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Cloning of Thymidine Kinase Gene of Duck Plague Virus Using Degenerate PCR

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Agricultural Sciences in China 2005年第8期4卷 634-640页

作者： han xian-jie WANG Jun-wei Department of Veterinary Medicine Northeast Agricultural University Harbin 150030 P.R.China Department of Animal Husbandry and Veterinary Science Laiyang Agricultural College Laiyang 265200 P.R.China

The DNA of duck plague virus （DPV） thymidine kinase （TK） gene was cloned and sequenced from a vaccine virus in the study. Degenerate oligonucleotide primers for the consensus site of herpesvirus UL24, TK, and glycoprotein H（gH） gene were used in the polymerase chain reaction （PCR） to amplify DNA product with 3 741-base-pairs （bp） in size. DNA sequence analysis revealed a 1 077-base-pairs （bp） open reading frame （ORF） encoding a 358 amino acid polypeptide homologous to herpesvirus TK proteins. The predicted TK protein shared 31.2, 41.3, 35.7, 37.4, and 28.4% identity with herpes simplex virus typel, equine herpesvirus type 4, Marek＇s disease virus 2, herpesvirus turkey, and infectious laryngotracheitis virus, respectively. Comparison of the amino acid sequences of other herpesvirus TK proteins showed that these proteins were not conserved on the whole, otherwise the portion of the TK proteins corresponding to the nucleotide binding domain and the nucleoside binding site were highly conserved among herpesvirus. Comparison with the amino acid sequences of the conserved nucleotide and nucleoside binding domains of other eleven herpesvirus TK proteins to the predicted DPV peptide confirmed its identity as the DPV TK protein.

关键词： Duck plague virus Degenerate PCR Thymidine kinase gene

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朝鲜民族教坊舞表演类型及其传承保存研究

北京舞蹈学院学报 2016年第2期 29-36页

作者：韩贤杰北京舞蹈学院中国民族民间舞系北京100081

历史上承传下来的朝鲜民族传统舞蹈,大体可分为宫廷舞、宗教舞、教坊舞和民俗舞这四类,教坊舞作为其中的一支,又被称为妓房舞,它起源发展于宫廷,后随着从宫廷流出的官妓,再进入民间盛行在各地券番(妓生组合)。对教坊的表演类型与传承保... 详细信息

历史上承传下来的朝鲜民族传统舞蹈,大体可分为宫廷舞、宗教舞、教坊舞和民俗舞这四类,教坊舞作为其中的一支,又被称为妓房舞,它起源发展于宫廷,后随着从宫廷流出的官妓,再进入民间盛行在各地券番(妓生组合)。对教坊的表演类型与传承保存进行分析和探究,不仅有利于该舞蹈的教学传承,也有利于以该舞蹈为资源的编创应用。

关键词：朝鲜民族教坊舞表演类型传承保存

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朝鲜半岛农乐舞巫戏构造与演绎程式

北京舞蹈学院学报 2024年第1期 100-109页

作者：韩贤杰郭罗乐北京舞蹈学院北京100081 南京艺术学院南京210013

农乐舞是朝鲜半岛古老的舞蹈形态,是农耕文化的产物。文章列举农耕仪礼中的祝愿仪礼、播种仪礼、成长仪礼、收获,分析农乐舞巫戏演绎程式与角色构造,阐述其蕴含的审美特征,体现执器而舞的审美传统,阐释朝鲜半岛传统舞蹈的文化价值。

关键词：传统农乐巫戏构造演绎程式

Storage time affects the level and diagnostic efficacy of plasma biomarkers for neurodegenerative diseases

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Neural Regeneration Research 2025年第8期20卷 2373-2381页

作者： Lifang Zhao Mingkai Zhang Qimeng Li Xuemin Wang jie Lu Ying han Yanning Cai Department of Clinical Biobank Xuanwu HospitalCapital Medical UniversityBeijingChina Beijing Geriatric Medical Research Center BeijingChina Department of Neurology Xuanwu HospitalCapital Medical UniversityBeijingChina Department of Radiology and Nuclear Medicine Xuanwu HospitalCapital Medical UniversityBeijingChina Beijing Key Laboratory of Magnetic Resonance Imaging and Brain Informatics BeijingChina Key Laboratory of Neurodegenerative diseases Ministry of EducationBeijingChina School of Biomedical Engineering Hainan UniversityHaikouHainan ProvinceChina Center of Alzheimer’s Disease Beijing Institute for Brain DisordersBeijingChina National Clinical Research Center for Geriatric Diseases BeijingChina Institute of Biomedical Engineering Shenzhen Bay LaboratoryGaoke Innovation CenterShenzhenGuangdong ProvinceChina Department of Neurobiology Xuanwu HospitalCapital Medical UniversityBeijingChina

