• Applied Microscopy
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• v.40 no.2
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• pp.73-80
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• 2010

Liver regeneration is a result of highly coordinated proliferation of hepatocytes and nonparenchymal liver cells. Partial hepatectomy (PH) is the most often used stimulus to study liver regeneration because, compared with other methods that use hepatic toxins, it is not associated with the tissue injury and inflammation, and the initiation of the regenerative stimulus is precisely defined. Granulocyte macrophage-colony stimulating factor (GM-CSF), which is a cytokine able to regulate the proliferation and differentiation of epithelial cells, was first identified as the most potent mitogen for bone marrow. Particularly, rrhGM-CSF, which is highly glycosylated and sustained longer than any other types of GM-CSF in the blood circulation, was specifically produced from rice cell culture. In this experiment, effects of rrhGM-CSF administration were evaluated in the regenerating liver after 78% PH of rats. Morphological changes induced by PH were characterized by destroyed hepatocyte plate around the central vein and enlarged nuclear cytoplasmic ratio and increased hepatocytes with two nuclei. And then, proliferation of liver cells (parenchymal and nonparenchymal) and rearrangement of plates and lobules seemed to be carried out during liver regeneration. These alterations in the experimental group preceded those of the control. Since proliferating cell nuclear antigen (PCNA) is known to be a nuclear protein maximally elevated in the S phase of proliferating cells, the protein was used as a marker of liver regeneration after PH in rats. PCNA levels by western blot analysis and immunohistology were compared between the two groups. PCNA protein expression of two groups at 12 hr and 24 hr after injury showed similar pattern. The protein expression showed the peak at 3 days in both groups, however, the protein level of the experimental group was higher than that of the control. On immunohistochemical observations, the reaction product of PCNA was localized at the nuclei of proliferating cells and the positive reaction in experimental group at 3 days was clearly stronger than that in control group. The results by Western blotting and immunohistology for PCNA showed similar pattern in terms of the protein levels. In conclusion, rrhGM-CSF administration during liver regeneration after 78% PH accelerated breakdown and restoration of the hepatic plate and expression of PCNA. These results suggest that rrhGM-CSF might play an important role during liver regeneration in rats.

• Korean Journal of Medicinal Crop Science
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• v.17 no.5
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• pp.311-320
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• 2009

The low quality fresh ginseng was fermented by Phelinus linteus or Hericium erinaceum mycelium. This fermented ginseng was extracted by water at $100^{\circ}C$ or water with ultrasonification at $60^{\circ}C$. Total phenolic compounds was improved by ultrasonification extraction process, compare to conventional water extraction. All extracts enhanced the growth of human B and T cells, showing 2.68 times and 3.43 times higher, respectively, than the control. The secretion of TNF-$\alpha$ and IL-6 from human immune cells was enhanced as $3.53{\times}10^{-4}\;pg/cell$, $3.40{\times}10^{-4}\;pg/cell$ by adding H. erinaceum mycelium fermented ginseng. H. erinaceum mycelium fermented ginseng yielded higher nitric oxide production from macrophage than Lipopolysaccharides (LPS). The cytotoxicity on human normal kidney cell (HEK293) was as low as 20.5% in adding the maximum concentration of $1.0\;mg/m{\ell}$ of fermented ginseng. Generally, the extracts from ultrasonification extraction process showed 10% lower toxicity than that by conventional process. H. erinaceum mycelium fermented ginseng had the highest anticancer activity on human lung cancer and stomach cancer cells as 69.33% and 75.32%, respectively at $1.0\;mg/m{\ell}$. It can be concluded that, in general, H. erinaceum mycelium fermented ginseng has relatively better immune and anticancer activities than P. linteus fermented ginseng. Expecially, the extracts treated with ultrasonification had higher activities than that from conventional extraction process.

• Journal of fish pathology
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• v.23 no.1
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• pp.47-55
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• 2010

This study was conducted to investigate the effects of dietary seleno-yeast on histological response in juvenile olive flounder Paralichthys olivaceus. Fish averaging $4.0{\pm}0.1g$ (mean ${\pm}$ SD) were fed one of the four semi-purified diets containing 0.56, 1.07, 2.86 and 4.56 mg Selenoyeast/kg diet ($Se_{0.56}$, $Se_{1.07}$, $Se_{2.86}$ and $Se_{4.56}$, respectively) in triplicates for 12 weeks. Swelling of blood cell in glomerulus, the rupture of some epithelial cell in the renal tubules and enlarged macrophage were observed in the $Se_{1.07}$ group. The hepatopancreas had hepatic cell, capillary and zymogen in the pancreas and was normal in the $Se_{0.56}$ group. Swelling of hepatocyte gradually decreased with increase in selenoyeast supplementation. The gill lamellae showed partial abnormal condition (terminal clubbing) in the $Se_{0.56}$ group, but there were little differences between these and other treatments. These results indicated that the optimum dietary supplementation level of selenoyeast in juvenile olive flounder, Paralichthys olivaceus, could be less than 1.07 mg selenoyeast/kg diet based on histological responses of the fish.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.16 no.1
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• pp.365-370
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• 2015

