• Title/Summary/Keyword: Differential staining

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Protein Expression of Mouse Uterus in Post-Implantation

  • Kim, Hong-Rye;Han, Rong-Xun;Kim, Myung-Youn;Diao, Yunfei;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • v.33 no.4
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    • pp.237-242
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    • 2009
  • Pregnancy is a unique event in which a fetus develops in the uterus despite being genetically and immunologically different from the mother, and the underlying mechanisms remain poorly understood. To analyze the differential gene expression profiles in nonpregnant and 7 days post coitus (dpc) pregnant uterus of mice, we performed a global proteomic study by 2-D gel electrophoresis (2-DE) and MALDI-TOF-MS. The uterine proteins were separated using 2-DE, Approximately 1,000 spots were detected on staining with Coomassie brilliant blue. An image analysis using Melanie III (Swiss Institute for Bioinformatics) was performed to detect variations in protein spots between pregnant and nonpregnant uterus. Twenty-one spots were identified as differentially expressed proteins, of which 10 were up-regulated proteins such as alpha-fetoprotein, chloride intracellular channel 1, transgelin, heat-shock protein beta-1, and carbonic anhydrase II, while 11 were down-regulated proteins such as X-box binding protein, glutathione S-transferase omega 1, olfactory receptor Olfr204, and metalloproteinase-disintegrin domain containing protein TECADAM. Most of the identified proteins appeared to be related with catabolism, cell growth, metabolism, regulation, cell protection, protein repair, or protection. Our results uncovered key proteins of mouse uterus involved in pregnancy.

A Case of Benign Fibrous Mesothelioma of the Pleura (흉막의 양성 섬유성 중피종 1예)

  • Lee, Sung-Keun;Kim, Doe-Min;Jan, Kyong-Sun;Park, Sae-Jong;Keun, Jae-Sung;Kim, Woing-Su;Kang, Jong-Yuel;Kim, Myung-Sun
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.3
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    • pp.432-437
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    • 1999
  • Benign fibrous mesothelioma of the pleura is a relatively rare neoplasm originated from pleural mesothelial cell, often asymptomatic or presenting with a specifical sign. One of the main problems, concerns the preoperative differential diagnosis, mainly because it is difficult to differentiate between benign and malignant type. A 62-year old woman presented with recurrent chest pain. The chest radiography in a patient was suspected localized pleural mesothelioma. The chest computed tomography scan showed that mass like lesion of well marginated ovoid shape with homogenous attenuation on anterior-basal segment of right lower lobe. After resection of a pleural mass by thoracoscopic extirpation from right hemithorax, Localized benign fibrous mesothelioma of the pleura was confirmed by pathology and immunohisto-chemical staining. We report here one case of pleural benign fibrous mesothelioma with some considerations on its diagnosis and treatment.

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Effect of cell-penetrating peptide-conjugated estrogen-related receptor ${\beta}$ on the development of mouse embryos cultured in vitro

  • Yang, Ning Jie;Seol, Dong-Won;Jo, Junghyun;Jang, Hyun Mee;Yoon, Sook-Young;Lee, Dong Ryul
    • Clinical and Experimental Reproductive Medicine
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    • v.41 no.1
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    • pp.1-8
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    • 2014
  • Objective: Estrogen related receptor ${\beta}$ (Esrrb) is a member of the orphan nuclear receptors and may regulate the expression of pluripotencyrelated genes, such as Oct4 and Nanog. Therefore, in the present study, we have developed a method for delivering exogenous ESRRB recombinant protein into embryos by using cell-penetrating peptide (CPP) conjugation and have analyzed their effect on embryonic development. Methods: Mouse oocytes and embryos were obtained from superovulated mice. The expression of Oct4 mRNA and the cell number of inner cell mass (ICM) in the in vitro-derived and in vivo-derived blastocysts were first analyzed by real time-reverse transcription-polymerase chain reaction and differential staining. Then 8-cell embryos were cultured in KSOM media with or without $2{\mu}g/mL$ CPP-ESRRB protein for 24 to 48 hours, followed by checking their integration into embryos during in vitro culture by Western blot and immunocytochemistry. Results: Expression of Oct4 and the cell number of ICM were lower in the in vitro-derived blastocysts than in the in vivo-derived ones (p<0.05). In the blastocysts derived from the CPP-ESRRB-treated group, expression of Oct4 was greater than in the non-treated groups (p<0.05). Although no difference in embryonic development was observed between the treated and non-treated groups, the cell number of ICM was greater in the CPP-ESRRB-treated group. Conclusion: Treatment of CPP-ESRRB during cultivation could increase embryos' expression of Oct4 and the formation rate of the ICM in the blastocyst. Additionally, an exogenous delivery system of CPP-conjugated protein would be a useful tool for improving embryo culture systems.

