• Journal of Microbiology
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• v.42 no.1
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• pp.25-31
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• 2004

Sf9 cells have obvious advantages for the conventional production technology of vaccine. They are useful tools for high concentration and large-scale cultures. Sf9 cells were grown to maximal concentration, 8${\times}$l0$\^$6/ cells/$m\ell$ in a 500$m\ell$ spinner flask, with a doubling time at the exponentially growing phase of 24.5 hours, using serum-free media. To explore the ability of Sf9 cells to be infected by the Japanese encephalitis (JE) virus Beijing-l strain, Sf9 cells were infected with the virus. By 4-5 days post-infection, 10-15 % of the Sf9 cells showed cytopathic effect (CPE), from granularity to the formation of syncytia and multinucleated giant cells continuously observed over a period of 35 days. Positive fluorescent reactions were detected in 30-40% of cells infected with the JE virus Beijing-l strain, and the uninfected Sf9 cells were completely negative. Virus particles, propagated in Sf9 and Vero cells, were concentrated by sedimentation on 40% trehalose cushions by ultracentrifugation, and showed identical patterns of viral morphogenesis. Complete virus particles, 40 to 50 nm in diameter, were observed, and JE virus envelope (E) proteins, at 53 kDa, were found in the western blot analysis to the anti-JE virus E protein monoclonal antibody and reacted as a magenta band in the same position to the glycoprotein staining. To evaluate whether the infectious virus was produced in Sf9 cells inoculated with the JE virus Beijing-l stain, Sf9 cells were inoculated with the virus, and sample harvested every 5 days. The titers of the JE virus Beijing-l strain rose from 1.0${\times}$l0$\^$5/ to 1.5${\times}$l0$\^$6/ pfu/$m\ell$. The infected Sf9 cells could be subcultured in serum-free medium, with no change in the plaque sizes formed by the JE virus Beijing-l strain in the plaque assay. It is suggested that the ability of the JE virus Beijing-l strain to infect Sf9 cells in serum-free media will provide a useful insect cell system, where the JE virus replication, cytopathogenicity and vaccine immunogen can be studied.

• Journal of dental hygiene science
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• v.19 no.4
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• pp.254-260
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• 2019

Background: The primary aims of periodontal disease treatment is to remove dental plaque and calculus, the main causes of tooth loss, and restore periodontal tissue destroyed by inflammation. Periodontal disease treatment should also help maintain the alveolar bone, alleviate inflammation, and promote periodontal ligament cell proliferation, which is essential for tissue regeneration. Conventional antibiotics and anti-inflammatories have adverse side effects, especially during long-term use, so there is a need for adjunct treatment agents derived from natural products. The purpose of this study was to investigate whether the herbal flavone baicalein has the osteogenic activity under inflammatory conditions, and assess the involvement of osteoblast immediate early response 3 (IER3) expression. Methods: Human osteoblastic MG-63 cells were cultured with the pro-inflammatory cytokines tumor necrosis factor α and interleukin 1β in the presence and absence of baicalein. Proliferation was assessed using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay, and expression of IER3 mRNA was assessed using real-time polymerase chain reaction. The expression of IER3 protein levels and activation of associated signal transduction pathways were assessed using western blotting. Results: Baicalein increased IER3 mRNA and protein expression synergistically. In addition, baicalein reversed the suppression of cell proliferation, and the downregulation of osteogenic transcription factor runt-related transcription factor 2 and osterix induced by pro-inflammatory cytokines. Baicalein also upregulated the phosphorylation of c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK 1/2). The upregulation of IER3 by pro-inflammatory cytokines was blocked by pretreatment with inhibitors of AKT, p38, JNK, and ERK 1/2. Conclusion: Baicalein mitigates the deleterious responses of osteoblasts to pro-inflammatory cytokines. Further, IER3 enhanced the effect of baicalein via activation of AKT, p38, JNK, and ERK pathways.

