• 대한한의학회지
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• 제24권4호
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• pp.71-81
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• 2003

So-Hap-Hyang-Won, a traditional oriental medicine used in the treatment of stroke patients, was examined for its ability to reverse the cell damage caused by lipid peroxidation products and oxidative stress in bovine aortic endothelial cells (BAEC). The effects of herbal medicine on cell proliferation and recovery of oxidative damaged situation were studied in BAEC, which was considered an appropriate in vitro model for stroke resulting from various vascular diseases prevalent in advanced age. In a clinical study of stroke patients, So-Hap-Hyang-Won appeared to improve considerably arm and leg movements as well as consciousness disturbance condition, compared with other traditional medicines used for stroke. When BAEC were treated with extracts of the lyophilized herbal medicines, only that of So-Hap-Hyang-Won stimulated cell proliferation and showed no toxicity even at high concentrations. In studies of BAEC treated with extracts of the lyophilized material of the 14 components of So-Hap-Hyang-Won, only the extract of Foeniculi Fructus stimulated cell growth at all concentrations tested. Moreover, when cells were treated with Foeniculi Fructus (10 and 100 mg/ml) extract after prior exposure to t-BHP ($l0\mu\textrm{M}$) or HNE ($0.2\mu\textrm{M}$), lipid peroxidation products which are known to be involved in aging and vascular diseases, or after the exposure to SIN-l ($500\mu\textrm{M}$), which generates nitric oxide (NO) and other reactive oxygen species, there was substantial recovery from the oxidative damage, presumably due to the radical-scavenging effect of Foeniculi Fructus extract. Foeniculi Fructus not only showed stimulatory effects on cell growth and cell damage repair in BAEC, but also appeared to show the most anti-aging activity among all the herbal components of So-Hap-Hyang-Won.

• Journal of Microbiology and Biotechnology
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• 제23권5호
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• pp.656-660
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• 2013

Pseudomonas fluorescens strain CL0145A was discovered at the New York State Museum Field Research Laboratory as an effective agent against the environmentally destructive zebra mussel, which has contaminated US waters. Dried cells of the microbe are being commercialized as an environmentally friendly solution to the problem. We found that antibiotic activity against the Gram-positive bacterium Bacillus subtilis is produced and excreted by this strain. We have carried out studies to optimize production of the antibiotic. Studies were begun in a complex corn meal medium. Activity was found in both cells and culture supernates and was maximal after one day of fermentation. Static fermentation conditions were found to be superior to shaken culture. Production of extracellular antibiotic in complex medium was found to be dependent on the content of sucrose and enzyme-hydrolyzed casein. Indeed, production was greater in sucrose plus enzyme-hydrolyzed casein than in the complex medium. Of a large number of carbon sources studied as improvements over sucrose, the best was glycerol. An examination of nitrogen sources showed that production was improved by replacement of enzyme-hydrolyzed casein with soy hydrolysates. Production in the simple glycerol-Hy-Soy medium was not improved by addition of an inorganic salt mixture or by complex nitrogen sources, with the exception of malt extract. In an attempt to keep the medium more defined, we studied the effect of amino acids and vitamins as replacements for malt extract. Of 21 amino acids and 7 vitamins, we found tryptophan, glutamine, biotin, and riboflavin to be stimulatory. The final medium contained glycerol, Hy-Soy, tryptophan, glutamine, biotin, and riboflavin.

• Applied Biological Chemistry
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• 제45권4호
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• pp.195-202
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• 2002

국내외에서 수집 재배한 29품종 유색미의 70% 에탄올 추출물로 세포독성, 변이원성, 화학적 직접변이원 mitomycin C, 4-nitroquinoline-N-oxide, 2,4,7-trinitro-9-fluorenone에 대한 항변이원성을 측정하였다. 세포 내재성 alkaline phosphatase활성을 지표로 유색미 추출물이 지시세포인 E. coli PQ 37의 성장에 미치는 영향을 조사한 결과 Jumlalocal, Jumlalocal-1을 포함하는 13종류의 유색미 추출물이 세포독성을 나타낸 반면 DK 1, SC-5, LK 1A-2-12-1-1및 wx 139-3-64-20-3-1등의 품종은 세포성장 촉진활성을 나타내었다. SOS chromotest 기법을 이용하여 유색미가 가지는 변이원성 및 항변이원성을 조사한 결과에서는 Jumlalocal-1, IR 17491-5-4-3-3 및 Jumlalocal 등의 품종이 변이원성을 유도하는 품종이었다. 화학적 직접변이원 mitomycin C, 2,4,7-trinitro-9-fluorenone 및 4-nitroquinoline-N-oxide 에 대해서 공통적으로 항변이원성 효과가 인정되는 유색미 품종은 LK1-3-6-12-1-1, Parnkhari 203, Jumlalocal, wx 139-3-64-20-3-1, Muthumanikam, HP 883-1-1-1-B-1-1, Jumla-local-1 등 7품종이었다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권3호
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• pp.1499-1506
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• 2016