Differential Effects of Nongenotoxic and Genotoxic Carcinogens on the Preneoplastic Lesions in the gat Liver

  • Kim, Dae-Joong;Lee, Kook-Kyung;Hong, Jin-Tae
    • Archives of Pharmacal Research
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    • v.21 no.4
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    • pp.363-369
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    • 1998
  • Glutathione S-transferase placental form (GST-P) positive foci development and its expression in liver exposed by nongenotoxic carcinogens phenobarbital (PB) and clofibrate (CF), and genotoxic carcinogen 2-amino-3-methylimidazo[4,5-f] quinoline (IQ) were investigated as a measure of carcinogenic potential of these chemicals. Male F344 rats were initially given a single intraperitioneal injection of diethyinitrosamine (200 mg/kg), and 2 weeks later, animals were fed diets containing 0.03% IQ or 0.5% CF or 0.05% PB or basal diet as a control for 6 weeks. All rats were subjected to two-thirds partial hepatectomy (PH) at week 3. Sequential sacrifice of rats was performed at 8 weeks or 52 weeks, and liver tissues were examined for immunohistochemical staining of GST-P positive foci. The numbers (No./$cm^2$) and areas ($mm^2$/ $cm^2$) of GST-P positive foci were increased by IQ or PB, but were decreased by CF compare to the control. Consistent with the development of GST-P positive foci, a time-related increase in the expression of GST-P mRNA was found in the rats treated with IQ, whereas CF decreased it. The incidence of hepatocellular carcinoma at 52 weeks was increased by all three chemicals. These results show that PB and IQ induced GST-P positive foci, but the peroxisome proliferator CF did not, which suggest that the prediction of carcinogenic potency based on the development of prenoplastic foci may cause false negative in a particular category compounds like peroxisome proliferators.

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Fine Needle Aspiration Cytology of Subacute Granulomatous Thyroiditis -A Clinico-Cytological Review of 10 Cases with Immunocytochemical Analysis- (아급성 육아종성 갑상샘염의 세침흡인 세포소견 -아급성 육아종성 갑상샘염 10예의 임상-세포소견 및 면역세포염색 소견 분석-)

  • Kim, Do-Kyung;Pyo, Ju-Yeon;Park, Jong-Pil;Jin, Lian-Hua;Jeong, Woo-Hee;Son, Eun-Ju;Hong, Soon-Won
    • 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.

Post-ischemic Time-dependent Activity Changes of Hippocampal CA1 cells of the Mongolian Gerbils

  • Won, Moo-Ho;Shin, Hyung-Cheul
    • The Korean Journal of Physiology and Pharmacology
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    • v.11 no.6
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    • pp.247-251
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    • 2007
  • Changes of single unit activity of CA1 hippocampus region were investigated in anesthetized Mongolian gerbils for six days following transient ischemia. Ischemia was produced immediately before the implantation of micro-wire recording electrodes. In control animals receiving pseudo-ischemic surgery, neither spontaneous neuronal activities ($5.70{\pm}0.4Hz$) nor the number of recorded neurons per animal changed significantly for six days. Correlative firings among simultaneously recorded neurons were weak (correlation coefficient > 0.6) in the control animals. Animals subjected to ischemia exhibited a significant elevation of neural firing at post-ischemic 12 hr ($9.95{\pm}0.9Hz$) and day 1 ($8.48{\pm}0.8Hz$), but a significant depression of activity at post-ischemic day 6 ($1.84{\pm}0.3Hz$) when compared to the activities of non-ischemic control animal. Ischemia significantly (correlation coefficient > 0.6) increased correlative firings among simultaneously recorded neurons, which were prominent especially during post-ischemic days 1, 2 and 6. Although the numbers of spontaneously active neurons recorded from control group varied within normal range during the experimental period, those from ischemic group changed in post-ischemic time-dependent manner. Temporal changes of the number of cells recorded per animal between control group and ischemic group were also significantly different (p = 0.0084, t = 3.271, df = 10). Cresyl violet staining indicated significant loss of CA1 cells at post-ischemic day 7. Overall, we showed post-ischemic time-dependent, differential changes of three characteristics, including spontaneous activity, network relationship and excitability of CA1 cells, suggesting sustained neural functions. Thus, histological observation of CA1 cell death till post-ischemic day 7 may not represent actual neuronal death.

