• The Korean Journal of Cytopathology
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• v.19 no.1
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• pp.27-33
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• 2008

Although subacute granulomatous thyroiditis(SGT) is usually diagnosed clinically, papillary carcinoma or other thyroid conditions must be considered in the differential diagnosis. We retrospectively reviewed the clinical and fine-needle aspiration(FNA) cytologic findings seen in 10 SGT cases to decide what are the most reliable cytologic findings and the most helpful molecular tools for reaching a confident cytologic diagnosis. The most representative smear slides were retrieved to perform immunocytochemistry for cytokeratin 19(CK19) and Ret protein. Five papillary carcinomas(PTCs) were included as controls. The constant and typical cytologic findings of SGT were multinucleated giant cells(MGCs) (100%), epithelioid granulomas(90%), an inflammatory dirty background(90%) and plump transformed follicular cells(80%) without fire-flare cells, oncocytic cells or transformed lymphocytes. The immunoreactivities for CK19(37.5%) and Ret(10%) of the follicular cells of SGT were less than those(CK19 and Ret:100%) of PTC. CK19 immunoreactivity of the MGCs was seen in only one case of PTC. There was no significant difference between CK19 and Ret immunocytochemical staining for the MGCs of both SGT and PTC. The results of this study demonstrate that the cytological diagnosis of SGT can be improved by employing a combination of the typical and constant diagnostic cytological features and immunocytochemical results.

• Development and Reproduction
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• v.10 no.1
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• pp.63-73
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• 2006

This study aimed to find out suitable culture conditions for the differentiation of human amniotic membrane-derived stem cells(HAM) into hepatocyte-like cells. Almost homogenous population of fibroblast-like cells was successfully isolated from the amniotic membrane. In comparison to the non-coated plates and in the absence of insulin/transferrin/selenium(ITS), HAM cultured on the fibronectin-coated plates and in the presence of ITS showed the more intense immunocytochemical staining against the albumin. Addition of both fibroblast growth factor(FGF)-1 and -2 to the differentiation medium gave stronger staining compared to the treatment with FGF-1 or -2 alone. Periodic acid Schiff's base staining of glycogen and morphological turnover of fibroblast-like appearance of HAM into round shape matched the results of immunocytochemical studies. When the efficiency of two-step culture method was examined on the differentiation of HAM into hepatocyte-like cells, all of the results of immunocytochemical staining, periodic acid Schiff's staining and morphological change exhibited effective hepatic differentiation of HAM compared to the continuous culture method. Immunoblot analyses of HAM- conditioned media against the albumin showed that the culture of HAM in the presence of both ITS and fibronectin always gave a stronger staining intensity than those in the absence of them, and that the addition of ether mixture of FGF-4 and either FGF-2 or transforming growth $factor(TGF)-{\alpha}$ to the culture medium significantly enhanced the albumin secretion by HAM. Based on these observations, it is suggested that HAM could differentiate into hepatocyte-like cells under a culture condition consisting of fibronectin and ITS, and addition of FGF-4 with either one of FGF-2 or $TGF-{\alpha}$ could enhance the hepatic differentiation of HAM.

• Animal cells and systems
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• v.3 no.1
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• pp.53-58
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• 1999

The immunocytochemical method was used to identify the existence of voltage-dependent $Ca^{2＋}$-channels in mouse follicular oocytes. Three types of voltage-dependent $Ca^{2＋}$-channels were shown to exist in the follicular oocytes for the first time, the P/Q-type $Ca^{2＋}$-channel, the N-type $Ca^{2＋}$-channel, and the L-type $Ca^{2＋}$-channel. Among proven $Ca^{2＋}$-channels distributions of the P/Q-type $Ca^{2＋}$-channel and L-type $Ca^{2＋}$-channel showed localized staining (clustered pattern) on the oolemma. The distribution of the P/Q-type $Ca^{2＋}$-channel showed all localized staining, and the range of localized staining was from 1 to 8 in staining intensity. As the staining intensity increased from 1 to 8, the number of localized staining decreased. The L-type $Ca^{2＋}$-channel are homogeneously stained (29.4%-54.2%), while some of them (around 28.7%-44.1%) showed localized staining on the oolemma. However, the rest of them showed no staining at all (17.1%- 26.5%). On the contrary, the N-type $Ca^{2＋}$-channel showed mostly homogeneous staining, while nonstaining oocytes were around 33.8%. The rest showed localized staining (10%). However, staining intensity was much weaker than those of the P/Q-type and L-type $Ca^{2＋}$-channel. In fact, the N-type $Ca^{2＋}$-channel has been known to exist only in neurons (from ectoderm origin), but it is unknown how the N-type $Ca^{2＋}$-channel exists in the follicular oocytes (from mesoderm origin). Further studies are needed to examine the expression of $Ca^{2＋}$-channels during the developmental stages of the oocytes.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.1
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• pp.13-24
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• 2001

