• 대한본초학회지
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• 제31권4호
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• pp.101-109
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• 2016

Objectives : HT070 is a mixture of herbal extracts from root of Scutellaria baicalensis and stem bark of Eleutherococcus senticosus , which have long been used for stroke therapy in traditional Korean Medicine. The purpose of this study was to investigate the neuroprotective effects of HT070 on global cerebral ischemia and its potential mechanisms.Methods : Transient global cerebral ischemia was produced by 10 min of four-vessel occlusion (4-VO) in male Wistar rats. HT070 was administered orally at a dosage of 200 mg/kg twice at 0 and 90 min after reperfusion. Hippocampal neuronal damage was measured 7 days after reperfusion. To explore the potential mechanisms, we used hydrogen peroxide (H₂O₂)-induced rat pheochromocytoma (PC12) cells as an in vitro model. PC12 cells were pretreated with HT070 for 1 h and then exposed to 100 μM H₂O₂ for 6 h in the presence of HT070. Cell viability was measured by MTT assay and the mRNA expression of Bax, Bcl-2, iNOS and COX-2 were measured by quantitative RT-PCR.Results : Oral administration of HT070 at a dose of 200 mg/kg significantly reduced neuronal death in the hippocampal CA1 region by 13.4% as compared to the vehicle-treated group. HT070 increased cell viability, reversed the down-regulated Bcl-2 mRNA level, and suppressed the up-regulated mRNA expressions of Bax, iNOS, and COX-2 in H₂O₂-treated PC12 cells.Conclusions : HT070 protects against delayed neuronal death after global cerebral ischemia and its neuroprotection properties might be attributed to the inhibition of mitochondrial apoptosis and ROS-generating enzymes.

• 미생물학회지
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• 제45권2호
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• pp.215-221
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• 2009

대장균에서 비천연 아미노산을 단백질 생합성시 특정 위치에 삽입하는 방법의 하나로 amber suppressor tRNA와 여기에 비천연 아미노산을 특이적으로 결합할 수 있는 변형된 aminoacyl-tRNA synthetase 쌍을 이용한다. 이러한 기술의 개발을 위해 필요한 여러 요소 중 하나는 이러한 시스템이 대장균에서 얼마나 잘 작동하는지를 확인할 수 있는 in vivo 보고시스템을 설정하는 것이다. 본 논문에서는 $\beta$-galactosidase 유전자의 N-말단에 amber 코돈을 삽입한 보고유전자를 제작하였으며, 이를 대장균(DH10B)의 chromosomal DNA에 삽입하여 DH10B(Tn:lacZam) 균주를 개발하였다. Genomic PCR과 Southern blot 분석을 통하여 lacZ amber 유전자가 대장균의 염색체에 삽입된 것을 확인하였으며, DH10B(Tn:lacZam)은 amber suppression을 유도할 수 있는 벡터가 형질 전환될 경우만 $\beta$-galactosidase 활성을 나타냈다. DH10B(Tn:lacZam)에 효모균의 amber suppressor $tRNA^{Tyr}$와 Tyrosyl-tRNA synthetase 쌍을 동시에 발현하는 벡터를 형질전환하였을 때, amber suppression에 의해서 $\beta$-galactosidase 활성이 나타났다. 하지만 이 활성은 대장균의 amber suppressor $tRNA^{Gln}$를 발현하는 pSupE2를 형질전환하였을 때와 비교 하여 매우 낮은 $\beta$-galactosidase 활성을 나타냈다. 이러한 결과는 DH10B(Tn:lacZam) 균주가 $\beta$-galactosidase 활성을 통하여 정성 및 정량적으로 in vivo amber suppression 활성을 비교 분석할 수 있는 특성을 가졌음을 나타낸다.