Schwannoma Originating from Infraorbital Nerve (안와하신경에서 기원한 신경초종의 치험례)

  • Ha, Won;Lee, Ji Won;Choi, Jae Il;Yang, Wan Suk;Kim, Sun Young
    • Archives of Craniofacial Surgery
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    • v.14 no.1
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    • pp.61-64
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    • 2013
  • Schwannomas are well-differentiated solitary benign tumors that originate from the schwann cells of the nerve sheath. They can readily occur in the head and neck regions, but the schwannoma originating from the infraorbital nerve is extremely rare and usually painless, slow-growing, and without specific symptoms. The author experienced a rare case of infraorbital schwannoma, which was completely removed through the intraoral approach. A 20-year-old woman was admitted to our hospital for a painless, solid and circular mass located on the right infraorbital region. The eyeball movement and visual field were normal. There was no globe displacement or proptosis. Preoperative computed tomography demonstrated $13{\times}10{\times}5mm$-sized soft tissue mass. On March 2011, the mass was removed through an intraoral approach. On histopathological examination, the gross specimen consisted of a smooth, well-encapsulated and light yellowish solid mass, measuring $12{\times}7{\times}5mm$. Microscopically, it presented a typical manifestation of schwannoma with Antoni A area with Verocay body, and Antoni B area on H&E stain. The result of the immunohistochemical staining was positive for the S-100 protein. The patient had hypoesthesia of the nasal septum and vestibule in the postoperative period, and this finding confirmed that the internal nasal branch of infraorbital nerve was the nerve in which the schwannoma originated. Infraorbital schwannomas are very rare and must be included in the differential diagnosis of the orbital masses inferior to the eyeball. In the case of early diagnosis, the small-sized infraorbital schwannomas can be completely removed without any scar through an intraoral approach.

The Usefulness of Immunocytochemistry of CD56 in Determining Malignancy from Indeterminate Thyroid Fine-Needle Aspiration Cytology

  • Cha, Hyunseo;Pyo, Ju Yeon;Hong, Soon Won
    • Journal of Pathology and Translational Medicine
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    • v.52 no.6
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    • pp.404-410
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    • 2018
  • Background: Fine-needle aspiration cytology serves as a safe, economical tool in evaluating thyroid nodules. However, about 30% of the samples are categorized as indeterminate. Hence, many immunocytochemistry markers have been studied, but there has not been a single outstanding marker. We studied the efficacy of CD56 with human bone marrow endothelial cell marker-1 (HBME-1) in diagnosis in the Bethesda System for Reporting Thyroid Cytopathology (TBSRTC) category III. Methods: We reviewed ThinPrep liquid-based cytology (LBC) samples with Papanicolaou stain from July 1 to December 31, 2016 (2,195 cases) and selected TBSRTC category III cases (n=363). Twenty-six cases were histologically confirmed as benign (six cases, 23%) or malignant (20 cases, 77%); we stained 26 LBC slides with HBME-1 and CD56 through the cell transfer method. For evaluation of reactivity of immunocytochemistry, we chose atypical follicular cell clusters. Results: CD56 was not reactive in 18 of 20 cases (90%) of malignant nodules and showed cytoplasmic positivity in five of six cases (83%) of benign nodules. CD56 showed high sensitivity (90.0%) and relatively low specificity (83.3%) in detecting malignancy (p=.004). HBME-1 was reactive in 17 of 20 cases (85%) of malignant nodules and was not reactive in five of six cases (83%) of benign nodules. HBME-1 showed slightly lower sensitivity (85.0%) than CD56. The specificity in detecting malignancy by HBME-1 was similar to that of CD56 (83.3%, p=.008). CD56 and HBME-1 tests combined showed lower sensitivity (75.0% vs 90%) and higher specificity (93.8% vs 83.3%) in detecting malignancy compared to using CD56 alone. Conclusions: Using CD56 alone showed relatively low specificity despite high sensitivity for detecting malignancy. Combining CD56 with HBME-1 could increase the specificity. Thus, we suggest that CD56 could be a useful preoperative marker for differential diagnosis of TBSRTC category III samples.

