• Journal of Acupuncture Research
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• 제17권2호
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• pp.169-186
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• 2000

To study anti-cancer effect and molecular biological mechanism of bee venom for aqua-acupuncture, the effects of bee venom on cell viability, apoptosis, and cell cycle were analyzed using MTT assay, tryphan blue assay, [3H]thymidine release assay, flow cytometric analysis, activity of caspase-3 protease activity assay, and immunocytometric analysis of PCNA. To explore whether anti-cancer effects of bee venom are associated with the transcriptional control of gene expression, quantitative RT-PCR analysis of apoptosis- and cell cycle-related genes was performed. The obtained results are summarized as follows: 1. The MTT assay demonstrated that cell viability was decreased by bee venom in a dose-dependant manner. 2. Significant induction of apoptosis was identified using tryphan blue assay, [$^3H$]thymidine release assay, and flow cytometric analysis of sub $G_1$ fraction. 3. In analysis of caspase-3 protease activity, the activity had increased significantly, in a dose-dependant manner. 4. Quantitative RT-PCR analysis of the apoptosis-related genes showed that Bcl-2 and $Bcl-X_L$ were down-regulated whereas Bax was up-regulated by bee venom treatment. 5. In flow cytometric analysis of cell cycle and immunocytometric analysis of PCNA expression, cell numbers of $G_1$ phase was increased by a dose-dependant manner. 6. In quantitative RT-PCR analysis of the cell cycle-related genes, p21, p27, and p57 were increased, while Cyclin D1, CDK4, c-Myc, c-Fos, and Histone H3 were decreased. In contrast, there were no remarkable changes in expression levels of CDC2 and c-Jun.

• Applied Chemistry for Engineering
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• 제22권2호
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• pp.178-184
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• 2011

In this study, the antibacterial, antioxidative and inhibitory effects of Allium cepa peel extracts on tyrosinase and elastase were investigated. MIC values of the ethyl acetate fraction of Allium cepa peel on especially, S. aureus among the skin resident flora (Staphylococcus aureus, S. aureus; Propionibacterium acnes, P. acnes; Pityrosporum ovale, P. ovale; Escherichia coli, E. coli) were 0.06%. The aglycone fraction showed more excellent free radical (1,1-diphenyl-2-picrylhydrazyl radical, DPPH) scavenging activity ($FSC_{50}=5.05{\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of the ethyl acetate fraction and aglycone fraction in the luminol-dependent $Fe^{3+}-EDTA/H_2O_2$ system were 0.05 and $0.03{\mu}g/mL$, respectively. The cellular protective effect of the aglycone fraction on the rose-bengal sensitized photohemolysis of human erythrocytes exhibited more prominent (${\tau}_{50}$, 480 min at $25{\mu}g/mL$). The inhibitory effects ($IC_{50}$) of the ethyl acetate fraction and aglycone fraction on tyrosinase were 9.16 and $8.68{\mu}g/mL$, the inhibitory effect ($IC_{50}$) of the aglycone fraction on elastase was $14.12{\mu}g/mL$ The transepidermal water loss of the cream containing 0.1% ethyl acetate fraction was decreased from $8.3g/m^2h$ in control to $6.8g/m^2h$ in the subjects applied with cream containing the ethyl acetate fraction. These results indicate that extract/fractions of Allium cepa peel can function as antioxidant in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS, and possibly as antiaging agents. Allium cepa peel extract could be used as a new cosmeceutical for whitening and anti-wrinkle products.

• Journal of the Korean Society of Food Science and Nutrition
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• 제40권12호
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• pp.1642-1647
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• 2011

