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In Vitro Evaluation of Anti-cancer Properties of Hongyoung on SNU-80 Anaplastic Thyroid Carcinoma Cell Line

  • Gaeun Kim;Eun-Jung Kim
    • Biomedical Science Letters
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    • v.29 no.4
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    • pp.321-329
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    • 2023
  • Anaplastic thyroid cancer has the highest mortality rate of all thyroid cancers and shows low responsiveness to most treatments. Hongyoung, a reddish-colored potato, is an excellent source of dietary polyphenol containing a large amount of anthocyanins, which has anti-cancer and anti-inflammatory effects. This study investigated the effects of Hongyoung extract on apoptosis and invasiveness in SNU-80 anaplastic thyroid cancer cells. The quantification of the total polyphenol content was done by spectrophotometric measurement. Cell growth was measured by using 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl) 2H tetrazolium, monosodium salt (MTS) assay. Cell cycle was analyzed through FACS analysis. Induction of apoptosis in cells was investigated by annexin V staining using flow cytometer and the expression of caspase-3 and Poly (ADP-ribose) polymerase (PARP) through western blot. mRNA expression and protein activation of matrix metalloproteinases (MMP)-2/-9 were examined by RT-PCR and zymography. As a result, the TPC of Hongyoung was 292.43±8.42 mg gallic acid equivalent (GAE)/100 g dry extract. Hongyoung showed a dose-dependent cell growth inhibition, and the IC50 values was 1,000 ㎍/mL. sub-G1 phase was more than doubled compared to the control group, and S and G2/M phase arrest were also induced. Hongyoung induced apoptosis by increasing FITC-Annexin V-positive cells and increased the activation of caspase-3 (cleaved caspase-3) and PARP (fragmented PARP). Hongyoung significantly inhibited mRNA expression and protein activation of MMP-2/-9 in phorbol 12-myristate 13-acetate (PMA)-treated SNU-80 cells. Therefore, this study suggests the possibility of development of Hongyoung extract as an anti-cancer agent.

Effects of Pyeonggangaeuljihyeol-tang(PGJT) Extract on Anti-inflammatory, Antioxidant and Anti-aging (평간개울지혈탕(平肝開鬱止血湯)의 항염증, 항산화 및 항노화에 미치는 영향)

  • Kyoung Won Lee;Young Chae Song;Hee Taek Kim
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.37 no.1
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    • pp.29-41
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    • 2024
  • Objectives : The purpose of this study is to confirm the anti-inflammatory, antioxidant and anti-aging efficacy of Pyeonggangaeuljihyeol-tang(PGJT) extract. Methods : In order to confirm the anti-inflammatory activity of PGJT extract, the inhibitory effect on NO PGE2 production were evaluated and the expression level of iNOS, COX-2 and IL-6 mRNA were measured through qRT-PCR, Antioxidant activity was evaluated for radical scavenging activity using DPPH and ABTS, anti-aging activity was evaulated for collagenase, elastase, tyrosinase and L-DOPA oxidation inhibitory activity. Results : PGJT extract shows anti-inflammatory effect by inhibiting the production of NO and PGE2 by reducing the expression level of iNOS, COX-2 and IL-6 mRNA, antioxidant effect by increasing DPPH and ABTS scavenging abillity in a concentration dependent manner. In addition, the anti-aging effect was confirmed by inhibiting collagenase, elastase, tyrosinase production and L-DOPA oxidation. Conclusion : This suggests that PGJT showed an overall excellent anti-inflammatory effect and an inhibitory effect on the activity of antioxidant and anti-aging related enzymes.

Phenotypic characterization of pre-harvest sprouting resistance mutants generated by the CRISPR/Cas9-geminiviral replicon system in rice

