• Title/Summary/Keyword: Growth-inhibitor

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Influence of Expression Plasmid of Connective Tissue Growth Factor and Tissue Inhibitor of Metalloproteinase-1 shRNA on Hepatic Precancerous Fibrosis in Rats

  • Zhang, Qun;Shu, Fu-li;Jiang, Yu-Feng;Huang, Xin-En
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7205-7210
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    • 2015
  • Background: In this study, influence caused by expression plasmids of connective tissue growth factor (CTGF) and tissue inhibitor of metalloproteinase-1 (TIMP-1) short hairpin RNA (shRNA) on mRNA expression of CTGF,TIMP-1,procol-${\alpha}1$ and PCIII in hepatic tissue with hepatic fibrosis, a precancerous condition, in rats is analyzed. Materials and Methods: To screen and construct shRNA expression plasimid which effectively interferes RNA targets of CTGF and TIMP-1 in rats. 50 cleaning Wistar male rats are allocated randomly at 5 different groups after precancerous fibrosis models and then injection of shRNA expression plasimids. Plasmid psiRNA-GFP-Com (CTGF and TIMP-1 included), psiRNA-GFP-CTGF, psiRNA-GFP-TIMP-1 and psiRNA-DUO-GFPzeo of blank plasmid are injected at group A, B, C and D, respectively, and as model control group that none plasimid is injected at group E. In 2 weeks after last injection, to hepatic tissue at different groups, protein expression of CTGF, TIMP-1, procol-${\alpha}1$ and PC III is tested by immunohistochemical method and,mRNA expression of CTGF,TIMP-1,procol-${\alpha}1$ and PCIII is measured by real-time PCR. One-way ANOVA is used to comparison between-groups. Results: Compared with model group, there is no obvious difference of mRNA expression among CTGF,TIMP-1,procol-${\alpha}1$, PC III and of protein expression among CTGF, TIMP-1, procol-${\alpha}1$, PC III in hepatic tissue at group injected with blank plasmid. Expression quantity of mRNA of CTGF, TIMP-1, procol-${\alpha}1$ and PCIII at group A, B and C decreases, protein expression of CTGF, TIMP-1, procol-${\alpha}1$, PC III in hepatic tissue is lower, where the inhibition of combination RNA interference group (group A) on procol-${\alpha}1$ mRNA transcription and procol-${\alpha}1$ protein expression is superior to that of single interference group (group B and C) (P<0.01 or P<0.05). Conclusions: RNA interference on CTGF and/or TIMP-1 is obviously a inhibiting factor for mRNA and protein expression of CTGF, TIMP-1, procol-${\alpha}1$ and PCIII. Combination RNA interference on genes of CTGF and TIMP-1 is superior to that of single RNA interference, and this could be a contribution for prevention of precancerous condition.

Optimization of Culture Conditions for Maintaining Pig Muscle Stem Cells In Vitro

  • Choi, Kwang-Hwan;Yoon, Ji Won;Kim, Minsu;Jeong, Jinsol;Ryu, Minkyung;Park, Sungkwon;Jo, Cheorun;Lee, Chang-Kyu
    • Food Science of Animal Resources
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    • v.40 no.4
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    • pp.659-667
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    • 2020
  • Muscle stem cells isolated from domestic animals, including cows and pigs, were recently spotlighted as candidates for the production of alternative protein resources, so-called cultured meat or lab-grown meat. In the present study, we aimed to optimize the in vitro culture conditions for the long-term expansion of pig muscle stem cells via the screening of various signaling molecules. Pig muscle stem cells were collected from the biceps femoris muscles of 3-d-old crossbred pigs (Landrace×Yorkshire×Duroc, LYD) and cultured in minimum essential medium-based growth media. However, the pig muscle stem cells gradually lost their proliferation ability and featured morphologies during the long-term culture over two weeks. To find suitable in vitro culture conditions for an extended period, skeletal muscle growth medium-2, including epidermal growth factor (EGF), dexamethasone, and a p38 inhibitor (SB203580), was used to support the stemness of the pig muscle stem cells. Interestingly, pig muscle stem cells were stably maintained in a long-term culture without loss of the expression of myogenic marker genes as determined by PCR analysis. Immunostaining analysis showed that the stem cells were capable of myogenic differentiation after multiple passaging. Therefore, we found that basal culture conditions containing EGF, dexamethasone, and a p38 inhibitor were suitable for maintaining pig muscle stem cells during expanded culture in vitro. This culture method may be applied for the production of cultured meat and further basic research on muscle development in the pig.

