• Title/Summary/Keyword: Protein stain

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Correlation of p53 Protein Overexpression, Gene Mutation with Prognosis in Resected Non-Small Cell Lung Cancer(NSCLC) Patients (비소세포폐암에서 p53유전자의 구조적 이상 및 단백질 발현이 예후에 미치는 영향)

  • Lee, Y.H.;Shin, D.H.;Kim, J.H.;Lim, H.Y.;Chung, K.Y.;Yang, W.I.;Kim, S.K.;Chang, J.;Roh, J.K.;Kim, S.K.;Lee, W.Y.;Kim, B.S.;Kim, B.S.
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.4
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    • pp.339-353
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    • 1994
  • Background : The p53 gene codes for a DNA-binding nuclear phosphoprotein that appears to inhibit the progression of cells from the G1 to the S phase of the cell cycle. Mutations of the p53 gene are common in a wide variety of human cancers, including lung cancer. In lung cancers, point mutations of the p53 gene have been found in all histological types including approximately 45% of resected NSCLC and even more frequently in SCLC specimens. Mutant forms of the p53 protein have transforming activity and interfere with the cell-cycle regulatory function of the wild-type protein. The majority of p53 gene mutations produce proteins with altered conformation and prolonged half life; these mutant proteins accumulate in the cell nucleus and can be detected by immunohistochemical staining. But protein overexpression has been reported in the absence of mutation. p53 protein overexpression or gene mutation is reported poor prognostic factor in breast cancer, but in lung cancer, its prognostic significance is controversial. Method : We investigated the p53 abnormalities by nucleotide sequencing, polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP), and immunohistochemical staining. We correlated these results with each other and survival in 75 patients with NSCLC resected with curative intent. Overexpression of the p53 protein was studied immunohistochemically in archival paraffin- embedded tumor samples using the D07(Novocastra, U.K.) antibody. Overexpression of p53 protein was defined by the nuclear staining of greater than 25% immunopositive cells in tumors. Detection of p53 gene mutation was done by PCR-SSCP and nucleotide sequencing from the exon 5-9 of p53 gene. Result: 1) Of the 75 patients, 36%(27/75) showed p53 overexpression by immunohistochemical stain. There was no survival difference between positive and negative p53 immunostaining(overall median survival of 26 months, disease free median survival of 13 months in both groups). 2) By PCR-SSCP, 27.6%(16/58) of the patients showed mobility shift. There was no significant difference in survival according to mobility shift(overall median survival of 27 in patients without mobility shift vs 20 months in patients with mobility shift, disease free median survival of 8 months vs 10 months respectively). 3) Nucleotide sequence was analysed from 29 patients, and 34.5%(10/29) had mutant p53 sequence. Patients with the presence of gene mutations showed tendency to shortened survival compared with the patients with no mutation(overall median survival of 22 vs 27 months, disease free median survival of 10 vs 20 months), but there was no statistical significance. 4) The sensitivity and specificity of immunostain based on PCR-SSCP was 67.0%, 74.0%, and that of the PCR-SSCP based on the nucleotide sequencing was 91.8%, 96.2% respectively. The concordance rate between the immunostain and PCR-SSCP was 62.5%, and the rate between the PCR-SSCP and nucleotide sequencing was 95.3%. Conclusion : In terms of detection of p53 gene mutation, PCR-SSCP was superior to immunostaining. p53 gene abnormalities either overexpression or mutation were not a significant prognostic factor in NSCLC patients resected with curative intent. However, patients with the mutated p53 gene showed the trends of early relapse.

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The Effect of Acori Graminei Rhizoma Pharmacopuncture at GV20 on Dementia in a Focal Cerebral Ischemia Mice Model

