• Journal of Periodontal and Implant Science
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• v.26 no.4
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• pp.841-858
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• 1996

Epidermal growth factor(EGF) is one of polypeptide growth factors. EGF has been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. The purposes of this study is to evaluate the effects of EGF on the human periodontal ligament cells and human gingival fibroblast cells that promote regeneration of periodntal tissue. The mitogenic effects of epidermal growth factor on human periodontal ligament cells and human gingival fibroblasts were evaluated by determining the incorporation of 5-Bromo-2'-deoxy-uridine into DNA of the cells in a dose dependent manner. The prepared cells were the primary cultured gingival fibroblast and periodontal ligament cells from humans, the fourth or sixth subpassages were used in the experiments. Cells were seeded in DMEM containing 10% FBS. 1, 10, 50, 100, $200{\eta}g/ml$ and epidermal growth factor were added to the quiescent cells for 24 hours, 48 hours and 72 hours. They were labeled with $10\{mu}l/200{\mu}l$ 5-Bromo-2'-deoxy-uridine for the last 6 hours of each culture. The results of the five determinants were presented as mean and S.D.. The results were as follows : The DNA synthetic activity of human gingival fibroblasts were increased dose dependently by epidermal growth factor at 24 hours, 48 hours and 72 hours. The mitogenic effects were similar at the 24 and 48 hours of epidermal growth factor, but the DNA synthetic activity of human gingival fibroblasts generally decreased at 72 hours. The DNA synthetic activity of human periodontal ligament cells were increased dose dependently by epidermal growth factor at 24 hours but the DNA synthetic activity decreased at $200{\eta}g/ml$ of each hour. Generally the maximum mitogenic effects were observed at the 48 hours application of epidermal growth factor. The DNA synthetic activity of human periodontal ligament cells generally decreased lower at 24, 72 hours than at 48 hours the application of epidermal growth factor. In the comparison of DNA synthetic activity between human gingival fibroblasts and human periodontal ligament cells, human periodontal ligament cells had slightly higher proliferation activity than human gingival fibroblasts for a longer time at the high dosage of the epidermal growth factor. In conclusion, epidermal growth factor have important roles in the stimulation of DNA synthesis in human periodontal ligament cells and human gingival fibroblasts, and thus may be useful for clinical applications in periodontal regenerative procedures.

• The Korean Journal of Food And Nutrition
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• v.27 no.1
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• pp.43-49
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• 2014

Panax ginseng C.A. Meyer has been used as a traditional medicinal ingredient and the ginseng flower-buds also proved to have good medicinal properties. In this study, in order to enhance immune activities of ginseng flower-bud, the ginseng flower-bud extract was being fermented by Bacillus subtilis KCTC 1022 (BS), Lactobacillus plantarum KCTC3 and Saccharomyces cerevisiae strain CHY1011 (SC). Mice were orally administered daily for two weeks at two different concentrations (100 and 200 mg/kg B.W.). Treatment samples were water extracts of ginseng flower-buds (FD), water extracts of fermented ginseng flower-buds (FM) and controls for saline solution. Cytokine production (IL-6, TNF-${\alpha}$) either stimulated with LPS or not stimulated with LPS was detected by the ELISA assay when using the cytokine kit. Cytokine was statistically increased at supplemented groups with LPS in both the 100 and the 200 mg/kg B.W. and treatment with FM significantly decreased the LPS-induced TNF-${\alpha}$ and IL-6 production more than the treatment with FD. The results of this study may suggest that supplementation with FM increases the immune function by regulating cytokine production capacity for activated macrophages.

