• Title/Summary/Keyword: ITS marker

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A Study on 𝛽-glucan, Ginsenoside Content, 2,2-diphenyl-1-picrylhydrazyl Free Radical Scavenging Activity, Anti-inflammatory Activity and Safety of Herbal Medicine Mix - Iksooyoungjingogami with Scutellariae Radix and Houttuynia cordata Thunb (황금, 어성초를 배합한 익수영진고가미 한약재배합물의 베타글루칸, 진세노사이드 함량, 2,2-diphenyl-1-picrylhydrazyl Free Radical 소거 활성, 항염 활성 및 안전성 연구)

  • Kim, Myeong-Hun;Moon, Yang-Seon;Kang, Sang-Mi;Kim, Heyong-Seok;Kim, Seon-Jong;Na, Chang-Su
    • Journal of Korean Medicine Rehabilitation
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    • v.32 no.2
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    • pp.1-17
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    • 2022
  • Objectives This study was conducted to investigate the beta-glucan & ginsenoside content, antioxidant activity, anti-inflammatory effect and safety of herbal medicine mix. Methods The marker compounds contents, antioxidant activity and safety of herbal medicine mix were tested. The contents of beta-glucan and ginsenoside Rg3 were measured, the antioxidant activity was measured using 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, anti-inflammatory and a safety test was conducted via single dose toxicity assessment. Results Analyzing the contents of marker compounds showed 362.3 mg/g of beta-glucan, and 0.4184 mg/g of ginsenoside Rg3. In the DPPH free radical scavenging activity, the IC50 of herbal medicine mix, was 0.146%. The scavenging activity of herbal medicine mix was 88.28% activity at 0.5% concentration, and 90.61% activity at 5% concentration. In the lipopolysaccharides (LPS) anti-inflammatory test, the herbal remix showed a significant decrease in tumor necrosis factor-alpha (TNF-𝛼) and interleukin-6 (IL-6) compared to the LPS-induced group. In the single dose toxicity test of herbal medicine mix, a dose of 2,000 mg/kg body weight (BW) was set at its highest capacity and observed after oral administration to female and male rats. No toxicological findings were recognized. It was observed that the resulting lethal dose can be set to 2,000 mg/kg BW or higher for both females and males. Conclusions The results of the experiment on herbal medicine mix showed that the marker compounds contents were beta-glucan and ginsenoside Rg3, that antioxidant activity was observed through the DPPH free radical scavenging activity, anti-inflammatory effect was observed through TNF-𝛼 and IL-6 measurement, and safety was confirmed through the single dose toxicity assessment.

PCR-based markers to select plastid genotypes of Solanum acaule (Solanum acaule 색소체 유전자형 선발을 위한 특이적 분자마커 개발)

  • Park, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.178-186
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    • 2022
  • The tetraploid Solanum acaule is a wild potato species from Bolivia widely used for potato breeding because of its diverse attractive traits, including resistance to frost, late blight, potato virus X, potato virus Y, potato leafroll virus, potato spindle tuber viroid, and cyst nematode. However, the introgression of useful traits into cultivated potatoes via crossing has been limited by differences in endosperm balance number between species. Somatic fusion could be used to overcome sexual reproduction barriers and the development of molecular markers is essential to select proper fusion products. The chloroplast genome of S. acaule was sequenced using next-generation sequencing technology and specific markers for S. acaule were developed by comparing the obtained sequence with those of seven other Solanum species. The total length of the chloroplast genome is 155,570 bp, and 158 genes were annotated. Structure and gene content were very similar to other Solanum species and maximum likelihood phylogenetic analysis with 12 other species belonging to the Solanaceae family revealed that S. acaule is very closely related to other Solanum species. Sequence alignment with the chloroplast genome of seven other Solanum species revealed four InDels and 79 SNPs specific to S. acaule. Based on these InDel and SNP regions, one SCAR marker and one CAPS marker were developed to discriminate S. acaule from other Solanum species. These results will aid in exploring evolutionary aspects of Solanum species and accelerating potato breeding using S. acaule.