• Korean Chemical Engineering Research
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• 제49권1호
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• pp.105-108
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• 2011

유가의 급등과 화석 연료에 의한 온난화 현상은 재생 가능한 대체 연료에 대한 필요성이 요구되었다. 수송용 바이오 연료를 비교하였을 때 에탄올보다 높은 알코올 경우 휘발유와 비슷한 장점을 갖는데 그 이유는 높은 에너지 밀도와 낮은 흡습성을 갖기 때문이다. 이러한 이유로 미생물의 발효는 지속적인 에너지를 얻을 수 있는 잠재적 생산자라 할 수 있다. 본 연구에서는 생물학적으로 생산되는 알코올 성분에 대하여 두 종의 세균과 한종의 효모인 Escherichia coli와 Clostridium acetobutylicum 그리고 Saccharomyces cerevisiae를 이용하여 바이오 알코올에 대한 세포 성장 정도와 함께 미생물내에 스트레스 반응 유전자들의 분석을 실시하였다. 분석한 알코올은 에탄올과 부탄올이며, 이들의 농도별 세균의 성장속도와 산화적 손상에 민감하게 반응하는 katG 유전자, 생물막 손상에 민감하게 반응하는 fabA 유전자, 단백질 손상에 민감하게 반응하는 grpE 유전자, 유전자 손상에 민감하게 반응하는 recA 유전자의 반응여부를 분석하였다. 그 결과, 에탄올과 부탄올 중 부탄올의 세포 독성이 더 높게 관찰되었으며, 부탄올의 경우 생물막 손상을 유발하는 세포내 독성효과를 지니고 있음을 확인하였다.

• 한국발생생물학회지:발생과생식
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• 제20권4호
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• pp.297-304
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• 2016

Lectins belong to the pattern-recognition receptors (PRRs) class and play important roles in the recognition and elimination of pathogens via the innate immune system. Recently, it was reported that lily-type lectin-1 is involved when a pathogen attacks in the early immune response of fish. However, this study is limited to information that the lectin is involved in the innate immune response against viral infection. In the present study, the lily-type lectin-2 and -3 of Oplegnathus fasciatus (OfLTL-2 and 3) have been presented to be included B-lectin domain and two D-mannose binding sites in the amino acid sequence that an important feature for the fundamental structure. To investigate the functional properties of OfLTLs, the tissue distribution in the healthy rock bream and temporal expression during early developmental stage analysis are performed using quantitative real-time PCR. OfLTL-2 and 3 are predominantly expressed in the liver and skin, but rarely expressed in other organ. Also, the transcripts of OfLTLs are not expressed during the early developmental stage but its transcripts are increased after immune-related organs which are fully formed. In the challenge experiment with RBIV (rock bream iridovirus), the expression of OfLTLs was increased much more strongly in the late response than the early, unlike previously known. These results suggest that OfLTLs are specifically expressed in the immune-related tissues when those organs are fully formed and it can be inferred that the more intensively involved in the second half to the virus infection.

• Journal of Animal Science and Technology
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• 제57권6호
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• pp.25.1-25.7
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• 2015

Rumen bypass fat is commonly added to increase energy intake in dairy cattle. The objective of this study is to examine the addition of rumen bypass fat during finishing period on performance and carcass characteristics in grain fed steers. This study was conducted as a completely randomized block design with 126 cross-bred steer calves (initial BW $471.5{\pm}7.5kg$) randomly assigned to pens with 9 steers/pen (n = 7 pens/treatment). Each pen was randomly assigned to one of two treatment groups; rumen bypass fat treatment (CCS, calcium soap of palm fatty acids) and control diet (CT, tallow). The diets were formulated to be isonitrogenous and isocaloric. Animals were fed twice daily at 110 % of the previous daily ad libitum intake. Blood from each sample was taken from the jugular vein. Muscle and adipose samples were collected from the longissimus dorsi regions. Feedlot performance and carcass characteristics were assessed. To examine adipogenic gene expression, quantitative real-time PCR was completed. Steers fed the CT had a greater level of performance for most of the parameters measured. The CT group had greater DMI (P < 0.05) and tended to have greater ADG (P < 0.10). Marbling score (P < 0.05) and quality grade (P < 0.05) were greater for steers fed the CT diet than those fed CCS. The longissimus muscle area tended to be greater (P < 0.10) in steers fed CT ($87.60cm^2$) than those fed CCS (84.88 cm2). The leptin mRNA expression was down-regulated (P < 0.05) in adipose tissue of steers fed a CCS when compared to those fed CT. These data suggest that calcium soap of palm fatty acids can be added to finishing diets without significant reduction in final body weight, although there may be modest reductions in marbling and quality scores.