Plasminogen Activator Inhibitor Type 1 (PAI-1) A15T Gene Polymorphism Is Associated with Prognosis in Patients with EGFR Mutation Positive Pulmonary Adenocarcinoma

  • Lim, Ju Eun;Park, Moo Suk;Kim, Eun Young;Jung, Ji Ye;Kang, Young Ae;Kim, Young Sam;Kim, Se Kyu;Shim, Hyo Sup;Cho, Byoung Chul;Chang, Joon
    • Tuberculosis and Respiratory Diseases
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    • v.75 no.4
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    • pp.140-149
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    • 2013
  • Background: Plasminogen activator inhibitor type 1 (PAI-1), an important regulator of plasminogen activator system which controls degradation of extracellular membrane and progression of tumor cells, and PAI-1 gene polymorphic variants have been known as the prognostic biomarkers of non-small cell lung cancer patients. Recently, experimental in vitro study revealed that transforming growth factor-${\beta}1$ initiated PAI-1 transcription through epithelial growth factor receptor (EGFR) signaling pathway. However, there is little clinical evidence on the association between PAI-1 A15T gene polymorphism and prognosis of Korean population with pulmonary adenocarcinoma and the influence of activating mutation of EGFR kinase domain. Methods: We retrospectively reviewed the medical records of 171 patients who were diagnosed with pulmonary adenocarcinoma and undergone EGFR mutation analysis from 1995 through 2009. Results: In all patients with pulmonary adenocarcinoma, there was no significant association between PAI-1 A15T polymorphic variants and prognosis for overall survival. However, further subgroup analysis showed that the group with AG/AA genotype had a shorter 3-year survival time than the group with GG genotype in patients with EGFR mutant-type pulmonary adenocarcinoma (mean survival time, 24.9 months vs. 32.5 months, respectively; p=0.015). In multivariate analysis of 3-year survival for patients with pulmonary adenocarcinoma harboring mutant-type EGFR, the AG/AA genotype carriers had poorer prognosis than the GG genotype carriers (hazard ratio, 7.729; 95% confidence interval, 1.414-42.250; p=0.018). Conclusion: According to our study of Korean population with pulmonary adenocarcinoma, AG/AA genotype of PAI-1 A15T would be a significant predictor of poor short-term survival in patients with pulmonary adenocarcinoma harboring mutant-type EGFR.

The Effect of Hybridoma Growth Rate on the Production of Monoclonal Antibodies (하이브리도마 세포의 증식속도와 단일클론 항체의 생산)

  • 최태부;조보연
    • KSBB Journal
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    • v.4 no.1
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    • pp.31-33
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    • 1989
  • The effect of growth rate change on glucose consumption and Ammonia production rate in batch culture of hybridoma was studied. The methods regulating growth rate of hybridoma were 1) decrease of serum concentration, 2) decrease of culture temperature and 3) addition of growth inhibitor (thymidine). The experimental results showed that hybridoma growth rate was dropped by 20~50%, while glucose consumption and ammonia production rate was decreased up to 40% On the other hand, the final concentration of monoclonal antibody was shown to be increased as high as 100% when the concentration of serum was decreased from 2% to 0.2%.

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Inhibitory Effect of Garlic on the Growth of Aspergillus parasiticus (한국산 마늘에 의한 Aspergillus parasiticus의 성장 억제 효과)

