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Effect of cDNA fragments in different length derived from Potato Virus Y coat protein gene on the induction of RNA-mediated virus resistance

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Science China(Life Sciences) 2004年第4期47卷 382-388页

作者： ZHU Junhua ZHU Xiaoping WEN Fujiang BAI Qingrong ZHU Changxiang song yunzhi Laboratory of Plant Genetic Engineering College of Plant ProtectionShandong Agricultural UniversityTaian 271018China

We have reported that cDNA derived from entire coat protein (CP) gene of potato virus Y (PVY) could induce resistance to PVY infection in transgenic tobacco plants, and the resistance was further demonstrated to be RNA-mediated rather than coat protein-mediated. In this study, we cloned cDNA fragments of 202 bp, 417 bp, and 603 bp in length derived from the 3′ end of the PVY CP gene, and the cDNA fragments were introduced into tobacco (var. NC89) plants via Agrobacterium-mediated transformation system. The results of resistance assay showed that the CP cDNA fragments of 417 bp and 603 bp could confer resistance of the trans-genic plants to PVY infection, but the fragment of 202 bp in length could not. Molecular analysis revealed that the resistance was RNA-mediated, which is believed to be a result of post-transcriptional gene silencing. The results indicate that the length of cDNA fragments needed for resistance induction was located somewhere between 202 bp and 417 bp from the 3′ end of PVY CP gene.

关键词： PVY partial length of CP gene fragments transgenic tobacco plants RNA-mediated resistance post-transcrip-tional gene silencing.

Dimeric artificial microRNAs mediate highly efficient RSV and RBSDV resistance in transgenic rice plants

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Dimeric artificial microRNAs mediate highly efficient RSV an...

马铃薯Y病毒复制酶基因不同位置cDNA区段介导对PVY的不同抗性

中国植物病理学会第十二届青年学术研讨会

作者： Lin Sun Chao Lin yunzhi song Jinwen Du Fujiang Wen Changxiang Zhu State Key Laboratory of Crop Biology Shandong Key Laboratory of Crop BiologyCollege of Life SciencesShandong Agricultural University

Infection of Rice stripe virus（RSV） and Rice black streaked dwarf virus（RBSDV） on rice results in severe economic losses on crop *** the natural resistance resources against these viruses are limited,it is imperative to elaborate a biotechnological approach that will provide effective and safe immunity to RSV and *** this study,we generated three dimeric amiRNA precursor expression vectors（pamiR-M,pamiR-3 and pamiR-U） that simultaneously targeting RSV and RBSDV based on the structure of rice osa-MIR528 *** transgenic rice plants were obtained by Agrobacterium tumefaciens-mediated transformation and proved to express amiRNAs *** viral challenge assays revealed that these transgenic plants could be conferred highly specific resistance against RSV and RBSDV infection *** amiR-RSVU and the amiR-RBSDVU induced the highest virus resistance against RSV and RBSDV,*** target viral RNAs could be cleavaged by the amiRNAs and there was a relationship between the resistance level and the accumulation of amiRNA expression.A Northern blot assay verified that silencing was induced by the original amiRNAs and could be bilaterally extended by the siRNA *** amiRNA together with the secondary siRNAs mediated the degradation of viral *** stability assay showed that transgene and amiRNAmediated virus resistance can be stably inherited toT2.

关键词： artificial microRNA Rice stripe virus Rice black streaked dwarf virus multiple virus resistances transgenic rice

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植物保护学报 2010年第3期37卷 227-233页

作者：徐丽宋云枝王文俊朱常香温孚江山东农业大学植物保护学院泰安271018 山东农业大学作物生物学国家重点实验室泰安271018 山东农业大学作物生物学国家重点实验室泰安271018 山东农业大学生命科学学院泰安271018

为探究靶序列位置对RNA介导的病毒抗性产生的影响，利用聚合酶链式反应（polymerase chain reaction，PCR）技术扩增马铃薯Y病毒（Potato virus Y, PVY）复制酶基因（nuclear inclusion b，NIb）不同位置的cDNA区段，反向插入双元载体pR... 详细信息

为探究靶序列位置对RNA介导的病毒抗性产生的影响，利用聚合酶链式反应（polymerase chain reaction，PCR）技术扩增马铃薯Y病毒（Potato virus Y, PVY）复制酶基因（nuclear inclusion b，NIb）不同位置的cDNA区段，反向插入双元载体pROKII中，构建了发夹RNA（hairpin RNA，hpR—NA）结构的植物表达载体。将构建的植物表达载体采用冻融法转入农杆菌LBA4404，叶盘法转化烟草NC89，获得转基因植株。攻毒试验表明：PVYNIb基因不同位置cDNA区段介导的对PVY的抗性存在显著差异；3’端1／2处和中间位置的序列可介导高水平的病毒抗性，抗性植株的比例在50％以上，而5‘端、5’端1／2处和3’端的序列介导的抗性效率较低，抗性植株的比例仅为10％～30％。Northern杂交显示：抗病植株中RNA的积累量明显低于同类型的感病植株，抗性与RNA积累量呈负相关；抗病转基因植株中有siRNA存在，表明病毒抗性是由RNA介导的。

