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The Effects of Platelet- Derived Growth Factor-BB on the Expression of Bone Matrix Protein in the MC3T3-E1 Cells (MC3T3-E1 세포의 골기질 단백질 발현에 대한 혈소판유래성장인자-BB의 효과)

  • Kim, Myo-Sun;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.30 no.2
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    • pp.347-360
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    • 2000
  • Bone remodeling results from the combined process of bone resorption and new bone formation which is regulated in part by some of the polypeptide growth factors such as platelet derived growth factor(PDGF), which has been known to be an important local regulator of bone cell activity and participate in normal bone remodeling. This process includes strictly regulated gene expression of several bone matrix proteins such as type I collagen and osteopontin, a 44 kDa phosphorylated glycoprotein, which has important roles in bone formation. The purpose of this study is to evaluate the effecs of PDGF-BB on the mRNA expression of bone matrix protein, type I collagen and osteopontin, in MC3T3- E1 cell culture. Cells were seeded at $5{\times}10^5$ cells in 10 ml of minimum essential medium alpha(${\alpha}-MEM$) containig 10% fetal bovine serum, 10 mM beta glycerophosphate. 0.1, 1, 10 ng/ml PDGF-BB were added to the cells for the day 3, 7, 14, 21, 28 and cultured for 24 hours. Type I collagen cDNA, Hf677, and osteopontin cDNA were used as probes for northern blot analysis. Total cellular RNA was purified at indicated day and northern blot analysis was performed. The results were as follows : Type I collagen mRNA expressions were higher at the day 3 and 7, and lower in the day 14, 21 in the control groups. In the experimental groups, mRNA expressions were increased when 0.1 ng/ml PDGF-BB were added on the day 3, 7, 21, and decreased in dose-dependent manner on the day 14, decreased at all added dose on the day 28. Osteopontin mRNA expressions were highest in the day 21 groups and lowest in the day 14 groups in the control groups. Interesting results were shown in the day 14 and 21 groups. We found that osteopontin mRNA level was increased in dose dependent manner in the day 14 groups, and decreased dose dependent manner in the day 21 groups. In conclusion, PDGF-BB may have various control effects on type I mRNA expression in the growth and differentiation process of MC3T3-E1 cells and may have contrary regulatory effects on osteopontin mRNA expression. For examples, when the baseline level of osteopontin mRNA was low, as in the day 14, PDGF-BB up-regulated osteopontin mRNA expression in dose dependent manner, and when the baseline level was high as in the day 21, PDGF-BB down-regulated dose dependent manner. Thus, it may be useful for clinical application in periodontal regeneration procedure if further study were performed.

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Antioxidant activities, production of reactive oxygen species, and cytotoxic properties of fractions from aerial parts of glasswort (Salicornia herbacea L.) (퉁퉁마디 지상부 분획의 산화방지 활성, 활성산소종 생성과 세포독성 조절작용)

  • Kang, Smee;Hong, Jungil
    • Korean Journal of Food Science and Technology
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    • v.48 no.6
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    • pp.574-581
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    • 2016
  • Glasswort (Salicornia herbacea L.) is an edible halophyte that grows in salt marshes. In the present study, anti- and pro-oxidant activities and cytotoxic properties of glasswort were investigated. Solvent fractions, including fractions of hexane, ethylether (Fr.E), ethylacetate (Fr.EA), butanol and water, were prepared from a 70% methanol extract of glasswort aerial parts. Fr.EA contained the highest levels of total polyphenols and flavonoids, showing the strongest scavenging activities against DPPH and ABTS radicals, and nitrite. In addition Fr.EA showed the most potent cytotoxic effects on HCT-116 colon cancer cells and INT-407 normal intestinal cells, followed by Fr.E. Most fractions also decreased the level of reactive oxygen species in the treated cells, but generated $H_2O_2$ in the medium. The cytotoxic activity of Fr.EA was more pronounced in the presence of ascorbic acid or N-acetylcysteine. These results indicate that the fractions from aerial parts of glasswort exhibit both anti- and pro-oxidant activities, and these activities modulate cytotoxic properties.

