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Journal of Integrative Agriculture 2014年第6期13卷 1355-1362页

作者： LI Yu-bao LI Juan WANG Zhi-liang liu si-dang School of Agriculture Liaoche ng University School of Veterinary Medicine Shandong Agricultural University China Animal Health and Epidemiology Center

To investigate the dynamic change of heat shock protein 90 （Hsp90） in the genesis and development of tumor, we successfully established tumor animal model using Marek’s disease and then determined the location of Hsp90 in the tumor tissue using immunohistochemistry method, the antibody titer level of Hsp90 in the serum and the expression level in the tissue using enzyme-linked immunosorbent assay （ELISA） method. Our result showed that Hsp90 location in the tumor tissue was signiifcantly associated with the tumor cell and most in the cytoplasm of the tumor cell, and Hsp90 expression level in the tissue and the antibody titer level in the serum was most signiifcantly increased with the development of tumor. This is the ifrst report to show the presence of Hsp90 in tumor tissues induced by the Marek’s disease, with its expression correlated to the tumoral grading. These data may also be valuable for developing new molecular anti-cancer therapies.

关键词： heat shock protein 90 Marek's disease immunohistochemistry expression antibody titer level

Phylogenetic and epidemiological characteristics of H9N2 avian influenza viruses in Shandong Province, China from 2019 to 2021

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Journal of Integrative Agriculture 2023年第3期22卷 881-896页

作者： ZHAO Yi-ran ZHAO Yu-zhong liu si-dang XIAO Yi-hong LI Ning liu Kui-hao MENG Fan-liang ZHAO Jun liu Meng-da LI Bao-quan College of Animal Science and Veterinary Medicine Shandong Agricultural University/Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and PreventionShandong Agricultural UniversityTai’an 271018P.R.China Division of Zoonoses Surveillance China Animal Health and Epidemiology CenterQingdao 266033P.R.China

H9N2 avian influenza virus(AIV) has widely circulated in poultry worldwide and sporadic infections in humans and mammals. During our surveillance of chicken from 2019 to 2021 in Shandong Province, China, we isolated 11 H9N2AIVs. Phylogenetic analyses showed that the eight gene segments of the 11 isolates were closely related to several sublineages of Eurasian lineage: BJ/94-like clades(HA and NA genes), G1-like clades(PB2 and M genes), and SH/F/98-like clades(PB1, PA, NP and NS genes). The isolates showed mutation sites that preferentially bind to humanlike receptors(HA) and mammalian fitness sites(PB2, PB1 and PA), as well as mutations in antigen and drug resistance sites. Moreover, studies with mice revealed four isolates with varying levels of pathogenicity. The average antibody titer of the H9N2 AIVs was 8.60 log2. Based on our results, the epidemiological surveillance of H9N2 AIVs should be strengthened.

关键词： influenza virus phylogenetic analyses mutation sites pathogenicity antibody titer epidemiological surveillance

Cloning, Prokaryotic Expression, and Antigenicity Analysis of NS1 Gene of H9N2 Swine Influenza Virus

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Agricultural Sciences in China 2008年第7期7卷 895-900页

作者： WANG Fang-kun YUAN Xiu-fang WANG Yi-cheng ZHANG Cun XU Li-huan liu si-dang College of Animal Sciences and Veterinary Medicine Shandong Agricultural University Tai＇an 271018 P.R.China Insititute of Animal Sciences and Veterinary Medicine Zhejiang Academy of Agricultural Sciences Hangzhou 310021 P.R.China

To obtain the NS1 gene of swine influenza virus H9N2 subtype expressed efficiently in E. coli, to develope the effective diagnostic methods for swine influenza virus H9N2 subtype, the NS 1 gene of swine influenza virus H9N2 subtype was amplified by reverse transcriptase polymerase chain reaction （RT-PCR） and cloned into a prokaryotic expression vector, pET-28a（＋）, and overexpressed in E. coli BL21-DE3 after induction with 5 mmol L-1 lactose. The recombinant protein was purified by Ni-NTA and identified by western-blotting. An indirect enzyme-linked immunosorbent assay （ELISA） was used to analyze the antigenicity of the recombinant protein. The recombinant protein of NS1 was about 26 kD. The Western-blotting test showed that the recombinant protein reacted specifically with positive sera. The results of the ELISA test showed that the recombinant protein had good antigenicity.