• The Korean Journal of Mycology
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• v.24 no.2 s.77
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• pp.142-148
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• 1996

To find compounds of immunomodulating and anti-allergic function, effects of protein-bound polysaccharides extracted from Phellinus igniarius (PI), Fomitella fraxinea (FF) and Agrocybe cylindracea (AC) on hemagglutinin titer (HA), hemolysin titer (HY), plaque forming cell (PFC), rosette forming cell (RFC) and phagocytosis were investigated in BALB/C mice. The oral administration of the protein-bound polysaccharides of PI, FF and AC for 10 days resulted in the enhanced phagocytic activity of peritoneal exudate cells (PEC), spleen cells (SC) and blood lymphocyte cells (BLC). Moreover, PI showed the activating effect on the phagocytosis of PEC and AC in SC. In the experiment of PFC and RFC, the results of the experimental group which was given each samples as compared to the control group, showed the enhanced level of activity such as PI 130%, FF 90% and AC 70%. Generally, HY and HA showed from ten to hundred times of level in each sample groups, as compared to the control group.

• Biomolecules & Therapeutics
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• v.2 no.4
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• pp.376-382
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• 1994

Repeated injections of low-doses of mercuric chloride in rats or mice induce polyclonal activation which includes the induction of anti-glomerular basement membrane (GBM) antibodies and circulating immune complex and it results in nephritis. Because this disease is autoimmune mediated disease resulted from immune dysfunction, immunomodulators are used to control the symptoms or to cure the disease. Irpex lacteus Fr. is a kind of new medicinal fungus. The polysaccharide fraction extracted from submerged fermentation of Irpex lacteus Fr. decreased the serum agglutinin, serolysin and IgM plaque forming cells in normal mice. The hitherto obtained clinical results suggested that it significantly improved the oligourea, edema, and hypertension in patients who have nephritis. To elucidate the action-mechanisms of Irpex lacteus Fr., we established the experimental model of HgCl$_2$induced polyclonal activation by intraperitoneal administrations of HgCl$_2$to mice. To assess the immunomodulating effect of Irpex lacteus fraction, we Investigated its effects on the mitogen induced proliferation and IgM PFC counts of splenic lymphocytes in mice during the treatment of HgCl$_2$. The Irpex lacteus polysaccharide reduced the abnormally increased mitogen induced Iymphocyte proliferation and IgM PFCs to almost normal levels. And the Irpex lacteus polysaccharides prevented the increasement of serum immunoglobulin level induced by HgCl$_2$. These data suggested that the Irpex lacteus polysaccharides might have the immunomodulating activity to prevent and /or improve the HgCl$_2$ induced autoimmune disease.

• Journal of Life Science
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• v.26 no.7
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• pp.835-846
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• 2016

Chemo-resistance is the biggest issue of effective cancer therapy. ABCG2 is highly correlated with multi-drug resistance, and represent a typical phenotype of multiple cancer stem-like cells. Accumulating evidence recently reported that oncolytic viruses represent a new strategy for multiple aggressive cancers and drug resistant cancers including cancer stem cell-like cells and ABCG2 expressing cells. In this study, we generated an evolutionally engineered vaccinia virus, SLJ-496, for drug-resistant cancer therapy. We first showed that SLJ-496 treatment enhanced tumor affinity using cytopathic effect assay, plaque assay, as well as cell viability assay. Next, we clearly demonstrated that in vitro SLJ-496 treatment represents significant cytotoxic effect in multiple cancers including colorectal cancer cells (HT-29, HCT-116, HCT-8), gastric cancer cells (AGS, NCI-N87, MKN-28), Hepatocellular carcinoma cells (SNU-449, SNU-423, SNU-475, HepG2), as well as mesothelioma cell (NCI-H226, NCI-H28, MSTO-221h). Highly ABCG2 expressing HT-29 cells represent cancer stem like phenotype including stem cell marker expression, and self-renewal bioactivities. Interestingly, we demonstrated that in vitro treatment of SLJ-496 showed significant cytotoxicity effect, as well as viral replication capacity in ABCG2 overexpressing cell. In addition, we also demonstrated the cytotoxic effect of SLJ-496 in Adriamycin-resistant cell lines, SNU-620 and ADR-300. Taken together, these findings provide us a pivotal clue that cancer therapy using SLJ-496 vaccinia virus might be new therapeutic strategy to overcome ABCG2 expressing cancer stem-like cell and multiple chemo-resistance cancer cells.

