• The Korean Journal of Zoology
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• v.25 no.1
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• pp.9-28
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• 1982

Cyclical changes in the fine structures of the surface epithelial, stroma and glandular cells of guinea pig endometrium during the estrous cycle were studied by transmission and scanning electron microscopy. Cytochemical studies were made in order to investigate the ultrastructural localization of the acid phosphatase, alkaline phosphatase and ATPase in these cells. The results obtained are as follows: 1. The endometrial surface epithelium was pseudostratified columnar during estrus and meterstrus, and simple columnar during proestrus and diestrus. The characteristic features observed in these cells include increased nucleocytoplasmic ratio at proestrus, elongated shapes of both the nucleus and the entire cell, increased volume of the cytoplasm and cytoplasmic bulding into the lumen during estrus, and smaller surface epithelial cells during metestrus. 2. In the cytoplasm of surface epithelial cells, the numbers of mitochondria and free ribosomes were increased, and rough endoplasmic reticulum and Golgi complex appeared during estrus, and the degenerated cells, lipid droplets, multilamellated bodies and lysosomes appeared during diestrus. 3. During estrus, scanning electron microscopic observations of endometrial surface showed a regular arrangement with polygonal outlines of epithelial cells, distinct intercellular border, and bulged surface into the lumen, whereas flat surface and indistinct cell border were characteristic during meterstrus and diestrus. 4. Microvilli which aligned on the surface were longer and most abundant during estrus while short and aparse during other phases. 5. Cytochemical studies indicated that during metestrus acid phosphatase activities were localized in the microvilli and vacuoles, and alkaline phosphatase activities were significant around luminal surface and lateral cell membrane in the surface epithelial cells. ATPase activities were present on the microvilli and cell membrane during proestrus and estrus.

• BMB Reports
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• v.43 no.6
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• pp.432-437
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• 2010

LBH is a transcription factor as a candidate gene for CHD associated with partial trisomy 2p syndrome. To identify potential LBH-interacting partners, a yeast two-hybrid screen using LBH as a bait was performed with a human heart cDNA library. One of the clones identified encodes ${\alpha}B$-crystallin. Co-immunoprecipitation and GST pull-down assays showed that LBH interacts with ${\alpha}B$-crystallin, which is further confirmed by mammalian two-hybrid assays. Co-localization analysis showed that in COS-7 cells, ${\alpha}B$-crystallin that is cytoplasmic alone, accumulates partialy in the nucleus when co-transfected with LBH. Transient transfection assays indicated that overexpression of LBH or ${\alpha}B$-crystallin reduced the transcriptional activities of p53 and p21, respectively, Overexpression of both ${\alpha}B$-crystallin and LBH together resulted in a stronger repression of the transcriptional activities of p21 and p53. These results showed that the interaction of LBH and ${\alpha}B$-crystallin may inhibit synergistically the transcriptional regulation of p53 and p21.

• Journal of Life Science
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• v.19 no.10
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• pp.1484-1488
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• 2009

Familial hypokalemic periodic paralysis (HOKPP) is an autosomal dominant muscle disorder characterized by episodic attacks of muscle weakness with concomitant hypokalemia. Mutations in either a calcium channel gene (CACNA1S) or a sodium channel gene (SCN4A) have been shown to be responsible for this disease. The combination of sarcolemmal depolarization and hypokalemia has been attributed to abnormalities of the potassium conductance governing the resting membrane potential. To understand the pathophysiology of this disorder, we examined both mRNA and protein levels of delayed rectifier potassium channel genes, KCNQ3 and KCNQ5, in skeletal muscle fibers biopsied from patients with HOKOur results showed an increase in the cytoplasmic level of KCNQ3 protein in patients' cells exposed to 50 mM external concentration of potassium. However, mRNA levels of both channel genes did not show significant change in the same condition. Our results suggest that long term exposure of skeletal muscle cells in HOKPP patients to high extracellular potassium alters the KCNQ3 localization, which could possibly hinder the normal function of this channel protein. These findings may provide an important clue to understanding the molecular mechanism of familial hypokalemic periodic paralysis.

