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Studies on the regulation of Hematopoietic enhancement of Brassica campestris var narinosa., Canavalia gladiata DC semen and their combinational prescription via Jak2/STAT5/GATA1 Pathway in Sca-1+ hematopoietic stem cells (Sca-1+골수조혈세포에서 JAK2/STAT5/GATA-1 신호전달 경로를 통한 다채, 도두 그리고 두 조합물에 의한 조혈증진 조절에 관한 연구)

  • Kim, Kunhoae;Kim, Seung-Hyung;Cho, In-Sik;Kim, Han-Young;Kim, Dong-Seon;Lee, Young-Cheol
    • The Korea Journal of Herbology
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    • v.28 no.4
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    • pp.7-16
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    • 2013
  • Objectives : Brassica campestris var narinosa (BCN), Canavalia gladiata DC semen (CGD) and their combinational prescription (BCN+CGD) have been use to demonstrate to regulate hematopoiesis. In the current study, we investigated whether Brassica campestris var narinosa, Canavalia gladiata DC semen and their combinational prescription is related to hemato-potentiating function using Sca-$1^+$ hematopoietic stem cells (Sca-$1^+HSCs$) as a testing system. Methods : Sca-$1^+HSCs$ isolated from femur in C57bl/6 mice with leukopenia and thrombocytopenia induced by cyclophosphamide (CTX). Then, Real-time PCR was performed to measure the mRNA expression, ELISA and haematopoiesis-related gene (EPO, TPO, IL-3, SCF, c-kit, GM-CSF), the phosphorylation of JAK2, GATA-1 and STAT-5a/b were observed by western blot, and the numbers of $CD117^+/Sca-1^+$ cell and the number of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E), semisolid clonogenic assay was performed. Result : When Sca-$1^+HSCs$ were treated with Brassica campestris var narinosa, Canavalia gladiata DC semen and their combinational prescription with rIL-3/rSCF, the expression of haematopoiesis-related (EPO, TPO, IL-3, SCF, c-kit, and GM-CSF) were significantly increased at the levels of mRNA as well as production in Sca-$1^+HSCs$. Additionally, CGS enhanced phosphorylation of JAK2, GATA-1, and signal transducer and activator of transcription-5a/b (STAT-5a/b) in Sca-$1^+HSCs$. Furthermore, their combinational prescription (BCN+CGD) significantly enhanced the growth rate of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E) in vitro. Conclusion : These result suggest that Brassica campestris var narinosa (BCN) and Canavalia gladiata DC have hematopoietic enhancement via hematopoietic cytokine-mediated JAK2/GATA-1/STAT-5a/b pathway, and their combinational prescription (BCN+CGD) has superior hematopoietic enhancement to those of individual extracts.

Comparison of Angelica Species Roots Using Taste Sensor and DNA Sequencing Analysis (미각센서와 DNA 염기서열을 이용한 당귀류 비교)

  • Kim, Young Hwa;Choi, Goya;Lee, Hye Won;Lee, Gwan Ho;Chae, Seong Wook;Kim, Yun Hee;Lee, Mi Young
    • The Korea Journal of Herbology
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    • v.27 no.6
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    • pp.37-42
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    • 2012
  • Objectives : Angelica Gigantis Radix is prescribed as the root of different Angelica species on the pharmacopoeia in Korea, Japan and China. Chemical components and their biological activities were also different according to their species. A study for the development of simple method to compare Angelica roots was needed. In order to classify them, the methods such as DNA sequencing analysis and taste sensor were applied to three Angelica species like Angelica gigas, Angelica acutiloba and Angelica sinensis. Methods : PCR amplification of intergenic transcribed spacer (ITS) region was performed using ITS1 and ITS4 primer from nine Angelica roots, and then nucleotide sequence was determined. Taste pattern of samples were measured using the taste-sensing system SA402B equipped with a sensing unit, which consists of artificial lipid membrane sensor probes of anionic bitterness, astringency, saltiness, umami, and cationic bitterness (C00, AE1, CT0, AAE, and AN0, respectively). Results : As a result of comparing the similarity of the ITS region sequences, A. sinensis was discriminated from the others (A. gigas and A. acutiloba). Equally this genetic result, A. gigas and A. acutiloba showed similar taste pattern as compared to A. sinensis. Sourness, bitterness, aftertaste of bitterness, astringency, and aftertaste of astringency of A. sinensis were significantly high as compared with A. gigas and A. acutiloba. In contrast, richness was significantly low. Conclusions : These taste pattern can be used as a way of comparison of Angelica species and this technic could be applied to establish a taste pattern marker for standardization of herbs in various purposes.

