• Herbal Formula Science
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• v.26 no.3
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• pp.223-236
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• 2018

Objectives : We compared the inhibitory effects of herb-combined remedies, which were recorded on "Yooam" prescription of Dongeuibogam, on cell migration and invasion, two critical cellular processes that are often deregulated during metastasis, in B16F10 melanoma cells. For this purpose, water extracts of Sipyukmiryukieum (SYMRKU), Danjacheongpitang (DJCPT), Cheongganhaeultang (CGHUT) and Jipaesan (JPS) were used. Methods : Cytotoxicity was assessed by an MTT assay. Wound healing and matrigel transwell assays were used to examine on B16F10 cell migration and invasion. The levels of mRNA and protein expression of matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMPs) were analyzed by RT-PCR and Western blotting. Results : Our data showed that DJCPT showed the strongest inhibitory effect among the four prescriptions in inhibiting cell motility of B16F10 melanoma cells within the concentration range that was not cytotoxic. The inhibitory potential of colony formation was higher in DJCPT and SYMRKU compared to the other two types of prescriptions, and the inhibitory effect of invasiveness is shown in order of DJCPT, SYMRKU, CGHUT and JPS. DJCPT, and SYMRKU strongly inhibited the activity and expression of MMP-2 and MMP-9, which are important mediators in cancer invasion, compared to CGHUT and JPS, and the increased expression of TIMP-1 and TIMP-2 was also more effective in these two prescriptions. In conclusion, DJCPT is expected to exhibit the most potent blocking effect on migration and invasion among four herb-combined remedies compared in B16F10 melanoma cells. Conclusion : Overall, the results of this study will be used as an important source to validate these prescriptions in animal models and to understand the mechanism of action of herbal remedies recorded in Dongeuibogam.

• Journal of Periodontal and Implant Science
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• v.33 no.3
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• pp.359-372
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• 2003

Among objectives of periodontal therapy. the principal one is the morphological and functional reconstruction of lost periodontal supporting tissues. This includes de novo formation of connective tissue attachment and the regrowth of alveolar bone. The use of enamel matrix derivative(EMD) may be a suitable means of regeneration new periodontal attachment in the infrabony defects. Implant used to replace lost tooth but, implantitis occurred after installation. The purpose of this study was to investigate the effects of EMD on differentiation and growth of osteoblast in titanium disc. Twentyfive millimeter diameter and 1mm thick Ti disc which was coated 25, 50, 100, 200${\mu}g$/ml of EMD(Emdogain(R)) used as experimental group, 25, 50, 100, 200ng/d of rhBMP-2 as positive control group, and no coat as negative control group. A human osteosarcoma cell line Saos-2 was cultured in Ti disc and cell proliferation and Alkaline phosphatase (ALP) activity were measured at 1 and 6 days. PCR was performed at 2 and 8 hours. Semi-quantitative RT-PCR for mRNA expressions of various osteoblastic differentiation markers -type I collagen, ALP, osteopontin, and bone sialoprotein - were performed at appropriate concentrations based upon the results of MTT and ALP assay. Cultured cell-disc complexes were prepared for scanning electron microscopy (SEM) at 2 hour. Data were analyzed using Mann-Whitney and repeated- measures 1-way analysis of variance(SPSS software version 10,SPSS. Chicago. IL). After culture, there was more osteoblast in EMD100${\mu}g$/ml than in EMD50, 200${\mu}g$/ml on day 6. There was significant difference in experimental and positive control group compared control group, as times go by(1 and 6 days). Alkaline phosphatase activity was different significantly in EMD100, 200${\mu}g$/ml and BMP100, 200${\mu}g$/ml on day 6. The results of reverse transcriptase-polymerase chain reaction (RT-PCR) showed that expression of mRNA for ALPase, collagen type I, osteopontin. hone sialoprotein and BMP-2 was detected at 2 hour and 8 hour in EMI 200${\mu}g$/ml subgroup and BMP100ng/ml subgroup. The results of this study suggest that application of enamel matrix derivative on osteoblast attached to titanium surface facilitate the expression of bone specific protein and the differentiation and growth of osteoblast.