• Tuberculosis and Respiratory Diseases
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• v.69 no.2
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• pp.81-94
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• 2010

Background: Autophagy is an important adaptive mechanism in normal development and in response to changing environmental stimuli in cancer. Previous papers have reported that different types of cancer underwent autophagy to obtain amino acids as energy source of dying cells in nutrient-deprived conditions. However, whether or not autophagy in the process of lung cancer causes death or survival is controversial. Therefore in this study, we investigated whether nutrient deprivation induces autophagy in human H460 lung cancer cells. Methods: H460, lung cancer cells were incubated in RPMI 1640 medium, and the starved media, which are BME and RPMI media without serum, including 2-deoxyl-D-glucose according to time dependence. To evaluate the viability and find out the mechanism of cell death under nutrient-deprived conditions, the MTT assay and flow cytometry were done and analyzed the apoptotic and autophagic related proteins. It is also measured the development of acidic vascular organelles by acridine orange. Results: The nutrient-deprived cancer cell is relatively sensitive to cell death rather than normal nutrition. Massive cytoplasmic vacuolization was seen under nutrient-deprived conditions. Autophagic vacuoles were visible at approximately 12 h and as time ran out, vacuoles became larger and denser with the increasing number of vacuoles. In addition, the proportion of acridine orange stain-positive cells increased according to time dependence. Localization of GFP-LC3 in cytoplasm and expression of LC-3II and Beclin 1 were increased according to time dependence on nutrient-deprived cells. Conclusion: Nutrient deprivation induces cell death through autophagy in H460 lung cancer cells.

• Journal of Periodontal and Implant Science
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• v.49 no.3
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• pp.138-147
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• 2019

Purpose: Several studies have shown that the oral cavity is a secondary location for Helicobacter pylori colonization and that H. pylori is associated with the severity of periodontitis. This study investigated whether H. pylori had an effect on the periodontium. We established an invasion model of a standard strain of H. pylori in human periodontal ligament fibroblasts (hPDLFs), and evaluated the effects of H. pylori on cell proliferation and cell cycle progression. Methods: Different concentrations of H. pylori were used to infect hPDLFs, with 6 hours of co-culture. The multiplicity of infection in the low- and high-concentration groups was 10:1 and 100:1, respectively. The Cell Counting Kit-8 method and Ki-67 immunofluorescence were used to detect cell proliferation. Flow cytometry, quantitative real-time polymerase chain reaction, and western blots were used to detect cell cycle progression. In the high-concentration group, the invasion of H. pylori was observed by transmission electron microscopy. Results: It was found that H. pylori invaded the fibroblasts, with cytoplasmic localization. Analyses of cell proliferation and flow cytometry showed that H. pylori inhibited the proliferation of periodontal fibroblasts by causing G2 phase arrest. The inhibition of proliferation and G2 phase arrest were more obvious in the high-concentration group. In the low-concentration group, the G2 phase regulatory factors cyclin dependent kinase 1 (CDK1) and cell division cycle 25C (Cdc25C) were upregulated, while cyclin B1 was inhibited. However, in the high-concentration group, cyclin B1 was upregulated and CDK1 was inhibited. Furthermore, the deactivated states of tyrosine phosphorylation of CDK1 (CDK1-Y15) and serine phosphorylation of Cdc25C (Cdc25C-S216) were upregulated after H. pylori infection. Conclusions: In our model, H. pylori inhibited the proliferation of hPDLFs and exerted an invasive effect, causing G2 phase arrest via the Cdc25C/CDK1/cyclin B1 signaling cascade. Its inhibitory effect on proliferation was stronger in the high-concentration group.

• Tuberculosis and Respiratory Diseases
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• v.46 no.2
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• pp.185-194
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• 1999