  • Jong Hee Kim;Jihyeon Yu;Jin Young Kim;Yong Jin Park;Sangsu Bae;Kwon Kyoo Kang;Yu Jin Jung
    • BMB Reports
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    • v.57 no.2
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    • pp.79-85
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    • 2024
  • Pre-harvest sprouting is a critical phenomenon involving germination of seeds in the mother plant before harvest under relative humid conditions and reduced dormancy. In this paper, we generated HDR mutant lines with one region SNP (C/T) and an insertion of 6 bp (GGT/GGTGGCGGC) in OsERF1 genes for pre-harvest sprouting (PHS) resistance using CRISPR/Cas9 and a geminiviral replicon system. The incidence of HDR was 2.6% in transformed calli. T1 seeds were harvested from 12 HDR-induced calli and named ERF1-hdr line. Molecular stability, key agronomic properties, physiological properties, and biochemical properties of target genes in the ERF1-hdr line were investigated for three years. The ERF1-hdr line showed significantly enhanced seed dormancy and pre-harvest sprouting resistance. qRT-PCR analysis suggested that enhanced ABA signaling resulted in a stronger phenotype of PHS resistance. These results indicate that efficient HDR can be achieved through SNP/InDel replacement using a single and modular configuration applicable to different rice targets and other crops. This work demonstrates the potential to replace all genes with elite alleles within one generation and greatly expands our ability to improve agriculturally important traits.

Expression of TASK-1 channel in mouse Leydig cells

  • Min Seok Woo;Eun-Jin Kim;Anjas Happy Prayoga;Yangmi Kim;Dawon Kang
    • Journal of Animal Reproduction and Biotechnology
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    • v.38 no.4
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    • pp.291-299
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    • 2023
  • Background: Leydig cells, crucial for testosterone production, express ion channels like ANO1 that influence hormone secretion. This study investigates the expression and role of the Tandem of P domains in a weak inward rectifying K+ channel-related Acid-Sensitive K+-1 (TASK-1) channel in these cells, exploring its impact on testicular function and steroidogenesis. Methods: TASK-1 expression in Leydig cells was confirmed using immunostaining, while RT-PCR and Western Blot (WB) validated its expression in the TM3 Leydig cell line. The effect of a TASK-1 channel blocker on cell viability was assessed through live/dead staining and MTT assays. Additionally, the blocker's effect on testosterone secretion was evaluated by measuring testosterone levels. Results: Immunohistochemical analysis revealed a predominant presence of TASK-1, along with c-Kit and ANO-1, in Leydig cells adjacent to seminiferous tubules and also in Sertoli and spermatogenic cells. Expression levels of TASK-1 mRNA and protein were significantly higher in TM3 Leydig cells compared to TM4 Sertoli cells. In addition, blocking TASK-1 in TM3 cells with ML365 induced cell death but did not affect LH-induced testosterone secretion. Conclusions: These findings suggest that TASK-1 in Leydig cells is crucial for their viability and proliferation, highlighting its potential importance in testicular physiology.

IL-17 Imbalance Promotes the Pyroptosis in Immune-Mediated Liver Injury Through STAT3-IFI16 Axis

  • Wenfang Xu;Yanan Wang;Changzhong Jin;Weiyang Zhang;Jiangnan Chen;Xuefang Chen;Junli Gao;Junshun Gao;Hong Wang
    • IMMUNE NETWORK
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    • v.23 no.6
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    • pp.46.1-46.16
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    • 2023
  • Autoimmune hepatitis (AIH) affects all age group and occurs mainly in women. Pyroptosis is a novel programmed cell death featured with cell bursting and release of proinflammatory cytokines. A deeper understanding of AIH pathogenesis will contribute to novel therapy for AIH patients. Here, we aimed to investigate the role of IL-17 in immune-mediated liver injury. The levels of cytokines were measured by ELISA, and mRNA levels of STAT3 and IFN gamma-inducible protein 16 (IFI16) were detected by PCR. Expressions of STAT3, IFI16, gasdermin D and cleaved caspase-1 were measured by western-blotting. Immunohistochemical staining and transmission electron microscopy were applied to evaluate liver histopathological changes of the treated mice. Our results showed that the levels of IFI16 was increased in hepatocytes treated with IL-17 protein, and further elevated after STAT3-overexpressed (STAT3-OE) lentivirus treatment. The levels of IFI16 were reduced in hepatocytes treated with IL-17 neutralizing Ab (nAb), but were significantly increased after STAT3-OE treatment. Pyroptosis was observed in hepatocytes treated with IL-17 protein, and further cell damage was observed after STAT3-OE lentivirus treatment. Liver damage was alleviated in mice treated with IL-17 nAb, however sever damage was experienced after STAT3-OE lentivirus treatment. A binding interaction between IFI16 and STAT3 was detected in IL-17 treated hepatocytes. Glutathione transaminase activity was enhanced in concanavalin A-induced AIH mice compared to the control group (p<0.01). IL-17 plays an important role in activating STAT3 and up-regulating IFI16, which may promote the pyroptosis in AIH-related liver injury through STAT3-IFI16 axis.