Objective: In muscle and neuronal cells, calcium channels have been classified by electrophysiological and pharmacological properties into (1) voltage-dependent $Ca^{2+}$-channel (1) P/Q-type $Ca^{2+}$-channel (2) N-type $Ca^{2+}$-channel (3) L-type $Ca^{2+}$-channel (4) T-type $Ca^{2+}$-channel (5) R-type $Ca^{2+}$-channel. The present study was done in order to investigate whether there is any difference in $Ca^{2+}$-channel distribution between activated and normally fertilized embryos. Methods: The immunocytochemical method was used to identify the existence of voltage-dependent $Ca^{2+}$-channels in parthenogenetically activated 2-cell embryos by ethanol and $SrCl_2$ treatment. These 2-cell embryos were obtained by exposure to 6% ethanol for 6 min and to 10 mM $SrCl_2$ for 2h. Results: P/Q-type $Ca^{2+}$-channels and L-type $Ca^{2+}$-channels have been identified. Whereas, three type of $Ca^{2+}$-channel P/Q-type, N-type, L-type have been identified in 2-cell embryos fertilized in vivo. Conclusion: Activation by ethanol was faster than those by $SrCl_2$. However, there was difference in DAB staining of the embryos between ethanol and $SrCl_2$ treatment (87.7% and 54.1 %). Intensity of staining was also different between ethanol- and $SrCl_2$-treated group. However, it has not been known why there was some difference in DAB staining and staining intensity in the present study.

• Development and Reproduction
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• v.12 no.2
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• pp.183-194
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• 2008

A variety of stem cells has been emerging as therapeutic cells that can replace organ transplantation in human liver diseases. The present study focused on whether human eyelid adipose-derived stem cells (HAD) might differentiate into functional hepatocyte-like cells in vitro. HAD were isolated from human eyelid adipose tissue. Effect of dimethyl sulfoxide (DMSO), fibroblast growth factor (FGF)-2 and FGF-4 on the hepatic differentiation of HAD have been examined in vitro. Immunocytochemical analysis and PAS staining showed that HAD cultured in both DMSO and FGF-4 exhibited the most intense staining than HAD of the other experimental groups. These HAD expressed numerous hepatocyte-related genes. Immunoblotting analyses showed that HAD cultured in the presence of DMSO and FGF-4 secreted higher amount of human albumin than HAD cultured in other conditions. Urea analysis also demonstrated that these HAD produced higher amount of urea than any other groups of HAD. In conclusion, combined treatment of DMSO and FGF-4 could effectively induce the functional differentiation of HAD into hepatocyte-like cells.

• Journal of Korean Ophthalmic Optics Society
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• v.16 no.1
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• pp.91-95
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• 2011

Purpose: In this study, we investigated the dye to staining for selective accumulation in rabbit retina. Methods: Rhodamine B was injected into the vitreous body in rabbit. After 24 h, the isolated retina was checked condition of cell staining on the microscope. We used conventional immunocytochemical techniques for recognizing cell type. Results: Well-labeled nuclei were seen in the middle of the inner nuclear layer of the rabbit retina. The number and distrbution of the accumulating cells were similar to those of the m$\ddot{u}$ller glia. To identify m$\ddot{u}$ller cell, we used antibody directed against vimentin. Rhodamine B-immunoreactive nuclei also were labeled with antivimentin antibody. We found that Rhodamine B was accumulated selectively in retinal m$\ddot{u}$ller cell. Conclusions: Specific accumulation in rabbit retinal m$\ddot{u}$ller cell occurred when Rhodamine B was applied to living retina.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.36 no.3
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• pp.177-185
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• 2010

Introduction: Epidermal growth factor receptor (EGFR) is expressed in human epithelial tumors including salivary cancers, and known to be correlated with tumor progression and poor clinical courses in some epithelial tumors. In this study, we determined the therapeutic effect of the anti-EGFR monoclonal antibody Erbitux (C225, cetuximab) in combination with the DNA topoisomerase I inhibitor irinotecan (CPT-11) on human salivary adenoid cystic carcinoma (SACC) cells growing in nude mice. Materials and Methods: At first, immunocytochemical analysis for the expression of EGFR and phosphorylated EGFR (pEGFR) on a human salivary ACC cell line (ACC3). To determine the in vivo effects of Erbitux and CPT-11, nude mice with orthotopic parotid tumors were randomized to receive intraperitoneal Erbitux (1 mg) two times per week, intraperitoneal Irinotecan (50 mg/kg) once per week, Erbitux plus CPT-11, or placebo. (control) Tumor volume and weight were measured. And mechanisms of in vivo activity of Erbitux and/or CPT-11 were determined by immunohistochemical/ immunofluorescent analyses. Results: Immunocytochemical staining of ACC3 demonstrated that EGFR was expressed and phosphorylated. CPT-11 inhibited ACC tumor growth in nude mice. Tumors of mice treated with CPT-11 and CPT-11 plus Erbitux exhibited increased tumor cell apoptosis and decreased microvessel density, which correlated with a decrease in the tumor volume in nude mice. But, CPT-11 seems not to be synergistic with Erbitux in our ACC3 model system. Conclusion: These results suggest that anti-EGFR monoclonal antibody and the DNA topoisomerase I inhibitor will be effective in the treatment of recurred or metastatic lesions of salivary ACC.

