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β-Xylosidase and β-mannosidase in combination improved growth performance and altered microbial profiles in weanling pigs fed a corn-soybean meal-based diet

  • Liu, Shaoshuai;Ma, Chang;Liu, Ling;Ning, Dong;Liu, Yajing;Dong, Bing
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.11
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    • pp.1734-1744
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    • 2019
  • Objective: In this study, two glycosidases (XMosidases), ${\beta}$-xylosidase and ${\beta}$-mannosidase, were investigated on their in vitro hydrolysis activities of feed and on the improvement of growth performance in vivo in weanling pigs. Methods: Enzyme activities of XMosidases in vitro were evaluated in test tubes and simulation of gastric and small intestinal digestion, respectively, in the presence of NSPase. In vivo study was performed in 108 weaned piglets in a 28-d treatment. Pigs were allotted to one of three dietary treatments with six replicate pens in each treatment. The three treatment groups were as follows: i) Control (basal diet); ii) CE (basal diets+CE); iii) CE-Xmosidases (basal diets+ CE+${\beta}$-xylosidase at 800 U/kg and ${\beta}$-mannosidase at 40 U/kg). CE was complex enzymes (amylase, protease, xylanase, and mannanase). Results: In vitro XMosidases displayed significant activities on hydrolysis of corn and soybean meal in the presence of non-starch polysaccharide degrading enzymes (xylanase and ${\beta}$-mannanase). In vitro simulation of gastric and small intestinal digestion by XMosidases showed XMosidases achieved $67.89%{\pm}0.22%$ of dry matter digestibility and $63.12%{\pm}0.21%$ of energy digestibility at $40^{\circ}C$ for 5 hrs. In weanling pigs, additional XMosidases to CE in feed improved average daily gain, feed conversion rate (p<0.05), and apparent total tract digestibility of crude protein (p = 0.01) and dry matter (p = 0.02). XMosidases also altered the gut bacterial diversity and composition by increasing the proportion of beneficial bacteria. Conclusion: Addition of a complex enzyme supplementation (contained xylanase, ${\beta}$-mannanase, protease and amylase), XMosidases (${\beta}$-xylosidase and ${\beta}$-mannosidase) can further improve the growth performance and nutrient digestion of young pigs.

Hypoxia Mediates Runt-Related Transcription Factor 2 Expression via Induction of Vascular Endothelial Growth Factor in Periodontal Ligament Stem Cells

  • Xu, Qian;Liu, Zhihua;Guo, Ling;Liu, Rui;Li, Rulei;Chu, Xiang;Yang, Jiajia;Luo, Jia;Chen, Faming;Deng, Manjing
    • Molecules and Cells
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    • v.42 no.11
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    • pp.763-772
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    • 2019
  • Periodontitis is characterized by the loss of periodontal tissues, especially alveolar bone. Common therapies cannot satisfactorily recover lost alveolar bone. Periodontal ligament stem cells (PDLSCs) possess the capacity of self-renewal and multilineage differentiation and are likely to recover lost alveolar bone. In addition, periodontitis is accompanied by hypoxia, and hypoxia-inducible $factor-1{\alpha}$ ($HIF-1{\alpha}$) is a master transcription factor in the response to hypoxia. Thus, we aimed to ascertain how hypoxia affects runt-related transcription factor 2 (RUNX2), a key osteogenic marker, in the osteogenesis of PDLSCs. In this study, we found that hypoxia enhanced the protein expression of $HIF-1{\alpha}$, vascular endothelial growth factor (VEGF), and RUNX2 ex vivo and in situ. VEGF is a target gene of $HIF-1{\alpha}$, and the increased expression of VEGF and RUNX2 proteins was enhanced by cobalt chloride ($CoCl_2$, $100{\mu}mol/L$), an agonist of $HIF-1{\alpha}$, and suppressed by 3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole (YC-1, $10{\mu}mol/L$), an antagonist of $HIF-1{\alpha}$. In addition, VEGF could regulate the expression of RUNX2, as RUNX2 expression was enhanced by human VEGF ($hVEGF_{165}$) and suppressed by VEGF siRNA. In addition, knocking down VEGF could decrease the expression of osteogenesis-related genes, i.e., RUNX2, alkaline phosphatase (ALP), and type I collagen (COL1), and hypoxia could enhance the expression of ALP, COL1, and osteocalcin (OCN) in the early stage of osteogenesis of PDLSCs. Taken together, our results showed that hypoxia could mediate the expression of RUNX2 in PDLSCs via $HIF-1{\alpha}$-induced VEGF and play a positive role in the early stage of osteogenesis of PDLSCs.