  • Jang, Yeo jin;Kwak, Min Kyung;Jeong, Sang Jun;Kim, Hye Hwa;Kim, Tae Gwang;Kim, Jae Hong
    • Journal of Acupuncture Research
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    • v.34 no.3
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    • pp.1-11
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    • 2017
  • Objectives : The purpose of this study was to examine the effects of Acori Graminie Rhizoma Pharmacopuncture (PA-AG) at GV20 on cerebral ischemia-induced dementia in Mice. Methods : Mice were divided into the five following groups: normal, control, acupuncture, PA-AG (17 mg/kg), and PA-AG (34 mg/kg). All groups, except the normal group, had cerebral ischemia induced by occlusion of middle cerebral artery. The control group was not treated. The acupuncture, PA-AG (17 mg/kg), and PA-GA (34 mg/kg) groups were treated every other day with a total of 6 treatments. The effect of treatment was observed by Bax, Bcl-2, Bax/Bcl-2 ratio, cytochrome c, cresyl violet, and choline acetyltransferase staining. Results : In the PA-AG (34 mg/kg) group, the intensity of Bax was decreased and the intensity of Bcl-2 was increased. The Bax/Bcl-2 ratio also decreased in the PA-AG (34 mg/kg) group. The intensity of cytochrome c protein stain was decreased in the PA-AG (17 mg/kg) group. The density of neurons stained by cresyl violet and choline acetyltransferase (ChAT) was increased in the AT, PA-AG (17 mg/kg), and PA-AG (34 mg/kg) groups when compared with that of the control group. Conclusion : PA-AG at GV20 was effective on cerebral ischemia-induced dementia in mice.

Erdheim-Chester Disease with Hepatitis, Glomerulonephritis, Aplastic Anemia and Lung Involvement

  • Park, Ji Won;Chung, Chae Uk;Shin, Ji Young;Jung, Sun Young;Yoo, Su Jin;Lee, Jeong Eun;Jung, Sung Soo;Kim, Ju Ock;Kim, Sun Young;Park, Hee Sun
    • Tuberculosis and Respiratory Diseases
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    • v.67 no.4
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    • pp.345-350
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    • 2009
  • Erdheim-Chester disease (ECD) is a proliferative non-Langerhans cell histiocytosis of multiple organs. This is a rare disease of unknown etiology with a high mortality. We present the case report of a 26-year-old man diagnosed with ECD. He was referred to our hospital with elevated levels of aminotransferases. Although the diagnosis was uncertain, the patient was lost to follow up at that time. One year later, the patient returned to the hospital with generalized edema. Although a specific bone lesion was not found, the patient was experiencing the following: glomerulonephritis, aplastic anemia, hepatitis, and lung involvement. A lung biopsy was performed: the immunohistochemical stain were positive for CD68 and negative for S-100 protein and CD1a. We diagnosed as the patient as havinf ECD. Approximately 50% of ECD cases present with extraskeletal involvement. ECD should be considered as part of the differential diagnosis when multiple organs are involved.

Two Cases of Pulmonary Eosinophilic Granuloma (폐 호산구성 육아종 2예)

  • Choi, Wan-Young;Yoo, Yong-Keol;Park, Dong-Il;Shin, Dong-Ho;Park, Sung-Soo;Lee, Jung-Hee;Jeon, Seok-Chul;Chung, Won-Sang;Park, Chan-Pil;Park, Moon-Hyang;Lee, Jung-Dal
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.2
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    • pp.186-193
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    • 1992
  • Eosinophilic granuloma (histiocytosis $\times$) is an uncommon granulomatous disorder affecting the mononuclear phagocytic cell system caused by unknown etiology. When confined to the lungs, it is also known as pulmonary eosinophilic granuloma or pulmonary histiocytosis $\times$. After the first case was described in 1951 by Farinacci et ai, more 250 cases were reported in the English literature. However only 4 cases has been introduced in our country. We describe two additional cases. In both cases, open lung biopsies are performed for definite diagnosis and examined by immunohistochemical stain for S-100 protein and electron microscopy for ultrastructural identification of Birbeck granule.

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Effect of Paternal DNA Damage on Paternal DNA Degradation and Early Embryonic Development in Mouse Embryo: Supporting Evidence by GammaH2AX Expression (마우스 수정란에 있어서 부계 DNA 손상이 부계 DNA 퇴화 및 초기 배발달에 미치는 영향)

  • Kim, Chang Jin;Lee, Kyung-Bon
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.3
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    • pp.197-204
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    • 2019
  • This study was investigated to test whether the zygote recognized the topoisomerase II beta (TOP2B) mediated DNA fragmentation in epididymal spermatozoa or the nuclease degradation in vas deferens spermatozoa by testing for the presence of gammaH2AX (γH2AX). The γH2AX is phosphorylation of histone protein H2AX on serine 139 occurs at sites flanking DNA double-stranded breaks (DSBs). The presence of γH2AX in the pronuclei of mouse zygotes which were injected with DNA broke epididymal spermatozoa was tested by immunohistochemistry at 5 and 9 h post fertilization, respectively. Paternal pronuclei that arose from epididymal spermatozoa treated with divalent cations did not stain for γH2AX at 5 h. On the other hand, in embryos injected with vas deferences spermatozoa that had been treated with divalent cations, γH2AX was only present in paternal pronuclei, and not the maternal pronuclei at 5 h. Interestingly, both pronuclei stained positively for γH2AX for all treatments and controls at 9 h after sperm injection. In conclusion, the embryos recognize DNA that is damaged by nuclease, but not by TOP2B because H2AX in phosphorylated in paternal pronuclei resulting from spermatozoa treated with fragmented DNA from vas deferens spermatozoa treated with divalent cations, but not from epididymal spermatozoa treated the same way.

