tumor regions in the paraffin tissue mass of prostate cancer.

AN IMMUNOHISTOCHEMICAL LOCALIZATION OF TENASCIN IN PERIODONTAL POCKET TISSUES (치주낭 조직내 tenascin의 분포에 관한 면역조직화학적 연구)

  • Han, Kyung-Yoon;Lee, Kang-Jin
    • Journal of Periodontal and Implant Science
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    • v.24 no.3
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    • pp.607-617
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    • 1994
  • To determine the effect of tenascin on forming periodontal pocket and pseudopocket, the ginival tissues were surgically obtained from the patients with adult periodontitis(10) and non-inflammatory phenytoin-associated gingival hyperplasia(5). The excised tissue specimens were fixed in neutral formalin for $6{\sim}24$ hours, embedded with paraffin, sectioned at 4-6m in thickness, mounted on glass slides coated with 3-aminopropyltriethoxysilane(Sigma Chemical Co., St. Louis, MO, U.SA.) and immunohistochemically processed by Avidin-Biotin peroxidase complex method for the localization of tenascin, using monoclonal mouse anti-human tenascin antiboday(Chemicon-International Inc., Temecula, CA, U.S.A., 1: 5,000) as the primary antibody. Regardless of periodontal pocket and pseudopocket, tenascin was localized along the connective tissue subjacent to basement membrane of gingival epithelium, and strong positive reactivity was obviously noted in the papillary projections of gingival connective tissue. The results suggest that tenascin may affect the development of papillary projections and the proliferation of epithelial cells.

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Prenatal diagnosis of the spinal muscular atrophy type I using genetic information from archival slides and paraffin-embedded tissues

  • Choi, Soo-Kyung;Cho, Eun-Hee;Kim, Jin-Woo;Park, So-Yeon;Kim, Young-Mi;Ryu, Hyun-Mee;Kang, Inn-Soo;Jun, Jung-Young;Chi, Je-G.
    • Journal of Genetic Medicine
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    • v.2 no.2
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    • pp.53-57
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    • 1998
  • Spinal muscular atrophy (SMA) type I is a common severe autosomal recessive inherited neuromuscular disorder that has been mapped to chromosome 5q11.2-13.3. The survival motor neuron (SMN) gene, a candidate gene, is known to be deleted in 96% of patients with SMA type I. Presently, PCR and single strand conformation polymorphism (PCR-SSCP) analyses have been made possible for application to both archival slides and paraffin-embedded tissues. Archival materials represent valuable DNA resources for genetic diagnosis. We applied these methods for the identification of SMN gene of SMA type I in archival specimens for the prenatal diagnosis. In this study, we performed the prenatal diagnosis with chorionic villus sampling (CVS) cells on two women who had experienced neonatal death of SMA type I. DNA extraction was done from archival slide and tissue materials and PEP-PCR was performed using CVS cells. In order to identify common deletion region of SMN and neuronal apoptosis-inhibitory protein (NAIP) genes, cold PCR-SSCP and PCR-restriction site assay were carried out. Case 1 had deletions of the exons 7 and 8, and case 2 had exon 7 only on the telomeric SMN gene. Both cases were found to be normal on NAIP gene. These results were the same for both CVS and archival biopsied specimens. In both cases, the fetuses were, therefore, predicted to be at very high risk of being affected and the pregnancy were terminated. These data clearly demonstrate that archival slide and paraffin-embedded tissues can be a valuable source of DNA when the prenatal genetic diagnosis is needed in case any source for genetic analysis is not readily available due to previous death of the fetus or neonate.

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AN EXPERIMENTAL STUDY ON THE CHANGES OF RAT MOLAR PERIODONTIUM INCIDENT TO INTERMITTENT FORCE (간헐적 교정력에 의한 백서 구치 치주조직의 변화에 관한 실험적 연구)

  • Kim, Sun-Hae;Suhr, Cheong-Hoon
    • The korean journal of orthodontics
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    • v.19 no.2
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    • pp.57-73
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    • 1989
  • The purpose of this study was to investigate the tissue response of the rat molar periodontium incident to intermittent orthodontic force. The author intended to observe the healing process of injured periodontium and the response of injured tissue to the resumed force. Oxytetracyclin 50mg/Kg was given to each rat intraperitonially. 5 days later, maxillary 1st molars were moved mesially from the incisors with closed coil spring of 100gram. 7 days later, the appliances were removed and 20mg/Kg of calcein were given intraperitonially to each rat. At the same time, maxillary left 1st molars of 15 rats were moved by the same method, but force was lowered to 20 gram. After 1 day, maxillary left 1st molars of another 15 rats were moved by the same method and 50mg/Kg of oxytetracycline was given intraperitonially. After 4 days, another 15 rats were treated as above. After 7 days, another 15 rats were treated as above. 1,4,7,10 and 14 days after change of force, 3 rats were sacrificed in each group respectively. 2 rats were decalcified, embedded in paraffin, and stained with hematoxylin-eosin stain and with Masson's trichrome stain. Another rat was embedded in polyester resin and undecalcified specimen were made. Microradiograms were taken with the undecalcified sections. Observations were made with light and fluorescence microscope. Following conclusions were made. 1. Connective tissue cells and vessels were infiltrated into the hyalinized tissue from the bony cleft and along the border of the hyalinized tissue with bone and root surface. At the same time, elimination of hyalinized tissue, bone and root resorption occurred. 2. Bone and root were resorbed directly and indirectly. 3. Hyalinized tissue was removed within 5 days after force removal. 4. Hyalinized zone was less extensive and easily removed as the rest period prolonged. 5. Hyalinized tissue developed more rapidly and extensively and lasted over 10 days as the force resumed on the already formed hyalinized tissue.