Effect of keratinase on ileal amino acid digestibility in five feedstuffs fed to growing pigs

  • Huang, Chengfei;Ma, Dongli;Zang, Jianjun;Zhang, Bo;Sun, Brian;Liu, Ling;Zhang, Shuai
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.12
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    • pp.1946-1955
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    • 2018
  • Objective: This study was conducted to evaluate the effect of keratinase (KE) on the apparent ileal digestibility (AID) and standardized ileal digestibility (SID) of amino acids (AA) in rice bran, cottonseed meal (CSM), rapeseed meal (RSM), corn distillers dried grains with solubles (DDGS), and peanut meal (PNM). Methods: Twelve crossbred barrows (Duroc${\times}$Landrace${\times}$Yorkshire, $50.5{\pm}1.4kg$ body weight [BW]) fitted with T-cannulas at the terminal ileum were allotted to a $12{\times}6$ Youden Square design with 12 diets and 6 periods. The treatment diets included rice bran, CSM, RSM, corn DDGS, PNM, or corn-soybean meal (cSBM) supplemented with 0.05% KE or not. Diets were given to pigs at a level of 3% BW in two equal meals. The endogenous AA losses were the mean results of three previously experiments determined by a same nitrogen-free diet fed to pigs. Pigs had free access to water during the experiment. Results: The KE supplementation improved (p<0.05) the AID and SID of Met, Thr, Val, Asp, Cys, and Tyr in rice bran. Inclusion of KE increased (p<0.05) the AID and SID of Met and Val in CSM. The KE supplementation decreased (p<0.05) the AID and SID of His in RSM and all measured AA except for Arg, Met, Trp, Val, Gly, and Pro in corn DDGS. There was an increase (p<0.05) in AID and SID of Leu, Ile, Met, Ala, Cys, Ser, and Tyr in PNM supplemented with KE compared with that without KE. Inclusion of KE increased (p<0.05) the AID and SID of crude protein, Leu, Ile, Phe, Thr, Asp, and Ser in cSBM. Conclusion: This study indicated that KE had different effects on ileal AA digestibility of feedstuffs for growing pigs, which can give some usage directions of KE in swine feed containing those detected feedstuffs.

Usability Test of Serious Game Robot App for Hospitalized Children (입원 아동 환자를 위한 로봇용 기능성 게임 앱 사용성 평가)

  • Jin, Mei-Ling;Kim, Jeong-Eun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.2
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    • pp.228-234
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    • 2019
  • This study was conducted to evaluate the usability of serious-game apps for robots that are being developed for hospitals to add familiarity to hospital life for hospitalized patients. We conducted a usability test of 10 experts and 12 elementary school students with inpatient experience. The usability evaluation of the developed apps was based on the professional (MARS) evaluation tool and the user (uMARS) evaluation tool. The results of the usability test of the serious game robot app were $3.67{\pm}0.342$ for professionals and $3.68{\pm}0.592$ for children. The expert group obtained the highest score in the aesthetics category, and the user group obtained the highest score in the functionality category. In the subjective comments, the experts pointed out the game layout and the consistency of the style. According to the children, the methods were easy to learn, and the screen movement was mainly described. Both groups received low ratings in terms of engagement through participation. This study will provide useful reference material when the functional game app is actually installed in the robot and the usability test is conducted again after the results obtained in this study are considered.