Amelanotic Melanoma on Fingertip: A Case Report (수지첨부에 발생한 멜라닌결핍흑색종의 치험례)

  • Paik, Hye Won;Kim, Sang Wha;Byeon, Jun Hee
    • Archives of Plastic Surgery
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    • v.35 no.3
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    • pp.312-315
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    • 2008
  • Purpose: Amelanotic melanoma represents a melanoma with an absence or a small number of melanin pigments and comprises 2% of all melanomas. These melanomas are frequently misdiagnosed, probably because of its nonspecific clinical features and difficulty in diagnosis, resulting in delayed diagnosis and treatment. We report a patient with amelanotic melanoma, who underwent surgical treatment with sentinel lymph node biopsy using gamma probe. Methods: A 32-year-old female was presented with a slowly growing ill-defined, hypopigmented nonerythematous lesion with nail defect on right index finger tip. Preoperative punch biopsy was performed, showing an amelanotic melanoma. Sentinel lymph node biopsy was done using gamma probe(Crystal probe system, CRYSTAL PHOTONICS GmbH, Germany) and confirmed no evidence of regional lymph node metastases. The patient underwent amputation at the proximal interphalangeal joint. Results: Histopathologic findings showed superficial spreading melanoma. There were no melanin pigments in Hematoxylin & Eosin stain but positive immunohistochemical stainings for S-100 protein and Hmb45, which were consistent with amelanotic melanoma. Patient's postoperative course was uneventful without any complication and had no evidence of recurrence of tumor in 6 months follow-up period. Conclusion: Amelanotic melanoma is extremely rare subtype of malignant melanoma with histopathologic findings of atypical melanocytes without melanin pigments. Early detection is crucial since survival is strongly related to tumor thickness and tissue invasion at the time of diagnosis. Wide excision is the treatment of choice and other conjunctive therapy has not been successful.

Expression of PTEN, Tumor Suppressor Protein, in Aggressive Fibromastosis (공격성 섬유종증에서 PTEN 종양 억제 인자의 발현)

  • Kim, Jung-Ryul
    • The Journal of the Korean bone and joint tumor society
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    • v.9 no.1
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    • pp.77-83
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    • 2003
  • Aggressive fibromatosis is a rare soft tissue tumor with locally invasive and infiltrative characteristics. The mechanism of this invasive nature was not reported until now. Mutations or reduction of PTEN, tumor suppressor gene, in cancer tissues, have been found to be associated with invasiveness and metastatic properties of cancer cells. To know the pattern of expression of PTEN in aggressive fibromatosis, we analysed the expression of PTEN with immunohistochemical stain and immunoblotting. PTEN was homogeneously expressed in the normal musculoaponeurotic tissues, but absent or very faint in tissues of patients with aggressive fibromatosis as evidenced by western blot analysis and immunohistochemical examinations. Although the meaning of decreased PTEN expression in aggressive fibromatosis is not certain, it might be involved in the growth of the aggressive fibromatosis, and associated with phenotype of aggressive fibromatosis.

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Expression of Human Serum Albumin in Milk of Transgenic Mice Using Goat β-casein/Human Serum Albumin Fusion Gene

