• Applied Biological Chemistry
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• 제23권4호
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• pp.199-205
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• 1980

인삼의 유효약리성분으로 밝혀진 saponin중의 각 ginsenosides를 효과적이고 능률적으로 분리하기 위하여 대량분취전용 고속액체 chromatograph인 preparative HPLC의 응용을 검토하였다. 조(粗) saponin획분을 preparative HPLC인 Prep LC/system-500를 사용하여 부분분획을 하고 각 획분에 함유되어 있는 ginsenosides의 조성을 Analytical HPLC로 동정한 후 Semi-preparative HPLC를 사용하여 인삼주성분 saponin을 단리했다. 그 결과 인삼 주성분 saponin인 $ginsenoside-Rb_1,\;-Rb_2,\;-Rc,\;-Rd,\;-Re$ 및 $-Rg_1$은 약 20 mg / 2.0 ml / injection으로 chromatography를 행하여 $300{\sim}400mg/day$로 대량분취가 가능하였다. 따라서 ginsenosides의 약리 및 임상효능 연구에 크게 기여하게 될 것이다.

• Applied Biological Chemistry
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• 제23권4호
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• pp.222-227
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• 1980

고려인삼근(자경종)(高麗人蔘根(紫莖種))의 중심부(형성층내부)(中心部(形成層內部))와 외피십피층(外皮十皮層)에 있는 ginsenoside를 고속액 체크로마토그라피로 분석(分析)하고 경(莖)의 자색정도(紫色程度)와의 관계(關係)를 검토(檢討)하였다. Ginsenoside의 단순상관(單純相關)에 의(依)한 saponin 양상(樣相)의 유사도(類似度)를 경색도군간(莖色度群間) 같은 뿌리 또는 다른 뿌리간(間)에 두부위(部位)에서 비교(比較)한 결과(結果) 경색도(莖色度)는 saponin 양상(樣相)과 관련(關聯)되지 않는 것으로 보였다. Saponin 양상(樣相)은 부위(部位)의 출처(出處)에 관계(關係)없이 서도 다른 부위간(部位間)에 약간 달랐다. 각(各) ginsenoside 함량순위(含量順位)는 표피십피층(表皮十皮層)에서 $Rg_1>Re>Rb_1>Rb_2>Rc>Rg_2{\geq}Rd>Rf$이고 중심부(中心部)에서는 $Rg_1>Re{\geq}Rg_2{\geq}Rb_1{\gg}Rb_2>Rc{\geq}Rd>Rf$였다.

• Journal of Microbiology and Biotechnology
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• 제24권12호
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• pp.1744-1754
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• 2014

The hepatitis C virus (HCV) RNA genome is replicated by an RNA replicase complex (RC) consisting of cellular proteins and viral nonstructural (NS) proteins, including NS5B, an RNA-dependent RNA polymerase (RdRp) and key enzyme for viral RNA genome replication. The HCV RC is known to be associated with an intracellular membrane structure, but the cellular components of the RC and their roles in the formation of the HCV RC have not been well characterized. In this study, we took a proteomic approach to identify stomatin, a member of the integral proteins of lipid rafts, as a cellular protein interacting with HCV NS5B. Co-immunoprecipitation and co-localization studies confirmed the interaction between stomatin and NS5B. We demonstrated that the subcellular fraction containing viral NS proteins and stomatin displays RdRp activity. Membrane flotation assays with the HCV genome replication-competent subcellular fraction revealed that the HCV RdRp and stomatin are associated with the lipid raft-like domain of membranous structures. Stomatin silencing by RNA interference led to the release of NS5B from the detergent-resistant membrane, thereby inhibiting HCV replication in both HCV subgenomic replicon-harboring cells and HCV-infected cells. Our results identify stomatin as a cellular protein that plays a role in the formation of an enzymatically active HCV RC on a detergent-resistant membrane structure.

