• Journal of Life Science
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• v.29 no.10
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• pp.1104-1110
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• 2019

Recently, there has been an increase in the elderly population of the world. Consequently, bone metabolic diseases such as osteoporosis are emerging as a social problem. Osteoclasts play a role in bone resorption, and osteoporosis is induced when bone resorption occurs excessively. Because currently used bone resorption inhibitors may cause side effects when used for a long period of time, it is necessary to develop a new material that effectively inhibits osteoclast differentiation. This study aimed to confirm the inhibitory effect of ethanol extract of Locusta migratoria on RANKL-induced osteoclast differentiation and its mechanism. The toxicity and proliferation effects of LME on RAW264.7 osteoclasts were measured by an MTS assay. There was no cytotoxicity or proliferation when the osteoclasts were treated with up to $2,000{\mu}g/ml$ of LME. In order to confirm the effect of LME on the differentiation of osteoclasts, osteoclasts were treated with RANKL alone or with LME for 3 days. As a result of a TRAP (tartrate-resistant acid phosphatase) assay, the increasing osteoclast differentiation by RANKL decreased in a concentration-dependent manner with the treatment of LME. In addition, LME suppressed the expression of differentiation-related marker genes (TRAP, RANK, NFATc1, and CK) and proteins (NFATc1 and c-Src) that had been increased by RANKL. Also, LME influenced the $NF-{\kappa}B$, ERK and JNK signaling pathways, resulting in the inhibition of osteoclast differentiation. These results suggest that LME may be used as a novel functional material for the prevention and treatment of osteoporosis by playing a role in inhibiting bone absorption.

• Journal of Life Science
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• v.31 no.2
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• pp.209-218
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• 2021

This study aimed to evaluate the anti-obesity effects of Cudrania tricuspidata leaf extract in the order of leaf development on the shoot (L0, L1, L2, L3, L4, and L5). The leaves at the apex of a Cudrania tricuspidata shoot were classified as L0; the next leaves of the apex were classified as L1, L2, L3, and L4 from highest to lowest; and the lowest leaf was classified as L5. A series of 70% ethyl alcohol leaf extracts were screened for the inhibitory effects of adipogenesis in 3T3-L1 preadipocytes. We found that the apical leaf extract of Cudrania tricuspidata (CTL0) was the most effective. Next, a study was conducted on the inhibitory action mechanism of CTL0. Treatment with CTL0 significantly suppressed the differentiation of 3T3-L1 preadipocytes in a dose-dependent manner, as confirmed by the decrease in lipid droplet content observed with Oil Red O staining. Treatment with 12.5 ㎍/ml, 25 ㎍/ml, and 50 ㎍/ml of CTL0 significantly reduced the lipid droplet content. Glucose and cellular triglyceride concentrations were reduced in the 3T3-L1 cells on the CTL0-treated medium compared to the differentiation medium (DM control, DMEM + insulin + dexamethasone + rosiglitazone). Compared with DM, CTL0 significantly inhibited the expression of key pro-adipogenic transcription factors, including peroxisome proliferator-activated receptor γ (PPARγ), LPL, A-FABP, and Glut4. These findings show that CTL0 extract has potent anti-obesity effects.

• Journal of Digital Convergence
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• v.12 no.4
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• pp.419-425
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• 2014

The objective of this study is to estimate the inhibition of skin melanin formation by extract of Angelica koreana and Cnidium monnieri and the possibility of functional cosmetic materials through anti-irritation and stability test. The extract used in this experiment is White-$AK^{TM}$ and the INCI name is Osthole. The main component of White-$AK^{TM}$ was identified as coumarin and EC50 value was 2.7ppm by mouse melanoma B 16 cell test. White-$AK^{TM}$ showed inhibitory effects 100 times lower concentration than arbutin. The main mechanism for skin whitening effect thought to be inhibition of tyrosinase-related gene expression. The basic essence formulation of White-$AK^{TM}$ 5% solution applied to the skin showed the effect of relieving skin irritation. White-$AK^{TM}$ in an opaque container, under UV conditions for 4 weeks, and showed close to 100% recovery and 97% recovery under $50^{\circ}C$ for 4 weeks. Therefore, it is thought that White-$AK^{TM}$ which helps skin whitening, relieving skin irritation and stable in UV condition is able to be used as the functional component in the cosmetic formulation.