Determination of the energy contents and nutrient digestibility of corn, waxy corn and steam-flaked corn fed to growing pigs

  • Ma, Dongli;Li, Juntao;Huang, Chengfei;Yang, Fengjuan;Wu, Yi;Liu, Ling;Jiang, Wei;Jia, Zhicheng;Zhang, Peijun;Liu, Xuezhen;Zhang, Shuai
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.10
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    • pp.1573-1579
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    • 2019
  • Objective: The research was conducted to determine the digestible (DE) and metabolizable energy (ME) contents as well as the apparent total tract digestibility (ATTD) of nutrients in corn, waxy corn and steam-flaked corn fed to growing pigs. Methods: Eighteen growing pigs with initial body weight of $15.42{\pm}1.41kg$ were randomly allotted to three diets including a corn diet, a waxy corn diet and a steam-flaked corn diet in a completely randomized design. Each treatment contained six replicates. The experiment lasted for 12 days, which comprised 7-d adaptation to diets followed by a 5-d total collection of feces and urine. The energy contents and the nutrient digestibility in three ingredients were calculated using direct method. Results: Compared to normal corn, both the amylose and dietary fiber contents in waxy corn were numerically lower, but the starch gelatinization degree was numerically greater. Moreover, the DE and ME contents as well as the ATTD of neutral detergent fiber and acid detergent fiber (ADF) in waxy corn were significantly greater (p<0.05) than those in normal corn when fed to growing pigs. Furthermore, the steam-flaked corn had greater (p<0.05) DE and ME contents, and ATTD of ether extract and ADF compared to normal corn. Conclusion: Both variety and processing procedure have influence on chemical compositions, energy contents and nutrient digestibility of corn. The waxy corn and steam-flaked corn had greater degree of starch gelatinization and DE and ME contents compared to normal corn when fed to growing pigs.

Determination and prediction of the digestible and metabolizable energy contents of corn germ meal in growing pigs

  • Shi, Meng;Liu, Zhaoyu;Wang, Hongliang;Shi, Chuanxin;Liu, Ling;Wang, Junjun;Li, Defa;Zhang, Shuai
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.3
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    • pp.405-412
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    • 2019
  • Objective: This experiment was conducted to determine the chemical composition, digestible energy (DE) and metabolizable energy (ME) contents of corn germ meals (CGM) and to develop equations to predict the corresponding energy contents based on the chemical characteristics of individual CGM. Methods: Sixty-six barrows (initial body weight = $51.3{\pm}4.6kg$) were allotted to 11 diets including a basal diet and 10 CGM test diets in a completely randomized design. In the test diets, CGM was included in replacement of 30% of the energy-providing ingredients in the basal diet, resulting in a final inclusion rate of 29.1%. Each diet was fed to 6 barrows housed in individual metabolism crates for a 7-d acclimation period followed by a 5-d total but separate collection of feces and urine. Results: Considerable variation was observed in acid-hydrolyzed ether extract, ether extract, ash, calcium (Ca) and total phosphorus contents among the CGM samples. On dry matter (DM) basis, the DE and ME contents of the CGM ranged from 10.22 to 15.83 MJ/kg and from 9.94 to 15.43 MJ/kg, respectively. The acid detergent fiber (ADF) contents were negatively correlated with the DE and ME contents of CGM samples. The best-fit prediction equations for the DE and ME values (MJ/kg DM) of the 10 CGM were: DE = 26.85-0.28 insoluble dietary fiber (%)-17.79 Ca (%); ME = 21.05-0.43 ADF (%)-11.40 Ca (%). Conclusion: The chemical compositions of CGM vary depending on sources, particularly in ether extract and Ca. The DE and ME values of CGM can be predicted based on their chemical composition in growing pigs.

