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Ovalicin attenuates atopic dermatitis symptoms by inhibiting IL-31 signaling and intracellular calcium influx

The Journal of Biomedical research 2021年第6期35卷 448-458页

作者： Sung-Hyun Hwang Yeseul Yang Yeji Jeong Yongbaek Kim Laboratory of Clinical Pathology College of Veterinary MedicineSeoul National UniversitySeoul 08826Republic of Korea The Brain Korea 21 Future veterinary medicine Leading Education and research center College of Veterinary MedicineSeoul National UniversitySeoul 08826Republic of Korea research Institute for veterinary Science College of Veterinary MedicineSeoul National UniversitySeoul 08826Republic of Korea.

Atopic dermatitis(AD)is a common skin disorder difficult to be treated with medication.This study investigated the potential of ovalicin extracted from Cordyceps militaris for the treatment of AD using in vitro and in vivo models.We found that,in canine macrophage cell line DH82,lipopolysaccharide(LPS)upregulated the expression of genes associated with inflammation and pruritic responses through activating calcium and interleukin-31(IL-31)signaling,and the upregulation could be suppressed by ovalicin,with an effect significantly stronger than dexamethasone.Ovalicin also reduced the expression of IL-31 downstream genes,including JAK2(Janus kinase 2),TRPV1(transient receptor potential vanilloid receptor-1),and HRH2(histamine receptor H2).Ovalicin significantly alleviated the allergic symptoms in the AD mouse model.Histologically,the number of macrophages and mast cells infiltrated in the dermis was significantly reduced by ovalicin treatment.In the skin tissue of AD mice,reduction of IL-31 receptor was observed in the ovalicin treated group compared to the group without ovalicin treatment.To our knowledge,this is the first study to elucidate the anti-atopic mechanism of ovalicin,which could be an alternative to steroidal drugs commonly used for AD treatment.

关键词： ovalicin IL-31 pruritus inflammation calcium atopic dermatitis

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First Isolation and Characterization of Ochrobactrum anthropi from Pig

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Engineering 2020年第1期6卷 49-55页

作者： Shijiang Gu Ruiqing Hou Shengguo Gao Zhe Sun Xiangdong Li Lufeng Zhai Yunyun Jin Qiaoyan Zhu Yonghong Liao Kegong Tian National Research Center for Veterinary Medicine Luoyang 471000China College of Animal Science and veterinary medicine Henan Agricultural UniversityZhengzhou 450002China

One Gram-negative Bacillus was isolated from a brain sample of a pig with neurological symptoms.Pathological examination showed meningitis at necropsy. Ochrobactrum anthropi (O. anthropi) was successfully isolated from the brain sample and was confirmed by biochemical reaction results (API 20 NE) and gene sequencing. The strain was highly resistant to b-lactam antibiotics. Mice were experimentally infected with O. anthropi and showed typical meningitis. This is the first report on O. anthropi isolated from a pig, and indicates that O. anthropi may have a broader host spectrum of infection.

关键词： Ochrobactrum anthropi Pig brain

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RR1 and RR2 gene deletion affects the immunogenicity of a live attenuated pseudorabies virus vaccine candidate in natural pig host

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Frontiers of Agricultural Science and Engineering 2016年第1期3卷 81-86页

作者： Shijun YAN He YAN Chaolin ZHANG Tongyan WANG Qingyuan YANG Zhe SUN Yan XIAO Feifei TAN Xiangdong LI Kegong TIAN National Research Center for Veterinary Medicine College of Animal Science and veterinary medicine Henan Agricultural University

As virulence-determining genes, RR1 and RR2 encode the small subunit and large subunit of viral ribonucleotide reductase(RR) in pseudorabies virus which have been extensively studied in mice. However,their role in pigs has not been adequately investigated. In this study, we deleted RR1 and RR2 genes based on a TK/g E/g I triple gene-deleted pseudorabies virus and tested its efficacy in pigs as a vaccine candidate. The rescued virus showed similar growth properties and plaque size in vitro as its parent strain. In an animal study, the virus could elicit humoral immune responses shown by generation of g B-specific antibodies and virus neutralizing antibodies.However, vaccination could not provide protection against virulent pseudorabies virus challenge since vaccinated pigs showed clinical pseudorabies-specific syndromes. The deficiency in protection may due to the generation of late and low levels of gB antibodies and virus neutralizing antibodies.

