• Journal of Applied Biological Chemistry
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• 제53권3호
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• pp.132-138
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• 2010

B. pseudomycoides로 부터 alanine racemase로 추정되는 유전자를 분리한 다음 6xHistidine 이 결합된 pET-21 운반체를 이용하여 E. coli BL21(DE3)에서 발현시키고 생화학 특성을 조사하였다. 재조합된 alanine racemase는 affinity chromatography를 이용하여 분리하였으며 SDS-PAGE 분석에서 약 46 kDa의 단일밴드를 나타내었다. 분리된 효소는 여러 아미노산 중에서 L-alanine에 대하여 가장 높은 활성도를 보이고 D-alanine에 대하여 두번째로 높은 활성도를 보였다. 따라서 분리된 효소는 alanine racemase로 판단되었다. 분리된 효소는 D-cysteine에 하여 상당히 저해가 되었다. 효소의 최적 활성도는 pH 9.0 근처에서 관찰되었고 산성의 조건보다는 알칼리의 조건에서 보다 안정하였다. 보효소인 PLP 0.3mM의 첨가에 의해 효소의 활성도는 약 70% 가량 증대되었다.

• Preventive Nutrition and Food Science
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• 제13권4호
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• pp.269-275
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• 2008

Dandelion (Taraxacum officinale) has been widely used as an anti-inflammatory agent in oriental medicine. In the current study, we investigated the protective effect, and the possible mechanism, of dandelion extracts against ethanol-induced acute hepatotoxicity in C57BL/6 mice. Dandelion water and ethanol extract was administered at 2 g/kg body weight (BW) once daily for 7 consecutive days, whereas control and ethanol groups received water by gavage. Ethanol (50% ethanol; 6 g/kg BW) was administered 12 hr before sacrificing the mice in order to generate liver injury. Significantly increased serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities as well as liver triglyceride (TG) and cholesterol levels were attenuated by dandelion supplementation. In addition, dandelion extracts not only enhanced alcohol dehydrogenase (ADH) and anti-oxidative enzyme activities, but reduced lipid peroxidation. Cytochrome P450 2E1 (CYP 2E1), one of the critical enzymes xenobiotic metabolism, expression was lower with ethanol treatment but restored by dandelion supplementation. These results were confirmed by improved histopathological changes in fatty liver and hepatic lesions induced by ethanol. In conclusion, dandelion could protect liver against ethanol administration by attenuating of oxidative stress and inflammatory responses.

• Journal of Microbiology and Biotechnology
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• 제20권3호
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• pp.542-549
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• 2010

In the present study, overexpression, purification, and characterization of Aeropyrum pernix K1 chaperonin B in E. coli were investigated. The chaperonin $\beta$-subunit gene (ApCpnB, 1,665 bp ORF) from the hyperthermophilic archaeon A. pernix K1 was amplified by PCR and subcloned into vector pET21a. The constructed pET21a-ApCpnB (6.9 kb) was transformed into E. coli BL21 Codonplus (DE3). The transformant cell successfully expressed ApCpnB, and the expression of ApCpnB (61.2 kDa) was identified through analysis of the fractions by SDS-PAGE (14% gel). The recombinant ApCpnB was purified to higher than 94% by using heat-shock treatment at $90^{\circ}C$ for 20 min and fast protein liquid chromatography on a HiTrap Q column step. The purified ApCpnB showed ATPase activity and its activity was dependent on temperature. In the presence of ATP, ApCpnB effectively protected citrate synthase (CS) and alcohol dehydrogenase (ADH) from thermal aggregation and inactivation at $43^{\circ}$ and $50^{\circ}$, respectively. Specifically, the activity of malate dehydrogenase (MDH) at $85^{\circ}$ was greatly stabilized by the addition of ApCpnB and ATP. Coexpression of pro-carboxypeptidase B (pro-CPB) and ApCpnB in E. coli BL21 Codonplus (DE3) had a marked effect on the yield of pro-CPB as a soluble and active form, speculating that ApCpnB facilitates the correct folding of pro-CPB. These results suggest that ApCpnB has both foldase and holdase activities and can be used as a powerful molecular machinery for the production of recombinant proteins as soluble and active forms in E. coli.