Analysis of Microsatellite Patterns in the Genome of Red Sea Cucumber (홍해삼 유전체 분석에 의한 microsatellite의 분포도 연구)

  • Lee, Tae Wook;Kim, Sam Woong;Kim, Jung Sun;Chi, Won-Jae;Bang, Woo Young;Kim, Jang Hyeon;Yang, Chul Woong;Bang, Kyu Ho;Gal, Sang Wan
    • Journal of Life Science
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    • v.32 no.9
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    • pp.690-697
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    • 2022
  • This study was conducted to analyze genome of red sea cucumber and to use it as basic data for the development of genetic markers for red sea cucumber. Microsatellite marker analysis of Ulleungdo_normal and Ulleungdo_native red sea cucumbers revealed that dinucleotide simple sequence repeats (SSRs) had the highest ratio, at 81.3~81.4%, and the number of the detected SSRs tended to decrease as the number of repeating sequence units in SSRs increased. In general, microsatellites with between 5 and 10 iterations were most common. As the size of the SSR repeating sequence units increased, the SSR iterations gradually decreased. The di-, tri-, and tetra-nucleotides in SSRs were detected in the highest numbers as (AT)5, (AAT)5, and (AAAT)5, respectively. (CG) and (CCG) had very low frequencies compared to the numbers of other repeating SSR units. The numbers of di-and tri-nucleotide repeats were up to 35 and 32, respectively, and then increased discontinuously up to 44 and 43 repeats, respectively. Tetra-, penta-, and hexa-nucleotides in SSRs occurred in numbers up to 25, 21 and 14, respectively. This analysis of red sea cucumber indicated that it maintains its own repetition sequence and repetition number; therefore, we suggest that using it as basic data for molecular marker will be possible in future research.

Evaluation of usefulness of the Gated Cone-beam CT in Respiratory Gated SBRT (호흡동조 정위체부방사선치료에서 Gated Cone-beam CT의 유용성 평가)

  • Hong sung yun;Lee chung hwan;Park je wan;Song heung kwon;Yoon in ha
    • The Journal of Korean Society for Radiation Therapy
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    • v.34
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    • pp.61-72
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    • 2022
  • Purpose: Conventional CBCT(Cone-beam Computed-tomography) caused an error in the target volume due to organ movement in the area affected by respiratory movement. The purpose of this paper is to evaluate the usefulness of accuracy and time spent using the Gated CBCT function, which reduces errors when performing RGRT(respiratory gated radiation therapy), and to examine the appropriateness of phase. Materials and methods: To evaluate the usefulness of Gated CBCT, the QUASARTM respiratory motion phantom was used in the Truebeam STxTM. Using lead marker inserts, Gated CBCT was scaned 5 times for every 20~80% phase, 30~70% phase, and 40~60% phase to measure the blurring length of the lead marker, and the distance the lead marker moves from the top phase to the end of the phase was measured 5 times. Using Cedar Solid Tumor Inserts, 4DCT was scanned for every phase, 20-80%, 30-70%, and 40-60%, and the target volume was contoured and the length was measured five times in the axial direction (S-I direction). Result: In Gated CBCT scaned using lead marker inserts, the axial moving distance of the lead marker on average was measured to be 4.46cm in the full phase, 3.11cm in the 20-80% phase, 1.94cm in the 30-70% phase, 0.90cm in the 40-60% phase. In Fluoroscopy, the axial moving distance of the lead marker on average was 4.38cm and the distance on average from the top phase to the beam off phase was 3.342cm in the 20-80% phase, 3.342cm in the 30-70% phase, and 0.84cm in the 40-60% phase. Comparing the results, the difference in the full phase was 0.08cm, the 20~80% phase was 0.23cm, the 30~70% phase was 0.10cm, and the 40~60% phase was 0.07cm. The axial lengths of ITV(Internal Target Volume) and PTV(Planning Target Volume) contoured by 4DCT taken using cedar solid tumor inserts were measured to be 6.40cm and 7.40cm in the full phase, 4.96cm and 5.96cm in the 20~80% phase, 4.42cm and 5.42cm in the 30~70% phase, and 2.95cm and 3.95cm in the 40~60% phase. In the Gated CBCT, the axial lengths on average was measured to be 6.35 cm in the full phase, 5.25 cm in the 20-80% phase, 4.04 cm in the 30-70% phase, and 3.08 cm in the 40-60% phase. Comparing the results, it was confirmed that the error was within ±8.5% of ITV Conclusion: Conventional CBCT had a problem that errors occurred due to organ movement in areas affected by respiratory movement, but through this study, obtained an image similar to the target volume of the setting phase using Gated CBCT and verified its usefulness. However, as the setting phase decreases, the scan time was increases. Therefore, considering the scan time and the error in setting phase, It is recommended to apply it to patients with respiratory coordinated stereotactic radiation therapy using a wide phase of 30-70% or more.