关键词： swine influenza virus NS1 antigenicity

Identification of an H1N1 subtype of swine influenza virus and serological analysis

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Journal of Integrative Agriculture 2019年第7期18卷 1436-1442页

作者： SUN Fa-chao TAN Min ZHANG Yuan-chao WANG Yu-chao CAO Sheng-liang DING Guo-fei CONG Fang-yuan GUO Li-hong liu si-dang XIAO Yi-hong Department of Basic Veterinary Medicine College of Animal Science and Veterinary Medicine/Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention/Shandong Provincial Engineering Technology Research Center of Animal Disease Control and PreventionShandong Agricultural University

To investigate the epizootic of swine influenza virus(siV),60 nasal swabs were collected from a clinical cases of pig farm in Tai’an City,Shandong Province of China in April *** was isolated by inoculating into 10-day-old Special Pathogen Free embryonated eggs and the whole genome was *** H1N1 subtype siV was isolated and designated as A/swine/Shandong/TA04/2017(H1N1).Phylogenetic analysis showed that apart from the polymerase A(PA) fragment belonging to the 2009 pandemic H1N1 branch,seven genome segments belonged to avian-like H1N1 influenza virus *** cleavage site sequence of the hemagglutinin(HA) protein was PsiQSR↓G,which is a typical molecular biological *** potential N-glycosylation sites(N14,N26,N277,N484 and N543) were found in the HA *** further investigate the epidemiology of siV in this farm,the 995 serum samples were assessed with EAH1N1 2009 pandemic H1N1 and H3 N2 *** results showed that the total positive rate was 65.43%.The positive rates of single virus infection detected by EAH1N1,2009 pdmH1N1 and H3 N2 for serum HI(Hemagglutination inhibition) were 48.35,30.85 and 7.47%,*** results showed that siV in Shandong Province has been reassorted in some segments and the siV-positive rate was high on the siV outbreak *** data provide evidence of an epizootic of siV.

关键词： siV H1N1 subtype genetic analysis molecular feature serology

Unraveling brain aging through the lens of oral microbiota

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Neural Regeneration Research 2025年第7期20卷 1930-1943页

作者： Qinchao Hu si Wang Weiqi Zhang Jing Qu Guang-Hui liu State Key Laboratory of Membrane Biology Institute of ZoologyChinese Academy of SciencesBeijingChina Hospital of Stomatology Guanghua School of StomatologyGuangdong Provincial Key Laboratory of StomatologySun Yat-sen UniversityGuangzhouGuangdong ProvinceChina Advanced Innovation Center for Human Brain Protection and National Clinical Research Center for Geriatric Disorders Xuanwu HospitalCapital Medical UniversityBeijingChina Aging Translational Medicine Center International Center for Aging and CancerBeijing Municipal Geriatric Medical Research CenterXuanwu HospitalCapital Medical UniversityBeijingChina CAS Key Laboratory of Genomic and Precision Medicine Beijing Institute of GenomicsChinese Academy of Sciences and China National Center for BioinformationBeijingChina University of Chinese Academy of Sciences BeijingChina Institute for Stem Cell and Regeneration Chinese Academy of SciencesBeijingChina Aging Biomarker Consortium BeijingChina State Key Laboratory of Stem Cell and Reproductive Biology Institute of ZoologyChinese Academy of SciencesBeijingChina Beijing Institute for Stem Cell and Regenerative Medicine BeijingChina

The oral cavity is a complex physiological community encompassing a wide range of *** of oral microbiota can lead to various oral infectious diseases,such as periodontitis and tooth decay,and even affect systemic health,including brain aging and neurodegenerative *** studies have highlighted how oral microbes might be involved in brain aging and neurodegeneration,indicating potential avenues for intervention *** this review,we summarize clinical evidence demonstrating a link between oral microbes/oral infectious diseases and brain aging/neurodegenerative diseases,and dissect potential mechanisms by which oral microbes contribute to brain aging and *** also highlight advances in therapeutic development grounded in the realm of oral microbes,with the goal of advancing brain health and promoting healthy aging.