Ginseng-derived nanoparticles induce skin cell proliferation and promote wound healing

  • Song Yang;Shuyan Lu;Limei Ren;Shuai Bian;Daqing Zhao;Meichen Liu;Jiawen Wang
    • Journal of Ginseng Research
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    • v.47 no.1
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    • pp.133-143
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    • 2023
  • Background: Past studies suggested that ginseng extracts and ginseng-derived molecules exerted significant regulatory effects on skin. However, no reports have described the effects of ginseng-derived nanoparticles (GDNPs) on skin cell proliferation and wound healing. In this study, we investigated whether GDNPs regulate the proliferation of skin cells and promote wound healing in a mouse model. Methods: GDNPs were separated and purified via differential centrifugation and sucrose/D2O gradient ultracentrifugation. GDNP uptake, cell proliferation and cell cycle progression were measured by confocal microscopy, CCK-8 assay and flow cytometry, respectively. Cell migration and angiogenic effects were assessed by the wound scratch assay and tube formation assay, respectively. ELISA was used to detect extracellular matrix secretion. The relevant signaling pathway was confirmed by western blotting. The effects of GDNPs on skin wound healing were assessed by wound observation, HE staining, and western blotting. Results: GDNPs possessed the essential features of exosomes, and they were accumulated by skin cells. Treatment with GDNPs notably enhanced the proliferation of HaCaT, BJ and HUVECs. GDNPs also enhanced the migration in HaCaT cells and HUVECs and angiogenesis in HUVECs. GDNPs increased the secretion of MMP-1, fibronectin-1, elastin-1, and COL1A1 in all three cell lines. GDNPs regulated cell proliferation through the ERK and AKT/ mTOR pathways. Furthermore, GDNPs facilitated skin wound healing and decreased inflammation in a mouse skin wound model. Conclusion: GDNPs can promote skin wound healing through the ERK and AKT/mTOR pathways. GDNPs thus represent an alternative treatment for chronic skin wounds.

Differential antiangiogenic and anticancer activities of the active metabolites of ginsenoside Rg3

  • Maryam Nakhjavani;Eric Smith;Kenny Yeo;Yoko Tomita;Timothy J. Price;Andrea Yool;Amanda R. Townsend;Jennifer E. Hardingham
    • Journal of Ginseng Research
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    • v.48 no.2
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    • pp.171-180
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    • 2024
  • Background: Epimers of ginsenoside Rg3 (Rg3) have a low bioavailability and are prone to deglycosylation, which produces epimers of ginsenoside Rh2 (S-Rh2 and R-Rh2) and protopanaxadiol (S-PPD and R-PPD). The aim of this study was to compare the efficacy and potency of these molecules as anti-cancer agents. Methods: Crystal violet staining was used to study the anti-proliferatory action of the molecules on a human epithelial breast cancer cell line, MDA-MB-231, and human umbilical vein endothelial cells (HUVEC) and compare their potency. Cell death and cell cycle were studied using flow cytometry and mode of cell death was studied using live cell imaging. Anti-angiogenic effects of the drug were studied using loop formation assay. Molecular docking showed the interaction of these molecules with vascular endothelial growth factor receptor-2 (VEGFR2) and aquaporin (AQP) water channels. VEGF bioassay was used to study the interaction of Rh2 with VEGFR2, in vitro. Results: HUVEC was the more sensitive cell line to the anti-proliferative effects of S-Rh2, S-PPD and R-PPD. The molecules induced necroptosis/necrosis in MDA-MB-231 and apoptosis in HUVEC. S-Rh2 was the most potent inhibitor of loop formation. In silico molecular docking predicted a good binding score between Rh2 or PPD and the ATP-binding pocket of VEGFR2. VEGF bioassay showed that Rh2 was an allosteric modulator of VEGFR2. In addition, SRh2 and PPD had good binding scores with AQP1 and AQP5, both of which play roles in cell migration and proliferation. Conclusion: The combination of these molecules might be responsible for the anti-cancer effects observed by Rg3.