Background: Crocus sativus and its major constituent crocin are well established to have anti-cancer properties in breast cancer cells (MCF-7). However the role of C. sativus extract (CSE) and crocin on caspase signaling mediated MCF-7 cell death at molecular level is remains unclear. In this study, we tried to unravel role of CSE and crocin on caspase mediated MCF-7 cells death and their in vivo preclinical toxicity profiling and immune stimulatory effect. Materials and Methods: CSE extract was fractionated by HPLC and crocin was isolated and characterized by NMR, IR, and MS. MCF-7 cells were treated with both CSE and crocin and expression of Bcl-2 and Bax was assessed after 24 and 36 hours. Furthermore, caspase 3, caspase 8 and caspase 9 expression was determined by Western blotting after 24 hours of treatment. DNA fragmentation analysis was performed for genotoxicity of CSE and crocin in MCF-7 cells. The in vivo toxicity profile of CSE (300 mg/kg of b.wt) was investigated in normal Swiss albino mice. In addition, peritoneal macrophages were collected from crocin (1, 1.5 and 2 mg/kg body weight) treated mice and analyzed for ex vivo yeast phagocytosis. Results: Immunoblot analysis revealed that there was time dependent decline in anti-apoptotic Bcl-2 with simultaneous upregulation of Bax in CSE and crocin treated MCF-7 cells. Further CSE and crocin treatment downregulated caspase 8 and 9 and cleaved the caspase 3 after 24 hours. Both CSE and crocin elicited considerable DNA damage in MCF-7 cells at each concentration tested. In vivo toxicity profile by histological studies revealed no observable histopathologic differences in the liver, kidney, spleen, lungs and heart in CSE treated and untreated groups. Crocin treatment elicited significant dose and time dependent ex vivo yeast phagocytosis by peritoneal macrophages. Conclusions: Our study delineated involvement of pro-apoptotic and caspase mediated MCF-7 cell death by CSE and crocin at the molecular level accompanied with extensive DNA damage. Further we found that normal swiss albino mice can tolerate the maximum dose of CSE. Crocin enhanced ex vivo macrophage yeast phagocytic ability.

• Clinical and Experimental Reproductive Medicine
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• 제29권1호
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• pp.45-56
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• 2002

Objectives: Recently, recombinant FSH (rFSH) has been manufactured using a Chinese hamster ovary cell line transfected with the gene encoding human FSH. Both rFSH and urinary gonadotropin (uFSH) could be used for controlled ovarian hyperstimulation (COH). However, uFSH implies a number of disadvantages, such as batch-to-batch inconsistency, no absolute source control, dependence on large amounts of urine, low specific activity, and low purity. The purpose of this study was to evaluate the efficacy of rFSH in human IVF-ET program. Materials and Methods: A total of 508 infertile women was enrolled in this study. They are classified into rFSH group (n=177) or uFSH group (n=331), and all of them were matched by age and cause of infertility in same period. The $Puregon^{(R)}$ (Organon, Holland) was used as rFSH, and the Metrodin-$HP^{(R)}$ (Serono, Switzeland) and $Humegon^{(R)}$ (Organon, Holland) was used as uFSH. We subdivided the patients into three age groups. The outcomes of IVF-ET program were analyzed using the statistical package for social sciences (SPSS). Results: There was no significant differences in the level of estradiol on hCG injection day, the numbers of retrieved oocytes, matured oocytes, fertilized oocytes, transferred embryos, frozen embryos between the two groups. The total dose (IU) of gonadotropin for COH was significantly lower in the rFSH group compared to uFSH group ($1339{\pm}5491.1$ vs $2527.8{\pm}1075.2$ IU, p<0.001). Clinical pregnancy rate per embryo transfer in the rFSH group showed increasing tendency, compared to the uFSH group, but there was no statistical significance (35.2% vs 29.3%). Our results demonstrated that the relative efficiency of rFSH compared with uFSH is higher in older patients. Conclusions: The ovarian stimulatory effect and clinical outcome of recombinant FSH was similar to that of the urinary gonadotropin. The IVF-ET cycles with significantly lower dose of gonadotropin in rFSH group showed comparable results. Therefore, we suggest that recombinant FSH is more potent and effective than urinary gonadotropin.

