• Journal of Periodontal and Implant Science
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• v.34 no.4
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• pp.759-769
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• 2004

Periodontal therapy has dealt primarily with attempts at arresting progression of disease, however, more recent techniques have focused on regenerating the periodontal ligament having the capacity to regenerate the periodontium. The effect of chitosan, a carbohydrate biopolymer extracted from chitin, on periodontal ligament regeneration is of particular interest. The purpose of this study was to evaluate the effect of chitosan on the expression of extracellular matrix proteins in primary rat calvarial cells in Vitro. In the control group, cells was cultured with BGjb media. In the experimental groups, cells were cultured with chitosan in concentration of 0.01, 0.1, 1.0 and 2.0 mg/ml. Then each group was characterized by examining alkaline phosphatase activity at 3 and 7 days, and the ability to produce mineralized nodules of rat calvarial cells at 14 and 21 days. Synthesis of type I collagen (COL-I), osteocalcin (OCN), bone sialoprotein (BSP) was evaluated by RT-PCR at 14 days. The results were as follows: 1. Alkaline phosphatase activity was significantly higher in the concentration of chitosan 0.01mg/ml, 0.1mg/ml and 1.0mg/ml compared to control (p<0.05). 2. The percentage of mineralized bone nodule was more in the concentration of chitosan 0.1mg/ml and 1.0mg/ml than the control. 3. At 14 day culture, the expression of OCN was increased by chitosan in concentration of 1.0 mg/ml and 2.0 mg/ml. These results suggested that chitosan in concentration of 0.1 and 1,0 mg/ml stimulate the extracellular matrix of primary rat calvarial cells and may facilitate the formation of bone.

• The Journal of Korean Obstetrics and Gynecology
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• v.15 no.4
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• pp.1-16
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• 2002

This work examines the effect of treatment with Gyukhachukeotang on the growth of uterine myomal cells. Comparisons of cell growth, MAP kinase activity and expression of bcl-2 (apoptosis-related gene) were made between the control and experimental samples. The results as fallows; 1. Any concentration of Gyukhachukeotang above 0.01% yielded growth inhibition. Concentrations of 5% and 10% stopped all cell growth, demonstrating the effectiveness of Gyukhachukeotang as a growth inhibitor on uterine myomal cells. 2. The MAP kinase activity in uterine myomal cells treated with Gyukhachukeotang was decreased to a high degree at the concentration of 10%, and some inhibition of activity was detected at a concentration of 5%. 3. The expression of bcl-2, a Cell Apoptosis-related gene, in uterine myoma cells treated with Gyukhachukeotang was gradually increased with increasing concentration of Gyukhachukeotang. These results indicate the ability of Gyukhachukeotang to control uterine myomal cell growth, with concurrent reduction of MAP kinase activity. Treatment with Gyukhachukeotang appears to trigger a normal apoptosis response, as indicated by increased bcl-2 expression. This observed increase in apoptosis indicates that Gyukhachukeotang is an appropriate prescription to treat uterine myomal cells.

• Journal of Korean Medicine Rehabilitation
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• v.19 no.4
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• pp.59-78
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• 2009