• The Korean Journal of Cytopathology
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• v.1 no.1
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• pp.74-84
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• 1990

Herpes simplex virus type 1 and 2(HSV-1, HSV-2) are the ubiquitous human pathogens responsible for a variety of afflictions. HSV-2 is one of the viruses that were suspected of promoting carcinogenesis in the uterine cervix. Certainly, there is a need for the more sensitive and accurate laboratory techniques for HSV detection. We examined total 80 cases of smears including 17 Tzanck smears of skin and 63 cases of Papanicolaou smears from total 77 patients with clinical impression of herpetic infections, from September, 1985 through August, 1989. Immunohistochemical typings for HSV-1 and HSV-2 were performed together with routine cytologic findings and compared. The results are as follows : 1) Patients were 9 males and 33 females, and age distribution was between 5 and 71 years. 2) Subjective symptoms such as ulceration, vesicle, vaginal discharge, pruritus, and pain were complained in 36 patients and 38 cases were genital herpes. Recurrence was noted in 11 cases. 3) Positive results were obtained in 42 among 80 cases. 4) Both routine cytology and immunohistochemical staining were positive in 13 cases and in 24 cases only immunohistochemical staining were positive. 5 cases were positive only in routine cytologic smears. 5) The cases that immunocytochemical stain had been performed were 37 cases, which were all positive in type 2. Among the above 37 cases, type 1 also were positive in 5 cases. The results show that the immunoperoxidase technique is one of the rapid and reliable method to confirm the herpetic infection when suspected and that it is particularly useful when the Papanicolaou smear findings are equivocal.

• IMMUNE NETWORK
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• v.6 no.4
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• pp.204-210
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• 2006

Background: Nitric oxide (NO) in articular chondrocytes regulates dedifferentiation and inflammatory responses by modulating MAP kinases. In this study, we investigated whether the Src kinase in chondrocytes regulates NO-induced dedifferentiation and cyclooxygenase-2 (COX-2) expression. Methods: Primary chondrocytes were treated with various concentrations of SNP for 24 h. The COX-2 and type II collagen expression levels were determined by immunoblot analysis, and prostaglandin $E_2\;(PGE_2)$ was determined by using a $PGE_2$ assay kit. Expression and distribution of p-Caveolin and COX-2 in rabbit articular chondrocytes and cartilage explants were determined by immunohistochemical staining and immunocytochemical staining, respectively. Results: SNP treatment stimulated Src kinase activation in a dose-dependent manner in articular chondrocytes. The Src kinase inhibitors PP2 [4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo(3,4-d)pyrimidine], a significantly blocked SNP-induced p38 kinase and caveolin-1 activation in a dose-dependent manner. Therefore, to determine whether Src kinase activation is associated with dedifferentiation and/or COX-2 expression and $PGE_2$ production. As expected, PP2 potentiated SNP-stimulated dedifferentiation, but completely blocked both COX-2 expression and $PGE_2$ production. And also, levels of p-Caveolin and COX-2 protein expression were increased in SNP-treated primary chondrocytes and osteoarthritic and rheumatoid arthritic cartilage, suggesting that p-Caveolin may playa role in the inflammatory responses of arthritic cartilage. Conclusion: Our previously studies indicated that NO caused dedifferentiation and COX-2 expression is regulated by p38 kinase through caveolin-1 (1). Therefore, our results collectively suggest that Src kinase regulates NO-induced dedifferentiation and COX-2 expression in chondrocytes via p38 kinase in association with caveolin-1.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.32 no.6
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• pp.495-505
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• 2006

In this study we evaluate the effects of bFGF-BB and PDGF on in vitro proliferation, differentiation and mineralization of mesenchymal stem cells (MSCs) from rat. MSCs were prepared from the bone marrow of 6 or 7-week-old male rats with a technique previously described by Maniatopoulos et al. in 1988. Lineage differentiation to osteogenesis, chondrogenesis and adipogenesis were performed. At first, we characterized the cultured cell on passage 1, 3, 5, 7 with immunocytochemical staining using CD29, 44, 34, 45, ${\alpha}$-SMA and type I collagen. And to study the effects of bFGF and PDGF-BB on proliferation, differentiation and mineralization, we seeded the expanded cell at a density of 6 $6{\times}10^3\;cells/cm^2$ to 100-mm dish for evaluation of cell proliferation and MTT assay was carried out on day 2, 4, 7, 9. We also resuspended the cells with same density $(6{\times}10^3\;cells/cm^2)$ to 24 well plates for subculture. On the following day, the attached cells were exposed to 2.5ng/ml bFGF and/or 25ng/ml PDGF-BB daily during 5 days. The osteocalcin (OC) level was assessed and mineral contents were evaluated with alizarin red S staining on subculture day 2, 7, 14, 21. We identified the mesenchymal stem cell from the bone marrow derived cells of rat through their successful multi-differentiation and stable display of its phenotype. And bFGF and PDGF-BB showed the effect that inhibited osteoblastic differentiation and mineralization mildly in above concentration at in vitro culture. This study was supported by grant 04-2004-0120 from the Seoul National University Hospital Research Fund.