Isolation and Identification of Antimicrobial Compounds against Helicobacter pylori from Rosemary (Rosmarinus officinalis L.) Extracts (로즈마리(Rosmarinus officinalis L.) 추출물로부터 Helicobacter pylori에 대한 항균물질 분리 및 동정)

  • Yoon, So-Jung;Kim, Jin-Sung;Jo, Bun-Sung;Kim, Jeung-Hoan;Lee, Sun-Ho;Ahn, Bong-Jeun;Cho, Young-Je
    • Journal of Applied Biological Chemistry
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    • v.54 no.3
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    • pp.159-165
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    • 2011
  • Rosmarinus officinalis extracts had a significant antimicrobial activity against Helicobacter pyori. Total phenolic contents and inhibition zone of rosemary extracts were estimated to be 25.7 mg/g and 14 mm at $200{\mu}g/mL$ of phenolic contents, respectively. The Sephadex LH-20 and MCI-gel CHP-20 column chromatographic separations for the phenolic extracts from R. officinalis leaves led to isolation of five acids, whose structures were determined as protocatechuic acid (A), coumaric acid (B), caffeic acid (C), chlorogenic acid (D), and rosmarinic acid (E), from interpretation of spectroscopic data including nagative fast atom bombardment (FAB)-mass, $^1H$-NMR, $^{13}C$-NMR, and IR. All isolated compounds were tested for antimicrobial activity against H. pyori. The purified single compound showed less antimicrobial activity against H. pylori than the mixed purified compounds, which generate A+B, A+E, C+D, C+E (each $200{\mu}g/disc$) excellent as large clear zone by synergy effect. These results indicate rosemary extracts are preventive agents against H. pyori.

Effect of Corni Fructus on Testosterone Deficiency Syndrome in In vitro and In vivo (In-vitro와 In-vivo에서 산수유의 남성갱년기 개선효과)

  • Kim, Tae Muk;Jung, Ho Kyung;Jang, Ji Hun;Sim, Mi Ok;Lee, Mu Jin;Cho, Jung Hee;Cho, Hyun Woo
    • Korean Journal of Pharmacognosy
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    • v.47 no.3
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    • pp.264-272
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    • 2016
  • This study was carried out to evaluate the preventive effect of the Corni Fructus (SSU) 50 % EtOH extract (SSU-E50) against bisphenol A (BPA) toxicity in Leydig cells and improving testosterone deficiency syndrome in orchidectomized Sprague-Dawly (SD) rats. Antioxidant properties were measured by radical scavenging activity of SSU-E50 in ABTS assay and DPPH assay. Also, real-time polymerase chain reaction(real-time PCR) was performed to quantify the mRNA expression levels of antioxidant enzyme. SD rats were divided into eight group: normal, sham operation (Sham), orchidectomized (ORX), ORX treated with testosterone 1 mg/kg (Tes. 1), ORX treated with SSU water extract 100 mg/kg (SSU-A 100) and 300 mg/kg (SSU-A 300), ORX treated with SSU 50 % EtOH extract 100 mg/kg (SSU-E 100) and 300 mg/kg (SSU-E 300). On a comparative basis, the SSU showed better activity quenching ABTS with an IC50 value of 0.29 mg/ml and DPPH with an IC50 value of 0.33 mg/ml. Cell viability was evaluated by MTS assay as described not cytotoxic at the highest concentration of $500{\mu}g/ml$. Cytotoxicity of BPA showed in $200{\mu}M$, but definitely survived by treatment with SSU in Leydig cells. In addition, SSU increased the mRNA expression levels of antioxidant enzyme in BPA induced Leydig cells. Superoxide dismutase (SOD) level was slightly increased and malondialdehyde (MDA) level was decreased with SSU-A 100 in in-vivo. These results suggest that Corni Fructus extracts have the greatest property as a natural anti-oxidative and improves testosterone deficiency syndrome source.

Functional Properties of 50% Methanol Extracts from Different Parts of Acanthopanax sessiliflorus (오가피 부위별 50% 메탄올 추출액의 기능적 특성)

  • Choi, Jae-Myoung;Ahn, Jun-Bae
    • Korean Journal of Food Science and Technology
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    • v.44 no.3
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    • pp.373-377
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    • 2012
  • This study was carried out in order to investigate the functional properties of 50% methanol extracts from four parts (root, stem, leaf and fruit) of Acanthopanax sessiliflorus by means of measuring the contents of eleutheroside B, E, and total polyphenols as well as determining electron donating ability (EDA), nitrite scavenging ability (NSA), and anticancer activity. The highest contents of eleutheroside B and E were found in the fruit (538.99 ${\mu}g/g$) and the stem (556.00 ${\mu}g/g$). The root extract demonstrated the highest polyphenol content (2.97 mg/g). EDA of the stem and root extracts were 90.21% and 85.71%. All of the extracts showed 81.5-93.0% of NSA at pH 1.2. In addition all extracts indicated no cytotoxicity to normal cell line (DC2.4). The root extract had a 23% inhibitory effect against the stomach cancer cell line (SNU-719). These results revealed that 50% methanol extracts from A. sessiliflorus can be used as a potential resource of nutraceuticals.