Objective : This study was designed to examine the effects of stem bark extracts of Cornus walteri Wanger on the lipid lowering, anti-oxidative activity and concentration of proinflammatory cytokines in hyperlipidemic rat. Methods : Male rats weighing $195.21{\pm}5.85g$ fed high fat diet for 8 weeks and 40 rats(above 400 g) were divided into 4 groups. Each groups were divided into a control group and 3 experimental groups. We fed a control group of rats a basal diet and administered normal saline(100 mg/kg, 1 time/1 day) for 4 weeks. And we fed each experimental group of rats Basal diet and administered an extract of Cornus walteri Wanger(100 mg/kg, 200 mg/kg, 300 mg/kg, 1 time/1 day) for 4 weeks. At the end of the experiment, the rats were sacrificed to determine their chemical composition. We measured lipid of plasma and liver, concentration of proinflammatory cytokines, anti-oxidative activity and gene expression. Results : 1. Concentration of plasma free fatty acid, LDL-cholesterol showed a tendency to decrease in Cornus walteri Wanger ext. groups. Concentration of plasma triglyceride, total cholesterol showed a significantly decrement in all Cornus walteri Wanger ext. group than that of control group. HDL-cholesterol showed a significantly increment in the 300 mg/kg Cornus walteri Wanger ext. group. 2. Concentration of liver total cholesterol showed a tendence to decrease in Cornus walteri Wanger ext. groups. Concentration of triglyceride liver showed a significantly decrement in all Cornus walteri Wanger ext. group than that of control group. 3. Concentration of plasma and liver TBARS showed a tendence to decrease in Cornus walteri Wanger ext. groups. The values of GSH-Px, SOD and CAT activity showed a significantly increment in the 300 mg/kg Cornus walteri Wanger ext. group than that of control group. 4. The values of plasma AST and ALT activity showed no significantly different in all treatment groups. 5. Concentration of plasma $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ showed a tendency to decrease in the Cornus walteri Wanger ext. groups. However the concentration of IL-10 in the 200 and 300 mg/kg Cornus walteri Wanger ext. groups showed a significantly increment than that of control group. Concentration of liver $IL-1{\beta}$, $TNF-{\alpha}$ and IL-10 showed no significantly difference in all treatment groups. However concentration of IL-6 in the 200 and 300 mg/kg Cornus walteri Wanger ext. groups showed a significantly decrement than that of control group. 6. In the analysis of RT-PCR, gene expression of $TNF-{\alpha}$, Apo-B, Apo-E and leptin in the Cornus walteri Wanger ext. groups showed a lower expression than that of control group. 7. The ratio of $TNF-{\alpha}$, Apo-E and leptin expression per $\beta$-actin expression in the 200 and 300 mg/kg Cornus walteri Wanger ext. showed a significantly decrement than that of control group. The ratio of Apo-B expression per $\beta$-actin expression in the 300 mg/kg Cornus walteri Wanger ext. showed a significantly decrement than that of control group. Conclusions : According to above results, in lowering lipid effect, antioxidative activity and antiinflammatory effect, the Cornus walteri Wanger ext. gives positive effect.

• BMB Reports
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• v.36 no.5
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• pp.493-498
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• 2003

The scFv antibody towards the Burkholderia pseudomallei exotoxin was previously constructed by phage display and exhibited good specificity towards the exotoxin. We report here the optimization of the scFv expression in an E. coli expression system. Four different E. coli strains (ER2537, TG1, HB2151, and XL1-Blue) were examined for optimal expression of the scFv protein. Two types of carbon source (i.e. 0.2% glucose and 0.2% glycerol) were also tested for their ability to induce the scFv expression. Cells that carried the scFv construct were grown at $30^{\circ}C$ and induced with 0.05 mM IPTG. The expression was then monitored by SDS-PAGE, Western blotting, and indirect ELISA. The Western blot profile showed different levels of the scFv expression among the host strains; XL1-Blue exhibited the highest level of the scFv protein expression. Glycerol at a concentration of 0.2% (v/v) significantly increased the scFv protein expression level when compared to 0.2% (w/v) glucose. Further optimization demonstrated that the scFv protein expression in XL1-Blue was the most optimal with a glycerol concentration as low as 0.05%. However, by indirect ELISA, only the scFv protein that was expressed in 0.2% (v/v) glycerol exhibited high specificity towards the Burkholderia pseudomallei exotoxin.