Molecular authentication of Lepidii seu Descurainiae Semen by the development of matK amplification primers and analysis of sequences (matK 증폭용 primer 개발 및 염기서열 분석을 통한 정력자(葶藶子) 유전자 감별)

  • Moon, Byeong Cheol;Kim, Wook Jin;Yang, Sungyu;Park, Inkyu;Yeo, Sang Min;Noh, Pureum
    • The Korea Journal of Herbology
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    • v.33 no.3
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    • pp.25-35
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    • 2018
  • Objectives : Lepidii seu Descurainiae Semen has been frequently adulterated with the seeds of several inauthentic plant species. However, the accurate identification of these plant seeds is very difficult. To develop a reliable genetic authentication tool for Lepidii seu Descurainiae Semen, we analyzed matK sequence. Methods : To obtain the matK sequences of plant materials, genomic DNA was extracted from 24 samples and PCR amplification was carried out using matK-AF/matK-8R universal primer set and matK-LDSF/matK-LDSR primer set. For identifying species-specific nucleotides and phylogenetic analysis, matK regions were sequenced and comparatively analyzed by the ClustalW and Maximum Likelihood method. Results : We developed a new primer set to amplify matK region in Lepidii seu Descurainiae Semen and closely related plant samples. From the comparative analysis of matK sequences, we identified species-specific marker nucleotides for D. sophia, L. apetalum, L. latifolium, E. cheiranthoides, E. macilentum, and D. nemorosa, respectively. Furthermore, phylogenetic analysis revealed clear classification depending on the species. These results indicated that the matK sequence obtained a new primer set in this study was useful to identify Lepidii seu Descurainiae Semen in species level. Conclusions : We developed a primer set and identified species-specific marker nucleotides enough to distinguish authentic Lepidii seu Descurainiae Semen and adulterants at the species level based on the matK sequences. These genetic tool will be useful to prevent adulteration and to standardize the quality of Lepidii seu Descurainiae Semen.

Effect of Paeonia Lactiflora Pallas on Atopic Dermatitis-Related Inflammation in HaCaT Cell (작약이 HaCaT 세포에서 아토피 피부염 관련 염증 억제에 미치는 영향)