• Herbal Formula Science
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• v.15 no.1
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• pp.163-173
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• 2007

Jeungaektang (JAT) is the herbal formula, has the effect of moistening the dryness by activating lung Qi and by nourishing Yin, has being used for dryness syndromes. Generally the herbal formulae for moistening dryness are used for exogenous or endogenous dryness syndromes. JAT has been clinically used for the treatment of endogenous dryness syndromes. It is composed of Scrophulariae Radix. Rehmanniae Radix and Liriopis Tuber. Recent studies showed that JAT has a protective effect against $CCl_{4}-induced$ hepatotoxicity and anti-inflammatory effects against ear swelling of mouse induced by Crotonis Fructus. However, the effect of JAT on the immunological activity was rarely studied. Therefore, this study evaluated the effects of JAT the regulatory mechanism of nitric oxide (NO) and cytokines in the lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. After the treatment of JAT water extract, cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. Cyclooxygenase-2 (COX -2) and inducible nitric oxide synthase (iNOS) were determined by immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. Results provided evidence that JAT inhibited the production of nitrite and nitrate ($0.1{\sim}1.0$ mg/ml), iNOS ($0.1{\sim}1.0$ mg/ml), $interleukin-1{\beta}$ ( $0.1{\sim}1.0$ mg/ml) and tumor necrosis $factor-{\alpha}$ ($0.1{\sim}1.0$ mg/ml) in RAW 264.7 cells activated with LPS. Furthermore, JAT inhibited the expression of COX-2 expression and production of prostagladin E2 ($0.1{\sim}1.0$ mg/ml). These findings suggest that JAT can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.3
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• pp.880-886
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• 2004

Phellinus igniarius has been clinically used for the treatment of hemorrhoidal fistula, dysmenorrhea and the prevention of cancer in traditional oriental medicine. Recent studies showed that Phellinus igniarius produced anti-cancer, anti-metastasis and immuno-modulatory effects, There is lack of studies regarding the effects of Phellinus igniarius on the immunological activities. The present study was conducted to evaluate the effect of Phellinus igniarius on the regulatory mechanism of cytokines and nitric oxide (NO) for the immunological activities in Raw 264,7 cells. After the treatment of Phellinus igniarius water extract, cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. COX-2 and iNOS were determined by Immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. Results provided evidence that Phellinus igniarius inhibited the production of nitrite and nitrate (NO), inducible nitric oxide synthase (iNOS), interleukin-1β (IL-1β) and interleukin-6 (IL-6), and the activation of phospholylation of inhibitor κBα (p-IκBα) in Raw 264.7 cells activated with lipopolysaccharide (LPS). These findings suggest that Phellinus igniarius can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections.

• The Korean Journal of Physiology
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• v.15 no.1
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• pp.61-66
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• 1981