The Effect of Hwangryunhaedok-tang and its Major Component Coptidis Rhizoma, Berberine on Nicotine Addiction in Rats (황련해독탕(黃連解毒湯)과 그 주성분인 황련(黃連) 및 황련(黃連)의 주성분인 berberine이 니코틴 중독 흰쥐에 미치는 영향)

  • Hong, Eui-Jae;Min, Sang-Yeon;Kim, Jang-Hyun
    • The Journal of Korean Medicine
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    • v.29 no.4
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    • pp.55-67
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    • 2008
  • Objectives: The purpose of this study was to investigate the effect of Hwangryunhaedok-tang, its major component Coptidis Rhizoma, and the major component of Coptidis Rhizoma, berberine, on the expression of behavioral sensitization induced by the repeated administration of nicotine. Methods: Experimental ratswere divided into 3 groups and each group treated with saline, nicotine, and Hwangryunhaedok-tang. Experiments were performed on 4 further groups, each group treated with saline, nicotine, Coptidis Rhizoma, and berberine. To obtain rats addicted todrugs, nicotine was injected repeatedly. After 3 days of withdrawal phase, on the 11th day, rats were re-exposed to nicotine and thus able to induce the expression of the activity sensitization phenomenon successfully. To examine the change of c-Fos in the brain of experimental rats with the nicotine-induced activity sensitization, their neurological activation rate was assessed by c-Fos staining with Fos-like immunihistochemistry of the dopamine releasing area, the striatum and the nucleus accumbens. Results: The result demonstrate that Hwangryunhaedok-tang, its major component Coptidis Rhizoma and berberine,the major component of Coptidis Rhizoma, significantly reduced the walking activity rate in the drug addict models induced by nicotine administration, and Coptidis Rhizoma and its major component berberine reduce the marker of the neurological activation c-Fos, and thus revealed an effect of suppressing drug addiction. Conclusions: These results suggest the possibility of Hwangryunhaedok-tang, its major component Coptidis Rhizoma and the major component of Coptidis Rhizoma, berberine,as therapeutic for nicotine addiction.

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Microsatellite Instability in Non-Small Cell Lung Cancer (비소세포폐암에서 Microsatellite Instability)

  • Jeon, Hyo-Sung;Kim, Jeong-Ran;Son, Ji-Woong;Park, Sun-Ha;Park, Tae-In;Kim, Chang-Ho;Kim, In-San;Jung, Tae-Hoon;Park, Jae-Yong
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.1
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    • pp.24-32
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    • 2000
  • Purpose: Microsatellite instability(MSI) is frequently used as an indicator of microsatellite mutator phenotype(MMP) tumors. MSI has been observed in a percentage of non-small cell lung cancer(NSCLC). However, its role in tumorigenesis of NSCLC remains unknown. The frequency and pattern of MSI in NSCLC were evaluated and clinical parameters of MSI-positive tumors with those of MSS(microsatellite stable) tumors were compared. Materials and Methods: Twenty surgically resected NSCLCs were analyzed for 15 microsatellite markers located at chromosomes 3p and 9p. The peripheral blood lymphocytes of patients were used as the source of the normal DNA. Results: 1) Of 20 cases, 8(40%) demonstrated MSI. 2) Instability was observed more frequently in tri- and tetra-nucleotide repeats than in dinucleotide repeats. In all cases, instability appeared as a shift of individual allelic bands. 3) LDH was observed in 10(50%) of 20 tumors analyzed. 4) Of 20 cases, MSI-H tumor(showing MSI in the majority of markers) was absent. There were 5 MSI-L tumors(showing MSI in a greater than 10% of markers). 5) No significant difference was observed between MSI-L tumors and MSI-negative tumors in terms of clinicopathologic features such as pack-year history of smoking, histologic subtype, and(delete) stage of disease. There was also no significant difference in the incidence of LDH in relation to the status of MSI. Conclusion: These data strongly suggest that MSI plays different roles in lung and colon cancer. MMP pathway appears to be far less important in the tumorigenesis of NSCLC, caused mainly by cigarette smoke, with little familial tendency.

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Phylogenetic Relationships of the Korean Trigonotis Steven (Boraginaceae) Based on Chloroplast DNA (cpDNA) and Nuclear Ribosomal Markers (nrDNA) Region

  • Trinh, Ngoc Ai;Nguyen, Hien Thi Thanh;Park, Seon Joo
    • Korean Journal of Plant Resources
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    • v.25 no.6
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    • pp.753-761
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    • 2012
  • We performed phylogenetic analyses of a total of 21 acessions covering 5 species in the Korean Trigonotis and one outgroup species using nuclear ribosomal ITS and chloroplast rbcL, matK, ndhF sequences. Outgroup were chosen from the closely related genus Lithospermum zollingeri. Both parsimony and Bayesian Inference methods were used to reconstruct the evolutionary history of the group. The evidence collected indicated that phylogenetic relationships among Korean Trigonotis species are unresolved based on nuclear marker (ITS), as the same as based on separated chloroplast sequences. While the phylogenetic relationships of Korean Trigonotis species almost clearly were resolved in combined chloroplast sequences. Thus, the members of Trigonotis coreana can be distinguished to the members of Trigonotis peduncularis in combined cpDNA sequences and Trigonotis nakaii was treated as a synonymed to Trigonotis radicans var. sericea. In addition, the MP and BI analysis showed Trigonotis icumae as sister of the remained Korean Trigonotis species based on combined molecular markers (BI: PP = 1).