• The Plant Pathology Journal
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• 제21권4호
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• pp.334-342
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• 2005

The prototype Chlorella virus PBCV-l encodes 11 tRNA genes and over 350 protein-encoding genes in its 330 kbp genome. Initial attempts to overexpress the recombinant A189/192R protein, a putative virus attachment protein, in E. coli strain BL21(DE3) SI were unsuccessful, and multiple protein bands were detected on Western blots. However, the full-length A189/192R recombinant protein or fragments derived from it were detected when they were expressed in E. coli BL21 CodonPlus (DE3) RIL, which contains extra tRNAs. Codon usage analysis of the a189/192r gene showed highly biased usage of the AGA and AVA codons compared to genes encoded by E. coli and Chlorella. In addition, there were biases of XXA/U($56\%$) and XXG/ C($44\%$) in the codons recognized by the viral tRNAs, which correspond to the codon usage bias in the PBCV-1 genome of XXA/U ($63\%$) over those ending in XXC/G ($37\%$). Analysis of the codon usage in the major capsid protein and DNA polymerase showed preferential usage of codons that can be recognized by the viral tRNAs. The Asn (AAC) and Lys (AAG) codons whose corresponding tRNA genes are duplicated in the tRNA gene cluster were the most abundant (i.e., preferred) codons in these two proteins. The tRNA genes encoded in the PBCV-l genome seem to play a very important role during the synthesis of viral proteins through supplementing the tRNAs that are frequently used in viral proteins, but are rare in the host cells. In addition, these tRNAs would help the virus to adapt to a wide range of hosts by providing tRNAs that are rare in the host cells.

• 한국가축번식학회지
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• 제21권2호
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• pp.131-137
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• 1997

본 연구는 초자화 동결된 생쥐 팽창, 탈출, 완전탈출 배반포기배의 체내 발달율을 조사하기 위해 실시하였다. 체외수정하여 얻어진 생쥐 배반포기배는 EFS40(40% ethylene glycol, 30% Ficoll, 0.3M sucrose)으로 초자화 동결하였다. 팽창, 탈출 배반포기배는 20% ethylene glycol에 5분동안 평형시킨 다음, EFS40 용액에 1분간 노출후 액체질소에 침지하여 초자화 동결하였다. 완전탈출 배반포기배는 0.4% BSA가 첨가된 m-CR1 배양액에서 5일동안 배양하여 얻었으며, 10% EG에 5분, EFS40에 30초동안 노출하여 초자화 동결시켰다. 융해후 재팽창이 이루어진 배반포기배는 가임신 3일된 대리모의 한쪽 또는 양쪽 자궁각에(6∼8개/자궁각) 이식하였다. 대리모의 임신율과 착상율은 임신 15일째 외과적 해부로 판정하였다. 그 결과를 요약하면 다음과 같다. 1) 임신율과 정상 산자율은 초자화 동결된 팽창 배반포기배의 경우 77.8과 25.0%이었고, 탈출 배반포기배의 경우는 77.8과 26.4%로서 각각의 대조군에 있어서 66.7과 42.9%, 83.3과 40.4%에 비해 유의차가 없었다. 2) 완전탈출 배반포기배의 체외 발달율은 34.0%였고, 3) 체내 발달율은 33.3%였다. 이러한 결과는 본 실험에 사용된 EFS40 동결액을 이용한 초자화 동결방법이 생쥐 팽창, 탈출 배반포기배의 초자화 동결은 물론, 완전탈출 배반포기의 초자화 동결에도 유용하게 이용될 수 있다는 가능성을 시사하였다.

• 대한한방내과학회지
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• 제21권1호
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• pp.169-180
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• 2000

Objectives : To satisfy the demand of good treatment of constipation Methods : we investigated the literatures of Oriental Medicine about Constipation. Results: 1. There are three categories of etimological factors of constipation, that is, endogenous, exogenous and non-exo-endogenous factor. The endogenous factor is caused by seven emotions, called depression of Ki and stagnation of Ki. The exogenous factor is six excessive atmospheric influences, for example, wind, cold, dampness, heat and dryness. And the non-exo-endogenous factors are overfatigue, improper diet, stagnated blood and deficiency of Ki and blood that comes from old age, long disease and after delivery. 2. Classification: According to cause of disease it is classified by constipation due to cold, heat, wind, dryness, retention of undigested and phlegm. According to Internal Organs there are constipation due to deficiency syndrome of the stomach, excess syndrome of the stomach, deficiency syndrome of kidney and splenic constipation. And Differentiation of syndromes according to Ki and blood, there are constipation of deficiency type and excess type. There are constipation due to stagnation and deficiency of Ki, deficiency of blood and stagnated blood. 3. Principles and Methods of treatment 1) Herbal Medicine (1) Excess type [1] Constipation due to heat : Seunggitang(承氣湯) and Majainwhan(麻子仁丸) [2] Constipation due to stagnation of Ki : Samatang(四磨湯) and Yukmatang(六磨湯) (2) Deficiency type [1] Constipation due to deficiency of Ki : Whanggitang(黃?湯) [2] Constipation due to deficiency of blood: Yunjangwhan(潤腸丸) [3] Constipation due to cold : Jechunjun(濟川煎) and Banyuwhan(半硫丸) 2) Enema therapy: It is a method to induce defecation by honey or pig's bile juice for weak people. 3) Acupuncture and Moxibustion: Acupoints used to treat constipation are BL25, ST25 and TE6. Moxibustion at CV8, CV6 is good for constipation due to cold. (4) Diet therapy: It is very important that we eat meals regularly and defecate on the same time even if you don't. And we have to eat food like fruits, vegetables and beans. (5) finger pressure: Finger pressing around these points like ST25, SP25, BL25, BL31, BL32, BL33 and BL34 is good for it. (6) Kigong therapy: Abdominal breathing (7) Old man' s constipation: Hip bath or diet therapy is commended. Laxation with lubricant like Supungyunjangwhan(搜風潤腸丸) is used. (8) Women' s constipation: After delivery, we have to administer tonifying prescription Sipjundaebotang(十全大補湯) and enema can be used. Conclusion : We have to examine the cause of disease and bowl movement carefully. After comprehensive analysis of the data gained by the four methods of diagnosis, we diagnose and treat patients on the base of overall of symptoms and signs.