• Development and Reproduction
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• v.17 no.3
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• pp.275-287
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• 2013

Although, one of the etiologies of localized lipodystrophy of the subcutaneous connective tissue (cellulite) is the histological alternation of adipose tissue, the characteristics of expression of the components of extracellular matrix (ECM) components during adipogenesis are not uncovered. In this study, the effects of caffeine and Ishige okamurae originated diphlorethohydroxycarmalol (DPHC) on the expression of extracellualr fibers was analyzed with quantitative RT-PCR during differentiation induction of mouse subcutaneous adipose derived stem cells (msADSC) into adipocyte. The expression levels of Col1a, Col3a1, and Col61a were decreased by the adipogenci induction in a time-dependent manners. However, Col2a mRNA and Col4a1 mRNA expressions were oposit to them. Caffeine and DPHC stimulated the changes of the expression of these collagens. Eln mRNA expression was increased by induction. DPHC stimulated the expression of it. Mfap5 mRNA expression was deceased in both adipogenic cell and matured adipocytes. Caffeine suppressed the expression of Mfap5 but the effect of DPHC was different by the concentration. The expression of bioglycan, decorin, and lumican were also modified by caffeine and DPHC in a concentration-dependent manner. Based on this study, we revealed firstly the effects of caffeine and DPHC on the expression of collagens, elastin, and glycoproteins during adipogenesis of msADSCs. Those results suggest that DPHC may have antiadipogenic effect and has more positive effets on normal adipose tissue generation and work as suppressor the abnormality of ECM structure. Such results indicate that DPHC can be applied in keeping the stability of the ECM of adipogenic tissues.

• The Korea Journal of Herbology
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• v.34 no.1
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• pp.91-98
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• 2019

Objectives : While inducing inflammatory response due to LPS it will investigate mechanism associated with anti-inflammatory effects from macrophages and provide basic data for the possible use as anti-inflammatory materials. Methods : We investigated cell viability, NO, $TNF-{\alpha}$ and IL-6 by ELISA and expressions of iNOS, COX-2, MAPKs and $NF-{\kappa}B$ were measured in RAW 264.7 cells induced by LPS. Results : The cell viability of Parthenocissus tricuspidata extracts(PTE) identified in macrophages showed that cell viability rate was more than 99% at the concentration of 8, 40, and $200{\mu}g/mL$. NO generated amounts revealed that it relied on concentration and was significantly reduced compared to the control. The expression of iNOS was restrained by the control at the concentration of 200 and $400{\mu}g/mL$. In addition, the expression of COX-2 was found to be significantly reduced to the untreated control at the concentration of $400{\mu}g/mL$. $TNF-{\alpha}$ relied on concentration and showed a significant decreased compared to the control. In contrast, IL-6 relied on concentration, reduced compared to the control. Phosphorylation of ERK, JNK, and p38 mediated by LPS were restrained by relying on concentration. Phosphorylation and decomposition of $I{\kappa}B{\alpha}$ as well as p65 nuclear transmission of $NF-{\kappa}B$ subunit were restrained. Conclusions : By restraining the activation of $NF-{\kappa}B$, anti-inflammatory effects were revealed by reducing phosphorylative activation of MAPKs, restraining the expression of iNOS and COX-2 and restraining the creation of NO, IL-6, and $TNF-{\alpha}$. Therefore, it can be assumed that they can be used as a variety of anti-inflammatory materials.

• The Journal of Korean Obstetrics and Gynecology
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• v.21 no.2
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• pp.17-26
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• 2008

Purpose: This study was conducted to investigate the effects of Cervi Pontotrichum Cornu extract solution on the cell cycle regulation and apoptosis in human leiomyoma cell. Methods: The leiomyoma cell of patients was used in the study, and we administered the extract solution of Cervi Pontotrichum Cornu concentration at 1, $10mg/m{\ell}$ to the leiomyoma cell for 48 hours. We used flow cytometry and western blotting to confirm cell cycle and apoptosis. Results: In flow cytometry, G1 phase of the $1mg/m{\ell}$ group prolonged. But G1 phase of $10mg/m{\ell}$ group was shortened and S phase was increased. Cyclin D1 expression increased in higher concentration group. And Bax expression that regulates cell apoptosis increased in $1mg/m{\ell}$ and $10mg/m{\ell}$ group than control group. Bcl-2 expression decreased in 1, $10mg/m{\ell}$ groups than control group. VEGF expression rised in higher Cervi Pontotrichum Cornu concentration group. Conclusion: This study means that Cervi Pontotrichum Cornu could induce the apoptosis of leiomyoma cell by increasing Bax and decreasing Bcl-2 expression. But Cervi Pontotrichum Cornu could increase Cyclin D1 and VEGF expression, so more detailed studies would be needed.