  • Lee, Hye-In;Kim, Eom Ji;Son, Dongbin;Joo, Byung Duk;Sohn, Youngjoo;Kim, Eun-Young;Jung, Hyuk-Sang
    • Korean Journal of Acupuncture
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    • v.39 no.2
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    • pp.43-53
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    • 2022
  • Objectives : Paeonia lactiflora Pallas (PLP) have been reported to have pharmacological effects such as anti-inflammatory and analgesic. However, it is not yet known whether PLP extract has anti-inflammatory effect on HaCaT cells, human keratinocyte. Methods : To confirm the anti-inflammatory effect of PLP on keratinocyte, TNF-𝛼/IFN-𝛾-stimulated HaCaT cells were used. HaCaT cells were pre-treated with PLP for 1h before stimulation with TNF-𝛼/IFN-𝛾. Then HaCaT cells were stimulated with TNF-𝛼/IFN-𝛾 for 24 h, the cells and media were harvested to measure the inflammatory cytokines levels. Granulocyte-macrophage colony stimulating factor (GM-CSF), monocyte chemoattractant protein-1 (MCP-1), interleukin 1 beta (IL-1𝛽), and TNF-𝛼 were analyzed by enzyme-linked immunosorbent assay (ELISA), and the mRNA expression of thymus and activation-regulated chemokines (TARC), IL-6, and IL-8 were measured by reverse transcription-polymerase chain reaction (RT-PCR). We also investigated the inhibitory mechanism of the mitogen-activated protein kinase (MAPKs) including ERK, JNK, and p38 and nuclear factor-kappaB (NF-𝜅B) by PLP using western blot. Results : PLP did not show cytotoxicity in HaCaT cells. In TNF-𝛼/IFN-𝛾-stimulated HaCaT cells, PLP significantly inhibited the expression of GM-CSF, MCP-1 IL-1𝛽, TNF-𝛼, TARC and IL-6. PLP inhibited the phosphorylation of ERK and translocation of NF-𝜅B into the nucleus. Conclusions : These results indicate that PLP could ameliorate the TNF-𝛼/IFN-𝛾-stimulated inflammatory response through inhibition of MAPK and NF-kB signal pathway. This suggests that PLP could be used beneficial agent to improve skin inflammation.

Cholesterol inhibitory activities of kaempferol and quercetin isolated from Allium victorialis var. platyphyllum (산마늘로부터 단리한 kaempferol과 quercetin의 콜레스테롤 저하 활성)

  • Lee, Sung-Suk;Moon, Seo-Hyun;Lee, Hak-Ju;Choi, Don-Ha;Cho, Myung-Haing
    • Journal of the Korean Wood Science and Technology
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    • v.32 no.1
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    • pp.17-27
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    • 2004
  • Cholesterol inhibitory activity was investigated to develop the functional food from edible forest resources such as Allium victorialis var. platyphyllum and other 12 species. Among tested samples by enzyme-linked immunosorbant assay (ELISA), leaf extracts of A. victorialis var. platyphyllum inhibited 73.9% of the activities of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase) which is the highly regulated and major rate-limiting of the cholesterol biosynthesis pathway. Moreover, those extracts inhibited 76.7% of squalene synthase which catalyzes the head-to-head condensation of two farnesyl pyrophosphate molecules to form squalene in the biosynthesis of cholesterol. In order to find out the compounds which would play a key role in inhibitory activity of cholesterol, kaempferol and quercetin were isolated from the dichloromethane soluble fraction of extracts of A. victorialis var. platyphyllum. Kampferol, quercetin and each soluble fraction was also subjected to the test of the mRNA expression of HMG-CoA reductase and squalene synthase by reverse transcriptase-polymerase chain reaction (RT-PCR) assay, respectively. By treating both enzymes with 10 ㎍/㎖ of kaempferol and quercetin for 24 hours, respectively, the mRNA expression was not observed, suggesting that both compounds inhibited the biosynthesis of cholesterol at mRNA level. In this regard, it could be inferred that cholesterol inhibitory activity of A. victorialis var. platyphyllum was derived from kaempferol and quercetin. Both compounds have already been found in many plant extracts including hardwood and softwood, but it might be first known that they have cholesterol inhibitory activity.

Occurrence of Fungal Species in Dried Ginseng Products from Retail Market (시판 건조 인삼류(백삼, 홍삼)의 곰팡이 발생)