Granulocyte-macrophage colony-stimulating factor (GM-CSF) is an important hematopoietic growth factor and immune modulator. The aim of this study was to evaluate the effects of GM-CSF on the development and cell number of porcine parthenotes, as well as on their expression of implantation-related genes. In the present study, porcine parthenogenatic activated embryos were cultured in a protein-free culture medium in the absence or presence of 5, 10 and 20 ng/ml GM-CSF for 7 days. The percentage of blastocyst formation, total cell number and gene expressions were evaluated. The results showed that the addition of 20 ng/ml GM-CSF to protein-free culture medium significantly increased the blastocoel formation ($26.14{\pm}2.03%$ vs. $3.55{\pm}0.51%$, p < 0.05). In addition, the cell number also increased when they were cultured in the presence of 20 ng/ml GM-CSF ($43.51{\pm}3.6%$ vs. $30.68{\pm}5.51%$, p < 0.05). A real time reverse transcripts polymerase chain reaction (RT-PCR) showed that GM-CSF enhances mRNA expression of the interleukin-6, but does not influence the leukemia inhibitory factor (LIF) receptor mRNA expression in blastocyst stage parthenotes. These results suggest that GM-CSF may enhance the viability of porcine embryos developing in vitro in a defined culture medium.

• The Korean Journal of Community Living Science
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• v.28 no.1
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• pp.29-43
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• 2017

We studied the antioxidant, the anti-inflammatory, antithrombotic, and antiobesity activities of seven different kinds of edible plants. 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ABTS radical scavenging activities were determined as a measurement of antioxidant activity. NO production inhibition by the macrophage cell line (Raw 264.7) treated with lipopolysaccharide (LPS) was carried out to assess anti-inflammatory activity. Thrombin inhibitory activity was measured for its antithrombotic function and inhibition of 3T3-L1 cell differentiation was evaluated as a measurement of antiobesity activity. Total phenolic components and total flavonoid contents were measured to determine functional materials in medicinal plants. Common sage, Japanese lady bell, and hairy agrimony showed high antioxidant activity ($IC_{50}$) of less than $100{\mu}g/mL$. All samples used in this study showed anti-inflammatory activity. Common sage, hairy agrimony, and hooker chives showed antithrombotic effects. Hairy agrimony showed the highest antithrombotic effect (98.1%). Common sage, Japanese lady belly, hairy agrimony, and hooker chives showed reduced 3T3-L1 cell differentiation and hooker chives strongly inhibited lipid accumulation in the cells compared to other medicinal plants. Common sage and hairy agrimony contained more than 1 mg GAE/g of phenolic compounds and more than 1 mg CE/g of flavonoids. Functional activities were different by plant part and extraction method from each sample. These results suggest that common sage, Japanese lady belly, hairy agrimony, and hooker chives may be used as healthy food sources with antioxidant, anti-inflammatory, antithrombotic, and antiobesity activities, and appropriate extracting methods from each plant need to be developed.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.42 no.5
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• pp.434-441
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• 2009

This study was conducted to compare the biological activities of 7 melania snails from the family Pleuroceridae (Semisulcospira coreana, Koreanomelania nodifila, Semisulcospira forticosta, Koreoleptoxis globus ovalis, Semisulcospira libertina, Semisulcospira tegulata and Semisulcospira gottschei) in Korea. Among the 7 species, S. coreana, Korean. nodifila, S. forticosta and S. gottschei showed over 80% cytotoxicities on three cancer cell lines (SNU-1, A549 and Hep 3B) compared to the non-treatment, whereas S. libertina and S. tegulata showed almost no growth inhibition activities on the same cancer cell lines. In relation to ACE inhibition activity, only S. coreana, Korean. nodifila, and S. forticosta showed over 60% ACE inhibition activities, whereas other melania snails exhibited inhibition activities of lower than 25%. DPPH radical scavenging activities were also determined, and used to categories melania snails into three groups based on Duncan's multiple range test at P<0.05. The amount of TNF-${\alpha}$ produced by in vitro mouse peritoneal macrophage was determined according to bioactivity on L-929 cells. Three melania snails, S. coreana, Korean. nodifila and S. gottschei, exhibited 95.2%, 89.7% and 93.7% cell death(%) on L-929 cells, respectively. Glucose-6-phosphate dehydrogenase inhibitory activity was also obtained in the extract of S. coreana (31.9%) and Korean. nodifila (28.1%), showing that these extracts can be used as supplemental dietary health foods. In conclusion, we believe that the extracts of melania snails should be given due consideration in functional health food development.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.43 no.1
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• pp.19-26
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• 2017

This study investigated the anti-inflammatory activities and cell viability of Amelanchier asiatica (A. asiatica) 70% ethanol extracts against RAW 264.7 cells induced by lipopolysaccharide (LPS). Cell toxicity test on macrophage cells (RAW 264.7) was performed by 3-[4,5-dimethyl-thiazol-2-yl]-2, 5-diphenyl-tetrazoliumbromide (MTT) assay and results showed 96% cell viability at $1,000{\mu}g/mL$ concentration. Anti-inflammatory activity was examined via the inhibitory tests on the production of LPS induced NO in RAW 264.7 cells by Griess assay. The result showed that the extract inhibited NO production in concentration dependent manner. The iNOS and COX-2 protein expression inhibitory effects were confirmed by western blot and by reverse transcription-polymerase chain reaction (RT-PCR). From the former they were decreased by 84.3%, 56.2% at $500{\mu}g/mL$ concentration, respectively, and from the latter decreased by 89.8%, 84.9% at $500{\mu}g/mL$, respectively. In conclusion, this study showed the anti-inflammatory effects of A. asiatica extracts. Thus, this could be applied to an anti-inflammatory agent.