• Journal of Acupuncture Research
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• 제18권1호
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• pp.157-169
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• 2001

Objective : The purpose of this study is to investigate the effects on WBC counts and each differential of neutrophils, lymphocytes, monocytes in whole blood sample of each experimental Aqua-acupuncture treated mice groups PCortex Ulmi Pumilae(CU), Ramulus Cinnamomum(RC), Radix Achyranthes(RA), Apitoxin(BV), Calculus Bovis Fel Ursi Moschus compound(BUM). Materials & Method : All the BALB/c mice used in this study were bred and maintained in our pathogen-free mouse facility and were 6 weeks of age at the start of the experiment. The experimental model of arthritis was induced by injecting 300${\mu}g$/kg LPS to all mice knee joint. The each of Aqua-acupuncture(Cortex Ulmi Pumilae, Ramulus Cinnamomum, Radix Achyranthes, Apitoxin, Calculus Bovis Fel Ursi Moschus compound) was injected into GB34(陽陵泉) of mice groups every other day for 6 times. And the WBC counts and each differential of neutrophils, lymphocytes, monocytes were measured at the 3rd, 7th and 14th day after LPS injection. Results : 1. The WBC counts were significantly decreased compared with the control(CON) group in every Aqua-acupuncture groups at all days. And at the 14th day, BV & BUM groups were more significantly decreased than RA group. 2. The Neutrophil's ratio was significantty increased compared with the CON group in CU & RC groups at the 3rd day and RC group was more significant than CU group. But at the 7th and 14th day, every Aqua-acupuncture groups were significantly increased compared with the CON group and at the 7th day, RC group was more significant than RA, BV & BUM groups and at the 14th day, RC, BV & BUM groups were more significant than RA group. 3. The Lymphocyte's ratio was significantly decreased compared with the CON group in RC group at the 3rd day. At the 7th day, CU, RC & BV groups were significantly decreased compared with the CON group. At the 14th day, every Aqua-acupuncture groups were significantly decreased compared with the CON group and RC group was more significant than RA group, 4. The Monocyte's ratio was significantly decreased compared with the CON group in every Aqua-acupuncture groups at the 7th day. At the 14th day, BV & BUM groups were significantly decreased compared with the CON group. Conclusion : According to the above results, it was concluded that CU & RC groups were more effective at the early period of this experiment, and at the latter period, BV & BUM groups were more effective than others. RA group was less effctive than others.

• 전자공학회논문지C
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• 제36C권7호
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• pp.1-9
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• 1999

본 논문에서는 칩 내부 회로 연결선의 모형으로 많이 사용되는 RC-class 회로에 대하여 시뮬레이션을 수행하지 않고 지연 시간을 계산할 수 있는 해석적 3차 근사 기법을 제시한다. 본 논문에서 제시하는 3차 근사 기법은 기존의 2차 근사 기법에 비해 크지 않은 수행 시간을 필요로 하면서도 보다 정확한 결과를 보장한다. 이 해석적 3차 근사 기법은 일반적인 q 차 AWE(Asymptotic Waveform Evaluation)기법의 계산 결과와 비교해 허용 가능한 수준의 오차를 보장하며, 계산 시간의 단축과 함께 수치적으로 안정된 값을 제공한다. 제안하는 기법의 첫 알고리즘은 3차의 근사를 위해 8개의 모멘트를 필요로 하며, 보다 정확한 지연 시간의 근사가 가능하다. 둘째 알고리즘은 3차의 근사를 위해 6개의 모멘트를 필요로 하며, 첫 알고리즘보다 정확도는 뒤지나 빠른 근사가 가능하다.

• Journal of Applied Biological Chemistry
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• 제62권4호
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• pp.315-322
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• 2019