Analysis of Molecular Diversity in Castanopsis sieboldii with Felt Disease Caused by Septobasidium sp. (Septobasidium sp.에 의한 구실잣밤나무 고약병의 분자학적 다양성 분석)

  • Geon-Woo Lee;Sang-Tae Seo;Byeongjin Cha;Sang-Sub Han
    • Research in Plant Disease
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    • v.29 no.4
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    • pp.420-424
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    • 2023
  • In 2020, within the Dongbaekdongsan area in Jeju Island, a Septobasidium sp. associated with a felt disease in Castanopsis sieboldii (Makino) Hatus. ex T. Yamaz. & Mashiba was identified. The symptom included the presence of brown, thin, and silk-like mycelial mats attached to the tree's bark, displaying variations in size from large to small. To induce hyphal growth, the samples collected were incubated in a moist chamber, and the newly formed hyphae were subjected to genomic DNA extractions. The nucleotide sequences of the internal transcribed spacer and small subunit rDNA genes were determined, and molecular characteristics among the isolates were investigated through polymerase chain reaction-based restriction fragment length polymorphism analysis. This Septobasidium sp. exhibited distinct morphological and phylogenetic features compared to those that were previously reported in South Korea. Consequently, this strain is taxonomically classified as a provisionally novel species of Septobasidium. Furthermore, the observed felt disease exhibited a high degree of host specificity, as it was exclusively identified in C. sieboldii without occurrence in other tree species at the time of observation.

Computational Evaluation on the Interactions of an Opaque-Phase ABC Transporter Associated with Fluconazole Resistance in Candida albicans, by the Psidium guajava Bio-Active Compounds

  • Mithil Vora;Smiline Girija Aseervatham Selvi;Shoba Gunasekaran;Vijayashree Priyadharsini Jayaseelan
    • Journal of Pharmacopuncture
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    • v.27 no.2
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    • pp.91-100
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    • 2024
  • Objectives: Candida albicans is an opportunistic pathogen that occurs as harmless commensals in the intestine, urogenital tract, and skin. It has been influenced by a variety of host conditions and has now evolved as a resistant strain. The aim of this study was thus detect the fluconazole resistant C. albicans from the root caries specimens and to computationally evaluate the interactions of an opaque-phase ABC transporter protein with the Psidium guajava bio-active compounds. Methods: 20 carious scrapings were collected from patients with root caries and processed for the isolation of C. albicans and was screened for fluconazole resistance. Genomic DNA was extracted and molecular characterization of Cdrp1 and Cdrp2 was done by PCR amplification. P. guajava methanolic extract was checked for the antifungal efficacy against the resistant strain of C. albicans. Further in-silico docking involves retrieval of ABC transporter protein and ligand optimization, molinspiration assessment on drug likeness, docking simulations and visualizations. Results: 65% of the samples showed the presence of C.albicans and 2 strains were fluconazole resistant. Crude methanolic extract of P. guajava was found to be promising against the fluconazole resistant strains of C. albicans. In-silico docking analysis showed that Myricetin was a promising candidate with a high docking score and other drug ligand interaction scores. Conclusion: The current study emphasizes that bioactive compounds from Psidium guajava to be a promising candidate for treating candidiasis in fluconazole resistant strains of C. albicans However, further in-vivo studies have to be implemented for the experimental validation of the same in improving the oral health and hygiene.

