• Title/Summary/Keyword: In Vivo and In Vitro Function

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A Review on Korean Traditional Medicine Research against Cervical Dysplasia - Focused on Studies of Cervical Cancer and Cervical Dysplasia- (자궁경부 이형증과 관련한 한의약 연구 고찰 - 자궁경부암 및 자궁경부 이형증 논문을 중심으로 -)

  • Yoo, Jeong-Eun;Lee, Ji-Yeon;Yoon, Young-Jin
    • The Journal of Korean Obstetrics and Gynecology
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    • v.26 no.4
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    • pp.191-203
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    • 2013
  • Purpose: This review is to figure out evidence that suggest effectiveness of Korean Medicine treatments against cervical dysplasia. Methods: Studies on cervical cancer and cervical dysplasia were searched through 6 databases: Korean Studies Information Service System(KISS), National Discovery for Science Leaders (NDSL), Korea Institute of Science and Technology Information (Korean TK), Oriental Medicine Advanced Searching Integrated System (OASIS), the Journal of Korean Medicine, and the Journal of Korean Obstetrics & Gynecology. After that, the articles were extracted with reference point of Korean Traditional Medicine. Results: 37 articles were included lastly according to selection criteria. 3 of them were case reports on cervical dysplasia, and 34 were in-vitro studies on Human Papilloma Virus (HPV) positive cancer cell. In case reports, acupuncture, moxibustion, medical herbs and pharmacoacupuncture were used for treatments of cervical dysplasia with about 3 months. Experimental studies on cervical cancer cell showed that several herbs function with clear heat, eliminate stasis (淸熱解毒, 化瘀消腫) have anti-cancer effects inducing apoptosis. Conclusions: The results of articles are not enough to use in practice. Therefore, we indicates more advanced research methodology as follows: development of Korean Medicine treatment protocol with oral and external, in-vivo experimental study, and evaluation immunity index.

Immune gene expression and protection effect against VHSV by injection of interferon regulatory factor 10 in zebrafish (Danio rerio) (제브라피쉬 interferon regulatory factor 10의 주사에 따른 면역 유전자 발현과 VHSV에 대한 방어 효과)

  • Kim, Hye Ji;Kim, Jin Young;Park, Jong Bin;Lee, Ji Hyun;Park, Jeong Su;Kim, Hyoung Jun;Kwon, Se Ryun
    • Journal of fish pathology
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    • v.34 no.1
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    • pp.23-29
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    • 2021
  • Interferon regulatory factors (IRFs) are a family of transcription factors essential to the control of antiviral immune response, cell growth, differentiation and apoptosis. IRF10 of zebrafish (Danio rerio) was negative regulation of the interferonΦ1 and 3 response in vitro. In this study, we analyze the induction of in vivo immune response activation from the IRF10 gene of zebrafish and the protective effect against VHSV. As the results, the group inoculated with IRF10 expression vectors, there was no expression of IFNΦ1, suggestion that IRF10 may function as a negative regulator of IRF3, which binds to the IFNΦ1 promoter. And other types of interferon genes (IFNΦ2-4) are thought to have been activated, inducing to the expression of pro-inflammatory cytokine and Mx genes. As the results of challenge test performed at 14 days after inoculation of the expression vectors, the maximum survival rate [50% (1㎍ DNA) and 42.5% (10㎍ DNA)] for IRF10 group were recorded. Meanwhile, the survival rates of pcDNA3.1 and PBS as the control groups were 10% and 15%, respectively. This study suggests that the possibility that activation of IRF10 molecule could be exploited as a VHS control method.

Effects of Hin Recombinase Dimer Interface Mutants on DNA Binding and Recombination

  • Lee, Hee-Jung;Lee, Sun-Young;Lee, Hee-Jin;Lim, Heon-Man
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2001.06a
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    • pp.27-31
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    • 2001
  • Previous biochemical assays and a structural model indicated that the dimer interface of the Hin recombinase is composed of the two a-helices. To elucidate the structure and function of the helix, amino acids in the N-terminal end of the helix, where the two helices contact most, were randomized, and inversion-incompetent mutants were selected. To investigate why the mutants lost their inversion activities, the DNA binding, hix-pairing, invertasome formation, and DNA cleavage activities were assayed using in vivo and in vitro methodologies. Results indicated that the mutants could be divided into 4 classes based on their DNA binding activity. We proposed that the a-helices might place a DNA binding motif of Hin properly to the minor DNA groove of the recombination site. All the mutants except the non-binders were able to perform hix-pairing and invertasome formation, suggesting that the dimer interface is not involved in the process of hix-pairing or invertasome formation. The inversion-incompetent phenotype of the binders was caused by the inability of mutants to perform the DNA cleavage activity. The less binders exhibited wild-type level of hix-pairing activity because the hix-pairing activity overcomes the DNA binding defect of the less binders. This phenotype of the less binders suggests that the binding domains of Hin could mediate Hin-Hin interaction during hix-pairing..