• Korean Journal of Food Science and Technology
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• v.38 no.1
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• pp.104-108
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• 2006

Water extracts of Jehotang were evaluated for their growth-promoting effects on Bifidobacterium longum, Lactobacillus sp., L. acidophilus, and Clostridium perfringens. Addition of Jehotang water extract to modified EG media at 0.1 mg/mL increased growths of B. longum, Lactobacillus sp., and L. acidophilus, with 1.8-fold increase in growth of L. acidophilus compared to that of control. Studies on these strains by agar diffusion method showed Lactobacillus sp. and L. acidophilus were activated by addition of Jehotang extract at 10 mg/disc. Proliferation responses of mice splenocytes and Peyer's patch cells to ConA by LPS-stimulation at 500 mg/kg B.W./day Jehotang extract were investigated in vitro. Upon treatment of 1 mg/mL Jehotang water extract to mice, proliferations of splenocytes and Peyer's patch cells increased 1.4- and 1.6-fold compared to control, respectively. In mice administered Jehotang extract, production of intestinal secretory IgA (sIgA) increased 2.4-fold compared to control. These results indicate water extract of Jehotang stimulated intestinal immune system of mice. In mice treated with Jehotang extract, production of lymphocytes was 4% lower, whereas those of granulocytes and platelets were 4% and slightly higher than control, respectively.

• Korean Journal of Veterinary Research
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• v.35 no.3
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• pp.543-552
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• 1995

This study was undertaken to investigate the eftects of Cd administered ad libitum for 6 weeks on the cellular immune responses of Balb/c mice. The results were summarized as follows; 1. The mice fed 25, 50 and 100ppm Cd drank as much as control, but the mice fed 200ppm Cd drank significantly less water after Cd exposure than did control. Increasing rates of body weight of Cd-fed mice for 6 weeks were as this, control group 27.0%, Cd administered groups(25, 50, 100 and 200ppm) 28.54%, 28.31%, 20.49% and 18.04%, respectively. 2. Absolute spleen to body weight(mg/g) of control, 25, 50, 100 and 200ppm Cd administered groups were $4.34{\pm}0.23$, $4.20{\pm}0.54$, $4.80{\pm}0.87$, $4.25{\pm}0.32$ and $4.40{\pm}0.32$, respectively. Splenic cellularity(${\times}10^7$) of control was $24.29{\pm}5.98$ but increased to $27.72{\pm}5.48$, $32.96{\pm}8.44$, $28.32{\pm}8.76$ and $29.64{\pm}4.08$ in 25, 50, 100 and 200ppm Cd-fed groups, respectively. 3. Total $CD_4{^+}$ cells(${\times}10^7$) of control, 25, 50, 100 and 200ppm Cd-fed groups were $9.15{\pm}2.24$, $10.40{\pm}2.04$, $12.04{\pm}3.08$, $10.20{\pm}3.16$ and $10.80{\pm}1.48$, respectively and total $CD_8{^+}$ cells(${\times}10^7$) of these groups were $2.32{\pm}0.56$, $2.54{\pm}0.27$, $3.12{\pm}0.80$, $2.25{\pm}0.70$ and $2.24{\pm}0.28$, in order. On the other hand, $CD_4{^+}/CD_8{^+}$ ratios in total cells were increased significantly except for 50ppm Cd-fed group($3.88{\pm}0.01$). And that of control was $3.97{\pm}0.02$, but those of 25, 100 and 200ppm were $4.35{\pm}0.01$, $4.54{\pm}0.03$ and $4.81{\pm}0.03$. 4. Phagocytosis rates of peritoneal macrophages were increased significantly in 25 and 50ppm Cd groups($36.34{\pm}9.45$ and $37.15{\pm}9.22$, respectively), but 100 and 200ppm groups showed similar rates($18.20{\pm}3.04$ and $19.48{\pm}3.22$ respectively) to that of control($21.43{\pm}3.62$). 5. In mitogen-induced splenocyte proliferation, various concentraions of $CdCl_2(10^{-4}-10^{-7}M)$ were added to mitogen-stimulated culture in vitro. Splenocyte proliferation induced by LPS was decreased dose dependently, but proliferation by Con-A was increased slightly in concentrations of $10^{-7}-10^{-6}M$. 6. Significant cytotoxicity of splenocytes with $CdCl_2$ were shown at $10^{-4}M$ treated group, especially at 24 hrs. From these results, it could be concluded that Cd might modulate the immune responses by modifying a distribution of T cell subpopulations.