A Study on Tradition Discourse in Korean Modern Drama between 1894 and 1910 (근대계몽기 한국연극사의 전통담론 연구 II)

  • Paek, Hyun-Mi
    • (The) Research of the performance art and culture
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    • no.18
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    • pp.347-377
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    • 2009
  • This paper examines the tradition discourses in Korean modern drama of the late 19th and the early 20th. It focuses particularly on how key intellectuals differentiated the definition and interpreted 'Theater-Improvement' while they insisted on improving traditional performance. Tradition discourse focused on how to improve its contents of performances. There were three categories in discussion. First, Confucian intelligent insisted that traditional performance should be discarded, because it led to customs disorder among the public. Second, patriotic and progressive intellectuals led by Sin Chae-Ho(신채호) and Bak Yen-Sik(박은식), preferred the reformed content of traditional Korean performance dealing with historical heroes. in order to stimulate patriotism in the Korean. Third, Lee In-Jik(이인직) who had worked at a popular newspaper company in Japan tried to make 'New Theater' criticizing decayed officials and the stupid nation. He did not concern about the colonialism of Japan. In the late 19th and the early 20th in Korean modern drama history, tradition discourses faced different directions, dependent on their political positions, familiar foreign cultures, and their view on traditional cultures.

Platelet-Derived Growth Factor Receptor-α Subunit Targeting Suppresses Metastasis in Advanced Thyroid Cancer In Vitro and In Vivo

  • Lin, Ching-Ling;Tsai, Ming-Lin;Chen, Yu-hsin;Liu, Wei-Ni;Lin, Chun-Yu;Hsu, Kai-Wen;Huang, Chien-Yu;Chang, Yu-Jia;Wei, Po-Li;Chen, Shu-Huey;Huang, Li-Chi;Lee, Chia-Hwa
    • Biomolecules & Therapeutics
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    • v.29 no.5
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    • pp.551-561
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    • 2021
  • Thyroid cancer is the most common endocrine malignancy. Patients with well-differentiated thyroid cancers, such as papillary and follicular cancers, have a favorable prognosis. However, poorly differentiated thyroid cancers, such as medullary, squamous and anaplastic advanced thyroid cancers, are very aggressive and insensitive to radioiodine treatment. Thus, novel therapies that attenuate metastasis are urgently needed. We found that both PDGFC and PDGFRA are predominantly expressed in thyroid cancers and that the survival rate is significantly lower in patients with high PDGFRA expression. This finding indicates the important role of PDGF/PDGFR signaling in thyroid cancer development. Next, we established a SW579 squamous thyroid cancer cell line with 95.6% PDGFRA gene insertion and deletions (indels) through CRISPR/Cas9. Protein and invasion analysis showed a dramatic loss in EMT marker expression and metastatic ability. Furthermore, xenograft tumors derived from PDGFRA geneedited SW579 cells exhibited a minor decrease in tumor growth. However, distant lung metastasis was completely abolished upon PDGFRA gene editing, implying that PDGFRA could be an effective target to inhibit distant metastasis in advanced thyroid cancers. To translate this finding to the clinic, we used the most relevant multikinase inhibitor, imatinib, to inhibit PDGFRA signaling. The results showed that imatinib significantly suppressed cell growth, induced cell cycle arrest and cell death in SW579 cells. Our developed noninvasive apoptosis detection sensor (NIADS) indicated that imatinib induced cell apoptosis through caspase-3 activation. In conclusion, we believe that developing a specific and selective targeted therapy for PDGFRA would effectively suppress PDGFRA-mediated cancer aggressiveness in advanced thyroid cancers.