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In vitro and Ex vivo Supplementation of Houttuynia cordata Extract and Immunomodulating Effect in Mice (어성초 추출물 첨가가 마우스 면역능 증진에 미치는 영향)

  • Kim, Jin;Ryu, Hye-Sook;Shin, Jung-Hee;Kim, Hyun-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.2
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    • pp.167-175
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    • 2005
  • Houttuynia cordata THNUB (He; Uh-Sung-Cho) is a medicinal plant which has been widely used as a component of blood-building decoctions. This study was performed to investigate the immunomodulative effect of He in mice. In vitro experiment, the mice splenocytes proliferation and three kinds of cytokines (IL-1$\beta$, IL-6, TNF- $\alpha$) production by mice peritoneal macrophages cultured with six (methanol, hexane, chloroform, ethylacetate, butanol and water) fractions of He were used to indicate the immunomodulative effect. Ex vivo experiment, the different concentrations of He water extract was orally administrated every other day for two weeks. The production of cytokines IL-1$\beta$, IL-6, TNF- $\alpha$) secreted by activated macrophages and the mice splenocytes proliferation were used as an index for the immunocompetence. The supplementation of all six fractions of He enhanced the splenocytes proliferation at the level of 6.58$\pm$1.23∼47.82$\pm$5.48 compared to that of control in the range of 1∼50 $\mu\textrm{g}$/mL. IL-1$\beta$ production was significantly increased with the supplementation of chloroform and water extract of He. Higher level of IL-6 production was detected by the supplementation of ethylacetate, butanol and water extract. TNF - $\alpha$ production was enhanced by the supplementation of all six fractions of He. From the ex vivo study, the highest proliferation of splenocytes was seen from the mice orally administrated with the He water extract at the concentration of 500 mg/kg bw In case of cytokines production, IL-1$\beta$, IL-6, and TNF- $\alpha$ release by activated peritoneal macrophages were augmented by the oral administration of He water extract. These results indicated that He may enhance the immune function by regulating the splenocytes proliferation and cytokines production capacity in mice.

Simple Methods for Production of Chimeric Mouse by Coculture with TT2 Embryonic Stem Cells (TT2 Embryonic Stem Cell 을 이용한 Chimeric Mouse 생산에 있어서 간단한 공배양방법)

  • Cho, Y.Y.;Moon, S.J.;Kang, M.J.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.451-455
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    • 2000
  • Gene targeting are very useful tools for the research on the gene function in vivo, mass production of foreign materials and biomedical approach of therapeutic process. But this process is very complicated and necessary highly skilled technique, because it is very different from ES cell origin, genetic background of embryo, and experimental conditions. We investigated the productivity ability of chimeric mouse after aggregation with TT2 ES cells. Increse of ES cell density caused gradual decrease in embryo development in vitro and in th $\varepsilon$ production of chimeric mice in vivo. One million ES cell density for the aggregation was very efficient to produce high percentage chimeric mice in their coat color. These results suggested that appropriate cell density plays a key role in the development and production of chimeric mice by a 8-cell aggregation method.

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Effects of Neutral Dammarane Saponin from Panax ginseng on the in vitro Function of Polymorphonuclear Leukocytes (인삼의 중성 Dammarane계 사포닌의 다형핵 백혈구 기능에 미치는 영향)