关键词： pseudorabies virus RR1 and RR2 ribonucleotide reductase vaccine candidate

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Effects of lysophosphatidic acid on human periodontal ligament stem cells from teeth extracted from dental patients

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The Journal of Biomedical research 2019年第2期33卷 122-130页

作者： Byung Cheol Kim Jae-In Song Kyoung-Ha So Sang-Hwan Hyun Laboratory of veterinary Embryology and Biotechnology Veterinary Medical Center and College of Veterinary MedicineChungbuk National University Institute of Stem Cell & Regenerative medicine Chungbuk National University

Despite their potential applications in future regenerative medicine, periodontal ligament stem cells(PDLSCs) are difficult to obtain in large amounts from patients. Therefore, maintaining sternness while expanding the cell numbers for medical use is the key to transitioning PDLSCs from the bench to the clinic. Lysophosphatidic acid(LPA), which is present in the human body and saliva, is a signaling molecule derived from phospholipids. In this study, we examined the effects of LPA on sternness maintenance in human PDLSCs. Several spindle-shaped and fibroblast-like periodontal ligament stem-like cell lines were established from PDLSC isolation. Among these cell lines, the most morphologically appropriate cell line was characterized. The expression levels of OCT4, NANOG(a stem cell marker), and CD90(a mesenchymal stem cell marker) were high. However, CD73(a negative marker of mesenchymal stem cells) expression was not observed. Notably, immunofluorescence analysis identified the expression of STRO-1, CD146(a mesenchymal stem cell marker), and sex determining region Y-box 2 at the protein level. In addition, lipid droplets were stained by Oil red O after the induction of adipogenesis for 21 days, and mineralized nodules were stained by Alizarin Red S after the induction of osteogenesis for 14 days. Alkaline phosphate staining also demonstrated the occurrence of osteogenesis. In summary, we established a human PDLSC line, which could be applied as a cell source for tissue regeneration in dental patients. However, further studies are needed to determine the detailed effects of LPA on PDLSCs.

关键词： periodontal ligament stem cell lysophosphatidic acid stemness primary cell culture

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In vitro antiproliferative and apoptosis inducing effect of a methanolic extract of Azadirachta indica oil on selected cancerous and noncancerous cell lines

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Asian Pacific Journal of Tropical medicine 2018年第10期11卷 555-561页

作者： Muhammad Kashif Dongwook Kim Gonhyung Kim veterinary Medical center College of Veterinary Medicine Chungbuk National University

Objective: To find the cytotoxic and apoptotic effects of neem oil extract on the selected cancerous(A-549, PC-3 and DU-145) and noncancerous(NIH3T3 and CCD-18Co) cell lines.Methods: Viability and cytotoxic effect induced by the extract was measured by using MTT assay and apoptotic effect of the extract was evaluated by using Hoechst 33342 and propidium iodide dual staining through a fluorescent microscope and activity of caspases 3, 8 and 9 through colorimetric assay kits. Results: The results showed that neem oil extract significantly reduced the viability in all selected cancer cells treated with varying concentrations of extract as compared with untreated cells and had less effect on noncancerous cell lines. It significantly increased the percentage of necrotic and apoptotic cells, and caspases 3, 8 and 9 activities in all cancer cells treated with extract as compared with untreated cells whereas no effect on noncancerous cell lines. It suggested that neem oil extract exerted a higher cytotoxic effect on cancer cells than normal cells and lower concentration induced apoptosis only in cancer cells.One of the apoptosis-inducing mechanism was through the activation of caspases signaling pathways. Conclusion: Conclusively, it implies that neem oil extract may contain one or more potential agents that can be used as a safe and effective anticancer therapy.