Seasonal ginsenoside flux in the leaves of 5-year-old Panax ginseng was analyzed from the field-grown ginseng, for the first time, to study possible biosynthesis and translocation of ginsenosides. The concentrations of nine major ginsenosides, Rg1, Re, Rh1, Rg2, R-Rh1, Rb1, Rc, Rb2, and Rd, were determined by UHPLC during the growth in between April and November. It was confirmed total ginsenoside content in the dried ginseng leaves was much higher than the roots by several folds whereas the composition of ginsenosides was different from the roots. The ginsenoside flux was affected by ginseng growth. It quickly increased to 10.99±0.15 (dry wt%) in April and dropped to 6.41±0.14% in May. Then, it slowly increased to 9.71±0.14% in August and maintained until October. Ginsenoside Re was most abundant in the leaf of P. ginseng, followed by Rd and Rg1. Ginsenosides Rf and Ro were not detected from the leaf. When compared to the previously reported root data, ginsenosides in the leaf appeared to be translocated to the root, especially in the early vegetative stage even though the metabolite translocated cannot be specified. The flux of ginsenoside R-Rh1 was similar to the other (20S)-PPT ginsenosides. When the compositional changes of each ginsenoside in the leaf was analyzed, complementary relationship was observed from ginsenoside Rg1 and Re, as well as from ginsenoside Rd and Rb1+Rc. Accordingly, ginsenoside Re in the leaf was proposed to be synthesized from ginsenoside Rg1. Similarly, ginsenosides Rb1 and Rc were proposed to be synthesized from Rd.

• Journal of Ginseng Research
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• 제39권4호
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• pp.338-344
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• 2015

Background: Ginseng (the roots of Panax ginseng Meyer) is a well-known traditional Oriental medicine and is now widely used as a health food. It contains several types of ginsenosides, which are considered the major active medicinal components of ginseng. It has recently been reported that the qualitative and quantitative properties of ginsenosides found in ginseng may differ, depending on cultivation regions, ages, species, and so on. Therefore, it is necessary to study these variations with respect to cultivation ages and regions. Methods: In this study, 3-6-yr-old roots of P. ginseng were collected from three different cultivation regions. The contents of five ginsenosides (Rb1, Rd, Rc, Re, and Rgl) were measured by rapid resolution liquid chromatography coupled with quadruple-time-of-flight mass spectrometry. The Kruskal-Wallis Rank sum test and multiple t test were used for comparative analysis of the data to evaluate the dynamic changes in the accumulation of these ginsenosides affected by cultivation regions and ages. Results: The content and composition of ginsenosides varied significantly among specimens collected from different cultivation regions and having different cultivation ages. For all samples, the content of Rg1 and Re ginsenosides increases with age and this rate of increase is different for each sample. The contents of Rb1, Rc, and Rd varied with cultivation ages in samples from different cultivation regions; especially, Rb1 from a 6-yr-old root showed approximately twofold variation among the samples from three cultivation regions. Furthermore, the content of Rb1 highly correlated with that of Rd (r = 0.89 across all locations and ages). Conclusion: In our study, only the contents of ginsenosides Rg1 and Re were affected by the root age. Ginsenosides Rb1, Rc, and Rd varied widely with ages in samples from different cultivation regions.

• Journal of Ginseng Research
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• 제39권3호
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• pp.221-229
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• 2015

Background: Minor ginsenosides, those having low content in ginseng, have higher pharmacological activities. To obtain minor ginsenosides, the biotransformation of American ginseng protopanaxadiol (PPD)-ginsenoside was studied using special ginsenosidase type-I from Aspergillus niger g.848. Methods: DEAE (diethylaminoethyl)-cellulose and polyacrylamide gel electrophoresis were used in enzyme purification, thin-layer chromatography and high performance liquid chromatography (HPLC) were used in enzyme hydrolysis and kinetics; crude enzyme was used in minor ginsenoside preparation from PPD-ginsenoside; the products were separated with silica-gel-column, and recognized by HPLC and NMR (Nuclear Magnetic Resonance). Results: The enzyme molecular weight was 75 kDa; the enzyme firstly hydrolyzed the C-20 position 20-O-${\beta}$-D-Glc of ginsenoside Rb1, then the C-3 position 3-O-${\beta}$-D-Glc with the pathway $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}C-K$. However, the enzyme firstly hydrolyzed C-3 position 3-O-${\beta}$-D-Glc of ginsenoside Rb2 and Rc, finally hydrolyzed 20-O-L-Ara with the pathway $Rb2{\rightarrow}C-O{\rightarrow}C-Y{\rightarrow}C-K$, and $Rc{\rightarrow}C-Mc1{\rightarrow}C-Mc{\rightarrow}C-K$. According to enzyme kinetics, $K_m$ and $V_{max}$ of Michaelis-Menten equation, the enzyme reaction velocities on ginsenosides were Rb1 > Rb2 > Rc > Rd. However, the pure enzyme yield was only 3.1%, so crude enzyme was used for minor ginsenoside preparation. When the crude enzyme was reacted in 3% American ginseng PPD-ginsenoside (containing Rb1, Rb2, Rc, and Rd) at $45^{\circ}C$ and pH 5.0 for 18 h, the main products were minor ginsenosides C-Mc, C-Y, F2, and C-K; average molar yields were 43.7% for C-Mc from Rc, 42.4% for C-Y from Rb2, and 69.5% for F2 and C-K from Rb1 and Rd. Conclusion: Four monomer minor ginsenosides were successfully produced (at low-cost) from the PPD-ginsenosides using crude enzyme.

