• Clinical and Experimental Reproductive Medicine
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• v.31 no.1
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• pp.67-74
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• 2004

Objective: This study was to examine in vitro neural cell differentiation pattern of the genetically modified human embryonic stem cells expressing tyrosine hydroxylase (TH). Materials and Methods: Human embryonic stem (hES, MB03) cell was transfected with cDNAs cording for TH. Successful transfection was confirmed by western immunoblotting. Newly transfected cell line (TH#2/MB03) was induced to differentiate by two neurogenic factors retinoic acid (RA) and b-FGF. Exp. I) Upon differentiation using RA, embryoid bodies (EB, for 4 days) derived from TH#2/MB03 cells were exposed to RA ($10^{-6}M$)/AA ($5{\times}10^{-2}mM$) for 4 days, and were allowed to differentiate in N2 medium for 7, 14 or 21 days. Exp. II) When b-FGF was used, neuronal precursor cells were expanded at the presence of b-FGF (10 ng/ml) for 6 days followed by a final differentiation in N2 medium for 7, 14 or 21 days. Neuron differentiation was examined by indirect immunocytochemistry using neuron markers (NF160 & NF200). Results: After 7 days in N2 medium, approximately 80% and 20% of the RA or b-FGF induced Th#2/MB03 cells were immunoreactive to anti-NF160 and anti-NF200 antibodies, respectively. As differentiation continued, NF200 in RA treated cells significantly increased to 73.0% on 14 days compared to that in b-FGF treated cells (53.0%, p<0.05), while the proportion of cells expressing NF160 was similarly decreased between two groups. However, throughout the differentiation, expression of TH was maintained ($\sim$90%). HPLC analyses indicated the increased levels of L-DOPA in RA treated genetically modified hES cells with longer differentiation time. Conclusion: These results suggested that a genetically modified hES cells (TH#2/MB03) could be efficiently differentiated in vitro into mature neurons by RA induction method.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.33 no.1
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• pp.10-18
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• 2011

Purpose: The purpose of this study is to find out the effects of bisphosphonates (BPs) on the proliferation and the alkaline phosphatase (ALP) activity of human bone marrow derived mesenchymal stem cells (hMSCs), and thus state its correlation with bisphosphonate related osteonecrosis of the jaw (BRONJ). Methods: hMSCs was obtained by collecting and culturing cancellous bone fragments from a patient undergoing iliac bone graft. Alendronate (Aln) and Pamidronate (Pam), Ibandronate (Ibn) were added to the culture media in the concentration from $10^{-3}$ M to $10^{-11}$ M and cell toxicity, viability were measured. For ALP activity evaluation, Aln and Pam were added to the culture media in the concentration from $5{\times}10^{-7}$ M to $1{\times}10^{-8}$ M and were cultured for 1 week, 2 weeks and 3 weeks. ALP activity data were standardized using protein assay. Control groups were prepared for each examination. Results: Aln, Pam and Ibn all failed to increase the proliferation of hMSCs. With 1 week, 2 weeks of $5{\times}10^{-8}$M of Aln treatment, the ALP activity increased. Pam treatment increased the ALP activity with 2 weeks of $5{\times}10^{-8}$ M and$1{\times}10^{-8}$M. Also Ibn treatment increased the ALP activity with 2 weeks of $5{\times}10^{-8}$ M and $1{\times}10^{-8}$ M. Conclusion: It is considered that BPs are not capable of improving the proliferation of hMSCs. Also, after a transient increase in the ALP activity with the lower concentration of BPs, the activity decreased again. Therefore, in patients on long-term medication of BPs, the proliferation and osteoblast differentiation of hMSCs are restrained, and thus delayed wound healing and increase in BRONJ complications may occur.

• Journal of the Ergonomics Society of Korea
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• v.16 no.2
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• pp.15-27
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• 1997

Brain potential is described using the mesocolumnar activity defined by averaged firings of excitatory and inhibitory neuron of neocortex. Lagrangian is constructed based on SMNI(Statistical Mechanics of Neocortical Interaction) and then Euler Lagrange equation is obtained. Excitatory neuron firing is assumed to be amplitude- modulated dominantly by the sum of two modes of frequency .omega. and 2 .omega. . Time series of this neuron firing is calculated numerically by Euler Lagrangian equation. I .omega. L related to low frequency distribution of power spectrum, I .omega. H hight frequency, and Sd(standard deviation) were introduced for the effective extraction of the dynamic property in the simulated brain potential. The relative behavior of I .omega. L, I .omega. H, and Sd was found by parameters .epsilon. and .gamma. related to nonlinearity and harmonics respectively. Experimental I .omega L, I .omega. H, and Sd were obtained from EEG of human in rest state and of canine in deep sleep state and were compared with theoretical ones.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.5
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• pp.1210-1214
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• 2008

The purpose of this study is to investigate the effect of water extract from Artemisiae Argi Folium (WAAF) on hepatotoxicity caused by acetaminophen (AAP) and acetaldehyde which are regarded as hepatotoxin. Artemisiae Argi Folium was known to have the antibacterial, immune-enhancing, and anticoagulative properties. In Korean Medicine, Artemisiae Argi Folium is supposed to be related with 'liver meridian' according to traditional medical theory. AAP and acetaldehyde reduce the intracellular production of hydrogen peroxide ($H_2O_2$) and nitric oxide (NO) production of human hepatocyte HepG2. The intracellular production of hydrogen peroxide ($H_2O_2$) was measured by dihydrorhodamine 123 (DHR) assay. NO production was measured with Griess test. WAAF increased the production of $H_2O_2$ and NO reduced by AAP and acetaldehyde in HepG2 cells. Therefore, It could be suggested that WAAF has the hepatoprotective activity against AAP and acetaldehyde.