  • Choi, Jang Nam;Kim, So Soo;Baek, Seul Gi;Park, Jin Ju;Choi, Jung Hye;Jang, Ja Yeong;Kim, Jeom-Soon;Lee, Theresa
    • Journal of Food Hygiene and Safety
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    • v.37 no.4
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    • pp.277-284
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    • 2022
  • To investigate the occurrence of fungi in dried ginseng products, we collected 24 white and 26 red ginseng samples from the retail market. Fungi were detected in 50% and 46% of white and red ginseng samples, respectively. The average level of fungal contamination was 0.5 and 0.2 log10 CFU/g in white and red ginseng, respectively. In white ginseng, Penicillium polonicum, P. chrysogenum, and Rhizopus microsporus dominated with each having an occurrence of 18.2%. In red ginseng, Aspergillus spp. was dominant with an occurrence of 87.6%, with A. chevalieri having the highest occurrence (50%). PCR screening for mycotoxigenic potential showed that the 13 isolates of 4 species (P. polonicum, P. chrysogenum, P. melanoconidium, and A. chevalieri) tested were negative for the citrinin biosynthetic gene. These results show that the samples tested in this study had low risk of mycotoxin contamination. However, there is a possibility of dried ginseng products, such as white and red ginseng, being contaminated with fungi.

Effects of Herbal Medicine Complex on Skin Inflammation and Atopic Dermatitis (한방 복합물이 피부 염증 및 아토피 피부염에 미치는 영향)

  • Ji-Hee, Choi;In-Hwan, Joo;Jong-Min, Park;Dong-Hee, Kim
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.36 no.5
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    • pp.187-192
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    • 2022
  • The purpose of this study is to examine the effect of herbal medicine complex (HMC) containing Camellia sinensis L., Duchoesna chrysantha, Houttuynia cordata Thunberg, Poncirus trifoliata Rafinesque on skin inflammation and atopic dermatitis. First, we examined the anti-inflammatory effect of HMC in TNF-α induced human keratinocytes (HaCaT cell). Real-time PCR and western blotting were performed to evaluate the expression of inflammatory cytokines (e.g., iNOS, COX-2, IL-6, IL-8) mRNA and protein. Four-weeks old male NC/Nga mice were treated with 1% 2,4-dinitrochlorobenzene (DNCB) solution and used as an atopic dermatitis mice model. And, HMC (200 mg/kg or 400 mg/kg) was administered directly into the stomach of mice for 4 weeks, and blood or serum analysis, tissue staining were performed after oral gavage. As a result HMC inhibited the mRNA expression of iNOS, COX-2, IL-6, and IL-8, which had been increased by TNF-α in HaCaT cells. In addition, the protein expression was also significantly suppressed in the same way as the mRNA expression results. The in vivo experiment results showed that, HMC administration reduced thickening of the epidermis and infiltration of eosinophil into the skin stratum basale compared to DNCB treatment. In addition, HMC administration significantly reduced the inflammatory cytokines (IL-4, IL-5, IL-6, and IL-13) production and immunocyte (white blood cell, lymphocyte, neutrophil, and eosinophil) count compared to DNCB treatment. Moreover, the serum IgE and histamine level was decreased by HMC administration. These results suggest that HMC can be used as effective herbal medicine extract for skin inflammation and atopic dermatitis. And this study may contribute to the development of the herbal medicine-based drug for the treatment of skin inflammation and atopic dermatitis.

Effects of Rearing Density Stress on Malformation and Stress and Immune Related Gene Expression of Juvenile Olive Flounder Paralichthys olivaceus (자어기 넙치(Paralichthys olivaceus)의 사육 밀도에 따른 기형 발생과 스트레스 및 면역 유전자 발현 분석)