The free radical scavenging activities of extracts from Ligustrum ovalifolium H. leaves (LOH) as well as various antioxidant activities such as ferric reducing antioxidant power (FRAP), 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity, reducing power and lipid peroxidation inhibition were evaluated by electron spin resonance (ESR). The total polyphenol and flavonoid contents of the water and ethanolic extracts from LOH were $105.5{\pm}1.31$ and $102.1{\pm}1.82$ mg gallic acid equivalent/g extract, respectively, and $84{\pm}1.72$ and $82.8{\pm}1.65$ mg catechin equivalent/g extract. In addition, $IC_{50}$ values for the 1,1-diphenyl-2-picryldrazyl (DPPH), alkyl, and hydroxyl radical scavenging activities of the water and ethanolic extracts were $0.021{\pm}0.002$ and $0.010{\pm}0.003$ mg/mL, $0.011{\pm}0.003$ and $0.012{\pm}0.002$ mg/mL, and $0.395{\pm}0.002$ and $0.443{\pm}0.002$ mg/mL, respectively. The ABTS radical scavenging activities of the water and ethanolic extracts from LOH and BHT were $0.073{\pm}0.12$, $0.130{\pm}0.06$ and $1.461{\pm}0.02$ mM Trolox equivalent/mg extract, respectively. The FRAP values of the extracts from LOH were higher than those of BHT, which was used as a positive control. The LOH extracts showed strong inhibitory effects on lipid peroxidation as measured by ferric thiocyanate (FTC) and thiobarbituric acid (TBA) assay compared to that of ${\alpha}$-tocopherol. Using MTT assay on human liver cells (Chang), extracts from LOH showed no toxicity at a concentration of 0.5 mg/mL. These results indicate that the LOH extracts possessed antioxidant activity.

• Journal of Life Science
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• 제19권11호
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• pp.1598-1604
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• 2009

In order to determine chemical components of onion flesh and peel, general nutrients, vitamin C, and total flavonoids were measured. Onion peel showed less moisture (14.3%) and no vitamin C compared to onion flesh. Onion peel contained more amounts of total flavonoids compared to onion flesh. In addition, the inhibitory effects of solvent extracts from onion flesh and peel on $H_2O_$-induced oxidative stress and growth of cancer cell lines (AGS human gastric adenocarcinoma and HT-29 human colon cancer cells) were investigated. Acetone with methylene chloride (A+M) and methanol (MeOH) extracts from onion flesh and peel appeared to significantly reduce the levels of intracellular reactive oxygen species (ROS) (p<0.05) and a greater antioxidant effect was observed in onion peel. Among fractions, 85% aq. methanol showed a higher protective activity against oxidative stress in both flesh and peel and there was no effect in the water and hexane fractions. The growth of cancer cells exposed to medium containing extracts and fractions from onion flesh and peel was inhibited dose-dependently. The growth of AGS was inhibited more in both flesh and peel compared to HT-29, and onion peel was more effective than onion flesh. Among fractions, 85% aq. methanol showed the greatest effect on growth inhibition in both flesh and peel. $IC_{50}$ values of 85% aq. methanol fraction from onion flesh and peel on AGS were 0.04 and 0.03 mg/ml, respectively, while those on HT-29 were 0.23 and 0.04 mg/ml. From our results, 85% aq. methanol fraction had an inhibitory effect against oxidative stress and growth of cancer cells, suggesting that it may contain biological active compounds.

• Korean Journal of Food Science and Technology
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• 제28권4호
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• pp.609-615
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• 1996

To study the effects of the fructooligosaccharide (FOS) intake on the intestinal microflora, the FOS (8g) was served to each of 5 volunteers (adult men $23{\sim}28$ years old) after every lunch for 4 weeks. Changes in fecal microflora, fecal moisture, and fecal pH were observed during the FOS intake and after the FOS intake, respectively. The fecal moisture content of the control period (4 weeks before the intake of FOS) was $81.77{\pm}1.18%$. The moisture content of the feces increased significantly at the end of the 4th week of FOS intake, and these effects lasted for 3 weeks after stopping FOS intake. The fecal pH before FOS intake was $6.56{\pm}0.09%$, while it decreased significantly (p<0.01) during the period of FOS intake. The pH reduction lasted for 4 weeks after stopping the intake of FOS. The log fecal number of Bifidobacteria during the period was $7.88{\pm}1.43%$ (CFU/g of wet feces) and it increased significantly during the FOS intake. After stopping the intake of FOS, the number of Bifidobacteria returned to those of control period. The log fecal number of Lactobacilli before the intake was $6.76{\pm}1.34%$ and significantly increased in 3 weeks due to the FOS intake. After stopping the intake of FOS, however, the number of Lactobacilli returned to those of control period. No remarkable changes were observed in the number of coliforms throughout all durations.