• Journal of the Korean Applied Science and Technology
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• v.36 no.4
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• pp.1303-1311
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• 2019

This study was to investigate the antioxidant, anti-inflammatory and neuronal cell protective effects of Poria cocos Wolf and Corni fructus extracted by water and 70% ethanol. Total polyphenol content in water extract of Poria cocos Wolf was significantly higher than those of other extracts. DPPH and ABTS free radical scavenging activity in water extract of Corni fructus was higher than those of other extracts. DPPH and ABTS free radical scavenging activity were increased in a dose-dependent manners. In order to effectively extract total polyphenol contents and anti-oxidant components in Poria cocos Wolf and Corni fructus, hot water extraction method is more efficient than ethanol extraction method. Poria cocos extracts were found to be a superior NO production inhibitory effect compared to Corni fructus extracts. In a neuronal cell viability assay using MPP⁺, the water extract of Poria cocos Wolf protected against MPP⁺-induced neurotoxicity than those of Corni fructus extract. It is considered to be a potential functional material with antioxidant, anti-inflammatory and neuronal protective effect against to oxidative stress according to the extract methods of extracting Poria cocos Wolf and Corni fructus.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.6
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• pp.1016-1024
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• 2000

The study was conducted to develop a rapid method for the detection of Listeria monocytogenes in foods via polymerase chain reaction (PCR) technique using hemolysin gene (hlyA) primers. Specificity and sensitivity of PCR, optimal conditions for PCR and application of hlyA gene primers for the detection of L. monocytogenes from milk and beef were investigeted. Each of the 20 L. monocytogenes strains gave a single 713 bp band, but other Listeria sup. and other bacteria did not show any bands. As few as 1 pg of L. monocytogenes DNA or 2.4$\times$10$^4$L. monocytogenes cells could be detected with hlyA gene primers. PCR product was most improved at 20~30 cycle in terms of removal of tailing and sensitivity. Also, the sensitivity was significantly improved by the further 10~15 cycle after 20 cycle PCR amplication. Milk (10 mL) and beef (10 g) samples were inoculated with L. monocytogenes at the concentrations ranging from 0 to 10$^{7}$ CFU/mL or g to determine the best sensitivity of PCR for the rapid detection of L. monocytogenes. PCR assay could detect 2 cells in milk with repeating PCR amplication and 2.6$\times$10$^2$cells in beef sample after 24 hr enrichment growth at 35$^{\circ}C$ in LEB.

• Bulletin of the Korean Chemical Society
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• v.30 no.12
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• pp.2999-3005
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• 2009

Among the more than 120 different types of human papillomavirus (HPV), types 16 and 18 have been known to be high risk agents that cause cervical cancer. We examined, in an immuno-chromatographic analysis, the potential of using the early gene product, E7 protein, as a diagnostic marker of cervical cancer caused by HPV. We developed monoclonal antibodies specific to HPV-16 and 18 E7 proteins that were produced from bacterial cells using gene recombinant technology. For each E7 protein, the optimal antibody pair was selected using the immuno-chromatographic sandwichtype binding system based on the lateral flow through membrane pores. Under these conditions, this rapid testing assay had a detection capability as low as 2 ng/mL of E7 protein. Furthermore, since viral analysis required the host cell to be lysed using chemicals such as detergents, it was possible that the E7 protein was structurally damaged during this process, which would result in a decrease in detection sensitivity. Therefore, we examined the detrimental effects caused by different detergents on the E7 protein using HeLa cells as the host. In these experiments, we found that the damage caused by the detergent, nonylphenylpolyethylene glycol (NP-40), was minimal relative to Triton X-100 commonly used for the cell lysis. Temperature also affected the stability of the E7 protein, and we found that the E7 protein was stabilized at 4$^{\circ}C$ for about 2 h, which was 4 times longer than at room temperature. Finally, a HPV-infected cervical cancer cell line, which was used as a real sample model, was treated using the optimized conditions and the presence of E7 proteins were analyzed by immuno-chromatography. The results of this experiment demonstrated that this rapid test could specifically detect HPV-infected samples.