  • Sanghyun Lee;Jong-Won Park;Minhwan Jeong;Hyo Sun Jung;Julan Kim;Woo-Jin Kim;Jeong-Ho Lee;Dain Lee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.56 no.4
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    • pp.380-387
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    • 2023
  • Stocking density is associated with Paralichthys olivaceus growth; thus, fish should be rapidly reared at high densities for commercial reasons. Studies have reported that high stocking density retards growth; however, few have investigated the malformations caused by stocking density stress. This study compared the growth and malformation rates of P. olivaceus at different densities and stress- and immune-related gene expression between malformed and normal fish. Forty days post-hatching, fish (total length, 1.49±0.02 cm) were reared at 800 (low density; LD), 1500 (medium density; MD), and 4000 (high density; HD) fish/m2, and the growth rate was measured weekly. On day 30, RNA was extracted from the kidneys, and the expression of stress-, immune-, and malformation-related genes was analyzed using qRT-PCR. The malformation rate in the HD groups was approximately three times higher (62%) than that in the LD and MD groups (approximately 20%), and growth was lower regarding length and weight. The stress-related (HSP70 and GPX) and immune-related (PIR and IgM) genes showed higher mRNA expression in the HD group and malformed fish than in the LD group and normal fish. However, TLR3 showed the opposite results. In summary, high stocking density suppressed growth and increased malformation risk in P. olivaceus.

Identification and Characterization of Polymorphic Microsatellite DNA Markers Using Next-generation Sequencing in Parapristipoma trilineatum (차세대 염기서열 분석법을 사용한 벤자리(Parapristipoma trilineatum)의 microsatellite 마커의 개발 및 유전학적 특성 분석)

  • Chun Mae Dong;Mi-Nan Lee;Jae Koo Noh;Jin Woo Park;Young-Ok Kim;Eun-Mi Kim
    • Journal of Life Science
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    • v.33 no.8
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    • pp.623-631
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    • 2023
  • This study was conducted to develop microsatellite markers in Parapristipoma trilineatum using next-generation sequencing. A total of 402,244,934 reads were generated on the Illumina Hiseq X Ten System, yielding 60,738,985,034 bp of sequences. The de novo assembly resulted in 1,320,995 contigs. A total of 952,326 contigs (0.016%) including 151 microsatellite loci were derived from the 1,320,995 contigs longer than 640 bp. A total of 34 primer sets were designed from the 151 microsatellite loci. As a result, 15 microsatellite loci were chosen and used for assuming population genetic parameters in the wild and farmed populations. The mean number of effective alleles was 12, ranging from 6 to 25. The observed heterozygosity (HO) and the expected heterozygosity (HE) ranged between 0.530 and 0.873, with an average of 0.750, and from 0.647 to 0.895, with an average of 0.793, respectively. According to these results, the developed set of 15 microsatellite markers is expected to be useful for the analysis of genetic characteristics in the population of P. trilineatum in Korea. There are requirements now for further genetic information, fishery resource management, breeding guidelines, support with the selection of breeds and studies on the effects of release, all of which will improve species conservation, and through future research, we aim to offer genetic foundational data with that goal.

The Role of Medical Technologists in Next-Generation Sequencing and Clinical Genetic Tests (임상유전자검사 및 차세대 염기서열분석을 위한 임상병리사의 역할)

  • Hyun-Seok JIN;Sangjung PARK;Mi-Sook AHN;Sangwook PARK
    • Korean Journal of Clinical Laboratory Science
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    • v.55 no.3
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    • pp.203-212
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    • 2023
  • Since the coronavirus disease-2019 (COVID-19) outbreak, it has been generally believed that a medical technologists (MTs) are supposed to perform polymerase chain reaction tests and next-generation sequencing (NGS) in the hospitals. However, many do not recognize that the duty of MT for clinical genetic testing has not been stated in the Medical Laws (72.5% for MT, N=200; 62.8% for students, N=123). In this regard, to evaluate the feasibility of MT's role for NGS genetic testing, we requested our subjects to fill out an online survey and analyzed the data. Among them, it shows that the scope of MT's role, including NGS performance should include clinical genetic testing (99.5% for MT, N=200; 86.8% for students, N=123). Also, questions on clinical genetics, which is associated with both cellular genetics and molecular genetic questions should be included in the National MT License Problem Bank (97.5% for MT; 71.4% for students). Based on these results, the Korean Association of Medical Technologists needs to cooperate synergically with the Academic Association of Biomedical Laboratory Science with respect to genetic education and legislation for the future benefit of both MTs and students.