关键词： Alzheimer's disease brain aging multiple sclerosis neurodegeneration neurodegenerative diseases oral microbiota Parkinson's disease periodontitis bacteria Porphyromonas gingivalis

Relationship between Tumor Lesion of Kidney Tissue and Expression of HSP60 in Chickens Infected by MD

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Animal Husbandry and Feed Science 2013年第5期5卷 210-215页

作者： LI Yu-bao LI Juan WU Xiao-dong liu Yu-tian ZHAO Yong-gang ZHANG Yong-qiang REN Wei-jie HAN Xiu-ju WANG Zhi-liang liu si-dang College of Veterinary Medicine Shandong Agricultural University College of Agriculture Liaocheng University China Animal Health and Epidemiology Center

[ Objective] The paper was to investigate the relationship between pathological lesion and transcription, expression and distribution of heat shock protein 60 （HSP60） in kidney of chickens infected by Marek＇s disease virus （MDV）. [ Method] Tumor animal model was successfully established with chickens infected by MDV. Real-time RT-PCR, histopathological and immunohistochemistrical methods were used to detect the pathological lesion, transcription level, expression level and distribution of HSP60 in the kidney. [ Result] Obvious pathological changes appeared in kidney tissue of chicken after injection of MDV for 21 d. HSP60 was mainly distributed in the cytoplasm of tumor cell and interstitial macrophages. The expression of HSP60 in challenge group was always higher than blank control group and vaccine control group, and the difference reached extremely significant level from 28 to 86 day-ages （ P 〈 0.01 ） ; the expression of HSP60 in challenge group at 28 day-age was about 8.608 and 12.752 times of blank control group and vaccine control group, respectively. The mRNA transcription levels of HSP60 in challenge group showed a downward parabola during 7 to 42 day-ages; the mRNA transcription level of HSP60 in challenge group was rising in early stage （7 -21 d）, and then gradually dropped; it reached the peak value at 21 d, being 1.222 and 1.179 times of blank control group and vaccine control group, respectively. The mRNA transcription level of HSP60 was higher than two control groups throughout the entire experiment, and the difference reached extremely significant level from 14 to 28 day-agas （P 〈0.01 ）. [ Conclusion] HSP60 expression in kidney tissue and its mRNA transcription level can reflect MDV infection, tumor formation and development progress, with close relationship with pathogenesis of tumor diseases, which is expected to be used as one of the determination indicators for diagnosis and morbidity process of tumor diseases in chicken.

关键词： Marek's disease Heat shock protein60 Immunohistochemistry FQ RT-PCR

鸡胚肺动脉内皮细胞与平滑肌细胞的分离培养及鉴定

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中国农业大学学报 2009年第2期14卷 17-21页

作者：韩燕燕刘思当杨鹰山东农业大学动物科技学院山东泰安271000 中国农业大学动物科学技术学院北京100193

探索简单快速实用的鸡胚肺动脉内皮细胞和平滑肌细胞培养方法：在严格无菌条件下对肺动脉内皮细胞和平滑肌细胞分离培养，待细胞融合度达约80％时进行传代，得到的内皮细胞利用差速消化法纯化，并观察2种细胞形态特征及培养特性，待细... 详细信息

探索简单快速实用的鸡胚肺动脉内皮细胞和平滑肌细胞培养方法：在严格无菌条件下对肺动脉内皮细胞和平滑肌细胞分离培养，待细胞融合度达约80％时进行传代，得到的内皮细胞利用差速消化法纯化，并观察2种细胞形态特征及培养特性，待细胞稳定生长后，采用荧光免疫组化技术鉴定细胞，并进行冻存和复苏，观察复苏后细胞活性。结果显示：24h后组织块周围即有少量细胞爬出，内皮细胞大量生长后呈突起状排列，立体感强，平滑肌细胞在培养后的第5天呈现“峰谷”样外观，经免疫荧光法证明是内皮细胞和平滑肌细胞的纯培养，冻存于液氮中1～2个月复苏后细胞的活性都较好，可以恢复旺盛生长。结论：本研究提出的组织贴块法能成功获得内皮细胞和平滑肌细胞的纯培养，比酶消化法操作简单，更适合小血管内皮细胞、平滑肌细胞的原代培养。

关键词：鸡胚肺动脉内皮细胞平滑肌细胞培养免疫组化鉴定

驴源猪链球菌2型的分离和鉴定及MLST分型

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中国兽医学报 2018年第10期38卷 1920-1925页

作者：刘朋程子龙陈萌刘思当山东农业大学动物科技学院

2017年4月,山东省济宁嘉祥县、梁山县和聊城东阿县的多个驴场从外地购买的驴相继发生体温升高、流鼻涕、咳嗽、呼吸困难等症状,病情严重的驴出现死亡。从5个驴场分别剖检5头病死驴,均表现化脓性肺炎及败血症病变,采集一典型病例的组织,... 详细信息