• Journal of Biomedical Engineering Research
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• 제33권2호
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• pp.72-77
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• 2012

Phase Contrast MR Angiography(PC MRA) is excellent MRA technique for measuring the velocity of vessels in the human body. PC MRA need to at least four images for angiogram reconstruction and it caused longer scan time. Therefore, we used keyhole imaging combined PC MRA to reduce the scan time. However, keyhole imaging can lead the erroneous effects as loss of phase information or frequency discontinuous. In this study, we applied the keyhole imaging combined 2D PC MRA for improving the temporal resolution and also measured the velocity to evaluate the accuracy of phase information. We used 0.32T MRI scanner(Magfinder II, Scimedix, Korea). Using the 2D PC MRA pulse sequence, the vascular images for a human brain targeted on the Superior Sagittal Sinus(SSS) were obtained. We applied tukey window function for keyhole images to minimize the ringing artifact and erroneous factors that are induced frequency discontinuous and phase information loss. We also applied zero-padded algorithm to peripheral missing k-space lines to compare keyhole imaging results and the artifact power(AP) value was measured on the complex difference images to validate the image quality. Consider as based on our results, heavy image distortions and artifacts were shown until using at least 50% keyhole factor. Using above the 50% keyhole factors are shown well reconstructed and matched for magnitude images and velocity information measurements. In conclusion, we confirmed the image quality and velocity information of keyhole technique combined 2D PC MRA. Especially, measured velocity information through the keyhole imaging combination was similar to the velocity information of full sampled k-space image despite of frequency discontinuous and phase information loss in the keyhole imaging reconstruction process. Consequently, the keyhole imaging combined 2D PC MRA will give some clinical usefulness and advantages as improving the temporal resolution and measuring the velocity information via selecting the appropriate keyhole factor at low tesla MRI system.

• Journal of Life Science
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• 제28권3호
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• pp.284-292
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• 2018

The melanin-concentrating hormone (MCH), a cyclic hypothalamic peptide composed of 17 amino acids, was initially identified in chum salmon (Oncorhynchus keta) as a regulator of pigmentation. Mammalian MCHs are cyclic hypothalamic peptides composed of 19 amino acids that regulate food intake and energy homeostasis. The present study examined not only MCH expression of different tissues but also the melanohore aggregation and intracellular $Ca^{2+}$ influx of fMCH and the other MCH. Real-time qPCR showed that MCH expressed specially in the brain, gonad, and ovary, and expression of MCH was observed during the developmental stages. In the application of synthetic fMCH and both types of synthetic fMCH, dN-fMCH and dC-fMCH, scale melanophore induced significant changes in aggregation activity with various concentrations of MCH. Also, compared to hMCH and sMCH, fMCH exhibited a 36~99.85% increase in relative potency (%), whereas aggregation of dN-fMCH and dC-fMCH remained in a high concentration. However, dispersion was induced rapidly according to be low concentration of dN-fMCH and dC-fMCH. We show that fMCH and its derivates were bound human MCHR1 and rat MCHR expressed in HEK293T cells with nano-molar affinity and are likely to be ligand-induced to mobilize intracellular $Ca^{2+}$. These results may provide new ligands for binding assay with MCHew ligands, as a structure similar to the mammalian MCH structure was discovered in fish. Once the fMCH receptor system is in place, it can be compared to the MCH system of mammals in terms of MCH function.

• Asian Pacific Journal of Cancer Prevention
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• 제15권24호
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• pp.10659-10663
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• 2015

Background: Rapamycin is an effective anti-angiogenic drug. However, the mode of its action remains unclear. Therefore, in this study, we aimed to elucidate the antitumor mechanism of rapamycin, hypothetically via apoptotic promotion, using MCF-7 breast cancer cells. Materials and Methods: MCF-7 cells were plated at a density of $1{\times}10^5$ cells/well in 6-well plates. After 24h, cells were treated with a series of concentrations of rapamycin while only adding DMEM medium with PEG for the control regiment and grown at $37^{\circ}C$, 5% $CO_2$ and 95% air for 72h. Trypan blue was used to determine the cell viability and proliferation. Untreated and rapamycin-treated MCF-7 cells were also examined for morphological changes with an inverted-phase contrast microscope. Alteration in cell morphology was ascertained, along with a stage in the cell cycle and proliferation. In addition, cytotoxicity testing was performed using normal mouse breast mammary pads. Results: Our results clearly showed that rapamycin exhibited inhibitory activity on MCF-7 cell lines. The $IC_{50}$ value of rapamycin on the MCF-7 cells was determined as $0.4{\mu}g/ml$ (p<0.05). Direct observation by inverted microscopy demonstrated that the MCF-7 cells treated with rapamycin showed characteristic features of apoptosis including cell shrinkage, vascularization and autophagy. Cells underwent early apoptosis up to 24% after 72h. Analysis of the cell cycle showed an increase in the G0G1 phase cell population and a corresponding decrease in the S and G2M phase populations, from 81.5% to 91.3% and 17.3% to 7.9%, respectively. Conclusions: This study demonstrated that rapamycin may potentially act as an anti-cancer agent via the inhibition of growth with some morphological changes of the MCF-7 cancer cells, arrest cell cycle progression at G0/G1 phase and induction of apoptosis in late stage of apoptosis. Further studies are needed to further characterize the mode of action of rapamycin as an anti-cancer agent.