Several promising plasma biomarker proteins,such as amyloid-β(Aβ),tau,neurofilament light chain,and glial fibrillary acidic protein,are widely used for the diagnosis of neurodegenerative ***,little is known about the long-term stability of these biomarker proteins in plasma samples stored at-80°*** aimed to explore how storage time would affect the diagnostic accuracy of these biomarkers using a large *** samples from 229 cognitively unimpaired individuals,encompassing healthy controls and those experiencing subjective cognitive decline,as well as 99 patients with cognitive impairment,comprising those with mild cognitive impairment and dementia,were acquired from the Sino Longitudinal Study on Cognitive Decline *** samples were stored at-80°C for up to 6 years before being used in this *** results showed that plasma levels of Aβ42,Aβ40,neurofilament light chain,and glial fibrillary acidic protein were not significantly correlated with sample storage ***,the level of total tau showed a negative correlation with sample storage ***,in individuals without cognitive impairment,plasma levels of total protein and tau phosphorylated protein threonine 181(p-tau181)also showed a negative correlation with sample storage *** was not observed in individuals with cognitive ***,we speculate that the diagnostic accuracy of plasma p-tau181 and the p-tau181 to total tau ratio may be influenced by sample storage ***,caution is advised when using these plasma biomarkers for the identification of neurodegenerative diseases,such as Alzheimer's ***,in cohort studies,it is important to consider the impact of storage time on the overall results.

关键词： Alzheimer’s disease amyloid-β diagnostic ability glial fibrillary acidic protein neurodegeneration neurofilament light chain plasma biomarkers single molecule array storage time tau

Trinuclear gold-catalyzed site-selective alkylation of peptides

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Science China Chemistry 2025年第1期68卷 249-256页

作者： Qing-Yun Fang Chengyihan Gu Yinghan Chen Wencheng Yan Yong Liang Weipeng Li Chengjian Zhu jie han Jin Xie State Key Laboratory of Coordination Chemistry Jiangsu Key Laboratory of Advanced Organic Materials Chemistry and Biomedicine Innovation Center (Chem BIC) School of Chemistry and Chemical Engineering Nanjing University Green Catalysis Center College of Chemistry Zhengzhou University State Key Laboratory of Natural Medicines China Pharmaceutical University

Selective modification of peptides has always been a challenging issue. The selective alkylation of peptides can alter the biological activity and physical properties of molecules, and it has gained significant research interest. We herein disclose an intriguing gold-catalyzed alkylation of peptides using bench-stable unactivated alkyl bromides under photoredox catalysis. A wide range of structurally diverse primary, secondary and tertiary alkyl bromides serve as effective coupling partners to precisely connect with the α-C(sp3)–H in glycine residue of peptides. This protocol demonstrates an exceptionally broad scope for alkyl halides and excellent tolerance for a wide range of useful functional groups. A series of relay transformations to construct cyclic peptides, to proceed click reaction and to realize peptide fluorescence labeling further enhance its synthetic practicality. In addition, mechanistic studies and density functional theory(DFT) calculations reveal an inner-sphere and subsequent outersphere single electron transfer(SET) mechanism.

关键词： gold catalysis peptide alkylation C-C coupling radical

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獭兔源铜绿假单胞菌的分离鉴定及药敏试验

动物医学进展 2016年第8期37卷 128-131页

作者：孙鑫鹏高善颂韩先杰青岛农业大学动物科技学院山东青岛266109

为了确定某獭兔养殖场獭兔发病的主要病原菌,分析该病原菌的耐药性,本试验对青岛市某獭兔养殖场送检的病死兔进行病理剖检,并从肺脏中分离得到1株细菌,随后进行分离菌形态观察、生化试验、细菌16S rRNA基因测序。该株分离菌为革兰阴性杆... 详细信息

为了确定某獭兔养殖场獭兔发病的主要病原菌,分析该病原菌的耐药性,本试验对青岛市某獭兔养殖场送检的病死兔进行病理剖检,并从肺脏中分离得到1株细菌,随后进行分离菌形态观察、生化试验、细菌16S rRNA基因测序。该株分离菌为革兰阴性杆菌,博检革兰阴性细菌鉴定系统的生化鉴定结果为铜绿假单胞菌,细菌的16S rRNA基因序列经同源性比较,与铜绿假单胞菌同源性为100%。药敏试验结果表明该株分离菌对多种头孢类、喹诺酮类药物高度敏感。细菌分离鉴定和药敏试验结果为临床用药治疗该病提供参考依据。

关键词：獭兔铜绿假单胞菌分离鉴定药敏试验

猪圆环病毒2d亚型衣壳蛋白在昆虫细胞中的表达

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农业生物技术学报 2016年第9期24卷 1346-1353页

作者：黄娟袁飞韩先杰李桂梅单虎青岛农业大学山东省高校预防兽医学重点实验室

为了在昆虫细胞中表达猪圆环毒病2型(Porcine circovirus type 2,PCV2)衣壳蛋白,本研究设计合成引物从临床病料中扩增获得猪圆环病毒(Porcine circovirus,PCV)山东株衣壳蛋白基因缺失核定位区的序列,并进行序列分析。将该片段克隆到杆... 详细信息