Electron Microscopic Study on the Parafollicular Cells of the Thyroid Gland of the Head and Neck-Irradiated Rats (방사선이 흰쥐 갑상샘 소포곁세포에 미치는 영향에 대한 미세구조적 연구)

  • Kim, Young-Sik;Yang, Nam-Gil;Ahn, E-Tay;Ko, Jeong-Sik;Park, Kyung-Ho;Kim, Jin-Gook
    • Applied Microscopy
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    • v.22 no.2
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    • pp.1-14
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    • 1992
  • This experiment was performed to study the morphological responses of the parafollicular cells of rat following X-ray irradiation. Male rats were divided into normal and experimental groups. The head and neck region of the rat, under sodium thiopental anesthesia, was exposed to 3,000 rads or 6,000 rads of radiation in a single dose, respectively. The source was a Mitsubishi Linear Accelerator ML-4MV. The target to skin distance was 80 cm, and the dose rate was 200 rads/min. The rate of experimental groups were sacrificed on the 6th hour, 2nd and 6th day after X-ray irradiation. Pieces of the tissue taken from the thyroid gland were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde (0.1M Millonig's phosphate buffer, pH 7.3), and in 1% osmium tetroxide (0.1M Millonig's phosphate buffer, pH 7.3), and embedded in araldite mixture. The ultrathin sections stained with uranyl acetate and lead citrate were observed with JEM 100 CX-II electron microscope. The results were as follow; 1. Two types of the parafollicular cells, according to their electron densities, were found, i. e., light cells and dark cells. 2. Three types of the parafollicular cells, according to their sizes of secretory granules were found, i.e., small granule cells ($85nm{\pm}20.1;64{\sim}102nm$), medium granule cells ($120nm{\pm}26.5;77{\sim}179nm$), and large granule cells ($165nm{\pm}25.7;128{\sim}189nm$). 3. The differential ultrastructural changes of the cells according to their cell types, i.e., dark and light cell, or small, medium and large granule cells, were hardly observed in the time and dose range covered by this study. 4. The morphological changes of the parafollicular cells were not pronounced after exposure to 3,000 rads of X-ray. 5. Swollen cisternae of the granular endoplasmic reticulum and partial cytolysis were observed after exposure to 6,000 rads of X-ray. 6. Above results suggest that the parafollicular cells showed the alterations of mitochondrial and granular endoplasmic reticular swelling, and partial cytolysis, but only in doses of 6,000 rads.

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Characterization of Two Novel mAbs Recognizing Different Epitopes on CD43

  • Kim, Soseul;Hong, Jeong Won;Cho, Woon-Dong;Moon, Yoo Ri;Yoon, Sang Soon;Kim, Min-Young;Hong, Kwon Pyo;Lee, Yong-Moon;Yi, Jae Hyuk;Ham, Young Jun;Rah, Hyung Chul;Kim, Seung Ryul;Song, Hyung Geun
    • IMMUNE NETWORK
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    • v.14 no.3
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    • pp.164-170
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    • 2014
  • JL1, a specific epitope on CD43, is a potential biomarker for the diagnosis of acute leukemia. Although qualitative assays for detecting leukemia-specific CD43 exist, there is a need to develop quantitative assays for the same. Here, we developed two novel monoclonal antibodies (mAbs), 2C8 and 8E10, recognizing different epitopes on CD43. These clones are capable of pairing with YG5, another mAb against JL1 epitope, because they were selectively obtained using sandwich ELISA. Antigens recognized by 2C8 and 8E10 were confirmed as CD43 by western blotting using the CD43-hFC recombinant protein. When expression on various leukemic cell lines was investigated, 2C8 and 8E10 displayed a disparity in the distribution of the epitope. Enzyme assays revealed that these mAbs recognized a sialic acid-dependent epitope on CD43. Using normal thymus and lymph node paraffin-embedded tissues, we confirmed a difference in the epitopes recognized by the two mAbs that was predicted based on the maturity of the cells in the tissue. In summary, we developed and characterized two mAbs, 2C8 and 8E10, which can be used with YG5 in a sandwich ELISA for detecting leukemia-specific CD43.

