• Title/Summary/Keyword: Immunohistochemical

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The Effects of Woohwangcheongsim-won on Reperfusion Following Middle Cerebral Artery Occlusion in Rats (우황청심원이 중대뇌동맥 결찰로 유발된 뇌허혈에 미치는 영향)

  • 조규선;정승현;신길조;이원철
    • The Journal of Korean Medicine
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    • v.22 no.1
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    • pp.78-89
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    • 2001
  • Objectives : The purpose of this investigation is to evaluate the effects of Woohwangcheongsim-won on reperfusion following MCA occlusion in rats. Methods : To evaluate the effect of Woohwangcheongsim-won on reperfusion following MCA occlusion, the volume of cerebral ischemia and edema were measured and the change of the CAI pyramidal neuron in the hippocampus was investigated by light microscopy. And the changes of several neurotransmitters and enzymes were investigated with the immunohistochemical methods. Results : 1. The volume of the control group, which was ischemic-damaged was 23.6%, and that of the sample group was 13.5%. 2. The voluminalratio of the right/left hemisphere was 116 in the control group, and that of the sample group was 107. 3. The pyramidal cells of CAI area in the control group were greatly damaged. The cells were changed into discontinuous and unsystematic forms, and nuclei, and cytoplasms were shrunk. On the other hand, the cells of the sample group were less damaged. 4. On the immunohistochemical methods, the sensitivities of GABA, NOS, DBH in the control group were increased, and those of synapsin and $eEF-l{\alpha}$ were decreased as compared with the normal group. NOS and DBH which were negative in the normal group showed positive reaction. On the other hand, the sensitivities of GABA, NOS and DBH in the sample group were decreased, but those of NPY, synapsin, CaMKII and $eEF-l{\alpha}$ were increased as compared with the control group. Conclusions : Woohwangcheongsim-won reduced the volume of cerebral ischemia and edema, and minimized the damage of pyramidal cells. The mechanism was related to protein synthesis, such as synapsin, ${\alpha}CaMKII$ and $eEF-l{\alpha}$, which resist neurotoxicity of glutamate receptors.

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Differentiation of Human Adult Adipose Derived Stem Cell in vitro and Immunohistochemical Study of Adipose Derived Stem Cell after Intracerebral Transplantation in Rats

  • Ko, Kwang-Seok;Lee, Il-Woo;Joo, Won-Il;Lee, Kyung-Jun;Park, Hae-Kwan;Rha, Hyung-Keun
    • Journal of Korean Neurosurgical Society
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    • v.42 no.2
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    • pp.118-124
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    • 2007
  • Objective : Adipose tissue is derived from the embryonic mesoderm and contains a heterogenous stromal cell population. Authors have tried to verify the characteristics of stem cell of adipose derived stromal cells (ADSCs) and to investigate immunohistochemical findings after transplantation of ADSC into rat brain to evaluate survival, migration and differentiation of transplanted stromal cells. Methods : First ADSCs were isolated from human adipose tissue and induced adipose, osseous and neuronal differentiation under appropriate culture condition in vitro and examined phenotypes profile of human ADSCs in undifferentiated states using flow cytometry and immunohistochemical study. Human ADSCs were transplanted into the healthy rat brain to investigate survival, migration and differentiation after 4 weeks. Results : From human adipose tissue, adipose stem cells were harvested and subcultured for several times. The cultured ADSCs were differentiated into adipocytes, osteoctye and neuron-like cell under conditioned media. Flow cytometric analysis of undifferentiated ADSCs revealed that ADSCs were positive for CD29, CD44 and negative for CD34, CD45, CD117 and HLA-DR. Transplanted human ADSCs were found mainly in cortex adjacent to injection site and migrated from injection site at a distance of at least 1 mm along the cortex and corpus callosum. A few transplanted cells have differentiated into neuron and astrocyte. Conclusion : ADSCs were differentiated into multilineage cell lines through transdifferentiation. ADSCs were survived and migrated in xenograft without immunosuppression. Based on this data, ADSCs may be potential source of stem cells for many human disease including neurologic disorder.