• 한국작물학회지
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• 제23권2호
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• pp.25-33
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• 1978

7 X 7이면교잡에 의한 수도의 양적형질에 대한 Heterosis정도와 조합능력을 검정하기 위하여 몇가지 실험을 한바 그 결과를 요약하면 다음과 같다. 1. 출수일수에 있어서는 교배친의 평균치에 비하여 F_1세대의 전조합의 평균치가 105%의 Heterosis정도를 보이고 BL-1과 밀양1005의 품종이 교배친으로 되어 있는 $_{x}\textrm{F}_1세대에서는 현저한 Heterosis를 나타내었다. 2. 간장에 있어서는 교배조합에 따라서 Heterosis 정도가 다르나 장간종인 Columbia II와 단간종인 Waito가 교배친으로 되어 있는 조합은 Heterosis정도가 높고, Waito가 교배친으로 되어 있는 $_{x}\textrm{F}_1세대에서는 초우성이 나타났으며, 어느조합에 있어서도 $_{x}\textrm{F}_2세대보다 $_{x}\textrm{F}_1세대에서 Heterosis정도가 크게 나타났다. 3. 수장에 있어서도 $_{x}\textrm{F}_1세대에서의 Heterosis정도가 $_{x}\textrm{F}_2세대보다 현저하고 Columbia II, Waito, BL-1, 밀양1005, YR675-153-2-2등이 교배친으로 되어 있는 조합에서는 Heterosis정도가 현저하고 또한 이들 조합의 $_{x}\textrm{F}_1세대에서는 대부분 초우성으로 나타났다. 4. 수수에 었어서도 $_{x}\textrm{F}_1세대에서의 Heterosis정도가 $_{x}\textrm{F}_2세대보다 현저하고, BL-1, Waito, YR67S-153-2-2, 밀양1005가 교배친으로 되어 있는 $_{x}\textrm{F}_1세대와 BL-1과 Waito가 교배친으로 되어 있는 $_{x}\textrm{F}_2세대에서 초우성으로 나타난 조합이 많았다. 5. 지엽장에 있어서는 조합에 따라 Heterosis 정도가 매우 다르나 BL-1과 Waito가 교배친으로 되어 있는 $_{x}\textrm{F}_1, $_{x}\textrm{F}_2세대에서 Heterosis정도가 높고, Waito가 교배친으로 되어 있는 $_{x}\textrm{F}_1세대에서는 초우성으로 나타났다. 6. 지엽폭도 지엽장과 같이 BL-1과 Waito가 교배친으로 되어 있는 조합에 한해서 Heterosis정도가 현저하다. 7. 추출도는 어느 조합에 있어서도 현저한 Heterosis 정도를 나타내었으며, $_{x}\textrm{F}_1세대에서 현저한 초우성을 나타내었다. 8. 천입중에 대한 Heterosis정도는 어느 조합에 있어서도 현저하지 아니하였다. 9. $_{x}\textrm{F}_2세대에서의 입중의 Heterosis정도는 영남조생과 Waito가 교배친으로 되어 있는 조합에 한해서 현저한 Heterosis를 보이고 그중에서도 영남조생\timesIR24 영남조생 \timesColumb II에서는 초우성을 나타내었다

• Korean Journal of Acupuncture
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• 제28권1호
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• pp.91-100
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• 2011