  • Park, Jeong-Yeong;Kim, Jong-Gyu
    • Journal of Environmental Health Sciences
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    • v.35 no.2
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    • pp.95-99
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    • 2009
  • This study was performed to investigate the possible effect of garlic produced in Korea on the inhibition/reduction of growth of A. parasiticus, a toxigenic strain. The effect was studied using different concentrations of freeze-dried garlic in potato-dextrose agar (PDA) and/or in yeast-extract sucrose (YES) broth at $25^{\circ}C$ for 15 days. While inhibition of the fungal growth due to increasing the concentration of garlic was observed, the more remarkable effect was observed on the ninth day. Reduction of fungal diameter as a result of addition of garlic on PDA was observed to range between 3.4% to 20.1 % while reduction of mycelial weight in YES broth ranged from 9.9% to 30.5%. The 0.5% and 1.0% concentrations of garlic significantly reduced fungal diameter in PDA on the 9th day, while 0.1 %, 0.5%, and 1.0% concentrations of garlic significantly reduced the mycelial weight in YES broth (p<0.05). Dose-response relationships were observed between the concentration of garlic and inhibition of growth both in solid culture and in liquid culture. This study indicates that garlic could be an effective inhibitor at a human consumption level of the growth of A. parasiticus. More research is needed to study the inhibitory effects of the main active component of garlic.

Studies on the inhibitory substance of yeast growth (Part II ) Effect on the nitrogen uptake (항효모성 물질에 관한 연구 (제II보) 질소대사와의 관계)

  • 서정훈;송방호;유춘발
    • Microbiology and Biotechnology Letters
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    • v.1 no.1
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    • pp.3-11
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    • 1973
  • The biochemical characteristics of Astradix -P, isolated from Astragalus membranaceus Bunge as yeaststatic substance, were reported on a previous paper. And on this report, some relation to the nitrogen metabolism of yeast was studied. Inorganic or organic source of nitrogen easily uptaking yeast did not show any antagonistic action to the inhibitory action of Astradix -P on the yeast growth. Especially an organic nitrogen source, arginine, histidine and lysine, classified to basic amino acid, was reacted as an antagonistic substance to the sample. But, ornithine, a basic amino acid, did not show any antagonistic action to the sample. In the mixed media containing neutral and acidic amino acids as a nitrogen source, yeast growth was inhibited strongly. If the basic amino acid was added to the same mixed media, the yeast growth was not inhibited by Astradix-P therefore, the antagonistic action of basic amino acid to the Astradix-p was readily observed. The yeast static action of Astradix-P was partially related to the isoelectric point of amino acid as a nitrogen source. Yeast cells which propagated under the media containing growth inhibitor, Astradix -p, did not bring any remarkable denaturation of cell structure by electro-microscopic observation.

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Effect of Inhibitors on cell growth and urease activity of Vibrio parahaemolyticus (저해제가 Vibrio parahzemolyticius 균주의 생육 및 요소분해효소의 활성에 미치는 영향)

  • 김종숙;김영희
    • Journal of Life Science
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    • v.10 no.6
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    • pp.558-563
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    • 2000
  • Effect of inhibitors on Vibrio parahaemolyticus cell growth and its urease activity was studied. The growth of the bacterium and the enzyme activity were inhibited by the addition of 0.02% p-hydroxymercuric benzoate, $HgCl_2$and $AgNO_3$. However, same concentration of boric acid, thallium acetate and $Pb(NO_3)_2$ did not affect the cell growth but inhibited urease activity by 25%, 29%, and 38%, respectively. Acetohydroxamic acid was the most potent inhibitor on cell growth by inhibiting 40% but did not affect urease activity. To investigate the effect of inhibitors on urease activity, urease was purified and confirmed on SDS-PAGE. The purified urease was inhibited 100% by the addition of 1 mM acetohydroxamic acid and $AgNO_3$but no inhibition was occurred by the addition of the same concentration of thallium acetate. and the addition of 0.01 mM of $HgCl_2$ and acetohydroxamic acid inhibited the purified urease activity by 39% and 24%, respectively. On 0.1 millimolar basic, acetohydroxamic acid and $HgCl_2$inhibited 4 times more active in urease inhibition than p-hydroxymercuric benzoate whereas no inhibition was occurred either thallium acetate or $Pb(NO_3)_2$.

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Inhibitory Effect of Korean Fermented Vegetable (Kimchi) on the Growth and Aflatoxin Production of Aspergillus parasicitus-Part 1.