  • Bridges Raymond B.;Park Ki Hyun;Han Byung Hoon;Han Yong Nam;Chung Soo Il
    • Proceedings of the Ginseng society Conference
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    • 1988.08a
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    • pp.115-121
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    • 1988
  • Although Saponin A from Panax ginseng has previously been shown to inhibit carageenin induced edema. a paucity of information exists on the effects of components from ginseng on the cellular inflammatory response. specifically polymorphonuclear leukocyte (PMNL) function. The purpose of this study was 10 determine the effects of isolated neutral dammarane saponins from ginseng (i.e..glycosidic derivatives of 20(S)-protopanaxadiol [ginsenoside $Rb_1,\;Rb_2$ and Rc] and 20(S)-protopanaxatriol [ginsenosides Re and $Rg_1$]) on in vivo PMNL function and to compare their effects with those produced by a steroidal anti-inflammatory agent (dexamethasone) and commercially available saponin. Dexamethasone. the ginsenosides and saponin were all shown to he potent inhibitors of PMNL chemotaxis using the $^{51}Cr$ assay with $5{\times}10^{-8}M$ f-met-leu-phe [FMLP] as the chemoattractant. Inhibition or PMNL chemotaxis by dexamethasone. the ginsenosides and saponin were all shown to be both time-and dose-dependent and these agents did not affect cellular viability at the concentrations tested Saponin and the ginsenosides were more potent inhibitors of chemotaxis than was dexamethasone. while oxidant generation (as measured by the luminol-enhaneed chemil-uminescence of PMNL using FMNL $[10^{-6}]$ as the stimulus) was inhibited by dexamethasone. the ginsenosides $(Rb_1\;Rb_2\;Rc\;Re\;and\;Rg_1)$ and saponin at a concentration of 1 ${\mu}M$ had no significant effect on PMNL chemiluminescence. Thus. the neutral dammarane saponins are potentially important modulators or PMNL function and their inhibitory effects may he differentiated from those of the Steroidal anti-inflammatory agents.

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Change of Antioxidative Activity at Different Harvest Time and Improvement of Atopic Dermatitis Effects for Persimmon Leaf Extract (감잎(柿葉) 추출물의 채취시기에 따른 항산화 활성 변화 및 아토피성 피부염 개선효과)

  • Jung, Woo-Young;Jeong, Jong-Moon
    • The Korea Journal of Herbology
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    • v.27 no.1
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    • pp.41-49
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    • 2012
  • Objectives : The purpose of this study was to investigate the anti-oxidative and anti-atopic dermatitis effects of persimmon leaf extract obtained from Cheongdo-gun, where more than 60% of Korean persimmon is produced. Methods : Anti-oxidative effects of the crude persimmon leaf extract harvested monthly between May and November were determined by in vitro assay using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide dismutase (SOD)-like reaction. Anti-atopic dermatitis effects of the crude persimmon leaf extract were determined by using collagenase type I inhibition assay and by quantitative assays including serum histamine, prostaglandin E metabolite and leukotriene $B_4$ levels in animal model of atopic dermatitis using Balb/c mice. Results : Persimmon leaf extract harvested in May had higher levels of total phenolic compounds (182.24 mg/g) and flavonoids (23.05 mg/g) than the ones of different month extract. Also, persimmon leaf extract harvested in May showed the most effective extract scavenging activities of DPPH free radical ($13.39{\pm}0.21\;{\mu}g/ml$) and superoxide anion radical ($40.52{\pm}2.32\;{\mu}g/ml$), leading to use the persimmon leaf extract harvested in May for the experiments hereafter. Persimmon leaf extract showed $326.71{\pm}4.6\;{\mu}g/ml$ of 50% inhibitory concentration ($IC_{50}$) for collagenase type I which is responsible for the degradation of extracellular matrix. In addition, persimmon leaf extract application group could decrease serum levels of histamine, prostaglandin E metabolite and leukotriene $B_4$ compared to the negative control in animal model of atopic dermatitis. Especially, persimmon leaf extract showed a significantly decreased serum leukotriene $B_4$ level relative to the levels of histamine and prostaglandin E metabolite. Conclusions : Persimmon leaf extract showed anti-oxidative and anti-atopic dermatitis effects in vitro and in vivo. These results suggest that persimmon leaf extract may have immunoregulatory function for alleviating atopic dermatitis by decreasing collagenase activity and mast cell activation.