• 대한한방내과학회지
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• 제27권2호
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• pp.379-393
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• 2006

Objectives: We are aimed to identify anti-tumor effects of Curcuma aromatics on some kinds of cancer cells through molecular biologic methods. Materials & Methods: We used 4 kinds of cancer cell lines such as lung cancer cells(AS49), cervical cancer cells(HeLa), glioma cancer cells(A172) and prostate cancer cells(PC3). We treated the boiled extract of Curcuma aromatica $5{\mu}g,\;10{\mu}g$ to cultural media(ml) for 24 hours. We measured the cytotoxicitv on 4 kinds of cancer cells through tryphan blue exclusion test and the suppressive effect on viability of 4 kinds of cancer cells via MTT assay. We measured change of mitochondria membrane potential via flow cytometry. The quantitative RT-PCR was used to examine the effect on the revelation of Bcl-2 and Bax which are genes related to apoptosis. We examined the effect on the revelation of Bcl-2 Protein and Bar protein by western blot analysis. Results : In the experiment of tryphan blue exclusion test, the extract of Curcuma aromatica showed more significant killing effect on AS49, HeLa than the control group with density dependent manner, which was statistically significant. In the experiment of MTT assay the extract of Curcuma aromatica showed more suppressive effect on viability of A549, HeLa than the control group with density dependent manner, which was statistically significant. Curcuma aromatica induced apoptosis by decreasing the membrane potential of mitochondria in A549, HeLa. In the experiment of the revelation of genes related to apoptosis, the revelation of Bcl-2 decreased and the revelation of Bax increased in A549, HeLa treated with Curcuma aromatica with dose dependent manner. In the experiment of the revelation of protein related to apoptosis, the protein levels of Bcl-2 decreased and the protein levels of Bax increased in AS49, HeLa treated with Curcuma aromatica with dose dependent manner. Conclusions: From this study, we can infer that Curcuma aromatica has anti-tumor effect on lung cancer cells and uterine carcinoma cells but not on glioma cells and prostate cancer cells.

• 대한한방내과학회지
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• 제27권2호
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• pp.429-443
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• 2006

Objectives : The Purpose of this study was to identify anti-tumor effects of Curcuma longa L. on some kinds of cancer cells through molecular biologic methods. Materials & Methods : We used 4 kinds of cancer cell lines such as glioma cells(A172), cervical cancer cells(HeLa), Prostate cancer cells(PC3), lung cancer cells(A549). We injected the boiled extract of Curcuma longa L. $5{\mu}g,\;10{\mu}g$ to culture media(ml) for 24 hours. We measured the cytotoxic effect on 4 kinds of cancer cells through trypan blue exclusion test and the suppressive effect on viability of 4 kinds of cancer cells via MTT assay. We measured the change of mitochondria membrane potential via flow cytometry. The quantitative RT-PCR was used to examine the effect on the revelation of Bcl-2 and Bax which genes are related to apoptosis. We examined the effect on the revelation of Bcl-2 protein and Bax protein by western blot analysis. Results: 1. Extract of Curcuma longa L. showed significant cytotoxic effect on A172, HeLa, PC3 compared to the control group with density dependent manner. 2. Extract of Curcuma longs L. showed significant suppressive effect on viability of A172, HeLa, PC3 compared to the control group with density dependent manner. 3. Curcuma longs L. induced apoptosis by decreasing the membrane potential of mitochondria in A172, HeLa, PC3. 4. In the test about the revelation of genes related to apoptosis, the revelation of Bcl-2 decreased and the revelation of Bax increased in A172. HeLa, PC3 treated with Curcuma longa L. with density dependent manner. 5. In the test about the revelation of protein related to apoptosis, the protein levels of Bcl-2 decreased and the protein levels of Bax increased in A172, HeLa, PC3 treated with Curcuma longa L. Conclusions: This experiment shewed that Curcuma longs L. has anti-tumor effect on glioma, cervical, Prostate cancer cells except on lung cancer. We hope that anti-tumor effects of Curcuma longa L. will be more Practically identified.