Blood-Brain Barrier Experiments with Clinical Magnetic Resonance Imaging and an Immunohistochemical Study

  • Park, Jun-Woo;Kim, Hak-Jin;Song, Geun-Sung;Han, Hyung-Soo
    • Journal of Korean Neurosurgical Society
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    • v.47 no.3
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    • pp.203-209
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    • 2010
  • Objective : The purpose of study was to evaluate the feasibility of brain magnetic resonance (MR) images of the rat obtained using a 1.5T MR machine in several blood-brain barrier (BBB) experiments. Methods : Male Sprague-Dawley rats were used. MR images were obtained using a clinical 1.5T MR machine. A microcatheter was introduced via the femoral artery to the carotid artery. Normal saline (group 1, n = 4), clotted autologous blood (group 2, n = 4), triolein emulsion (group 3, n = 4), and oleic acid emulsion (group 4, n = 4) were infused into the carotid artery through a microcatheter. Conventional and diffusion-weighted images, the apparent coefficient map, perfusion-weighted images, and contrast-enhanced MR images were obtained. Brain tissue was obtained and triphenyltetrazolium chloride (TTC) staining was performed in group 2. Fluorescein isothiocyanate (FITC)-labeled dextran images and endothelial barrier antigen (EBA) studies were performed in group 4. Results : The MR images in group 1 were of good quality. The MR images in group 2 revealed typical findings of acute cerebral infarction. Perfusion defects were noted on the perfusion-weighted images. The MR images in group 3 showed vasogenic edema and contrast enhancement, representing vascular damage. The rats in group 4 had vasogenic edema on the MR images and leakage of dextran on the FITC-labeled dextran image, representing increased vascular permeability. The immune reaction was decreased on the EBA study. Conclusion : Clinical 1.5T MR images using a rat depicted many informative results in the present study. These results can be used in further researches of the BBB using combined clinical MR machines and immunohistochemical examinations.

The immunohistochemical studies of Herbal drug-acupuncture of Radix paeoniae lactiflorae on Gastric ulcer induced by HCI-aspirin in Rats (백작약약침(白芍藥藥鍼)이 백서(白鼠)의 위궤양(胃潰瘍)에 미치는 면역조직화학적(免疫組織化學的) 연구(硏究))

  • Park Ki-Hyun;Han Sang-Won;Park Jae-Hyun
    • Journal of Acupuncture Research
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    • v.15 no.2
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    • pp.61-79
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    • 1998
  • In order to study the immunohistochemical effects of herbal drug-acupuncture of Radix paeoniae lactiflorae on gastric ulcer induced by HCl-aspirin in rats. this experiment was done by herbal drug-acupuncture to Wisu($B_{21}$). Chung-Wan($CV_{12}$), Chok-Samni($S_{36}$) loci to measure histological features of ulcer lesion, the change of numbers of parietal cell, chief celI, gastrin and somatostatin-immunoreactive cell. The obtained results were as follows; 1. The ulcerative lesions of gastric mucosa were decreased to WiSU($B_{21}$) loci followed by Chung-Wan($CV_{12}$) and Chok-Samni($S_{36}$) loci compared to the control groups. 2. In the numbers of parietal cell, the most remarkable decrease was observed to Wisu($B_{21}$) loci followed by Chung-Wan($CV_{12}$) and Chok-Samni($S_{36}$) loci compared to the control groups. 3. In the numbers of chief cell, the most remarkable increase was observed to Wisu($B_{21}$) loci followed by Chung-Wan($CV_{12}$) and Chok-Samni($S_{36}$) loci compared to the control groups. 4. In the numbers of gastrin-immunoreactive cell, the most remarkable decrease was observed to Wiu($B_{21}$) loci followed by Chung-Wan($CV_{12}$) and Chok-Samni($S_{36}$) loci compared to the control groups. 5. In the numbers of somatostatin-immunoreactive cell, the most remarkable decrease was observed to Wisu($B_{21}$) loci followed by Chung-Wan($CV_{12}$) and Chok-Samni($S_{36}$) loci compared to the control groups.