• Journal of Oral Medicine and Pain
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• 제34권3호
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• pp.227-235
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• 2009

Many of the protective functions of saliva can be attributed to the biological, physical, structural, and rheological characteristics of salivary glycoproteins. Therefore, the development of ideal saliva substitutes requires understanding of the rheological as well as biological properties of human saliva. In the present study, we investigated the changes of salivary enzymatic activities by saliva substitutes and compared viscosity of saliva substitutes with human saliva. Five kinds of saliva substitutes such as Moi-Stir, Stoppers4, MouthKote, Saliva Orthana, and SNU were used. Lysozyme activity was determined by the turbidimetric method. Peroxidase activity was determined with an NbsSCN assay. $\alpha$-Amylase activity was determined using a chromogenic substrate, 2-chloro-p-nitrophenol linked with maltotriose. The pH values of saliva substitutes were measured and their viscosity values were measured with a cone-and-plate digital viscometer at six different shear rates. Various types of saliva substitutes affected the activities of salivary enzymes in different ways. Stoppers4 enhanced the enzymatic activities of hen egg-white lysozyme, bovine lactoperoxidase (bLP), and $\alpha$-amylase. Saliva Orthana and SNU inhibited bLP activity and enhanced $\alpha$-amylase activity. MouthKote inhibited $\alpha$-amylase activity. Moi-Stir inhibited the enzymatic activities of bLP and $\alpha$-amylase. The pH values were very different according to the types of saliva substitutes. Stoppers4, MouthKote, and Saliva Orthana showed lower values of viscosity at low shear rates and higher values of viscosity at high shear rates compared with unstimulated and stimulated whole saliva. Moi-Stir and SNU displayed much higher values of viscosity than those of natural whole saliva. Collectively, our results indicate that each saliva substitute has its own biological and rheological characteristics. Each saliva substitute affects the enzymatic activity of salivary enzyme and finally oral health in different ways.

• Asian Pacific Journal of Cancer Prevention
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• 제15권16호
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• pp.6633-6638
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• 2014

The Pharmacological potential, such as antioxidant, anti-inflammatory, and antibacterial activities of Portulaca oleracea (PO) and Petroselinum sativum (PS) extracts are well known. However, the preventive properties against hepatocellular carcinoma cells have not been explored so far. Therefore, the present investigation was designed to study the anticancer activity of seed extracts of PO and PS on the human hepatocellular carcinoma cells (HepG2). The HepG2 cells were exposed with $5-500{\mu}g/ml$ of PO and PS for 24 h. After the exposure, cell viability by 3-(4,5-dimethylthiazol-2yl)-2,5-biphenyl tetrazolium bromide (MTT) assay, neutral red uptake (NRU) assay, and cellular morphology by phase contrast inverted microscope were studied. The results showed that PO and PS extracts significantly reduced the cell viability of HepG2 in a concentration dependent manner. The cell viability was recorded to be 67%, 31%, 21%, and 17% at 50, 100, 250, and $500{\mu}g/ml$ of PO, respectively by MTT assay and 91%, 62%, 27%, and 18% at 50, 100, 250, and $500{\mu}g/ml$ of PO, respectively by NRU assay. PS exposed HepG2 cells with $100{\mu}g/ml$ and higher concentrations were also found to be cytotoxic. The decrease in the cell viability at 100, 250, and $500{\mu}g/ml$ of PS was recorded as 70%, 33%, and 15% by MTT assay and 63%, 29%, and 17%, respectively by NRU assay. Results also showed that PO and PS exposed cells reduced the normal morphology and adhesion capacity of HepG2 cells. HepG2 cells exposed with $50{\mu}g/ml$ and higher concentrations of PO and PS lost their typical morphology, become smaller in size, and appeared in rounded bodies. Our results demonstrated preliminary screening of anticancer activity of Portulaca oleracea and Petroselinum sativum extracts against HepG2 cells, which can be further used for the development of a potential therapeutic anticancer agent.