Background: NF-$\kappa$B is a characteristic transcriptional factor whose functional activity is determined by post-translational modification of protein and subsequent change of subcellular localization. The involvement of the NF-$\kappa$B family of the transcription factors in the control of such vital cellular functions as immune response, acute phase reaction, replication of certain viruses and development and differentiation of cells has been clearly documented in many previous studies. Several recent observations have suggested that the NF-$\kappa$B might also be involved in the carcinogenesis of some hematological and solid tumors. Investigating the possibility that members of the NF-$\kappa$B family participate in the molecular control of malignant cell transformation could provide invaluable information on both molecular pathogenesis and cancer-related gene therapy. Method: To determine the expression patterns and functional roles of NF-$\kappa$B family transcription factors in human lung cancer cell lines NCI-H792, NCI-H709, NCI-H226 and NCI-H157 were analysed by western blot, using their respective antibodies. The nuclear and the cytoplasmic fraction of protein extract of these cell lines were subsequently obtained and NF-$\kappa$B expression in each fraction was again determined by western blot analysis. The type of NF-$\kappa$B complex present in the cells was determined by immunoprecipitation. To detect the binding ability of cell-line nuclear extracts to the KB consensus oligonucleotide, electrophoretic mobility shift assay(EMSA) was performed. Results: In the cultured human lung cancer cell lines tested, transcription factors of the NF-$\kappa$B family, namely the p50 and p65 subunit were expressed and localized in the nuclear fraction of the cellular extract by western blot analysis and immunocytochemistry. Immunoprecipitation assay showed that in the cell, the p50 and p65 subunits made NF-$\kappa$B complex. Finally it was shown by Electrophoretic Mobility Shift Assay(EMSA) that nuclear extracts of lung cancer cell lines are able to bind to NF-$\kappa$B consensus DNA sequences. Conclusion: These data suggest that in human lung cancer cell lines the NF-$\kappa$B p50/p65 complex might be activated. and strengthen the hypothesis that NF-$\kappa$B family transcription factors might be involved in the carcinogenesis of human lung cancer.

• Applied Microscopy
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• v.34 no.3
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• pp.171-178
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• 2004

Metallothionein (MT) is a family of ubiquitous, low molecular weight (6-7 kDa), cysteine-rich protein with a high affinity to metal ions and has no aromatic amino acids and histidine. Some of the known functions of MT include detoxification of heavy metals and alkylating agents and neutralization of free radicals. Also, this protein may affect a number of cellular processes including gene expression, apoptosis, proliferation and differentiation. But, its actual functions are still not clear. The present study was undertaken to examine immunocytochemically the localization of MT in developing rat liver. On the day 11 of gestation, the fetal rat liver has already been formed and contained numerous oval cells with high nuclear cytoplasmic ratio, which were the progenitors of hepatic parenchymal cells, but no reaction products of MT were detected at this time. And then, positive reactions against MT started to appear predominantly in the parenchymal cells of liver from the 13th day after gestation. Reaction products, immunogold particles or brown coloration, were localized at both the nucleus and the cytoplasm of the parenchymal cells, except mitochondria. The intensity of this reaction gradually increased, and exhibited the strongest at birth. The intensity of MT staining and immunogold labelling diminished with growth, and by the 15th day after birth weak positive reaction was observed in the cells. In brief, positive reactions for MT were observed in the oval cells and the parenchymal cells during fetal stage, meanwhile they were present only in the parenchymal cells after birth. The present results suggest that MT possibly involves parechymal cell proliferation and differentiation through the storage or the supply of various metal ions in the developing rat liver.

• Applied Microscopy
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• v.30 no.2
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• pp.173-183
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• 2000

This study was designed to observe the ultrastructural localization of synoviocytes, which are concerned with the function of phagocytic synovial cells (type A synoviocytes, macrophage-like synoviocytes), in the knee joint of the human for CD14 and CD105 by cryo-immune-electron microscopic technique. The synovium were dissected and fixed for two hours (in 4% paraformaldehyde and 0.1% glutaraldehyde mixture), and were immerged in 2.3 M sucrose and 20% PVP solution. Finally, they were cut with the cryoultramicrotome and labelled with primary antibodies (monoclonal mouse anti-human CD14, monoclonal mouse anti-human CD105 (endoglin) and secondary (donkey anti-mouse IgG) tagged with 6 nm colloidal gold particles. The tissues were observed under transmission electron microscope. This study was resulted as follows. 1. In the synovium of the human knee joint, CD14+ cells were identified. These cells showed phagocytic synovial cell's features. In the phagocytic synoviocyte, the distributions of CD14 were marked in the cytoplasm, around vacuoles, and in cytoplasmic process, but not detected inside of vacuoles. 2. In the synovium of the human knee joint, CD105+ cells were identified. These cells were recognized endothelial cells and phagocytic synovial cells. In the phagocytic synovial cells, the distributions of CD105 (endoglin) were marked in cytoplasic process, around vacuoles, and in cell membrane, but not detected inside of vacuoles. On the basis of above findings, it is obvious that phagocytic synovial cells were marked at CD 14 and CD 105, and might be play the role of activated macrophages or phagocytes in the synovial membrane.