  • Wu, H.T.;Chou, C.K.;Huang, M.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.6
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    • pp.743-749
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    • 2004
  • The gene encoding human serum albumin (HSA) was cloned from human liver cDNA library by PCR. The HSA cDNA in size of 2,176 bp, including 1,830 bp of open reading frame, was cloned into the plasmid carried with the 5'flanking sequence of goat $\beta$-casein gene (-4,044 to +2,025 bp) to get a tissue specific expression vector in mammary gland named pGB562/HSA (12.5 kb). A 9.6 kb DNA fragment in which the sequence is in order of goat $\beta$-casein gene regulatory sequence, HSA cDNA and SV40 polyadenylation signals was isolated from the pGB562/HSA by SacI and DraIII cutting, and used to microinject into the pronuclei of mouse fertilized eggs to produce transgenic mice. Three transgenic mice (2 female and 1 male) were identified by PCR and dot Southern blot analysis. The copy numbers of integrated transgene were more than 10 copies in line #21 and #26 as well as over 50 copies in line #31 of transgenic mice. HSA protein collected from the milk of lactating transgenic mice was confirmed by immuno-detection of Western and slot blot. The concentrations of HSA in the milk were from 0.05 to 0.4 mg/ml. An obvious antigen and antibody conjugate could be observed in immunohistochemical stain of mammary gland tissue from lactating day 11 of HSA transgenic mice. The transmission of transgene and its expression was recognized according to the results of RT-PCR and sequences analyses of their progeny.

Microanatomy and Histological Features of Central Myelin in the Root Exit Zone of Facial Nerve

  • Yee, Gi-Taek;Yoo, Chan-Jong;Han, Seong-Rok;Choi, Chan-Young
    • Journal of Korean Neurosurgical Society
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    • v.55 no.5
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    • pp.244-247
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    • 2014
  • Objective : The aim of this study was to evaluate the microanatomy and histological features of the central myelin in the root exit zone of facial nerve. Methods : Forty facial nerves with brain stem were obtained from 20 formalin fixed cadavers. Among them 17 facial nerves were ruined during preparation and 23 root entry zone (REZ) of facial nerves could be examined. The length of medial REZ, from detach point of facial nerve at the brain stem to transitional area, and the thickness of glial membrane of central myelin was measured. We cut brain stem along the facial nerve and made a tissue block of facial nerve REZ. Each tissue block was embedded with paraffin and serially sectioned. Slices were stained with hematoxylin and eosin (H&E), periodic acid-Schiff, and glial fibrillary acid protein. Microscopy was used to measure the extent of central myelin and thickness of outer glial membrane of central myelin. Thickness of glial membrane was examined at two different points, the thickest area of proximal and distal REZ. Results : Special stain with PAS and GFAP could be differentiated the central and peripheral myelin of facial nerve. The length of medial REZ was mean 2.6 mm (1.6-3.5 mm). The glial limiting membrane of brain stem is continued to the end of central myelin. We called it glial sheath of REZ. The thickness of glial sheath was mean $66.5{\mu}m(40-110{\mu}m$) at proximal REZ and $7.4{\mu}m(5-10{\mu}m$) at distal REZ. Conclusion : Medial REZ of facial nerve is mean 2.6 mm in length and covered by glial sheath continued from glial limiting membrane of brain stem. Glial sheath of central myelin tends to become thin toward transitional zone.

AN IMMUNOHISTOCHEMICAL STUDY ON CALCIFYING TISSUES OF THE FLORID CEMENTO-OSSEOUS DYSPLASIA EASILY INVOLVING CHRONIC DIFFUSE SCLEROSING OSTEOMYELITIS (만성 미만성 경화성 골수염을 동반한 개화성 백악질-골 이형성증의 면역조직화학적 연구)

  • Kim, Ji-Hyuck;Jo, Joung-Ae;Kim, Soung-Min;Park, Young-Wook;Huh, Jin-Young;Lee, Suk-Keun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.29 no.5
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    • pp.293-297
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    • 2003
  • Florid cemento-osseous dysplasia (FCOD) is a benign, non-neoplastic lesion characterized by multiple sclerosing masses only within jawbones. It is frequently confused with chronic diffuse sclerosing osteomyelitis (CDSO) in previous literatures. In our study, two cases of FCOD were examined to know the characteristics of their calcifying tissues. The first case was non-infected, while the second case was severely infected, displaying the typical features of CDSO in clinico-radiologic findings. The infected FCOD case showed a lot of bacterial colonies in the main lesion with relatively rare inflammatory reaction. The globular cementum-like materials of FCOD showed woven bone pattern and was positive for Alcian blue stain, and also positive for the antibodies of ameloblastin, bone morphogenetic protein (BMP) -2 and -4. On the other hands, in the immunostains of matrix metalloproteinase (MMP) -3, -9, -10, and $TNF-{\alpha}$, macrophage infiltrated in the FCOD lesion was rarely observed. These data suggest that the cementum-like materials of FCOD contain various matrix proteins, and that the cementum-like materials are relevant to the overgrowth of the bacterial colonies by inhibition of the regional inflammatory reactions.