Insulin activates EGFR by stimulating its interaction with IGF-1R in low-EGFR-expressing TNBC cells

  • Shin, Miyoung;Yang, Eun Gyeong;Song, Hyun Kyu;Jeon, Hyesung
    • BMB Reports
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    • v.48 no.6
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    • pp.342-347
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    • 2015
  • The expression of epidermal growth factor receptor (EGFR) is an important diagnostic marker for triple-negative breast cancer (TNBC) cells, which lack three hormonal receptors: estrogen and progesterone receptors as well as epidermal growth factor receptor 2. EGFR transactivation can cause drug resistance in many cancers including TNBC, but the mechanism underlying this phenomenon is poorly defined. Here, we demonstrate that insulin treatment induces EGFR activation by stimulating the interaction of EGFR with insulin-like growth factor receptor 1 (IGF-1R) in the MDA-MB-436 TNBC cell line. These cells express low levels of EGFR, while exhibiting high levels of IGF-1R expression and phosphorylation. Low-EGFRexpressing MDA-MB-436 cells show high sensitivity to insulinstimulated cell growth. Therefore, unexpectedly, insulin stimulation induced EGFR transactivation by regulating its interaction with IGF-1R in low-EGFR-expressing TNBC cells. [BMB Reports 2015; 48(6): 342-347]

Polymorphism of Growth Hormone Gene in 12 Pig Breeds and Its Relationship with Pig Growth and Carcass Traits

  • Wang, Wenjun;Huang, Lusheng;Gao, Jun;Ding, NengShui;Chen, Kefei;Ren, Jun;Luo, Ming
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.2
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    • pp.161-164
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    • 2003
  • The polymorphism of the growth hormone gene in 12 pig breeds (total n=475) was detected by PCR-Apa I-RFLP, and allele A (449 bp, 101 bp and 55 bp) or allele B (316 bp, 133 bp, 101 bp and 55 bp) were observed. In these pig breeds, we found that European pig breeds had high frequencies of allele B, while Chinese native pig breeds had high frequencies of allele A. In addition, the role of porcine GH was investigated in 117 Nanchang White pigs and 361 Large Yorkshire pigs. Eight traits about growth and carcass were recorded for analyzing associations between GH gene polymorphism and performance quantitative traits. In the Nanchang White pigs, no significant difference was observed between different genotypes and different growth and carcass traits. In Large Yorkshire pigs, those with BB genotype had more lean percentage than pigs with AA genotype (p<0.05). Based on these results, we conclude that the GH locus should be further investigated in commercial breeds to determine its suitability for use in marker-assisted selection programmes.

The Relative Identification of C. officinale and L. chuanxiong by PCR-Mediated Fingerprinting (천궁류(川芎類) 한약재의 유전자 감식 연구)

  • Choi, Ho-Young;Kim, Dong-Wook;Kim, Dong-Eun;Suh, Young-Bae;Ham, In-Hye
    • The Korea Journal of Herbology
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    • v.20 no.4
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    • pp.151-161
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    • 2005
  • Objectives : Our research purpose is to establish the standard identification analysis on C. officinale and L. chuanxiong in Korea and China by PCR-mediated fingerprinting. Methods : The Restriction Fragment Length Polymorphism (RFLP) and Randomly Amplified Polymorphic DNA (RAPD) method was used on Internal Transcribed Spacer (ITS) regions and rbcL regions to compare and discriminate genes extracted from crude drugs as C. officinale and L. chuanxiong in Korea and China. Results : L. chuanxiong Korea and China have very similar polymorphism, whereas L. chuanxiong in Korea and C. officinale have very different polymorphism in RFLP. And restriction enzymes AluI and SacI forms the specific fragment band only in C. officinale, they can be used as RFLP marker on ITS regions to discriminate among the species. Conclusions : The results could be applied in discriminating crude drugs among C. officinale and L. chuanxiong in Korea and China. Also they could be used in controlling drug quality, preserving medicinal plants, and improving plant description.

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