• Tuberculosis and Respiratory Diseases
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• v.45 no.4
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• pp.861-869
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• 1998

Backgrounds: The injury of a tissue results in the infalmmation, and the imflammed tissue is replaced by the normal parenchymal cells during the process of repair. But, constitutional or repetitive damage of a tissue causes the deposition of collagen resulting in the loss of its function. These lesions are found in the lung of patients with idiopathic pulmonary fibrosis, complicated fibrosis after diffuse alveolar damage (DAD) and inorganic dust-induced lung fibrosis. The tissue from lungs of patients undergoing episodes of active and/or end-stage pulmonary fibrosis shows the accumulation of inflammatory cells, such as mononuclear cells, neutrophils, mast cells and eosinophils, and fibroblast hyperplasia. In this regard, it appears that the inflammation triggers fibroblast activation and proliferation with enhanced matrix synthesis, stimulated by inflammatory mediators such as interleukin-1 (IL-1) and/or tumor necrosis factor (TNF). It has been well known that TGF-$\beta$ enhance the proliferation of fibroblasts and the production of collagen and fibronectin, and inhibit the degradation of collagen. In this regard, It is likely that TGF-$\beta$ undergoes important roles in the pathogenesis of pulmonary fibrosis. Nevertheless, this single cytokine is not the sole regulator of the pulmonary fibrotic response. It is likely that the balance of many cytokines including TGF-$\beta$, IL-1, IL-6 and TNF-$\alpha$ regulates the pathogenesis of pulmonary fibrosis. In this study, we investigate the interaction of TGF-$\beta$, IL-1$\beta$, IL-6 and TNF-$\alpha$ and their effect on the proliferation of fibroblasts. Methods: We used a human fibroblast cell line, MRC-5 (ATCC). The culture of MRC-5 was confirmed by immunofluorecent staining. First, we determined the concentration of serum in cuture medium, in which the proliferation of MRC-5 is supressed but the survival of MRC-5 is retained. Second, we measured optical density after staining the cytokine-stimulated cells with 0.5% naphthol blue black in order to detect the effect of cytokines on the proliferation of MRC-5. Result: In the medium containing 0.5% fetal calf serum, the proliferation of MRC-5 increased by 50%, and it was maintained for 6 days. IL-1$\beta$, TNF-$\alpha$ and IL-6 induced the proliferation of MRC-5 by 45%, 160% and 120%, respectively. IL-1$\beta$ and TNF-$\alpha$ enhanced TGF-$\beta$-induced proliferation of MRC-5 by 64% and 159%, but IL-6 did not affect the TGF-$\beta$-induced proliferation. And lNF-$\alpha$-induced proliferation of MRC-5 was reduced by IL-1$\beta$ in 50%. TGF-$\beta$, TNF-$\alpha$ and both induced the proliferation of MRC-5 to 89%, 135% and 222%, respectively. Conclusions: TNF-$\alpha$, TGF-$\beta$ and IL-1$\beta$, in the order of the effectiveness, showed the induction of MRC-5 proliferation of MRC-5. TNF-$\alpha$ and IL-1$\beta$ enhance the TGF-$\beta$-induced proliferation of MRC-5, but IL-6 did not have any effect TNF-$\alpha$-induced proliferation of MRC-5 is diminished by IL-1, and TNF-$\alpha$ and TGF-$\beta$ showed a additive effect.

• Journal of Periodontal and Implant Science
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• v.32 no.3
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• pp.457-473
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• 2002