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No Association between BRCA1 Immunohistochemical Expression and Tumor Grade, Stage or Overall Survival in Platinum-Treated Epithelial Ovarian Cancer Patients

  • Shawky, Abd El-Aty;El-Hafez, Amal Abd;El-Tantawy, Dina;Hamdy, Rasha
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.10
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    • pp.4275-4279
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    • 2014
  • Background: The aim of this work is to assess the frequency of BRCA1 protein immunohistochemical (IHC) expression in epithelial ovarian cancer (EOC) and to evaluate the association of BRCA1 expression with clinical and pathological characteristics and the overall survival (OS) of patients treated with postoperative platinum-based chemotherapeutic agents. Materials and Methods: This retrospective study was conducted on 35 cases of epithelial ovarian cancer selected from the files of the Pathology Department, Faculty of Medicine, Mansoura University, Egypt. Immunohistochemistry (IHC) was performed for BRCA1 gene protein. BRCA1 expression was compared to patient's age, tumor histology, grade, stage and OS time. Statistical analysis was carried out with the SPSS version 16.0 to assess significant associations. Results: BRCA1 nuclear expression was detected in 40% of EOC, in which a mild increase in the percentage of positive cases was observed with serous histology, stage IV, and grade 3 carcinomas. There was a significant statistical difference in BRCA1 expression with regard to histological subtypes of EOC (p=0.048), but not grade or stage. Mean OS and survival rate were slightly better for BRCA1 expressing group, but there was no statistically significant difference (p=0.528). Conclusions: No association between BRCA1 immunohistochemical expression and tumor grade, stage or overall survival was noted in platinum-treated epithelial ovarian cancer patients.

Stromal cell-derived factor-1 (SDF-1) expression in the oral squamous cell carcinoma (구강편평상피암종에서 stromal cell-derived factor-1의 발현)

  • Kim, Kyung-Wook;Han, Se-Jin;Roh, Kyu-Seob
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.1
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    • pp.1-6
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    • 2010
  • Purpose: Chemokines are structurally related, small polypeptide signaling molecules that bind to and activate a family of transmembrane G protein-coupled receptors, the chemokine receptors. Recently, interaction between the chemokine receptor CXCR4 and its ligand, stromal cell-derived factor 1 (SDF-1 or CXCL12), has been found to play an important role in tumorigenicity, proliferation, metastasis and angiogenesis in many cancers such as lung cancer, breast cancer, melanoma, glioblastoma, pancreatic cancer and cholangiocarcinoma. Hence, the goal of this study is to identify the correlation of clinicopathological factors and the up-regulation of SDF-1 expression in oral squamous cell carcinoma. Material and methods: We studied the immunohistochemical staining of SDF-1, quantitative RT-PCR (qRT-PCR) of SDF-1 gene in 20 specimens of 20 patients with oral squamous cell carcinoma. Results: 1. In the immunohistochemical study of poor differentiated and invasive oral squamous cell carcinoma, the high level staining of SDF-1 was observed. And the correlation between immunohistochemical SDF-1 expression and tumor nodes metastases (TNM) classification of specimens was significant.($x^2$ test, P < 0.05) 2. In the SDF-1 gene qRT-PCR analysis, SDF-1 expression was more in tumor tissue than in carcinoma in situ tissue. Paired-samples analysis determined the difference of SDF-1 mRNA expression level between the cancer tissue and the carcinoma in situ tissue.(Student's t-test, P < 0.05) Conclusion: These findings suggest that up-regulation of the SDF-1 may play a role in progression and invasion of oral squamous cell carcinoma.

Analyzing the factors that influence occult metastasis in oral tongue cancer

  • Shin, Jung-Hyun;Yoon, Hye-Jung;Kim, Soung-Min;Lee, Jong-Ho;Myoung, Hoon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.46 no.2
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    • pp.99-107
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    • 2020
  • Objectives: We accessed the various clinico-histopathological factors, and their association with occult metastasis (OM) in oral tongue squamous cell carcinoma (OTSCC). Materials and Methods: One hundred-nine patients with OTSCC were divided into the elective neck dissection (END) group and the watchful waiting (WW) group. Age, sex, T-stage, depth of invasion and differentiation were evaluated to determine the correlation between clinico-histopathological factors and OM. For immunohistochemical analysis, paraffin-embedded blocks of 41 OTSCC specimens were examined with antibodies (VEGF-c, c-Met, and ROR1). Results: The group with tumor thickness of oral tongue cancer ≥3 mm had higher incidence of OM than those with a thickness of <3 mm. The depth of invasion was statistically correlated with OM (P=0.022). Immunohistochemical analysis showed that high expression of VEGF-c (P=0.043), c-Met (P=0.009), and ROR-1 (P=0.003) were statistically correlated with OM. Conclusion: The analysis of these clinico-histopathological and immunohistochemical factors can help to determine neck dissection in clinically negative (cN0) patients.