为了在昆虫细胞中表达猪圆环毒病2型(Porcine circovirus type 2,PCV2)衣壳蛋白,本研究设计合成引物从临床病料中扩增获得猪圆环病毒(Porcine circovirus,PCV)山东株衣壳蛋白基因缺失核定位区的序列,并进行序列分析。将该片段克隆到杆状病毒供体载体p Fast Bac-1上,然后转化大肠杆菌(Escherichia coli)DH10-Bac感受态细胞,获得重组穿梭杆粒,将该杆粒转染到sf9昆虫细胞中,观察重组杆状病毒致细胞病变效应,并采用PCR、间接免疫荧光实验、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate polyacrylamide gel electrophoresis,SDS-PAGE)和Western blot鉴定衣壳蛋白的表达情况。结果表明,扩增获得的猪圆环病毒山东株属于PCV2d亚型,与我国目前使用的商品化疫苗株(PCV2a及PCV2b亚型)在160~180 aa抗原位点有差异。成功构建了重组供体质粒p Fast Bac1-PCV2d-Cap和重组穿梭杆粒Bacmid-PCV2d-Cap,获得的重组杆状病毒在昆虫细胞中成功表达了约24 k D的PCV2d-Cap,该蛋白可以与猪圆环病毒阳性血清及His标签抗体发生反应,证明具有良好的反应原性。本研究是我国首次采用杆状病毒系统表达PCV2d亚型衣壳蛋白,为制备PCV2d亚单位疫苗以及酶联免疫吸附测定(enzyme linked immunosorbent assay,ELISA)试剂盒、防范PCV2d亚型流行提供了理论依据。

关键词：猪圆环病毒2d亚型衣壳蛋白杆状病毒表达系统基因分型

Insights from untargeted metabolomics: revealing potential marker compounds into the maturation of fermented chili

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Food Science and Human Wellness 2025年

作者： Shiyu Liu Mingyuan Yang Jianhua Zhao Meijuan Lei Jinlin han Min Xu Ping Liu jie Tang Hongbin Lin School of Food and Bio-engineering， Xihua University China Agricultural University-Sichuan Advanced Agricultural & Industrial Institute Chongqing Key Laboratory of Speciality Food Co-Built by Sichuan and Chongqing Food Microbiology Key Laboratory of Sichuan Province

Chili fermentation is one of the most important processes in the production of Pixian Douban (PXDB)， which determines the flavor and product quality of PXDB. However， the maturity of fermented chili mainly depends on empirical judgment， which can not satisfy the need for standardized production of PXDB. Therefore， this study aimed to investigate volatile and non-volatile substances during the maturation process and to find markers related to the maturity of fermented chili. GC×GC-MS combined with multivariate statistical analysis revealed that 2-methoxy-3-isobutyl pyrazine， linalool， 3-(methylthio)propionaldehyde， myrcene， and decanal (ROAV ≥ 1， VIP > 1， P < 0.05) were regarded as potentially active aromatic markers for differentiating fermentation time. Additionally， UPLC-QTOF-MS combined with multivariate statistical analysis resulted that 14 compounds can serve as differentiated non-volatile compounds. The correlation of maturity-related physico-chemical indicators with volatile and non-volatile compounds revealed that three volatile compounds (2-methoxy-3-isobutylpyrazine， linalool， and decanal) along with fructose and adenosine could serve as markers for evaluating the maturity of chili. This study is expected to establish a standard for the determination of maturity the fermented chili and lay the foundation for intelligent production of PXDB.

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H9N2亚型禽流感病毒对豚鼠致病力的研究

黑龙江畜牧兽医 2015年第1期 31-34,213页

作者：王珂张传美杨海燕韩先杰乔健青岛农业大学山东省高校预防兽医学重点实验室山东青岛266109 中国农业大学动物医学院北京100094

为了研究用豚鼠建立禽流感动物模型的可能性,试验采用禽流感病毒Ck/HB/4/08(H9N2)鼻腔接种豚鼠,检测H9N2禽流感病毒对豚鼠的致病力。结果表明:感染后的豚鼠出现聚堆、精神不振、流鼻涕等感冒症状,体温及体重均发生显著变化,同时能够产... 详细信息

为了研究用豚鼠建立禽流感动物模型的可能性,试验采用禽流感病毒Ck/HB/4/08(H9N2)鼻腔接种豚鼠,检测H9N2禽流感病毒对豚鼠的致病力。结果表明:感染后的豚鼠出现聚堆、精神不振、流鼻涕等感冒症状,体温及体重均发生显著变化,同时能够产生相应抗体;感染豚鼠的肺脏出现严重的淤血及水肿,肺泡形态发生改变并伴有大量炎性细胞浸润;鼻甲骨和肺脏中可检测并分离到病毒,其他组织器官未检测到病毒。感染豚鼠的临床症状及病理、体重、体温、抗体、肺干湿重的变化和组织中病毒的检测和分离结果说明H9N2亚型禽流感病毒可以感染豚鼠发病,症状明显,但不引起豚鼠死亡,可以建立禽流感感染豚鼠的发病模型。

关键词： H9N2亚型禽流感病毒豚鼠致病力动物模型