The purposes of this study is to evaluate the combination effects of TGF-${\beta}_1$ and PDGF-BB on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM/100% FBS at the $37^{\circ}C$, 5% $CO_2$ incubator. Authors measured the DNA synthesis, total protein, collagen and noncollagenous protein synthesis according to the concentration of TGF-${\beta}_1$,(1,5ng/ml) and PDGF-BB (1,10 ng/ml) in combination. To explore further this delayed effect of TGF-${\beta}_1$, we preincubated human periodontal ligament cells with TGF-${\beta}_1$ for 4 or 24 hours before PDGF-BB stimulation. The results were as follows: The DNA synthetic activity was increased dose dependently by TGF-${\beta}_1$, PDGF-BB. The combination of TGF-${\beta}_1$ and PDGF-BB consistently enhanced the DNA synthetic activity to PDGF-BB alone. The ability of TGF-${\beta}_1$ to enhance DNA synthetic activity in PDGF-BB treated periodontal ligament cells was dose dependent. The maximum mitogenic effect was at the 5ng/ml of TGF-${\beta}_1$ and l0ng/ml of PDGF-BB. Preincubation of cell with TGF-${\beta}_1$ resulted in significantly greater response to PDGF-BB at all TGF-${\beta}_1$ concentration studied, and may be useful for clinical application in periodontal regenerative procedures. The total protein, collagen and noncollagen synthesis was increased dose pendently by TGF-${\beta}_1$, PDGF-BB. The % of collagen was slightly decreased according to the concentration of TGF-${\beta}_1$, PDGF-BB. The effect of TGF-${\beta}_1$, PDGF-BB were not specific for collagen synthesis since it also increased noncollagenous protein synthesis. This study demonstrates that PDGF-BB is major mitogens for human periodontal ligament cells in vitro, and supports a role for TGF-${\beta}_1$ as a regulation of the mitogenic and total protein formation to PDGF-BB in these cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.7
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• pp.948-957
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• 2016

We previously developed an herbal composition (HemoHIM) based on the water extracts of Angelica gigas radix, Cnidium officinale rhizoma, and Paeonia japonica radix to protect and recover hematopoietic and intestinal tissues against radiation injuries. In this study, to develop a composition with improved activities based on enhanced fat-soluble polyphenol contents, we prepared a new herbal composition, MH-30, from the above three herbs by 30% ethanol extraction and hot water extraction. HPLC analysis of the ethanol fractions of MH-30 and HemoHIM revealed that MH-30 had higher contents of many fat-soluble polyphenol compounds than HemoHIM (8.7-fold increase for decursin), whereas contents of water-soluble polyphenol compounds showed little differences between the two compositions. Then, we evaluated MH-30 and HemoHIM for their in vitro antioxidant and immune cell-stimulating activities as well as in vivo protective effects against radiation injuries in hematopoietic and self-renewal tissues. In antioxidant activity assays, MH-30 showed higher hydroxyl radical scavenging activity than HemoHIM (1.4- to 1.9-fold for compositions and 2.3- to 4.5-fold for ethanol fractions). On the other hand, MH-30 and HemoHIM exhibited similar immune cell-stimulating activities as measured by in vitro lymphocyte proliferation. MH-30 increased endogenous spleen colony formation, decreased bone marrow cell apoptosis, and enhanced survival of intestinal crypts in irradiated mice, demonstrating effective protection of MH-30 against radiation-induced injuries in hematopoietic and self-renewal tissues. The 30-day survival rate of lethally irradiated mice, a comprehensive index for radioprotective efficacy, was also elevated by MH-30. Noticeably, MH-30 showed higher protective effects than HemoHIM in all mouse experiments. These results demonstrate that MH-30 can protect hematopoietic and self-renewal tissues against radiation injuries more effectively than HemoHIM. Therefore, MH-30 can be a good candidate to reduce radiation injuries in hematopoietic and self-renewal tissues incurred by radiation accidents or cancer radiation therapy.

• Journal of Chest Surgery
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• v.37 no.8
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• pp.691-701
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• 2004