关键词： Neem oil extract Cell lines Cytotoxicity Apoptosis Caspase activity

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Interspecies transmission and evolution of the emerging coronaviruses: perspectives from bat physiology and protein spatial structure

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Frontiers of Agricultural Science and Engineering 2020年第2期7卷 218-226页

作者： Baicheng HUANG Kegong TIAN National Research Center for Veterinary Medicine Luoyang 471003China College of Animal Science and veterinary medicine Henan Agricultural UniversityZhengzhou 450046China

Emergent coronaviruses(CoVs)such as SARS-CoV and MERS-CoV have posed great threats to public health worldwide over the past two decades.Currently,the emergence of SARS-CoV-2 as a pandemic causes greater public health concern.CoV diversity is due to the large size and replication mechanisms of the genomes together with having bats as their optimum natural hosts.The ecological behavior and unique immune characteristics of bats are optimal for the homologous recombination of CoVs.The relationship of spatial structural characteristics of the spike protein,a protein that is critical for recognition by host receptors,in different CoVs may provide evidence in explaining the coevolution of CoVs and their hosts.This information may help to enhance our understanding of CoV evolution and thus provide part of the basis of preparations for any future outbreaks.

关键词： bat coronavirus evolution host receptor spike protein transmission

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Replication, pathogenicity, and transmission of SARS-CoV-2 in minks

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national Science Review 2021年第3期8卷 21-28页

作者： Lei Shuai Gongxun Zhong Quan Yuan Zhiyuan Wen Chong Wang Xijun He Renqiang Liu Jinliang Wang Qinjian Zhao Yuxiu Liu Ningning Huo Junhua Deng Jingjing Bai Hongchao Wu Yuntao Guan Jianzhong Shi Kegong Tian Ningshao Xia Hualan Chen Zhigao Bu State Key Laboratory of veterinary Biotechnology Harbin Veterinary Research Institute Chinese Academy of Agricultural Sciences national High Containment Laboratory for Animal Diseases Control and Prevention State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics National Institute of Diagnostics and Vaccine Development in Infectious DiseasesSchool of Public Health Xiamen University National Research Center for Veterinary Medicine

Minks are raised in many countries and have transmitted severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) to humans.However,the biologic properties of SARS-CoV-2 in minks are largely unknown.Here,we investigated and found that SARS-CoV-2 replicates efficiently in both the upper and lower respiratory tracts,and transmits efficiently in minks via respiratory droplets;pulmonary lesions caused by SARS-CoV-2 in minks are similar to those seen in humans with COVID-19.We further found that a spike protein-based subunit vaccine largely prevented SARS-CoV-2 replication and lung damage caused by SARS-CoV-2 infection in minks.Our study indicates that minks are a useful animal model for evaluating the efficacy of drugs or vaccines against COVID-19 and that vaccination is a potential strategy to prevent minks from transmitting SARS-CoV-2.

关键词： SARS-CoV-2 mink replication pathogenicity transmission animal model

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Temporal profiles of synaptic plasticity-related signals in adult mouse hippocampus with methotrexate treatment

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Neural Regeneration research 2012年第21期7卷 1651-1658页

作者： Miyoung Yang Juhwan Kim Sung-Ho Kim Joong-Sun Kim Taekyun Shin Changjong Moon Department of veterinary Anatomy College of Veterinary Medicine and Animal Medical Institute Chonnam National University Department of Experimental Radiation Research Center Dongnam Institute of Radiological & Medical Sciences (DIRAMS) Department of veterinary Anatomy College of Veterinary Medicine Jeju National University

Methotrexate, which is used to treat many malignancies and autoimmune diseases, affects brain functions including hippocampal-dependent memory function. However, the precise mechanisms underlying methotrexate-induced hippocampal dysfunction are poorly understood. To evaluate temporal changes in synaptic plasticity-related signals, the expression and activity of N-methyI-D-aspartic acid receptor 1, calcium/calmodulin-dependent protein kinase II, extracellular signal-regulated kinase 1/2, cAMP responsive element-binding protein, glutamate receptor 1, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor were examined in the hippocampi of adult C57BL/6 mice after methotrexate （40 mg/kg） intraperitoneal injection. Western blot analysis showed biphasic changes in synaptic plasticity-related signals in adult hippocampi following methotrexate treatment. N-methyI-D-aspartic acid receptor 1, calcium/calmodulin-dependent protein kinase II, and glutamate receptor 1 were acutely activated during the early phase （1 day post-injection）, while extracellular signal-regulated kinase 1/2 and cAMP responsive element-binding protein activation showed biphasic increases during the eady （1 day post-injection） and late phases （7-14 days post-injection）. Brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor expression increased significantly during the late phase （7-14 days post-injection）. Therefore, methotrexate treatment affects synaptic plasticity-related signals in the adult mouse hippocampus, suggesting that changes in synaptic plasticity-related signals may be associated with neuronal survival and plasticity-related cellular remodeling.