• Biomolecules & Therapeutics
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• 제18권1호
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• pp.39-47
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• 2010

Brain-derived neurotrophic factor (BDNF) is a neurotrophic factor involved in neuronal differentiation, plasticity, survival and regeneration. BDNF draws massive attention mainly due to the potential as a therapeutic target in neurological diseases such as depression and Alzheimer's disease. In a primary screening for the natural compounds enhancing BDNF release from cultured rat primary cortical neuron, we found that compounds such as baicalein, tanshinone IIa, cinnamic acid, epiberberine, genistein and wogonin among many others increased BDNF release. All the compounds at $0.1{\mu}M$ of concentration barely showed stimulatory effect on BDNF induction, however, their combination (mixture 1; baicalein, tanshinone IIa and cinnamic acid, mixture 2; epiberberine, genistein and wogonin) showed synergistic increase in BDNF release as well as mRNA and protein expression. The level of BDNF expression was comparable to the maximum BDNF stimulation attainable by a positive control oroxylin A ($20{\mu}M$) without cell toxicity as determined by MTT analysis. Both mixtures synergistically increased the phosphorylation of extracellular signal-regulated kinase (ERK) as well as cAMP response element binding protein (CREB), an immediate and essential regulator of BDNF expression. Similar to these results, mixture of these compounds synergistically inhibited the up-regulation of inducible nitric oxide synthase (iNOS) induced by lipopolysaccharide treatments in rat primary astrocytes. These results suggest that the combinatorial treatment of natural compounds in lower concentration might be a useful strategy to obtain sufficient BDNF stimulation in neurological disease condition such as depression, while minimizing potential side effects and toxicity of higher concentration of a single compound.

• 대한수의학회지
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• 제34권4호
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• pp.873-880
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• 1994

Progesterone catabolizing enzyme, the enzyme $20{\alpha}$-hydroxysteroid deltydrgenase($20{\alpha}$-HSD) is pivotal in the regulation of ovarian function in rodents, which catabolizes progesterone into biologically inactive $20{\alpha}$ hydroxypregn-4-en-3-one($20{\alpha}$-OHP). In this study was carried out the influence of $20{\alpha}$-HSD activity to ovarian function and regulation such as ovulation, formation of corpus luteum, maintenance of estrous cycles and pregnancy, we investigated changes in activities of the splenic macrophages and ovarian histological findings in rat. During the estrous cycles, the ratious of phagocytotic macrophage in splenic adherent cells were highest on the proestrous, but they were lowest on metestrous. During the pregnancy, the ratious of phagocytotic macrophages in splenic adhrent cells were lowest by 1 day and then significantly increased toward the pregnancy, which were highest pregnancy on day 12. On histological findings in rat ovary, the mean number of growing, antral and mature follicles were 15.9(72.9%), 3.4(15.9%), and 2.4(11.2%), respectively. Growing follicles reached to lowest number at diestrus and mature follicle reached to lowest number at metestrus. The numbers of corpus luteums per tissue section of the ovary were 14.4 and the number of normal and atretic follicles were 11.0(76.4%) and 3.4(23.6%), respectively. The number of corpus luteum with vacolated cell were 1.7(11.8%). In this study suggests splenic macrophages are as a source of the substance which maintainer progesterone secretion from luteal cells, and are recognized pass the vessels and reside in the fresh corpora lutea soon after ovulation via inflammatory reactions, and these macrophages are felt to have a stimulatory effect on the formation of cropora lutea. In view of the results include previous report, ovarian $20{\alpha}$-hydroxysteroid dehydrogenase enzyme and splenic macrophages consider play central role in the control and maintenance of estrous cycles and pregnancy, and also applicable to both clinical and research in a wide variety such as control of reproductive system.

• Asian-Australasian Journal of Animal Sciences
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• 제20권4호
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• pp.536-542
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• 2007