• The Journal of Internal Korean Medicine
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• v.15 no.1
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• pp.60-79
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• 1994

The studies were conducted to investigate the combined effects of Tonics and Mitomycin C(MMC). The effects of Tonics and MMC on the proliferation of Molt-4 cells, human leukemic cell line, and activation of human lymphocytes were estimated by MTT colorimetric assays. Selected medicines among 9 kinds of Tonics by results of MTT assays were treated with MMC in mice. The Tonics itself enhanced the proliferation of Molt-4, but the anti-proliferative effect of MMC was not intercalated by the combined treatment of Tonic and MMC. Inhibitory action of MMC was augmented by Sa Kun Ja Tang(SKT). This result was due to the inhibition of DNA synthesis. Among 9 kinds of Tonics, Sip Jean Dae Bo Tang(SDT), Saeng Maek San(SMS) and Kwi Bi Tang(KBT) did not inhibit the action of MMC, but activated lymphocytes. When the mice were treated by MMC, the number of leukocytes was decreased significantly at the 1st day, but recovered at the 7th day. In the groups of MMC treated with SDT or KBT, the number of leukocytes was increased significantly than the group of MMC treated only at the 3rd day. Combined treatment of the Tonics(SDT, SMS) and MMC retained the body weight of mice at the level of normal mice. SDT, SMS and KBT did not change the number of plaque forming cells(PFC), but MMC treated group decreased the number of PFC. The combined treatment of MMC and SDT increased the number of PFC significantly than the MMC treated group. SDT, SMS and KBT did not influence the proliferation of T cells, but MMC treated group decreased the proliferation of T cells. The combined treatment of MMC and those tonics increased the T cell proliferation significantly than the MMC treated group. In conclusion, the results presented in this paper suggest that SDT, SMS and KBT can recover the side effects of MMC, such as weight loss, leukopenia and immunosuppresion, without any intercalating the anti-proliferative action of MMC in vivo.

• The Journal of Korean Medicine
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• v.30 no.3
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• pp.1-14
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• 2009

Objectives : Alzheimer's disease (AD) is characterized by neuronal loss and extracellular senile plaque. Moreover, the cellular actions of ${\beta}$-amyloid (A${\beta}$ play a causative role in the pathogenesis of AD. This study was designed to determine whether Woo-Gui-Um, a commonly used Korean herbal medicine, has the ability to protect cortical and hippocampal neurons against A${\beta}_{25-35}$ neurotoxicity Methods : In the present study, the authors investigated the preventative effects of the water extract of Woo-Gui-Um in a mouse model of AD. Memory impairment was induced by intraventricularly (i.c.v.) injecting A${\beta}_{25-35}$ peptides into mice. Woo-Gui-Um extract was then administered orally (p.o.) for 14 days. In addition, A${\beta}_{25-35}$ toxicity on the hippocampus was assessed immunohistochemically, by staining for Tau, MAP2, TUNEL, and Bax, and by performing an in vitro study in PC12 cells. Results : Woo-Gui-Um extract had an effect to improve learning ability and memory score in the water maze task. Woo-Gui-Um extract had significant neuroprotective effects in vivo against oxidative damage and apoptotic cell death of hippocampal neurons caused by i.c.v. A${\beta}_{25-35}$. In addition, Woo-Gui-Um extract was found to have a protective effect on A${\beta}_{25-35}$-induced apoptosis, and to promote neurite outgrowth of nerve growth factor (NGF)-differentiated PC12 cells. Conclusions : These results suggest that Woo-Gui-Um extract reduces memory impairment and Alzheimer's dementia via an anti-apoptotic effect and by regulating Tau and MAP2 in the hippocampus.

• Microbiology and Biotechnology Letters
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• v.35 no.2
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• pp.118-127
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• 2007

Germanium-fortified yeast (GY) is a organic germanium-fortified yeast with potent immune modulating activities including anti-inflammatory effect. Through cell line studies, we observed that GY can modulate the diverse immune activity but little evidence was provided on the mechanism of GY in modulating immune activities in other higher animals. In this study, we investigated the effect of GY on modulation of immune function in mice. GY was administered in normal mice or tumor-bearing mice and then effect of GY on modulation of host immune system was analyzed by using ex vivo isolated macrophages, B cells, NK cells. Admistration of GY in mice induced macrophage activation thereby increased effector function of macrophage such as increased phagocytosis, chemotaxis, adherence, $O_2-release$, NO, $TNF-{\alpha}$ production. In addition, GY administration Increased B lymphocyte activation and plaque forming cells. Furthermore, GY administration increased NK-cell mediated cytotoxicity. Furthermore, GY administration suppressed progression of tumor in mice by increasing $TNF-{\alpha}$ production and effector function of NK cells. Our results showed that GY has a potent immunostimulatory function in vivo mice model. Proper modulation and administration of GY in human could be helpful to maintaining immunological homeostasis by modulating host immune system.