Whitening effects of fermented Trigonotis radicans var. sericea with Lactobacillus brevis in α-MSH-stimulated B16F10 melanoma cells

  • Da-Eun Jeong;Byung-Oh Kim;Young-Je Cho
    • Food Science and Preservation
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    • v.31 no.2
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    • pp.227-234
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    • 2024
  • This study was designed to compare the whitening effects of 60% ethanol extracts of Trigonotis radicans var. sericea (TR) and Lactobacillus brevis-fermented T. radicans var. sericea (FTR). Measurement of cytotoxicity in B16-F10 melanoma cells to confirm the whitening effect, FTR showed higher cell viability than TR. FTR showed inhibitory activity on melanin contents similar to the normal group at concentrations of 50 and 100 ㎍/mL. MITF expression was used to confirm the effect on melanogenesis-related protein expression. TR and FTR showed significant concentration-dependent decrease, and FTR showed lower expressions than the normal group at concentrations of 25, 50, and 100 ㎍/mL. Additionally, the mRNA expression of melanogenesis-related genes (MC1R, Rab27a, TGF-β1 and Myo5a) were measured by RT-qPCR to confirm the whitening effect. In MC1R expression at a concentration of 100 ㎍/mL in FTR showed effective inhibitory activities, and in TGF-β1 expression, TR and FTR both showed effective activities compared to normal groups even at low concentrations. Results of myo5a and Rab27a, a similar pattern was shown, and FTR showed effective inhibitory activities at a concentration of 100 ㎍/mL. As a result, FTR had higher whitening effects through bioconversion and is expected to be a good material for whitening functional cosmetics.

Effects of Fermented Achyranthes japonica Nakai, Angelica gigas Nakai, and Eucommia ulmoides Oliver Extracts on Regulation of Apoptosis in Articular Chondrocytes (Primary Chondrocytes에서 발효우슬, 당귀, 두충 복합물의 세포사멸 조절 효과)

  • Dakyung Kim;Wonhee Jo;Minhee Lee;Hyun Cheol Jeong;Sung-Jin Lee;Seunghun Lee;Jeongmin Lee
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.7-14
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    • 2023
  • The effects of fermented Achyranthes japonica Nakai, Angelica gigas Nakai, and Eucommia ulmoides Oliver extracts (FAAE) on regulation of inflammation and apoptosis were investigated in primary cultured rat cartilage cells. To identify the protective effects of FAAE against H2O2, cell survival was measured by MTT assay. Smad3, Collagen type I, MMP3, and MMP13 were measured by real-timpe PCR and westernbot and the inflammatory (NF-κB pathway, COX-2, iNOS) factors were determined by western blot. The apoptosis related factors (JNK, c-Fos, c-Jun, caspase 3, Bax, and Bcl-2) were determined by western blot. FAAE significantly increased the follwing: H2O2 treated cell survival, mRNA and protein expression of Smad 3, collagen type I. In addition, FAAE significantly decreased the protein expression of inflammatory and apoptosis related factors. This study suggests that FAAE have a protection effect of chondrocytes through inhibition of inflammation and apoptosis. Thus, FAAE is a therapeutic potential food componet in osteoarthritis.

Ginsenoside Rk1 inhibits HeLa cell proliferation through an endoplasmic reticulum signaling pathway

  • Qiuyang Li;Hang Sun;Shiwei Liu;Jinxin Tang;Shengnan Liu;Pei Yin;Qianwen Mi;Jingsheng Liu;Lei yu;Yunfeng Bi
    • Journal of Ginseng Research
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    • v.47 no.5
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    • pp.645-653
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    • 2023
  • Background: Changes to work-life balance has increased the incidence of cervical cancer among younger people. A minor ginseng saponin known as ginsenoside Rk1 can inhibit the growth and survival of human cancer cells; however, whether ginsenoside Rk1 inhibits HeLa cell proliferation is unknown. Methods and results: Ginsenoside Rk1 blocked HeLa cells in the G0/G1 phase in a dose-dependent manner and inhibited cell division and proliferation. Ginsenoside Rk1 markedly also activated the apoptotic signaling pathway via caspase 3, PARP, and caspase 6. In addition, ginsenoside Rk1 increased LC3B protein expression, indicating the promotion of the autophagy signaling pathway. Protein processing in the endoplasmic reticulum signaling pathway was downregulated in Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses, consistent with teal-time quantitative PCR and western blotting that showed YOD1, HSPA4L, DNAJC3, and HSP90AA1 expression levels were dramatically decreased in HeLa cells treated with ginsenoside Rk1, with YOD1 was the most significantly inhibited by ginsenoside Rk1 treatment. Conclusion: These findings indicate that the toxicity of ginsenoside Rk1 in HeLa cells can be explained by the inhibition of protein synthesis in the endoplasmic reticulum and enhanced apoptosis, with YOD1 acting as a potential target for cervical cancer treatment.