• Asian Pacific Journal of Cancer Prevention
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• 제16권16호
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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.

• 한국축산식품학회지
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• 제40권4호
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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.

• Tuberculosis and Respiratory Diseases
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• 제75권4호
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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.

• KSBB Journal
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• 제4권1호
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• pp.31-33
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• 1989

하이브리도마의 회분식 배양에서 세포증식 속도가 기질의 소비 속도나 단일클론 항체의 생산에 미치는 영향을 조사하였다. 세포의 증식속도를 조절하는 방법으로는 다음 3가지를 이용하여다. 1)배양액내 혈청의 농도를 낮추는 방법, 2) 배양온도를 낮추는 방법, 3)세포증식 억제제를 첨가하는 방법, 실험결과에 따르면 위의 세가지 방법에 의해 세포증식 속도가 20~50% 감소하는 동안 기질의 소비속도와 암모니아의 생성 속도는 40%까지 감소하였다. 반면에 단일클론 항체의 최종농도는 하이브리도마 세포의 증식이 저해되지 않았을 경우데 비해 100%까지 증가하는 것으로 나타났다. 따라서 이 결과는 하이브리도마 배양에서 단일크론 항체 생산의 증가에 유용한 방법으로 이용 될 수 있다.

• 한국환경보건학회지
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• 제35권2호
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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.

• 한국미생물·생명공학회지
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• 제1권1호
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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.

• 생명과학회지
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• 제10권6호
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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 $Pb(NO_3)_2$.

• 한국환경보건학회지
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• 제33권3호
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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.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제31권1호
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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.

• Applied Biological Chemistry
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• 제15권1호
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• pp.19-25
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• 1972

Astragalus membranaceus Bunge로 부터 항효모성물질(抗酵母性物質) Yeaststatic substance인 일종(一種)의 peptide (Astradix-P)를 얻었으며 이 물질(物質)의 특성(特性)은 다음과 같다. 1) 이 물질(物質)은 일종(一種)의 peptide이며 그 PI는 8.2로 나타났다. 2) 이 물질(物質)은 효모(酵母)의 증식(增殖)을 조해(阻害)하며 이 조해(阻害)는 정균효과(靜菌效果)에 기인(起因)한다. 3) 본(本) 물질(物質)은 열안정성(熱安定性)이 매우 커서 $100^{\circ}C$에서 10시간(時間), 혹은 $121^{\circ}C$에서 30분간(分間) 가열(加熱)하여도 거의 실활(失活)하지 않는다. 4) pH에 대(對)해서도 안정(安定)하며 $pH2{\sim}pH10$까지는 별(別) 영향(影響)이 없다. 5) 본(本) 물질(物質)의 작용(作用)에 있어서 glucose, sucrose, maltose등의 당류(糖類)는 그 농도(濃度)에 관계(關係)없이 전연(全然) 영향(影響)을 미치지 아니한다. 6) 본(本) 물질(物質)의 작용(作用)에 있어서 Mg, Ca, $PO_4$ 등 염류(鹽類)도 전연(全然) 영향(影響)을 미치지 않는다. 7) 본(本) 물질(物質)은 효모류(酵母類)의 생육(生育)을 억제(抑制)하며 그 유효농도(有效濃度)는 ${\mu}g/ml$의 범위(範圍)에 있으며 예(例)로서, Saccharomyces carlsbergensis, S. cerevisiae 등 균주(菌株)의 $G.I.\;_{50}$는 $4{\mu}g/ml$이며, S. coreanus, S. sake는 13, $18\;{\mu}g/ml$이며, Candida pulcherrima는 $3\;{\mu}g/ml$, C. tropicalis는 $38{\mu}g/ml$로 나타났다. 8) 본(本) 물질(物質)은 효모(酵母)의 alcohol 발효(醱酵)에 대(對)해서 전연(全然) 영향(影響)을 미치지 않는다.