In 1944 Cavallito and Bailey first extracted an essential oil, a powerful antibacterial principle, from the garlic and named it allicin. Later Stoll and Seeback elucidated that allicin was produced from alliin by the enzymatic action of arinase. Damaru observed the depressor responses following intraperitoneal administration of garlic juice in cats. And Thiersch presented evidence that garlic had a protective action against experimental arterosclerosis in cholesterol-fed animals. On the other hand it was also reported that anemias were caused by long-term ingestion of garlic as a result of reduction in hemoglobin and RBC. From the experiment in which the effect of garlic on the blood sugar level was studied, Lee insisted garlic elevated blood sugar level. However, August and Jain claimed that hypoglycemia was induced by garlic administration. Recently Bordia and Bansal suggested that essential oils extracted from onion and garlic have a strong preventive effect on hyperlipemia and prolonged coagulation time resulted from fat-feeding. Furthermore Bordia et al indicated that garlic exerted a strong fibrinolytic activity. In early 1920 s Sugihara reported that essential oil of garlic not only decreased arterial blood pressure but also had a paralytic effect on the isolated heart and intestinal strip of animals. The present study was proposed to investigate the effect of garlic juice and the mechanism of its action on the motility of the isolated rabbit duodenum. The motility of the isolated duodenum was recorded on polygraph by means of force transducer connected with Magnus apparatus. And the isolated duodenum was separtely pretreated with $acetylcholine(5{\times}10^{-7}\;gm/ml)$, $pilocarpine(2.5{\times}10^{-6}\;gm/ml)$, $histamine(5{\times}10^{-6}\;gm/ml)$ and barium $chloride(2.5{\times}10^{-5}\;gm/ml)$ in order to find out interations of these drugs with ASJ. The results obtained were as follows; At concentrations of 0.25%, 0.5% and 1.0% ASJ markedly inhibited contractions of isolated duodenum while tonus as well as contractility of the isolated intestine were decreased also with 0.5% and 1.0% ASJ. Since ASJ markedly abolished augmented motility of isolated intestine by histamine and partly reduced that by $BaCl_2$, it is strongly suggested that inhibitory action of ASJ on the intestinal motility is caused mainly by its antihistamine effect and partly by its direct action on the intestinal smooth muscle.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.4
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• pp.2301-2305
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• 2013

Ardisia crispa (Family: Myrsinaceae) is an evergreen, fruiting shrub that has been traditionally used as folklore medicine. Despite a scarcity of research publications, we have succeeded in showing suppressive effects on murine skin papillomagenesis. In extension, the present research was aimed at determining the effect of a quinone-rich fraction (QRF) isolated from the same root hexane extract on both initiation and promotion stages of carcinogenesis, at the selected dose of 30 mg/kg. Mice (groups I-IV) were initiated with a single dose of 7,12-dimethylbenz(${\alpha}$)anthracene (DMBA, $100{\mu}g/100{\mu}l$) followed by repeated promotion of croton oil (1%) twice weekly for 20 weeks. In addition, group I (anti-initiation) received QRF 7 days before and after DMBA; group II (anti-promotion) received QRF 30 minutes before each croton oil application; group III (anti-initiation/promotion) was treated with QRF as a combination of group I and II. A further two groups served as vehicle control (group V) and treated control (group VI). As carcinogen control, group IV showed the highest tumor volume ($8.79{\pm}5.44$) and tumor burden ($3.60{\pm}1.17$). Comparatively, group III revealed only 20% of tumor incidence, tumor burden ($3.00{\pm}1.00$) and tumor volume ($2.40{\pm}1.12$), which were significantly different from group IV. Group II also showed significant reduction of tumor volume (3.11), tumor burden (3.00) and tumor incidence (11.11%), along with prominent increase of latency period of tumor formation (week 12). Group I, nonetheless, demonstrated marked increment of tumor incidence by 40% with prompted latency period of tumor formation (week 7). No tumor formation was observed in groups V and VI. This study provided clear evidence of inhibitory effects of QRF during promotion period which was in agreement with our previous findings. The mechanism(s) underlying such effects have yet to be elucidated.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.3
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• pp.401-409
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• 2010