• Polymer(Korea)
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• v.38 no.6
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• pp.702-707
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• 2014

Corneal endothelium is mono-inner cell layer of cornea and lay on Descmet's membrane which comprised of various proteins called extracellular matrix such as fibronectin, collagen, laminin, and proteoglycan, etc. In this study, we fabricated transparent poly(lactic-co-glycolic acid) (PLGA) film because PLGA is widely used for tissue engineering based on their properties. We investigated the behaviors of rabbit corneal endothelial cells (rCEnCs) on PLGA film surfaces coated with various cell-adhesive molecules like fibronectin, laminin, collagen type I and IV and FNC coating mix. The morphologic images, proliferation and adhesion assay, immunofluorescence for ZO-1 and $Na^+/K^+-ATPase$ and RT-PCR for expression of specific markers were conducted. These results showed that PLGA film plays a role as CEnC carriers in vitro and the cell-adhesive molecules give positive effects on the behaviors of rCEnC.

• Exercise Science
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• v.21 no.3
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• pp.289-298
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• 2012

In the present study, we investigated the effect of treadmill exercise on apoptotic neuronal cell death in the retinas of streptozotocin-induced diabetic rats. Twenty-eight male Sprague-Dawley rats were used for this study. The animals were divided into four groups(n = 7 in each group):(1) control group, (2) exercise group, (3) diabetes-induced group, (4) diabetes-induced and exercise group. Diabetes mellitus(DM) was induced by intraperitoneal injection of streptozotocin. The rats in the exercise groups were forced to run on the treadmill for 30 minutes once a day, five times per a week, during 12 weeks. In this study, a terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) assay and western blot for the expressions of caspase-3, cytochrome c, Bax, and Bcl-2 in the retinas were conducted for the detection of apoptotic retinal cell death. The present results showed that the number of TUNEL-positive cells was increased in the retinas of the diabetic rats, whereas treadmill exercise suppressed this number. The expressions of pro-apoptotic factors caspase-3, cytochrome c, and Bax were enhanced and the expressions of anti-apoptotic factor Bcl-2 was decreased in the retinas of the diabetic rats. In contrast, treadmill exercise suppressed the expressions of caspase-3, cytochrome c, and Bax and increased the expression of Bcl-2. The present study demonstrated that treadmill exercise suppressed diabetes-induced apoptotic neuronal cell death in the retinas. Based on the present results, treadmill exercise may be effective therapeutic strategy for the alleviating complications of diabetes patients.

• Korean Journal of Agricultural Science
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• v.49 no.2
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• pp.175-184
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• 2022

The present study investigated the neuroprotective effects of Coreopsis lanceolate extract against hydrogen-peroxide (H₂O₂)-induced oxidative damage and cell death in pheochromocytoma 12 (PC12) cells. Reactive oxygen species (ROS), 2,2'-azinobis (3-ethylbebzothiazoloine-6-sulfonic acid) diammonium salt, and 1,1-diphenyl-2-picrrylhydrazyl radical scavenging activities, as well as the expression levels of proteins associated with oxidative damage and cell death were investigated. According to the results, C. lanceolate extract exhibited inhibitory activity against intracellular ROS generation and cell-damaging effects induced by hydroxyl radicals in a dose-dependent manner. Total phenolic and flavonoid contents were 22.3 mg·g^-1 gallic acid equivalent and 16.2 mg·g^-1 catechin equivalent, respectively. Additionally, a high-performance liquid chromatography (HPLC) assay based on the internal standard method used to detect phenolic compounds. The phenolic compounds identified in C. lanceolata extract contained (+)-catechin hydrate (5.0 ± 0.0 mg·g^-1), ferulic acid (1.6 ± 0.0 mg·g^-1), chlorogenic acid (1.5 ± 0.0 mg·g^-1), caffeic acid (1.2 ± 0.0 mg·g-1), naringin (0.9 ± 0.0 mg·g^-1), and p-coumaric acid (0.5 ± 0.0 mg·g^-1). C. lanceolata extract attenuated pro-apoptotic Bax expression levels and enhanced the expression levels of anti-apoptotic Bcl-2, caspase-3, and caspase-9 proteins. Therefore, C. lanceolata is a potential source of materials with neuroprotective properties against neurodegenerative disorders, such as Alzheimer's and Parkinson's diseases.