The responses of whole body protein and glucose kinetics and of nitrogen (N) metabolism to non-protein energy intake (NPEI) were determined using an isotope dilution approach and measurement of N balance in three adult male goats. The diets containing 1.0, 1.5 and 2.0 times ME maintenance requirement, with fixed intake of CP (1.5 times maintenance) and percentage of hay (33%), were fed twice daily for each 21 d experimental period. After an adaptation period of 11 d, N balance was determined over 3 d. On day 17, whole body protein synthesis (WBPS) and glucose irreversible loss rate (ILR) were determined during the absorptive state by a primed-continuous infusion of [$^2H_5$]phenylalanine, [$^2H_2$]tyrosine, [$^2H_4$]tyrosine and [$^{13}C_6$]glucose, with simultaneous measurements of plasma concentrations of metabolites and insulin. Ruminal characteristics were also measured at 6 h after feeding over 3 d. Nitrogen retention tended to increase (p<0.10) with increasing NPEI, although digestible N decreased linearly (p<0.05). Increasing NPEI decreased (p<0.01) ammonia N concentration, but increased acetate (p<0.05) and propionate (p<0.05) concentrations in the rumen. Despite decreased plasma urea N concentration (p<0.01), increased plasma tyrosine concentration (p<0.05), and trends toward increased plasma total amino N (p<0.10) and phenylalanine concentrations (p<0.10) were found in response to increasing NPEI. Increasing NPEI increased ILR of both glucose (p<0.01) and phenylalanine (p<0.05), but did not affect ($p{\geq}0.10$) that of tyrosine. Whole body protein synthesis increased (p<0.05) in response to increasing NPEI, resulting from increased utilization rate for protein synthesis (p<0.05) and unchanged hydroxylation rate of phenylalanine ($p{\geq}0.10$). These results suggest that increasing NPEI may enhance WBPS and glucose turnover at the absorptive state and improve the efficiency of digestible N retention in goats, with possibly decreased ammonia and increased amino acid absorption. In addition, simultaneous increases in WBPS and glucose ILR suggest stimulatory effect of glucose availability on WBPS, especially when sufficient amino acid is supplied.

• Asian-Australasian Journal of Animal Sciences
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• 제26권5호
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• pp.638-645
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• 2013

Interferon-tau (IFNT) is thought to be the conceptus protein that signals maternal recognition of pregnancy in ruminants. We and others have observed that OCT4 expression persists in the trophectoderm of ruminants; thus, both CDX2 and OCT4 coexist during the early stages of conceptus development. The aim of this study was to examine the effect of CDX2 and OCT4 on IFNT gene transcription when evaluated with other transcription factors. Human choriocarcinoma JEG-3 cells were cotransfected with an ovine IFNT (-654-bp)-luciferase reporter (-654-IFNT-Luc) construct and several transcription factor expression plasmids. Cotransfection of the reporter construct with Cdx2, Ets2 and Jun increased transcription of -654-IFNT-Luc by about 12-fold compared with transfection of the construct alone. When cells were initially transfected with Oct4 (0 h) followed by transfection with Cdx2, Ets2 and/or Jun 24 h later, the expression of -654-IFNT-Luc was reduced to control levels. OCT4 also inhibited the stimulatory activity of CDX2 alone, but not when CDX2 was combined with JUN and/or ETS2. Thus, when combined with the other transcription factors, OCT4 exhibited little inhibitory activity towards CDX2. An inhibitor of the transcriptional coactivator CREB binding protein (CREBBP), 12S E1A, reduced CDX2/ETS2/JUN stimulated -654-IFNT-Luc expression by about 40%, indicating that the formation of an appropriate transcription factor complex is required for maximum expression. In conclusion, the presence of OCT4 may initially minimize IFNT expression; however, as elongation proceeds, the increasing expression of CDX2 and formation of the transcription complex leads to greatly increased IFNT expression, resulting in pregnancy establishment in ruminants.

• 생명과학회지
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• 제23권8호
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• pp.984-988
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• 2013

꽃송이버섯(Sparassis crispa)은 항암 및 면역증강 효과가 뛰어나다고 알려져 있는 베타-글루칸의 함량이 건조중량의 40% 이상인 것으로 보고되고 있는 산림버섯이다. 일반적으로 버섯에서 추출한 베타-글루칸의 면역활성 및 항암 효과는 널리 알려져 있는 반면 꽃송이버섯 유래의 베타-글루칸에 의한 대식세포의 활성화 기작 및 자실체 추출물에 의한 항암 효과는 아직 보고되지 않았다. 본 연구에서는 꽃송이버섯 유래 베타-글루칸의 면역활성 효과를 확인하고자 마우스의 대식세포인 Raw 264.7 cell을 이용하였으며, 베타-글루칸 처리시 TNF-${\alpha}$와 interleukin-$1{\beta}$(IL-$1{\beta}$)와 같은 사이토카인의 발현 및 분비가 증가됨을 확인하였고, 이러한 과정에 필수적인 역할을 담당하는 전사 인자인 NF-${\kappa}B$가 활성화됨도 확인하였다. 뿐만 아니라 꽃송이버섯 추출물 처리 시 TNF-${\alpha}$의 분비가 증가됨과 함께 in vivo에서 암의 성장을 억제하는 기능이 있음을 확인하였다. 이러한 결과들은 꽃송이버섯이 면역활성을 증가시켜 암의 성장을 억제하는 건강 기능성 식품으로서의 개발 가능성이 높음을 의미하는 것이다.