• 대한약침학회지
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• 제13권1호
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• pp.63-77
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• 2010

Objectives: The aim of this experiment is to provide an objective differentiation of cultivated ginseng, cultivated wild ginseng, and wild ginseng through component analysis, and to know the change of ginsenoside components in the process for making red ginseng. Methods: Comparative analysis of ginsenoside $Rb_1,\;Rb_2$, Rc, Rd, Re, Rf, $Rg_1,\;Rg_3,\;Rh_1$ and $Rh_2$ from the cultivated ginseng 4 and 6 years, cultivated wild ginseng, and wild ginseng were conducted using High Performance Liquid Chromatography(hereafter HPLC). And the same analyses were conducted in the process of red ginseng. Results: 1. For content comparison of ginsenoside $Rb_1$, Rc, Rd, Rf, $Rg_1$ and $Rh_1$, wild ginseng showed high content, followed cultivated ginseng 4 and 6 years, cultivated wild ginseng showed low content than any other samples. 2. For content comparison of ginsenoside $Rb_2$ and Re, cultivated ginseng 4 years showed high content, followed wild ginseng and cultivated ginseng 6 years, cultivated wild ginseng showed low content than any other samples. 3. For content comparison of ginsenoside $Rg_3$, wild ginseng and cultivated wild ginseng were only showed low content. 4. For content comparison of ginsenoside $Rh_2$, cultivated wild ginseng was only showed low content. 5. In the process of red ginseng, ginsenoside $Rb_1,\;Rb_2$, Rc, Rd, $Rg_3$ and $Rh_1$ were increased, and ginsenoside Re and $Rg_1$ were decreased in cultivated wild ginseng. 6. In the process of red ginseng, ginsenoside $Rg_3$ and $Rh_1$ were increased, and ginsenoside $Rb_2$, Rc, and Re were decreased in cultivated ginseng 4 years. 7. In the process of red ginseng, ginsenoside $Rb_1,\;Rb_2$, Rf and $Rh_1$ were increased, and ginsenoside Rc and Rd were decreased in cultivated ginseng 6 years. Conclusions: Distribution of ginsenoside contents to the cultivated ginseng, cultivated wild ginseng, and wild ginseng was similar and was not showed special characteristics between samples. And the change of ginsenoside to the process of red ginseng, cultivated ginseng and cultivated wild ginseng were showed different aspect.

• Archives of Pharmacal Research
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• 제5권1호
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• pp.7-12
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• 1982

Ginsenosides Rb1, Rb2, Rc, Rd, Re, Rf and Rg1 were effectively isolated from ginseng root by preparative liquid chromatography (LC) on two PrepPAK-500/c18 cartridges in series and semipreparative LC on a .mu. Bondapak cabohydrate analysis column, a .mu.Bondapak C18 column or a .mu. Porasil column. The identities of the isolated ginsenosides were confirmed by analytical high-performance liquid chromatography (HPLC) and infrared spectrophotometry. By this method large scale isolation of pure ginsenosides was efficiently accomplished.