• The Journal of Korean Academy of Prosthodontics
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• v.44 no.3
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• pp.284-294
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• 2006

Statement of problem. Proliferation of Candida albicans is primarily within the plaque on the fitting surface of the denture rather than on the inflamed mucosa. Consequently, the treatment of the denture is equally important as treatment of the tissue. Cleansing and disinfection should be efficiently carried-out as the organisms can penetrate into the voids of the acrylic resin and grow in them, from which they can continue to infect and reinfect bearing tissues. Purpose. The purpose of this study was to evaluate the applicability of photocatalytic reaction to eliminate Candida albicans from acrylic resin denture base, and to investigate the anti-fungal effect with various UVA illumination time. Materials and Methods. The specimens were cured by the conventional method following the manufacturer's instruction using thermal polymerized denture base resin (Vertex RS: Dentimex, Netherlands). $TiO_2$ photocatalyst sol(LT), which is able to be coated at normal temperature, was made from the Ti-alkoxide progenitor. The XRD patterns, TEM images and nitrogen absorption ability of the $TiO_2$ photocatalyst sol(LT) were compared with the commercial $TiO_2$ photocatalyst P-25. The experimental specimens were coated with the mixture of the $TiO_2$ photocatalyst sol(LT) and binder material (silane) using dip-coater, and uncoated resin plates were used as the control group. Crystallinity of $TiO_2$ of the specimen was tested by the XRD. Size, shape and chemical compositions were also analyzed using the FE-SEM/ EDS. The angle and methylene blue degradation efsciency were measured for evaluating the photocatalytic activity of the $TiO_2$ film. Finally, the antifungal activity of the specimen was tested. Candida albicans KCTC 7629(1 ml, initial concentration $10^5$ cells/ ml) were applied to the experiment and control group specimens and subsequently two UVA light source with 10W, 353 nm peak emission were illuminated to the specimens from 15cm above. The extracted $2{\mu}l$ of sample was plated on nutrient agar plate ($Bacto^{TM}$ Brain Heart Infusion; BD, USA) with 10 minute intervals for 120 minute, respectively. It was incubated for 24 hours at $37^{\circ}C$ and the colony forming units (CFUs) were then counted. Results. Compared the characteristics of LT photocatalyst with commercial P-25 photocatalyst, LT were shown higher activity than P-25. The LT coated experimental specimen surface had anatase crystal form, less than 20 nm of particle size and wide specific surface area. To evaluate the photocatalytic activity of specimens, methylene blue degradation reaction were used and about 5% of degradation rate were measured after 2 hours. The average contact angle was less than $20^{\circ}$ indicating that the LT photocatalyst had hydrophilicity. In the antifungal activity test for Candida albicans, 0% survival rate were measured within 30 minute after irradiation of UVA light. Conclusion. From the results reported above, it is concluded that the UVA-LT photocatalytic reaction have an antifungal effect on the denture surface Candida albicans, and so that could be applicable to the clinical use as a cleaning method.

• Food Science and Preservation
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• v.14 no.2
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• pp.213-219
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• 2007

To develop an anti-dementia agent with potential therapeutic value in the protection of neuronal cells, we selected a water extract of Helianthus annuus seed for analysis. We measured acetylcholinesterase inhibitory activity in the extract, and analyzed the protective effect of the extract on neuronal cell death induced by hydrogen peroxide, or amyloid ${\beta}-peptide$, of SH-SY5Y neuroblastoma cells. The result showed that the extinct exerted protective effects of 83%, 72% and 53% respectively, on cell death induced by 100M, 200M, and 500M hydrogen peroxide. Also, when 50M of amyloid ${\beta}-peptide$ was added to the cells, the extract showed a protective effect (up to 80%) on cell death. Overall, the results showed that the H. annuus extract inhibited acetylcholinesterase activity in a dose-dependent manner, and the extract also strongly protected against cell death induced by hydrogen peroxide or amyloid ${\beta}-peptide$.