FLICE inhibitory protein (FLIP) is one of the important anti-apoptotic proteins in the Fas/FasL apoptotic path which has death effect domains, mimicking the pro-domain of procaspase-8. To reveal the intracellular signal transduction molecules involved in the process of follicular development in the bovine ovary, we cloned the c-FLIP(L) gene in bovine ovary tissue with the reverse transcription polymerase chain reaction (RT-PCR), deleted the termination codon in its cDNA, and directionally cloned the amplified c-FLIP(L) gene into eukaryotic expression vector pAcGFP-Nl, including AcGFP, and successfully constructed the fusion protein recombinant plasmid. After identifying by restrictive enzyme BglII/EcoRI and sequencing, pAcGFP-bFLIP(L) was then transfected into follicular granulosa cells, mediated by Lipofectamine 2000, the expression of AcGFP observed and the transcription and expression of c-FLIP(L) detected by RT-PCR and Western blot. The results showed that the cattle c-FLIP(L) was successfully cloned; the pAcGFPbFLIP(L) fusion protein recombinant plasmid was successfuly constructed by introducing a BglII/EcoRI cloning site at the two ends of the c-FLIP(L) open reading frame and inserting a Kozak sequence before the start codon. AcGFP expression was detected as early as 24 h after transfection. The percentage of AcGFP positive cells reached about 65% after 24 h. A 1,483 bp transcription was amplified by RT-PCR, and a 83 kD target protein was detected by Western blot. Construction of the pAcGFP-bFLIP(L) recombinant plasmid should be helpful for further understanding the mechanism of regulation of c-FLIP(L) on bovine oocyte formation and development.

• Journal of Acupuncture Research
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• v.20 no.6
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• pp.128-139
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• 2003

Juglans sinensis Dode has been reported to have antioxidant activity. However, the effect of Juglans sinensis Dode aquacupuncture(JS) on reactive oxygen species(ROS)-induced alterations in membrane transport function in renal tubular cells. This study was performed to evaluate the effect of JS on the organic hydroperoxide t-butylhydroperoxide(tBHP)-induced inhibition of $Na^+$-dependent phosphate($Na^+$-Pi) uptake in opossum kidney (OK) cells, an established renal proximal epithelial cell line. tBHP inhibited $Na^+$-Pi uptake in a time-dependent manner. The inhibitory effect of tBHP was prevented by JS over concentration range of 0.05-1mg/100ml in a dose-dependent manner. Kinetic studies showed that tBHP caused an decrease in Vmax for $Na^+$-Pi uptake without any a significant change in Km. $Na^+$-dependent phosphonoformic acid binding, a irreversible inhibitor of renal $Na^+$-Pi uptake, was decreased by tBHP treatment. The reduction in Vmax and phosphonoformic acid binding by tBHP was prevented by JS. tBHP induced lipid peroxidation and its effect was completely inhibited by JS and antioxidant N,N'-diphenyl-p-phenylenediamine. These data suggest that the oxidant inhibits phosphate uptake by a reduction in the number of active carrier across the membrane. JS may prevent oxidant-induced inhibition of membrane transport function by a mechanism similar to antioxidants in renal epithelial cells. Although the precise constituents remain to be explored, JS may be employed as a useful candidate herb for drug development to prevent and treat oxidant-mediated renal failure.

• Environmental Analysis Health and Toxicology
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• v.18 no.2
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• pp.95-100
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• 2003

Dichlorodiphenyltrichloroethane (DDT), is a widespread environmental pollutant. In this study, we investigated the effect of DDT on testosterone production through aromatase and investigated its molecular mechanism in testicular leydig cell, R2C. We investigated that the effects of DDT on testosterone production and its effects on aromatase activity in R2C cell by radio immunoassay (RIA). As the results, the potent leyding cell activator LH increased testosterone production compared to the control. DDT exposure significantly decreased testosterone production in R2C cell and DDT alone affected T reduction in a dose-dependent manner in R2C cell slightly. In addition, DDT was found to increase aromatase activity in R2C cell in a dose dependent manner. In order to assess whether the suppressive effects of DDT on LH-inducible testosterone production might be influenced by the ER, ICI 182.780, a pure antiestrogen, was used, and it was found that these inhibitory effects of DDT were antagonized by ICI 182.780, implying that the ER mediates the suppressive effects of DDT. Furthermore, the inducible effects of DDT on aromatase might be influenced by the ER, ICI 182.780 was used, and it was found that these enhancing effects of DDT were antagonized by ICI 182.780, implying that the ER mediates the inducible effects of DDT. Our results indicated that DDT inhibition of LH-inducible testosterone production in R2C is mediated through aromatase. However, the precise mechanisms by which DDT enhance in leyding cell remains unknown. The current study suggests the possibility that DDT might act as a modulator aromatase gene transcription.