2017年4月,山东省济宁嘉祥县、梁山县和聊城东阿县的多个驴场从外地购买的驴相继发生体温升高、流鼻涕、咳嗽、呼吸困难等症状,病情严重的驴出现死亡。从5个驴场分别剖检5头病死驴,均表现化脓性肺炎及败血症病变,采集一典型病例的组织,进行细菌分离培养,并通过病理组织学检查、革兰染色、生化鉴定、血清学鉴定、16S基因序列分析以及MLST（aroA、cpn60、dpr、gki、mutS、recA、thrA）分型研究,此外还分析了中国目前所流行的ST型以及不同ST型之间的关系。结果显示：病原为猪链球菌2型;16SDNA序列与国内外部分地区菌株序列的同源性达到99%～100%;MLST分型为ST2型;该分离菌株对林可霉素、青霉素、阿莫西林、头孢噻呋钠产生耐药性,对左氧氟沙星、氟苯尼考高度敏感。

关键词：驴猪链球菌分离和鉴定病理组织学 16S基因序列分析 MLST分型

NOX4 exacerbates Parkinson's disease pathology by promoting neuronal ferroptosis and neuroinflammation

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Neural Regeneration Research 2025年第7期20卷 2038-2052页

作者： Zhihao Lin Changzhou Ying Xiaoli si Naijia Xue Yi liu Ran Zheng Ying Chen Jiali Pu Baorong Zhang Department of Neurology The Second Affiliated HospitalZhejiang University School of MedicineHangzhouZhejiang ProvinceChina

Parkinson's disease is primarily caused by the loss of dopaminergic neurons in the substantia nigra ***,a novel form of regulated cell death characterized by iron accumulation and lipid peroxidation,plays a vital role in the death of dopaminergic ***,the molecular mechanisms underlying ferroptosis in dopaminergic neurons have not yet been completely *** oxidase 4 is related to oxidative stress,however,whether it regulates dopaminergic neuronal ferroptosis remains *** aim of this study was to determine whether NADPH oxidase 4 is involved in dopaminergic neuronal ferroptosis,and if so,by what *** found that the transcriptional regulator activating transcription factor 3 increased NADPH oxidase 4 expression in dopaminergic neurons and astrocytes in an 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine-induced Parkinson's disease *** oxidase 4 inhibition improved the behavioral impairments observed in the Parkinson's disease model animals and reduced the death of dopaminergic ***,NADPH oxidase 4 inhibition reduced lipid peroxidation and iron accumulation in the substantia nigra of the Parkinson's disease model ***,we found that NADPH oxidase 4 interacted with activated protein kinase Cαto prevent ferroptosis of dopaminergic ***,by lowering the astrocytic lipocalin-2 expression,NADPH oxidase 4 inhibition reduced 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine-induced *** findings demonstrate that NADPH oxidase 4 promotes ferroptosis of dopaminergic neurons and neuroinflammation,which contribute to dopaminergic neuron death,suggesting that NADPH oxidase 4 is a possible therapeutic target for Parkinson's disease.

关键词： dopaminergic neuron ferroptosis NADPH oxidase 4(NOX4) neuroinflammation Parkinson's disease

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鸡新城疫HI抗体水平及带毒状况调查

中国兽医杂志 2009年第6期45卷 12-14页

作者：孙洪磊王刚刘思当山东农业大学动物科技学院山东泰安271018

对2007年3月-2008年5月泰安及周边地区鸡群新城疫抗体水平检测结果进行了统计分析,统计结果显示：抗体效价超过正常免疫水平且离散度较大,疑似新城疫野毒感染的鸡群较多。其中肉仔鸡（或麻鸡）群、育成蛋鸡群分别为12.3%、21.5%,而产蛋... 详细信息

对2007年3月-2008年5月泰安及周边地区鸡群新城疫抗体水平检测结果进行了统计分析,统计结果显示：抗体效价超过正常免疫水平且离散度较大,疑似新城疫野毒感染的鸡群较多。其中肉仔鸡（或麻鸡）群、育成蛋鸡群分别为12.3%、21.5%,而产蛋鸡群高达51.2%。采用接种鸡胚分离病毒和间接免疫荧光试验验证HI试验检测结果的判定,探讨鸡群的带毒状况。调查结果表明,目前鲁中地区各种鸡群的新城疫感染带毒状况较为严重,高比例带毒鸡群的存在是我国控制新城疫的最大难题。

关键词：新城疫 HI抗体水平病毒分离间接免疫荧光试验