• Journal of Korean Ophthalmic Optics Society
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• v.20 no.1
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• pp.51-60
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• 2015

Purpose: To determine the effect of marketed multipurpose contact lens solutions (MPSs) on human corneal epithelial cells (HCEpiCs) toxicity by using image analysis. Methods: HCEpiCs were exposed six MPSs (product A-F) at 0.05~50% for 2h, 12h, 24h, and 48h respectively. HCEpiCs were fixed and stained with Draq5 after exposure with MPSs, and the cell viability and apoptosis were evaluated by using confocal microscope and ImageXpress UltraTM. Results: Viabilities of HCEpiCs exposed to MPS A-F for a 2h were not affected, while reductions (52~75%) in cell viability over a 12h exposure of MPS B, MPS C, MPS D and MPS F, and significant more reductions (29~73%) over a 24h and 48h-exposure. Apoptosis of HCEpiC was not affect over a 12h MPS exposure, however was significantly increased (199~526%) over 24h and 48h MPS exposure. Among the products MPS D, E and F reduced viability of HCEpiCs and apoptosis increased more than MPS A (p<0.05). Conclusions: Lower concentration of MPSs have not an cytotoxic effect on HCEpiCs, however higher concentration of MPSs induce apoptosis and reduce viability of HCEpiCs. Therefore, it need to develop MPS having antimicrobial effectiveness with low cytotoxicity.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1997.04a
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• pp.86-86
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• 1997

Human annexin I is a member of annexin family of calcium dependent phospholipid binding proteins, which have been implicated in various physiological roles including phospholipase A$_2$ (PLA$_2$) inhibition, membrane fusion and calcium channel activity. In this work, the structure of N-terminally truncated human annexin I (Δ-annexin I) and its interactions with Ca$\^$2+/, ATP and cAMP were studied at atomic level by using $^1$H, $\^$15/N, $\^$l3/C NMR (nuclear magnetic resonance) spectroscopy. The effect of Ca$\^$2+/ binding on the structure of Δ-annexin I was investigated, and compared with that of Mg$\^$2+/ binding. The addition of Ca$\^$2+/ to Δ-annexin I caused some changes in the high field and low field regions of $^1$H NMR spectra. Whereas, upon addition of Mg$\^$2+/ to Δ-annexin I, almost no change could be observed. Also we found that the binding ratio of ATP to Δ-annexin I is 1. Because Δ-annexin I is a large protein with 35 kDa molecular weight, site-specific (carbonyl-$\^$l3/C, amide-$\^$15/N) labeling technique was used to determine the interaction sites of Δ-annexin I with Ca$\^$2+/ and ATP. Assignments of all the histidinyl carbonyl carbon resonances have been completed by using Δ-annexin I along with its specific 1,2-subdomain. The carbonyl carbon resonances originating from His52 and His246 of Δ-annexin I were significantly affected by Ca$\^$2+/ binding, and some Tyr and Phe resonances were also affected. The carbonyl carbon resonances originating from His52 is significantly affected by ATP binding, therefore His52 seems to be involved in the ATP binding site of Δ-annexin I.

• Journal of Embryo Transfer
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• v.33 no.3
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• pp.149-157
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• 2018

The objective of this study was to evaluate the effect of treating dairy cattle with exogenous human chorionic gonadotrophin (hCG), five (5) days post artificial insemination (AI) on serum progesterone (P4) concentration and pregnancy rate. In this experiment, five days after AI, cows were assigned randomly to two groups namely: a) treated group (67) which were administrered with 1500 IU hCG (Chorulon) and b) control group (61), which received no treatment. On day 5, 10, 15 and 20 after the artificial insemination, blood samples from a total of 8 cows (4 from each group) were collected and were analyzed for serum P4 concentration. Cows were detected for estrus according to standing heat by visual observation. Cows that were detected still in estrus after days 18-24 were re-inseminated and recorded as not pregnant (open). Pregnancy diagnosis was conducted by ultrasonographic examination and transrectal palpation of the uterus on approximately 60 days in cows that observed to be not in estrus. The conception rate in hCG treated and control groups were 52.5 and 36.1%, respectively. The results proved that there were no significant differences in conception rate between two groups (p=0.0568). However, pregnancy rates were reduced by hCG treatment. Average serum P4 concentrations did not differ between Hcg-treated and control groups on day 5 (0.377 versus 0.375 ng/ml). On day 20 serum P4 concentrations were greater in the treated group compared with the control group (3.085 versus 2.010 ng/ml). The treatment with hCG seemed to increase P4 level compared with the control. In conclusion, the results of this study showed that 1500 IU of hCG administered on 5 day post AI increased conception rate in dairy cows. This was supported by the results on serum P4 concentration which was greater in hCG treated group.