An immunohistochemical study on the effects of low-level laser irradiation on expression of actin filaments of human gingival fibroblasts in vitro (저출력레이저조사가 배양치은섬유아 세포의 actin filaments발현에 미치는 영향에 관한 면역조직화학적 연구)

  • Kim, Hyung-Sung;Kim, Chun-Suk;Kim, Hyung-Soo;Kim, Hyun-Seop;Kim, Byung-Ock;Han, Kyung-Yoon
    • Journal of Periodontal and Implant Science
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    • v.26 no.4
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    • pp.1003-1012
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    • 1996
  • The induction of a phenotype with preoperties may have clinical significance in the acceleration of the wound-healing process. Wound contraction involves a specialized cell known as the myofibroblast. The myofibroblasts can be identified by their intense staining of actin bundles with anti-actin antibody. Tissue-specific actin distribution is correlated with the contractile activity of the myofibroblasts and smooth muscle etc. This study was performed to determine the expression of actin filaments in the cytoplasm of cultured human gingival fibroblsts after GaAs laser(BIOSAER, Korea) irradiation. Human gingival fibroblasts were cultured from explants of normal interdental gingival tissue. The third-generation fibroblasts were used for immunohistochemical study. The cultured fibroblasts were exposed $0.53joule/cm^2$(lmW, 7 mimutes) of energy density, and then observed by immunohistochemical method using, rabbit anti0gelsolin, hen smooth muscle polyclonal antibody(Chemicon international inc.), and biotinylated goat anti-rabbit IgG(Vectastain) 24-, 36-, 48-hour after laser irradiation Following results were obtained ; 1. In nonirradiated cultures, round shaped active fibroblasts with abundant cytoplasm and prominet nucleoli were observed. 2. In 24- and 36-hour cultures after laser irradiation, spindle shaped cells with long process were observed. The intensity of stain was seen in cytoplasm of these modified fibroblasts. 3. In 48-hoour cultures after laser irradiation, stained spindle shape cell were not observed. The results suggest that the effect of the galium-arsenide laser treatment on cultured gingival fibroblasts is the rapid development of cytoplasmic actin filaments.

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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.

Epidermal growth factor receptor overexpression and K-ras mutation detection in the oral squamous cell carcinoma (구강편평상피암종에서 상피성장인자 수용체의 과발현과 K-ras 유전자 변이)

  • Moon, Byeong-Chool;Han, Se-Jin;Jeong, Dong-Jun;Kim, Kyung-Wook
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.37 no.5
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    • pp.396-402
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    • 2011
  • Introduction: Epidermal growth factor is a single-chain polypeptide consisting of 53 amino acids with potent mitogenic activity that stimulates the proliferation of a range of normal and neoplastic cells through an interaction with its specific receptor (epidermal growth factor receptor, EGFR). This interaction plays a key role in tumor progression including the induction of tumor cell proliferation. An increased EGFR copy number have been associated with a favorable response to EGFR tyrosine kinase inhibitors therapy. In contrast, K-ras mutations tend to predict a poor response to such therapy. The aim of this study was to determine the correlation between the clinicopathological factors and the up-regulation of EGFR expression and Kras mutations in oral squamous cell carcinoma. Materials and Methods: This study examined the immunohistochemical staining of EGFR, K-ras mutation detection with peptide nucleic acid (PNA)-based real-time polymerase chain reaction (PCR) clamping in 20 specimens from 20 patients with oral squamous cell carcinoma. Results: 1. In the immunohistochemical study of poorly differentiated and invasive oral squamous cell carcinoma, a high level of EGFR staining was observed. The correlation between immunohistochemical EGFR expression and histological differentiation, as well as the tumor size of the specimens was significant (Pearson correlation analysis, significance [r] >0.5, P<0.05). 2. In PNA-based real-time PCR clamping analysis, a K-ras mutation was not detected in all specimens. Conclusion: These findings suggest that the up-regulation of the EGFR may play a role in the progression and invasion of oral squamous cell carcinoma that is, independent of a K-ras mutation.