Background: Tissue hypoxia is a characteristic of many human malignant neoplasms, and hypoxia inducible factor-1 (HIF-1) plays a pivotal role in essential adaptive response to hypoxia, and activates a signal pathway for the expression of the hypoxia-regulated genes, resulting in increased oxygen delivery or facilitating metabolic adaptation to hypoxia. Increased level of HIF-1 a has been reported in many human malignancies, but in esophageal squamous cell carcinoma, the influence of HIF-1 a on tumor biology, including neovascularization, is not still defined. Material and Method: The influence of HIF-1 a expression on angiogenic factors, correlation between the tumor proliferation and HIF-1 a expression, interaction of HIF-1 a expression and p53, and correlation between HIF-1 a expression and clinicopathological prognostic parameters were investigated, using immunohistochemical stains for HIF-1 a, VEGF, CD34, p53, and Ki-67 on 77 cases of resected esophageal squamous cell carcinoma. Result: HIF-1 a expression in cancer cells was found in 33 of 77 esophageal squamous cell carcinoma cases. The 33 cases (42.9%) showed positive stain for HIF-1 a. High HIF-1 a expression was significantly associated with several pathological parameters, such as histologic grade (p=0.032), pathological TMN stage (p=0.002), the depth of tumor invasion (p=0.022), regional lymph node metastasis (p=0.002), distant metastasis (p=0.049), and lymphatic invasion (p=0.004). High HIF-1 a expression had significant VEGF immunoreactivity (p=0.008) and Ki-67 labeling index (p<0.001), but was not correlated with microvascular density within tumors (p=0.088). The high HIF-1 a expression was correlated with aberrant p53 accumulation with a marginal significance (p=0.056). The overall 5-year survival rate was 34.9%. The survival rate of patients with a high HIF-1 a expression was worse than that of patients with low-expression tumors (log-rank test, p=0.0001). High HIF-1 a expression was independent unfavorable factors although statistical significance is marginal in multivariate analysis. Conclusion: It is suggested that (1) high HIF-1 a expression in esophageal squamous cell carcinoma is associated with tumor hypoxia, or with genetic alteration in early carcinogenesis and progressive stages, (2) high HIF-1 a expression may be associated with intratumoral neovascularization through HIF-VEGF pathway, and (3) high HIF-1 a expression is associated with poor prognosis in patients with esophageal squamous cell carcinoma and may playa role as biomarker for regional lymph node metastasis.

• Journal of Nutrition and Health
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• v.39 no.3
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• pp.225-235
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• 2006

Although it has traditionally known that deer antler and medicinal herbs extract contain some functional components for health promotion, the nutritional significance remains to be elucidated. This study examined the efficacy of deer antler extract (DA) , medicinal herbs extract (MH) and their mixture (DAMH) on serum IGF-I, bone growth with growing rats in vivo and splenocyte proliferation with spleen cells in vitro. Three week-old young female rats (Sprague-Dawley) were divided into 4 groups and then fed basal diet (AIN-93G) or experimental diets containing DA, MH, DAMH, respectively, for 7 weeks. We collected blood, liver, kidney, spleen, femur and tibia from rats. There was no significant difference in weight gain, but food intake increased in DA- and MH-fed groups. There were no signs of liver and kidney damage in the DA, MH and DAMH-fed groups compared to basal diet group. In femur and tibia, wet weights: breaking forces and bone minerals (Ca, Mg and Zn) were significantly higher in the DA-fed group than in the other groups. Serum alkaline phosphatase (ALP) , bone-specific alkaline phosphatase (BALP) activities were significantly lower in the DA, MH, DAMH-fed groups than in basal diet group. Also, serum insulin-like growth factor-I (IGF-I) concentrations were significantly increased in DA-fed group compared to the other groups. Therefore DA was shown to have an activity of bone growth promotion by increasing the IGF-I, a major bone growth factor. The deer antler extract showed an enhanced immune action on the primary cultured-cells from spleen of rats, representing that splenocytes were proliferated by lipopolysaccharide (LPS), but not by concanavalin A (Con A). These results indicate that deer antler extract has beneficial effects on bone growth via IGF-I and on splenocyte activation.

• Journal of Ginseng Research
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• v.33 no.1
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• pp.33-39
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• 2009

The effects of white ginseng and red ginseng ethanol extracts on the proliferation, nitric oxide (NO) production and tumor necrosis factor-$\alpha$ (TNF-$\alpha$) in human Band T lymphocytes isolated from blood cell of sasang constitution were examined in vitro. White ginseng extract known as medicine suitable for soeumin constitution showed the highest immune activities such as proliferation, NO production and TNF-$\alpha$ in soeumin constitution, and followed by taeeumin and soyangin, respectively (P<0.05). However, there were immunologically insignificant differences among sasang constitution lymphocytes treated by red ginseng ethanol extract. Both white and red ginseng ethanol extracts showed significantly higher immunological activities than LPS-induced sasang constitution lymphocytes on NO production and TNF-$\alpha$ release (P<0.05). The different immunological activities of white and red ginseng extracts in sasang constitution lymphocytes might be due to the component changes during the processing of ginseng. Thus, the findings of components suitable for all constitutions from red ginseng extract could be applied as a new constitution marker.