• Journal of Life Science
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• v.34 no.2
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• pp.105-112
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• 2024

In this study, the various whitening activities of Inonotus obliquus were assessed for potential use as functional cosmetic materials. When measuring electron donating ability to confirm the antioxidant ability of I. obliquus extract, increased activity was observed as the concentration increased, and it was found an outstanding antioxidant capacity of 82.1% at a 1,000 ㎍/ml concentration. Also, the tyrosinase inhibitory effect, related to a whitening effect, was found to have inhibitory activity that increased in a concentration-dependent manner. The results of verifying the viability of melanoma cells (B16F10) using an MTT assay showed cell viability of more than 80% at concentrations below 100 ㎍/ml. Therefore, cell-related experiments were conducted at 25, 50, and 100 ㎍/ml concentrations. By measuring protein expression related to melanin synthesis via treating B16F10 cells with I. obliquus extract, it was confirmed that protein expression was inhibited in all factors, depending on the concentration. TRP-1 and MITF appeared by 40.1% and 64.2% amount of expression, respectively, at 100 ㎍/ml concentrations, and tyrosinase and TRP-2 were verified as having better protein expression inhibition than arbutin. In measuring mRNA expression related to melanin synthesis by treating B16F10 cells with I. obliquus extract, it was confirmed that mRNA expression was suppressed as the concentration increased. Accordingly, it was confirmed that I. obliquus extract has excellent whitening activity and could be used as a cosmetic material.

• Journal of Microbiology
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• v.33 no.2
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• pp.120-125
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• 1995

.betha.-galactosidase activity of Escherichia coli cells containing operon fusion nhaA'-'lacZ was monitored to study the regulation of expression of nhaA gene under various conditions. The expression of the fusion was enhanced only by chemicals containing Na$^{+}$ or Li$^{+}$. This Na$^{+}$ or Li$^{+}$. This Na$^{+}$(Li$^{+}$)-specific enhancement of .betha.-galactosidase activity represented the increase in the rate of synthesis of .betha.-galactosidase rather than the decrease in the breakdown rate. The induction pattern was influenced by copy numbers of the gene. Induction by Na$^{+}$ or Li$^{+}$ was concentration and time dependent, reaching maximum 5-6 fold induction after 2 hours at 0.4-0.5 M for Na$^{+}$ or at 0.25-0.35 M for Li$^{+}$, Although the expression was induced at much lower concentration of Na$^{+}$ at alkaline pH values than at neutral pH in the presence of Na$^{+}$, alkaline pH itself did ot induced the expression of the fusion in the absence of Na$^{+}$. Temperature shift and growth phase of culture did not affect the level of induction.he level of induction.

• Genomics & Informatics
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• v.8 no.1
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• pp.50-57
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• 2010

Mercuric chloride, a model nephrotoxicant was used to elucidate time- and dose- dependent global gene expression changes associated with proximal tubular toxicity. Rat kidney cell lines NRK-52E cells were exposed for 2, 6 and 12 hours and with 3 different doses of mercuric chloride. Cell viability assay showed that mercuric chloride had toxic effects on NRK-52E cells causing 20% cell death (IC20) at $40{\mu}M$ concentration. We set this IC20 as high dose concentration and 1/5 and 1/25 concentration of LC20 were used as mid and low concentration, respectively. Analyses of microarray data revealed that 738 genes were differentially expressed (more than two-fold change and p<0.05) by low concentration of mercuric chloride at least one time point in NRK-52E cells. 317 and 2,499 genes were differentially expressed at mid and high concentration of mercuric chloride, respectively. These deregulated genes showed a primary involvement with protein trafficking (CAV2, CANX, CORO1B), detoxification (GSTs) and immunity and defense (HMOX1, NQO1). Several of these genes were previously reported to be up-regulated in proximal tubule cells treated with nephrotoxicants and might be aid in promoting the predictive biomarkers for nephrotoxicity.