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Studies on the rabbit viral hepatitis : Immunohistochemical observations (토끼의 바이러스성 간염에 관한 연구 : 면역조직화학적 관찰)

  • Lee, Cha-soo;Shin, Tae-kyun;Choi, Youn-ju;Jeong, Kyu-sik;Jyeong, Jong-sik
    • Korean Journal of Veterinary Research
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    • v.33 no.2
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    • pp.269-275
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    • 1993
  • Tissue distribution of RHDV in rabbits were examined by immunofluorescence and ABC methods. Tissues including liver, spleen, kidneys, lungs and brain were frozen, cut in a crycut, and fixed in 10% buffered formalin, embedded in paraplast, and cut $5{\sim}7{\mu}m$ thickness. Sections were immunostained Tissue distribution of RHDV in rabbits were examined by immunofluorescence and ABC methods. Tissues including liver, spleen, kidneys, lungs and brain were frozen, cut in a crycut, and fixed in 10% buffered formalin, embedded in paraplast, and cut $5{\sim}7{\mu}m$ thickness. Sections were immunostained with primary antiserum and conjugated second antibodies as recommended by manufacturer. None of the cultures tested showed virus-induced phenomena. Immunoreactive products were commonly found in the liver, in some cases there were also positive staining in the spleen and kidneys. Other organs showed weak or insignificant immunoreactions. By ABC method on the formalin-fixed, paraffin-embedded liver tissues, strong immunoreactivity was found in the periportal triad lesions and peripheral lesions of the hepatic lobules. Immunoreactive products showed diffuse fine granular in the cytoplasm of hepatocytes and sinusoidal cells. In some cells, immunoproducts marginate at the periphery of the cells. The intensive staining of the cytoplasm of infected cells allowed their exact differentiation from surrounding uninfected cells. The positive area involved coincided with histopathological lesion on serial liver sections. In conclusion, liver was proved to be a consistent target organ in RHD, and the immunoperoxidase method in the section of formalin-fixed, paraffin-embedded hepatic tissue could be broadly used for the routine diagnosis of the disease.

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Human Papilloma Virus Detection by INNOLiPA HPV in Prostate Tissue from Men of Northeast Mexico

  • Rodriguez, Martha I Davila;Morales, Cesar V Ignacio;Tovar, Anel R Aragon;Jimenez, Delia Olache;Maldonado, Edmundo Castelan;Miranda, Sandra Lara;Gutierrez, Elva I Cortes
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.11
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    • pp.4863-4865
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    • 2016
  • Background: Prostatic adenocarcinoma by Prosate cancer (PCa) is the most prevalent cancer and the second cause of cancer-related death among men in the Western world. Human papilloma virus (HPV) may be considered as a preventable risk factor. In this study, we assessed the frequencies of HPV infection in prostatic adenocarcinoma and benign prostatic hyperplasia (BPH) cases in Northeast Mexico. Materials and Methods: A total of 87 paraffin-embedded blocks (from 25 and 62 patients with definite diagnoses of BPH and adenocarcinoma, respectively) were selected and subjected to INNOLiPA HPV Genotyping to detect 28 high- and low-risk HPV types. The rates of infection were compared in the two studied groups. Results: INNOLiPA HPV demonstrated great sensitivity for HPV detection on paraffin-embedded tissue. Global prevalence was 14.9% (13/87). HPV infection was positive in 19.4% (12/62) of patients with adenocarcinoma and 4.0% (1/25) of patients with BPH. HPV-11, which is considered to be low risk, was more prevalent. Interestingly, one patient with BPH and six with prostate cancer showed examples considered to be high risk (HPV-18, -51, -52, and -66). Conclusion: A higher rate of HPV infection among Mexican patients with prostatic carcinoma than among those with BPH was observed. HPV infections may thus contribute to the risk of prostate cancer. Further studies are required to elucidate any roles of HPV infection in prostate disease in Mexico and the effect of prevention and treatment of HPV infection on prostatic adenocarcinoma.