• 생명과학회지
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• 제24권12호
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• pp.1284-1290
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• 2014

본 연구에서는 C2C12 근육 세포에서 IGF-I이 지방산 저장과 사용에 영향을 미치는 FATP1의 mRNA 및 단백질 발현에 미치는 영향에 대해 알아보았다. 그 결과 IGF-I이 FATP1의 단백질과 mRNA 발현을 유의성 있게 조절하였음을 알 수 있었다. 이는 골격근에서 IGF-I이 근육 관련 유전자들의 발현을 조절하여 근부피 유지 및 증대에 중심적인 역할을 한다는 기존의 연구 패턴들에서 벗어나, IGF-I이 골격근 세포의 분화에 있어 지방산의 수송을 담당하는 FATP1의 발현에도 영향을 미친다는 사실을 증명하였다는데 의의가 있다고 사료된다. 향후 IGF-I에 의한 FATP1의 지방산 저장의 수준과 산화 과정에 있어 상호작용하는 기타 매개체들과의 관계에 대한 연구가 필요할 것으로 사료된다.

• 한국식품영양학회지
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• 제26권3호
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• pp.485-491
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• 2013

본 연구에서는 피로 회복 또는 원기 회복에 효능이 있는 것으로 알려진 홍경천과 홍삼을 이용하여 홍경천-홍삼 복합 발효물의 산화적 손상 억제 효과를 평가하고자 $H_2O_2$로 산화적 스트레스를 유도시킨 C2C12 근육세포에 홍경천-홍삼 복합 발효물의 처리한 후, 세포의 morphology, cell viability 및 항산화 효소들의 유전자 발현 양상을 비교, 분석하였다. 홍경천-홍삼 복합 발효물은 C2C12 근육세포의 cell viability를 유의적으로 증가시켰으며, Cu/Zn-SOD, Mn-SOD 및 GPx 등과 같은 세포내 항산화 효소의 발현을 증가시킬 뿐만 아니라, 근육세포 분화의 주요 전사인자인 Myo D의 발현 또한 증가시키는 것으로 나타났다. 이상의 결과로, 홍경천-홍삼 복합 발효물은 세포내 항산화 효소 시스템을 증가시켜 외부로부터의 산화적 손상에 대한 방어효능을 갖는 것으로 나타났으며, 향후 in vivo 시스템 이용한 추가적인 연구가 수행된다면, 홍경천-홍삼 복합 발효물을 이용한 항피로 건강기능식품의 소재개발이 가능할 것으로 판단된다.

• 대한한방내과학회지
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• 제36권4호
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• pp.518-526
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• 2015

Objectives This study investigated the effect of purified safflower (Carthamus tinctorius Linne) and safflower seed (Carthamus tinctorius L. seed; CS) extract, using hot water and ethanol extract methods , on the osteogenic differentiation of MC3T3E1 cells.Methods The safflower and safflower seed were extracted with hot water and ethanol. The samples were concentrated by a rotary evaporator and then freeze-dried using a freeze-dryer. The MC3T3E1 cells were propagated and maintained in DMEM (Gibco) containing 10% FBS and a 1% antibiotic antimycotic solution. To induce osteogenic differentiation, the cells were treated for 14 days with DMEM with 10 mM β-glycerophosphate and 50 μM ascorbic acid. Extract doses were confirmed by the results of an MTT assay, and treatment of the extracts was performed in a differentiation medium every two days. The ALP staining and activity were tested after osteogenic differentiation for five days, and after 14 days, osteogenic differentiation was determined by alizarin red S staining. The mRNA expressions of osteogenic-related genes were quantified using quantitative real-time PCR.Results In the results of the MTT assay, all concentrations of safflower extracts had no toxicity in the MC3T3El cells. But in the groups of 100 ng/ml and 200 ng/ml concentrations of safflower seed extracts, the cell viability was significantly reduced by up to 40-50%. So we fixed the treatment concentration of the extract at 50 ng/ml. In the ALP and alizarin red S staining, all extract groups increased osteogenic differentiation compared with the control group. The water-safflower extract group showed the highest mRNA level of Alp, Runx2, and Dlx5 genes. The mRNA level of Ocn, an osteogenic gene related to late-stage differentiation, in the ethanol-safflower extract group increased the mineralization more significantly than in other groups.Conclusions These data suggest that the extract of safflower increases the osteoblastic differentiation activates of MC3T3E1 cells like the extract of safflower seed. The water-extract and ethanol-extract of safflower have effects on different stages of osteogenesis in MC3T3El. Not only safflower seed but also safflower will be useful therapeutic reagents for age-associated chronic diseases such as osteoporosis.