Influence of Water Temperature on Growth of Yearling Sea Bass, Lateolabrax japonicus in Indoor Tank (농어, Lateolabrax japonicus 유어의 성장에 있어 사육 수온의 영향)

  • 강덕영;한형균;전창영
    • Journal of Aquaculture
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    • v.17 no.4
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    • pp.240-245
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    • 2004
  • Two experiments were conducted to study the effect of temperature on growth and survival of yearling sea bass, Lateolabrax japonicus reared from 3$^{\circ}C$ to 33$^{\circ}C$. In the I st experiment, we used yearlings fish averaging 19.2$\pm$0.2 cm/fish (Mean$\pm$S.E.M.) of total length (TL) and 67.0$\pm$1.8 g/fish (Mean$\pm$S.E.M.) of body weight (BW), and we cultured the fish at 3, 6, 9, 12 and 17$^{\circ}C$ for 90 days. In the 2nd experiment, the experimental fish averaging 24.9$\pm$0.1 cm/fish (Mean$\pm$S.E.M) of TL and I 46.4$\pm$3.0 g/fish (Mean$\pm$S.E.M) of BW were reared at 21,24,27,30 and 33$^{\circ}C$ for 90 days. During these experiments, we measured food intake, feed efficiency, survival and growth (TL and BW) in the both experiments. Although food intake of the yearling increased with the temperature, the feed efficiency was only enhanced within the temperature range, from 21 to 27$^{\circ}C$. Growth of yearling was normal within the temperature range from 17 to 3$0^{\circ}C$, but it was stoped or reduced in other temperatures. Survival rate was significantly reduced in 3$^{\circ}C$ from the 1st experiment and in 30 and 33$^{\circ}C$ from the 2nd experiment, but there was no significant difference among other groups.

Enhancement of Anticancer Activities of Ephedra sinica Stapf Extracts by Nano-encapsulation (마황 추출물의 나노 입자화를 통한 항암 활성 증진)

  • Jeong, Hyang-Suk;Kim, Seoung-Seop;Oh, Sung-Ho;Jeong, Myoung-Hoon;Choi, Woon-Yong;Seo, Yong-Chang;Na, Chun-Soo;Kwak, Hyeong-Geun;Lee, Hyeon-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.18 no.3
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    • pp.143-150
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    • 2010
  • This study showed the increase of antitumor activities of water soluble E. sinica extract by nano-encapsulation process with lecithin. Five groups of lecithin only group (LO), lecithin nano-encapsulated E. sinica group (LE), E. sinica only group (EO), one negative control group (NCO) and positive control group (PCO) were set for several anticancer experiment and fed into Sarcoma-180 injected mice. The cytotoxicity of LE on the human normal kidney cell (HEK293) showed 14.8% lower than 19.2% of EO and 18.4% of LO. Growth of human liver carcinoma cell and human stomach carcinoma cell as representative of digestive system in vitro was inhibited up to about 85.1% and 87.3%, in adding 1.0 mg/$m{\ell}$ of LE, which values 15% higher than that from conventional EO. The survival rates of each mice group were 40%, 63%, 48%, 33% and 100%, respectively after 40 days of injecting Sarcoma-180. The increment of their body weights of the extract feeding groups was suppressed down to 10~15%, compared to the negative control. The nano-particles also reduced the hypertrophy of the internal organs such as spleen and liver down to 15~20%, compared to those as the other groups. Among them, LE effectively reduced the size of tumor form to 20%. From these results, in vitro and in vivo antitumor activities of E. sinica could be enhanced by using nano-encapsulation process with lecithin because of better permeation into the cancer cells by confocal observations.

Physiological and Anatomical Studies of Quinclorac Action (Quinclorac의 작용성(作用性)에 대한 연구(硏究))

  • Hong, S.Y.;Lee, I.J.;Kim, K.U.;Shin, D.H.;Lee, C.N.
    • Korean Journal of Weed Science
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    • v.13 no.1
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    • pp.62-70
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    • 1993
  • There was intraspecific variation in Echinochloa crus-galli var, crus-galli in response to quinclorac, showing that plan height and dry weight of a locally collected barnyardgrass(Chinjupi) from Chinju were 90.5 and 37.8% of the untreated control, while those of a locally collected one(Iripi) from Iri showed 19.1 and 14.4%, respectively. The normal distribution curve was obtained from frequency distribution of 89 rice cultivars as affected by the application rates of quinclorac at 30, 300, and 3,000g ai/ha. Protein patterns(SDS-PAGE) of two barnyardgrasses belonging to E, crus-galli var. crus-galli such as Iripi and Chinjupi were not affected by the quinclorac application, indicating that inhibition of enzyme and/or protein biosynthesis seems to be not the primary action target of quinclorac. Electronmicroscopic observation on the injured leaf of Iripi which is considered as a susceptible one showed prominent membrane disruption. Chuchungbyeo(rice variety) resulted in a greater inhibition of tomato growth than those from Chinjupi or Iripi, indicating a great amount of quinclorac discharged from rice root, Chinjupi which is relatively tolerant to quinclorac than Iripi, discharged more quinclorac causing a greater inhibition of tomato growth.

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