• Journal of the Korean Society of Radiology
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• v.85 no.2
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• pp.394-408
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• 2024

Purpose To develop models to predict programmed death ligand 1 (PD-L1) expression in pulmonary squamous cell carcinoma (SCC) using CT. Materials and Methods A total of 97 patients diagnosed with SCC who underwent PD-L1 expression assay were included in this study. We performed a CT analysis of the tumors using pretreatment CT images. Multiple logistic regression models were constructed to predict PD-L1 positivity in the total patient group and in the 40 advanced-stage (≥ stage IIIB) patients. The area under the receiver operating characteristic curve (AUC) was calculated for each model. Results For the total patient group, the AUC of the 'total significant features model' (tumor stage, tumor size, pleural nodularity, and lung metastasis) was 0.652, and that of the 'selected feature model' (pleural nodularity) was 0.556. For advanced-stage patients, the AUC of the 'selected feature model' (tumor size, pleural nodularity, pulmonary oligometastases, and absence of interstitial lung disease) was 0.897. Among these factors, pleural nodularity and pulmonary oligometastases had the highest odds ratios (8.78 and 16.35, respectively). Conclusion Our model could predict PD-L1 expression in patients with lung SCC, and pleural nodularity and pulmonary oligometastases were notable predictive CT features of PD-L1.

• International Journal of Stem Cells
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• v.16 no.1
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• pp.78-92
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• 2023

Background and Objectives: This study aims to clarify the systems underlying regulation and regulatory roles of hydrogen combined with 5-Aza in the myogenic differentiation of adipose mesenchymal stem cells (ADSCs). Methods and Results: In this study, ADSCs acted as an in vitro myogenic differentiating mode. First, the Alamar blue Staining and mitochondrial tracer technique were used to verify whether hydrogen combined with 5-Aza could promote cell proliferation. In addition, this study assessed myogenic differentiating markers (e.g., Myogenin, Mhc and Myod protein expressions) based on the Western blotting assay, analysis on cellular morphological characteristics (e.g., Myotube number, length, diameter and maturation index), RT-PCR (Myod, Myogenin and Mhc mRNA expression) and Immunofluorescence analysis (Desmin, Myosin and 𝛽-actin protein expression). Finally, to verify the mechanism of myogenic differentiation of hydrogen-bound 5-Aza, we performed bioinformatics analysis and Western blot to detect the expression of p-P38 protein. Hydrogen combined with 5-Aza significantly enhanced the proliferation and myogenic differentiation of ADSCs in vitro by increasing the number of single-cell mitochondria and upregulating the expression of myogenic biomarkers such as Myod, Mhc and myotube formation. The expressions of p-P38 was up-regulated by hydrogen combined with 5-Aza. The differentiating ability was suppressed when the cells were cultivated in combination with SB203580 (p38 MAPK signal pathway inhibitor). Conclusions: Hydrogen alleviates the cytotoxicity of 5-Aza and synergistically promotes the myogenic differentiation capacity of adipose stem cells via the p38 MAPK pathway. Thus, the mentioned results present insights into myogenic differentiation and are likely to generate one potential alternative strategy for skeletal muscle related diseases.