  • Kim, Jong-Gyu;Lee, Kyung-Min
    • Journal of Environmental Health Sciences
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    • v.33 no.3
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    • pp.190-194
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    • 2007
  • Aflatoxin $B_1$ is a mycotoxin produced by Aspergillus flavus and A. parasiticus and is a human carcinogen. This study was performed to investigate reduction of growth and aflatoxin production of A. parasiticus by kimchi. A. parasiticus was grown in a modified APT broth with the juice of kimchi (at a concentration of 7%) at $28^{\circ}C$ for 9 days. Aflatoxin $B_1$ was determined by use of high performance liquid chromatography (HPLC). The addition of the juice of kimchi significantly reduced mycelial growth and aflatoxin production during the incubation period (p<0.05). Reduction of mycelial growth of A. parasiticus as the result of addition of the juice of kimchi was observed to range between 64.8 to 83.4% while reduction of aflatoxin production ranged from 62.2 to 73.0%. This study indicates that kimchi could be an effective inhibitor of aflatoxin production although mycelial growth may be permitted. More research is needed to study the inhibitory effects of the metabolites of kimchi.

TARGETED MOLECULAR THERAPY IN A MURINE MODEL OF ORAL SQUAMOUS CELL CARCINOMA WITH AN EPIDERMAL GROWTH FACTOR RECEPTOR INHIBITOR (구강 편평상피세포암 마우스 모델에서 상피성장인자 수용체 억제제를 적용한 분자표적치료)

  • Park, Young-Wook
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.31 no.1
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    • pp.8-17
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    • 2009
  • Purpose: We determined the therapeutic effect of an epidermal growth factor receptor (EGFR)-specific monoclonal antibody (mAb), cetuximab (Erbitux) on the growth of oral squamous cell carcinoma (OSCC) xenografted in athymic nude mice. Experimental Design: We induced subcutaneous tumors by inoculating human tumor cell suspension into the right flank of nude mice. Nude mice with subcutaneous tumors were randomized to receive cetuximab alone, paclitaxel alone, cetuximab plus paclitaxel, or a placebo (control). Antitumor mechanisms of cetuximab were determined by immunohistochemical and apoptosis assays. Results: Cetuximab, paclitaxel, and cetuximab/paclitaxel combined therapy resulted in 50%, 52%, 67% in vivo inhibition of tumor proliferation, respectively. Tumors of mice treated with cetuximab plus paclitaxel demonstrated decreased PCNA-positive tumor cells and increased apoptotic tumor cells, which slowed growth of the murine tumors. Conclusion: These data show that EGFR can be a molecular target for the treatment of OSCC. And combination therapy with cetuximab and paclitaxel warrants further clinical study.

Studies on the inhibitory substance of yeast growth - (Part 1) Isolation and biochemical characteristics of yeast growth inhibitor - (항효모성물질(抗酵母性物質)에 관(關)한 연구(硏究) - (제 1 보)(第 1 報) 분리(分離) 및 그 생화학적작용(生化學的作用)과 성질(性質)에 대(對)해서 -)

  • Seu, Jung-Hwn;Kho, Yung-Hee
    • Applied Biological Chemistry
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    • v.15 no.1
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    • pp.19-25
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    • 1972
  • A kind of peptide which posseses an yeaststatic activity was isolated from Astragalus membranaceus Bunge and following characteristics was obtained. 1. The isoelectric pH of this peptide was 8.2 and histidine, an alkaline amino acid, was identified from this peptide. 2. This substance showes conspicuous heat stability and does not indicate any remarkable reduction of yeaststatic activity even for 5 hours treatment at $100^{\circ}C$. or for 30 minutes at $121^{\circ}C$. 3. The inhibitory activity of the yeast growth is not originated from the yeastcidal action but yeaststatic effect of this sample. 4. The sample shows strong stability ranging from pH 2 to 10. 5. The saccharide; glucose, sucrose, maltose, gives no effect on the yeaststatic activity of the sample even high concentration, 15 percent, and also no effect gives by magnesium, calcium and phosphate salts. 6. The available concentration of this sample on the inhibition of yeast growth was located at the ppm extent, for example, the concentration of fifty percent growth inhibition to Saccharomyces cerevisiae or S. carsbergensis was 4 ppm and 3 ppm to Candida pulcherrima, 13 ppm to S. coreanus, 18 ppm to S. sake and 38 ppm to C. tropicalis. 7. On the alcohol fermentation of S. coreanus, the peptide, an yeast growth inhibitor, gives no effect at all. 8. This substance is named as Astradix-P (Astragalus membranaceus, Radix, Peptide).

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