Ultrastructural Observations of Some Oral Mucosal Lesions II. Oral Leukoplakia (수종 구강점막질환에 관한 전자현미경적 연구 II. 구강백반증)

  • 정성창
    • Journal of Oral Medicine and Pain
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    • v.8 no.1
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    • pp.33-41
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    • 1983
  • The authors observed the ultrastructure of oral leukoplakia simplex of gingiva, buccal mucosa, tongue and alveolar ridge. For the purpose of clearly defining the lesions under investigation in this study, leukoplakias were cinsidered to be any white patches on the oral mucous membranes that could not be removed by rubbing and could not be classified clinically or microscopically as another diagnosable disease. The tissue to be examined were embedded in paraffin for light microscopic study. The tissue to be examined under the electronomicroscope were fixed in 2.5% glutaraldehyde in 0.1M cacodylate buffer and 1% osmic acid in 0.1M cacodylate buffer, dehydrated with guaded alchol, and treated with propylene oxide, and embedded in Epon.Ultrathin sections were obtained by LKB III ultrotome, stained with uranyl acetate/lead citrate, and examined with Corinth 500EM. The results were as follows : 1. Epithelium of leukoplakia consisted of stratum basale, stratum spinosum, stratum granulosum and stratum corneum. 2. There was hyperorthokerotosis or hyperparakeratosis. 3. Granular cells contained a lot number of membrane coating granule showing lamellar structure, clearing ot codensation, and a lot of keratohyaline granule varied in size. 4. An increased concentration of tonofilaments and an increased number of desmosomes were found in the stratum spinosum. 5. Basal lamina generally showed its continuity, but in some locatoins, its interreption and multiplication appeared.

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Immunohistochemical diagnosis of infectious pancreatic necrosis (어류 전염성훼장괴사증의 면역조직화학적 진단)

  • Kim, Soon-Bok
    • Korean Journal of Veterinary Pathology
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    • v.3 no.1
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    • pp.1-5
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    • 1999
  • This experiment was carried out to establish the immunohistochemical diagnostic method for infectious pancreatic necrosis in the monolayers of CHSE-214 cell cultures and paraffin-embedded tissue sections from rainbow trout infected with infectious pancreatic necrosis virus(IPNV). Specific identification of IPNV antigens was often demonstrated in the pancreatic exocrine cells, and less in the intestinal mucous epithelia and the renal hemopoietic tissues by the use of monoclonal antibodies against capsid protein VP2. The specific reaction was seen as a distinct red cytoplasmic color, often as small granules of various sizes. The result showed that streptavidin alkaline phosphatase immunohistochemisry specifically identified IPNV antigens in both infected cell cultures and tissue sections.

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The Relationship Between Expression of Matrix Metalloproteinases(MMPs)-2, 9 and Tissue Inhibitors of Metalloproteinase(TIMPs)-1, 2 and Survival Time in Resected Non-Small Cell Lung Cancer (비소세포폐암에서 Matrix Metalloproteinase(MMPs)-2, 9와 Tissue Inhibitor of Metalloproteinase(TIMPs)-1, 2의 발현과 생존율과의 관계)

  • Kim, Hak-Ryul;Yang, Sei-Hoon;Jeong, Eun-Taik
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.5
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    • pp.453-462
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    • 2002
  • Background : Matrix metalloproteinases(MMPs) are a large family of proteolytic enzymes, which are involved in the degradation of many different components of the extracellular matrix. There is increasing evidence indicating that individual MMPs have important roles in tumor invasion by inactivating the MMPs. In this study, the correlation between MMPs and TIMPs expression, and the clinical outcome was investigated. Materials and Methods : Immunohistochemical staining of MMP-2, 9 and TIMP-1,2 were performed on paraffin-embedded tumor sections from 74 resected primary non-small cell lung cancers. Results : In 74 patients, MMP-2, MMP-9, TIMP-1, and TIMP-2 immunoreactivity was demonstrated in 24(34%), 19(26%) and 32(43%) of the paraffin-embedded tumors, respectively. The median survival of the MMP-2 positive cases was significantly shorter than that of the negative cases(20 vs 34 months). The median survival of the TIMP-2 positive cases was also was significantly longer than that of the negative cases (34 vs 18 months). The MMP-2, and MMP-9 expression level had a positively correlation with a more advanced stage and lymph node metastasis. There was inverse correlation between TIMP-2 expression and tumor invasion. The median survival of the MMP-2 negative/TIMP-2 positive cases was higher than that of the other cases. Conclusion : These results suggest that tumor invasion and lymph node metastasis are closely related to MMP-2 and MMP-9 expression. There was an inverse correlation between TIMP-2 and MMP-9 expression, and tumor invasion.