Ginsenoside Ro, an oleanolic saponin of Panax ginseng, exerts anti-inflammatory effect by direct inhibiting toll like receptor 4 signaling pathway

  • Xu, Hong-Lin;Chen, Guang-Hong;Wu, Yu-Ting;Xie, Ling-Peng;Tan, Zhang-Bin;Liu, Bin;Fan, Hui-Jie;Chen, Hong-Mei;Huang, Gui-Qiong;Liu, Min;Zhou, Ying-Chun
    • Journal of Ginseng Research
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    • v.46 no.1
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    • pp.156-166
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    • 2022
  • Background: Panax ginseng Meyer (P. ginseng), a herb distributed in Korea, China and Japan, exerts benefits on diverse inflammatory conditions. However, the underlying mechanism and active ingredients remains largely unclear. Herein, we aimed to explore the active ingredients of P. ginseng against inflammation and elucidate underlying mechanisms. Methods: Inflammation model was constructed by lipopolysaccharide (LPS) in C57BL/6 mice and RAW264.7 macrophages. Molecular docking, molecular dynamics, surface plasmon resonance imaging (SPRi) and immunofluorescence were utilized to predict active component. Results: P. ginseng significantly inhibited LPS-induced lung injury and the expression of proinflammatory factors, including TNF-α, IL-6 and IL-1β. Additionally, P. ginseng blocked fluorescencelabeled LPS (LPS488) binding to the membranes of RAW264.7 macrophages, the phosphorylation of nuclear factor-κB (NF-κB) and mitogen-activated protein kinases (MAPKs). Furthermore, molecular docking demonstrated that ginsenoside Ro (GRo) docked into the LPS binding site of toll like receptor 4 (TLR4)/myeloid differentiation factor 2 (MD2) complex. Molecular dynamic simulations showed that the MD2-GRo binding conformation was stable. SPRi demonstrated an excellent interaction between TLR4/ MD2 complex and GRo (KD value of 1.16 × 10-9 M). GRo significantly inhibited LPS488 binding to cell membranes. Further studies showed that GRo markedly suppressed LPS-triggered lung injury, the transcription and secretion levels of TNF-α, IL-6 and IL-1β. Moreover, the phosphorylation of NF-κB and MAPKs as well as the p65 subunit nuclear translocation were inhibited by GRo dose-dependently. Conclusion: Our results suggest that GRo exerts anti-inflammation actions by direct inhibition of TLR4 signaling pathway.

Cardioprotective effect of ginsenoside Rb1 via regulating metabolomics profiling and AMP-activated protein kinase-dependent mitophagy

  • Hu, Jingui;Zhang, Ling;Fu, Fei;Lai, Qiong;Zhang, Lu;Liu, Tao;Yu, Boyang;Kou, Junping;Li, Fang
    • Journal of Ginseng Research
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    • v.46 no.2
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    • pp.255-265
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    • 2022
  • Background: Ginsenoside Rb1, a bioactive component isolated from the Panax ginseng, acts as a remedy to prevent myocardial injury. However, it is obscure whether the cardioprotective functions of Rb1 are related to the regulation of endogenous metabolites, and its potential molecular mechanism still needs further clarification, especially from a comprehensive metabolomics profiling perspective. Methods: The mice model of acute myocardial ischemia (AMI) and oxygen glucose deprivation (OGD)-induced cardiomyocytes injury were applied to explore the protective effect and mechanism of Rb1. Meanwhile, the comprehensive metabolomics profiling was conducted by high-performance liquid chromatography and quadrupole time-of-flight mass spectrometry (HPLC-Q/TOF-MS) and a tandem liquid chromatography and mass spectrometry (LC-MS). Results: Rb1 treatment profoundly reduced the infarct size and attenuated myocardial injury. The metabolic network map of 65 differential endogenous metabolites was constructed and provided a new inspiration for the treatment of AMI by Rb1, which was mainly associated with mitophagy. In vivo and in vitro experiments, Rb1 was found to improve mitochondrial morphology, mitochondrial function and promote mitophagy. Interestingly, the mitophagy inhibitor partly attenuated the cardioprotective effect of Rb1. Additionally, Rb1 markedly facilitated the phosphorylation of AMP-activated protein kinase α (AMPKα), and AMPK inhibition partially weakened the role of Rb1 in promoting mitophagy. Conclusions: Ginsenoside Rb1 protects acute myocardial ischemia injury through promoting mitophagy via AMPKα phosphorylation, which might lay the foundation for the further application of Rb1 in cardiovascular diseases.