关键词： hippocampus methotrexate neurotrophic factor synaptic plasticity-related signal neuralregeneration

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ASFV transcription reporter screening system identifies ailanthone as a broad antiviral compound

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Virologica Sinica 2023年第3期38卷 459-469页

作者： Yuhang Zhang Zhenjiang Zhang Fan Zhang Jiwen Zhang Jun Jiao Min Hou Nianchao Qian Dongming Zhao Xiaofeng Zheng Xu Tan College of veterinary medicine Hunan Agricultural UniversityChangsha 410128China State Key Laboratory of veterinary Biotechnology Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150009China Beijing Advanced Innovation center for Structural Biology Beijing Frontier Research Center for Biological StructureMOE Key Laboratory of Bioorganic Phosphorus Chemistry&Chemical BiologySchool of Pharmaceutical SciencesTsinghua-Peking Center for Life SciencesTsinghua UniversityBeijing 100084China Institute of Health and medicine Hefei Comprehensive National Science CenterHefei 230601 China

African swine fever(ASF)is an acute,highly contagious and deadly viral disease in swine that jeopardizes the worldwide pig industry.Unfortunately,there are no authoritative vaccine and antiviral drug available for ASF control.African swine fever virus(ASFV)is the etiological agent of ASF.Among the ASFV proteins,p72 is the most abundant component in the virions and thus a potential target for anti-ASFV drug design.Here,we con-structed a luciferase reporter system driven by the promoter of p72,which is transcribed by the co-transfected ASFV RNA polymerase complex.Using this system,we screened over 3200 natural product compounds and obtained three potent candidates against ASFV.We further evaluated the anti-ASFV effects and proved that among the three candidates,ailanthone(AIL)inhibits the replication of ASFV at the nanomolar concentration(IC_(50)=15 nmol/L).Our in vitro experiments indicated that the antiviral effect of AIL is associated with its inhibition of the HSP90-p23 cochaperone.Finally,we showed the antiviral activity of AIL on Zika virus and hepatitis B virus(HBV),which supports that AIL is a potential broad-spectrum antiviral agent.

关键词： African swine fever virus(ASFV) Replication inhibition Broad-spectrum antiviral drugs Ailanthone(AIL)

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research and development of a novel subunit vaccine for the currently circulating pseudorabies virus variant in China

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Frontiers of Agricultural Science and Engineering 2015年第3期2卷 216-222页

作者： Yuzhou WANG Tongyan WANG He YAN Fanli YANG Linghua GUO Qingyuan YANG Xule HU Feifei TAN Yan XIAO Xiangdong LI Kegong TIAN National Research Center for Veterinary Medicine Luoyang 471003China College of Animal Science and veterinary medicine Henan Agricultural UniversityZhengzhou 450002China

Pseudorabies(PR)is a devastating viral disease which leads to fatal encephalitis and respiratory disorders in pigs.Commercial gE-deleted live pseudorabies virus(PRV)vaccine has been widely used to control this disease in China.However,the new-emerging variants of PRV compromises the protection provided by current vaccines and lead to the outbreak of PR in vaccinated pig herds.Several killed and live vaccine candidates based on current PRV variants have been reported to be effective to control the disease.A subunit vaccine based on gB protein,one major PRV glycoprotein which elicits strong humoral and cellular immune responses,however,was never evaluated for protection against the current circulating PRV variants.In this study,full-length PRV gB protein was successfully expressed in baculovirus/insect cells in the soluble format and was tested on 3-week-old piglets as a subunit vaccine.Compared with unvaccinated pigs,the gB-vaccinated pigs developed specific antibody-mediated responses and were protected from the virulent PRV HN1201 challenge.All vaccinated pigs survived without showing any PRV-specific respiratory and neurological signs,but all unvaccinated pigs died within 7 days after HN1201 challenge.Hence,this novel gB-based vaccine could be applied as an effective subunit vaccine to control PRV variant in China.

关键词： pseudorabies virus glycoprotein B protein subunit vaccine

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