• Journal of Haehwa Medicine
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• v.15 no.2
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• pp.135-148
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• 2006

Sophorae Radix (SFR) is known as a therapeutic drug that has been used in Oriental traditional medicine for the treatment of skin and mucosal ulcers, gastrointestinal hemorrhage, diarrhea, inflammation and arrhythmia. In the present study, we examined the effects of the aqueous extract of SFR on anti-inflammation, anti-allergic and anti-oxidant effect in various cell lines; they include mouse lung fibroblast cells (hFCs), human mast cells (HMC-1), human monocytic cells (THP-1), and RAW 264.7 cells. Treatment with SFR extract at a concentration of 250 ${\mu}g$/ml for 24h showed no significant decrease in the survival rate of the hFCs. SFR decreased the mRNA expression of IL-8, TNF-$\alpha$, and IL-6 in HMC-1 cells. SFR extract treatment significantly inhi-bited the protein expression of IL-6 and, IL-8 induced by mite in THP-1 cells and it also did MCP-1 expression. We examined the alternation of histamine release in HMC-1 cells for investigating anti-allergic effect of SFR. Histamine secretion decreased after the treatment with SFR. In addition, SFR extract treatment at a concentration of 10 ${\mu}g$/ml, 100 ${\mu}g$ /ml, and 200 ${\mu}g$/ml lowered the $\beta$-hexosaminidase to 10.3%, 21.7%, and 50.8%, respectively. IC50 of SFR extract in RBL-2H3 cells was 196.85 ${\mu}g$/ml. Both activity of NF-$\kappa$B promoter in RBL-2H3 cells significantly diminished after the dose-dependent treatment of SFR. Therefore, our results indicate that SFR has anti-inflammatory and it may be useful for treating allergic diseases such as atopic dermatitis.

• Journal of Microbiology and Biotechnology
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• v.15 no.6
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• pp.1258-1266
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• 2005

The present study was carried out in order to assess the protective effects of calycosin-7-O-$\beta$-D-glucopyranoside, isolated from Astragali radix (AR), on hyaluronidase (HAase) and the recombinant human interleukin-$1\beta$ (IL-$1\beta$)-induced matrix degradation in human articular cartilage and chondrocytes. We isolated the active component from the n-butanol soluble fraction of AR (ARBu) as the HAase inhibitor and structurally identified as calycosin-7-O-$\beta$-D-glucopyranoside by LC-MS, IR, ${1}^H$ NMR, and ${13}^C$ NMR analyses. The $IC_{50}$ of this component on HAase was found to be 3.7 mg/ml by in vitro agarose plate assay. The protective effect of ARBu on the matrix gene expression of immortalized chondrocyte cell line C28/I2 treated with HAase was investigated using a reverse transcription polymerase chain reaction (RT-PCR), and its effect on HAase and IL-$1\beta$-induced matrix degradation in human articular cartilage was determined by a staining method and calculating the amount of degraded glycosaminoglycan (GAG) from the cultured media. Pretreatment with calycosin-7-O-$\beta$-D-glucopyranoside effectively protected human chondrocytes and articular cartilage from matrix degradation. Therefore, calycosin-7-O-$\beta$-D-glucopyranoside from AR appears to be a potential natural ant-inflammatory or antii-osteoarthritis agent and can be effectively used to protect from proteoglycan (PG) degradation.

• Journal of Applied Biological Chemistry
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• v.63 no.2
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• pp.131-136
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• 2020

When blood vessels are damaged, a fast hemostatic response should occur to minimize blood loss and maintain normal circulation. Platelet activation and aggregation are essential in this process. However, excessive platelet aggregation or abnormal platelet aggregation may be the cause of cardiovascular diseases such as thrombosis, stroke, and atherosclerosis. Therefore, finding a substance capable of regulating platelet activation and suppressing agglutination reaction is important for the prevention and treatment of cardiovascular diseases. 6,7-Dimethoxy-2H-chromen-2-one (Scoparone), found primarily in the roots of Artemisia or Scopolia plants, has been reported to have a pharmacological effect on immunosuppression and vasodilation, but studies of platelet aggregation and its mechanisms are still insufficient. This study confirmed the effect of scoparone on collagen-induced human platelet aggregation, TXA₂ production, and major regulation of intracellular granule secretion (ATP and serotonin release). In addition, the effect of scoparone on the phosphorylation of the phosphoproteins PI3K/Akt and mitogen-activated protein kinases (MAPK) involved in signal transduction in platelet aggregation was studied. As a result, scoparone significantly inhibited the phosphorylation of PI3K/Akt and MAPK, which significantly inhibited platelet aggregation through TXA₂ production and intracellular granule secretion (ATP and serotonin release). Therefore, we suggest that scoparone is an antiplatelet substance that regulates the phosphorylation of phosphoproteins such as PI3K/Akt and MAPK and is of value as a preventive and therapeutic agent for platelet-derived cardiovascular disease.