Objectives : To classify needling or moxibustion-prohibited acupoints in Emergency Formulas Worth a Thousand in Gold (EFWTG) and Supplement to the Formulas Worth a Thousand in Gold (SFWTG). Methods : We found needling or moxibustion-prohibited acupoints in EFWTG and SFWTG, then investigated the influences of needling or moxibustion-prohibited acupoints on A-B Classic of Acupuncture and Moxibustion. Results : In EFWTG, the needling-prohibited points were LI13, ST17, BL56, TE8, CV8, CV15 and GV24. Acupoints needed careful needling were LU2, ST12, KI2, KI7, TE19, GB3 and jwagak. The moxibustion-prohibited points were LU3, LU8, ST1, ST8, ST9, ST17, ST32, ST33, BL6, BL30, TE18, TE23, GB33, GB42, CV5, CV15, GV6, GV15, GV16 and GV17. Acupoints needed careful moxibustion were ST7, ST30, TE21 and GB22. In SFWTG, the needling-prohibited points were LU2, LI13, ST12, ST17, ST32, BL56, KI2, KI7, TE8, TE19, GB3, CV8, CV15, GV24 and jwagak. The moxibustion-prohibited points were LU3, LU8, ST1, ST7, ST8, ST9, ST17, ST30, ST32, ST33, BL6, BL30, TE18, TE23, GB22, GB33, GB42, CV5, CV15, GV6, GV15, GV16, GV17 and ijung. Conclusions : There were 7 needling-prohibited points, 7 acupoints needed careful needling, 20 moxibustion-prohibited points, and 4 acupoints needed careful needling in EFWTG, and 15 needling-prohibited points and 24 moxibustion-prohibited points in SFWTG. The needling or moxibustion-prohibited acupoints in A-B Classic of Acupuncture and Moxibustion had a strong influence on those in the two literatures.

• Journal of Acupuncture Research
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• 제21권6호
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• pp.63-84
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• 2004

Objective & Methods : To study the effects of anti-cancer, anti-metastasis and immune response improvement of herbal-acupuncture with Eclipta prostrata diffusae herba infusion solution(ELP-HAS), we injected ELP-HAS into Chung-wan(CV12) of C57BL/6 mice implanted intravenously with B16-F10 melanoma. We have reached the following conclusion through the effect on the percentage of $CD25^+/CD4^+$, $CD8^+/CD3e^+$, $CD69^+/B220^+$, $NK1.1^+/CD3e^+$cells in mouse PBMCs, the effect on the pulmonary colony formation number, and the effect on MST(Median Survival Time) and ILS(Increase of Life Span) of C57BL/6 mice implanted intravenously with B16-F10 melanoma. Results : The results were obtained as follows : 1. In the experiment groups treated with ELP(Ecliptae Herba) Herbal acupuncture, the spleen cell proliferation in BALB/c mouse was significantly increased compared with control group. 2. In the experiment groups treated with ELP(Ecliptae Herba) Herbal acupuncture, the percentage of $CD25^+/CD4^+$, $CD8^+/CD3e^+$, $CD69^+/B220^+$, $NK1.1^+/CD3e^+$cells in C57BL/6 mouse PBMCs was increased compared with control group. 3. In the experiment groups treated with ELP(Ecliptae Herba) Herbal acupuncture, the pulmonary colony formation number of C57BL/6 mice implanted intravenously with B16-F10 melanoma was decreased significantly compared with control group. 4. In the experiment groups treated with ELP(Ecliptae Herba) Herbal acupuncture, MST(Median Survival Time) and ILS(Increase of Life Span) of C57BL/6 mice implanted intravenously with B16-F10 melanoma were increased significantly compared with control group. Conclusions : ELP Herbal acupuncture injection into Chung-wan(CV12) of C57BL/6 mice is considered to be effective in anti-cancer, anti-metastasis and immune response improvement.

• 대한본초학회지
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• 제36권5호
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• pp.101-108
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• 2021

Objectives : A root of Dipsacus asperoides C. Y. Cheng et T. M. Ai (D. asperoides) has been traditionally used as a medicinal resource in several Asian countries, including Korean and traditional Chinese medicine that has been traditionally used for treating several medical conditions including pain, arthritis, and bone fractures in Korea. In the present study, we investigated potential subacute toxicities of D. asperoides extract. Methods : C57BL/6 mice (male, 7weeks) were randomly divided into 4 groups of 5 mice. Except for the control group, the mice were orally administrated D. asperoides extract at doses of 50, 150, or 450 mg/kg/day for 2 weeks. At the end of the treatment period, all mice were euthanized, and the following parameters were examined: mortality, body weight, clinical signs, gross findings, hematology, serum biochemistry, organ weight, and histopathology. Results : There were no abnormalities in mortality, clinical signs, body weight, gross findings, or organ weight after repeated administration of D. asperoides extract for 2 weeks, compared with the control group. In addition, there were no significant changes in hematological, serum biochemical, and histopathological parameters between the control group and D. asperoides extract administrated groups with doses of up to 450 mg/kg/day. Conclusion : In this study, D. asperoides extract showed no significant toxicities at a dose of up to 450 mg/kg/day in mice. Although we could not confirm the toxic dose of D. asperoides extract, it can be considered safe for further pharmacological use.