Comparison of Cellular Senescence Phenotype in Human Fibroblasts from New-born and Aged Donors. (신생아와 노인 유래 섬유아세포의 노화과정에서의 세포학적 성질의 비교)

  • Yi, Hye-Won;Hwang, Eun-Seong
    • Journal of Life Science
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    • v.18 no.3
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    • pp.344-349
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    • 2008
  • Normal somatic cells proliferate for a limited number of doublings in culture and then enter an irreversible growth-arrest state called replicative senescence. Replicative senescence has been believed a reason for the limited cellular turnover and deterioration of tissue function in aged animals. However, there is no experimental evidence supporting this assumption. Furthermore, cells from aged person have been poorly characterized with an exception of the cases of T cells. In this study, we examined cell biological changes occurring in replicative senescence of fibroblast strains originated from a new-born (NHF-NB) and a 87 year old man (NHF-87). NHF-87 (and the cells from a 75-year old) proliferated to smaller population doublings and with longer doubling times than NHF-NB did. At early passages, NHF-87 exhibited a low senescence-associated ${\beta}-Gal$ (SA ${\beta}-Gal$) activity and lipofuscin level, typical markers for cellular senescence. Furthermore, they maintained low levels of lysosome and reactive oxygen species (ROS). All of these levels increased dramatically in the late passage NHF-87 quite similarly as those in the late passaged NHF-NB did. These results indicate that most cells originated from the aged maintain a phenotype of the cells originated from new-born donors and undergo replicative senescence with the same kinetics as that of the cells from new-born. It is also indicated that not SA ${\beta}-gal$ activity but cell proliferation rate may be qualified as a biomarker for cells aged in vivo.

D99 Type I Signal Peptidase Implicated Stabilizing the Protein Structure (Type I 신호펩디드 가수분해효소에 존재하는 D99 아미노산 잔기의 구조적 역할 가능성)

  • Sung, Meesook;Eunyoung Han;Lee, Hoyoung
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.140-144
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    • 2003
  • Type Ⅰ signal peptidase is an integral membrane protein that functions to cleave signal peptides from secreted and membrane proteins. The enzyme serves as a potential target for the development of novel antibacterial agents due to its unique physiological properties. Despite being one of the best characterized enzymes, the catalysis of Type Ⅰ signal peptidase still remains controversy over the catalytic serine/lysine dyad mechanism. It appears that the dyad proteases are generally less efficient than the prototypical serine/histidine/aspartic acid triad found in most enzymes, although Type Ⅰ signal peptidase is an exception to this rule. In this paper, we have proposed that Type Ⅰ signal peptidase may act as the serine/lysine/aspartic acid triad cataltytic mechanism. Therefore, the aspartic acid 99 residue in the E. coli signal peptidase was chosen and mutated to an alanine to see if there is any possible role of the aspartic acid in the catalytic function. Type Ⅰ signal peptidase D99A protein was inactive in vitro assay using the procoat synthesized by in vitro transcription translation. However, the mutant was active using a highly sensitive in vivo assay. Pulse-chase experiments show that the replacement of aspartic acid 99 with alanine results in a very unstable signal peptidase molecule. Therefore, we conclude that it is unlikely that the residue is directly involved in catalysis, but rather plays an important role in stabilizing the protein structure.

Antithrombotic Activities of Cheongkookjang and Cheongkookjang Fermented with Green Tea or Mugwort (청국장 및 녹차, 쑥이 첨가된 청국장의 항혈전 활성)

  • Lee, Kyung-Ae;Jang, Jeong-Oak;Yoon, Hye-Kyung;Kim, Moo-Sung
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.298-303
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    • 2007
  • Antithrombotic activities of water extract of cheongkookjang and cheongkookjang fermented with green tea or mugwort were evaluated on some antithrombosis related activities in vitro and thrombotic death inhibition in vivo. Cheongkookjang made of white soybean (Glycine max) or black small soybean (Rhynchosia nulubilis) showed potent antioxidative activities. Addition of green tea or mugwort during cheongkookjang fermentation increased the antioxidative activity, cheongkookjang with green tea showed more drastic increase compared with cheongkookjang with mugwort. Nitrite scavenging effects of the cheongkookjang extracts were prominent but the addition of green tea or mugwort seldom increased the scavenging effects. All the cheongkookjang extracts showed strong inhibitory activities on platelet aggregation. The inhibitory activities of cheongkookjang were increased considerably by addition of green tea or mugwort even with low concentration. Plasmin unit as fibrinolytic activity was not affected considerably by addition of green tea. Addition of mugwort decreased the activity transiently at low concentration ($0.3{\sim}1.0%$) but increased again slowly at higher concentration ($1{\sim}3%$). In vitro thrombotic death inhibition test, the antithrombotic activity of cheongkookjang made of black small bean with green tea was higher by about 1.5 times compared to that without green tea. As results, cheongkookjang might inhibit antithrombosis not only by fibrinolytic action but also by inhibition of platelet aggregation and antioxidative action. The addition of functional materials such as green tea or mugwort could increase the antithrombotic function, even at low concentration.