• Polymer(Korea)
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• v.31 no.6
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• pp.505-511
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• 2007

Poly(lactide-co-glycolide)(PLGA) and hyaluronic acid (HA) has been widely used as biocompatible scaffold materials to regenerate tissue. In this present study, we fabricated microporous PLGA and HA loaded PLGA scaffolds by a emusion freeze-drying method. In order to confirm that the release profile of cytokine or water-soluble drugs, we manufactured the granulocyte macrophage colony stimulating factor(GM-CSF) loaded PLGA and HA-PLGA scaffold. All scaffolds were characterized using scanning electron microscope(SEM), mercury porosimeter and wettability measurement. Cell proliferation and viability were assessed by a 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium-bromide (MTT) test. The porosity of HA-PLGA scaffold was greater than 95% with the total pore area of $261\;m^2/g$. The HA-FLGA scaffold exhibited well interconnected pores to allow greater cell adhesion and prolixferation. It was proven by higher cell viability in the HA-PLGA scaffold than PLGA alone. This may be due to the enhanced natural properties and higher water retention capacity of HA.

• Journal of Physiology & Pathology in Korean Medicine
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• v.28 no.2
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• pp.178-185
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• 2014

Leonuri Fructus, a semen of Leonuri Herba, has been used for the treatment of menstrual disorders such as amenorrhea, dysmenorrhea and leukorrhea and for the remedy of hyperemia. The present study was conducted to evaluate the anti-inflammatory effects of the Leonuri Fructus extract (Leonurus japonicus Houtt. EtOH extract; LJE) in vivo and in vitro. In vitro study, the MTT assay for cell viability was conducted to determine the non-cytotoxic concentration of LJE treatment in media. The levels of NO were measured with Griess reagent. Pro-inflammatory cytokines were detected by ELISA method. The inflammation-related proteins of this study were detected by immunoblot anlaysis. The increases of NO production and iNOS expression were detected in LPS-treated cells compared with control, but LJE attenuated the increases of NO and iNOS by LPS. LJE reduced the production of TNF-${\alpha}$ and IL-$1{\beta}$ induced by LPS stimulation. LJE suppresses the signaling pathways of NF-${\kappa}B$ and MAPKs in LPS-induced macrophage cells. In vivo study, carrageenan-induced hind paw acute edematous inflammation rat model was used for evaluation of anti-inflammatory activity of LJE. LJE significantly inhibited the increases of hind paw swelling, skin thicknesses and inflammatory cell infiltrations, and decreased the numbers of mast cell induced by carrageenan injection. These results suggest that LJE has an anti-inflammatory therapeutic potential, which is mediated through modulating NF-${\kappa}B$ activation and MAPK phosphorylation. Inhibition of the rat paw edema induced by carrageenan is considered as direct evidence that LJE may be a useful source to treat inflammation.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.2
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• pp.83-90
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• 2002

Objective : The purpose of the current series of experiments were to assess the effect of GM-CSF, as a medium supplement, on the development of mouse embryos and the expression of LIF and IL-1? mRNA. Materials and Methods: Mouse 2-cell embryos were collected from the oviducts of 6 weeks old ICR mice at 48 hours after hCG injection. Embryos were cultured in P-1 medium supplemented with mouse GM-CSF (0, 1, 5, 10 ng/ml). The embryo development to blastocysts and hatching blastocysts was assessed and the cell number in blastocyst was also examined. Using RT-PCR, the expressions of LIF and IL-1? mRNA in blastocyst were evaluated in the GM-CSF supplemented group and control group. Results: In mouse, the addition of GM-CSF increased the percentage of blastocysts (65.5%, 68.6%, 73.0% and 76.1% for control and 1, 5 and 10 ng/ml, respectively), and increased the proportion of hatching blastocysts (35.2%, 36.4%, 43.2% and 53.0% for control and 1, 5 and 10 ng/ml, respectively). The mean cell numbers in blastocyst were significantly increased in GM-CSF supplemented groups compared to control group. LIF and IL-1? expression in blastocyst were significantly higher in GM-CSF supplemented group than in control group. Conclusion: The results of experiment by mouse embryos showed beneficial effects of GM-CSF as a medium supplement. Furthermore, the addition of GM-CSF significantly increased the expression of LIF and IL-1? in mouse embryos. These results suggest that GM-CSF might be a important molecule in embryo implantation.