关键词：马铃薯Y病毒复制酶基因 RNA介导的病毒抗性

水稻条纹病毒山东济宁分离物RSV-SD-JN2分子变异分析

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植物保护学报 2008年第5期35卷 415-420页

作者：马进宋云枝李开东朱常香温孚江山东农业大学作物生物学国家重点实验室泰安271018 山东农业大学生命科学学院泰安271018

为从分子水平探讨强致病性水稻条纹病毒山东济宁分离物RSV-SD-JN2的变异及其致病性,采用RT-PCR技术,克隆RSV-SD-JN2的RNA3、RNA4区段cDNA。结果显示,RSV-SD-JN2的RNA3和RNA4核苷酸序列长度分别为2487、2157bp;RNA3中,NS3基因长为636bp,基因间隔区(IR)为725bp,CP基因为969bp;RNA4中,SP基因长为537bp,基因间隔区(IR)为654bp,NSvc4基因为861bp。不同时期、不同区域和不同致病性的RSV分离物的序列比较发现,RNA3、RNA4序列5′和3′末端非编码区具有高度保守性,仅存在个别碱基的差异;基因编码区保守性较高,核苷酸和氨基酸序列同源性分别在93%和97%以上,且大部分碱基变异为无意义变异;基因间隔区(IR)易于变异。RSV的分子变异与其地理分布具有密切的关系。RNA4的IR的变异导致RNA二级结构——发夹结构的稳定性提高是病毒致病性增强的重要原因。

关键词：水稻条纹病毒 RNA3 RNA4 序列分析分子变异

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基于时序空间关系模型的交通标志识别

计算机时代 2019年第6期 63-66,70页

作者：翟孝威宋云志中国海洋大学计算机科学与技术系

道路交通标志识别是汽车无人驾驶技术的重要组成部分。通过调整卷积层和池化层数据输入方式,改进具有融合特征的多尺度卷积神经网络,提高识别准确率。依据视频图像的空间连续关系构建时序空间关系模型(Temporal-SpatialModel,TSM),结合... 详细信息

道路交通标志识别是汽车无人驾驶技术的重要组成部分。通过调整卷积层和池化层数据输入方式,改进具有融合特征的多尺度卷积神经网络,提高识别准确率。依据视频图像的空间连续关系构建时序空间关系模型(Temporal-SpatialModel,TSM),结合多尺度卷积神经网络减少识别数据量,提高处理的效率,实现视频图像交通标志的高效率和高准确率识别。实验结果表明,所提出的算法识别率保持90.36%,在原始图像上运行平均帧率为32fps,有效地改进了基于单帧的交通标志图像识别效率低的问题。

关键词：交通标志识别汽车无人驾驶多尺度卷积神经网络时序空间关系模型

Complete Genome Sequences of Three Chinese Isolates of Tobacco Vein Banding Mosaic Virus and Analysis of Genetic Diversity

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Complete Genome Sequences of Three Chinese Isolates of Tobac...

中国植物病理学会2011年学术年会

作者： Tongtong Yuan,yunzhi song,Kaidong Li,Quanjun Han, Changxiang Zhu~*,Fujiang Wen~* (State Key Laboratory of Crop Biology,Shandong Key Laboratory of Crop Biology,Shandong Agricultural University, Taian,271018,P.R.China)

＜正＞Three complete RNA genomic sequences of Tobacco vein banding mosaic virus （TVBMV） isolates of China,designed as TVBMV-SDYS1,SDFX1 and SDZC1,had been *** complete genomic RNA sequence of each isolate was 9570nt excluding the poly（A） *** identities of genomic sequences and predicted gene products of them were 97.58%～98.65%and 98.96%～99.32%.No matter the complete genomes or every region of the genomes compared,the three isolates shared much closer identities with isolate HN39 than isolate *** address the most likely part for genetic engineering,we compared the CP and 3’-UTR nucleotide sequences of Chinese TVBMV isolates from GenBank,and the 3’-UTR sequences were much more conserved that can be used to culture transgenic TVBMV-resistant *** study reported three new TVBMV isolates obtained from China besides HN39 and YND published before,and firstly found the potentially best genome sequence to develop transgenic.

关键词： Tobacco vein banding mosaic virus Chinese isolates genomic sequence genetic anlysis

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利用鱼腥藻作为饲料的研究

水生生物学报 1999年第5期23卷 425-433页

作者：沈银武刘永定朱运芝宋立荣中国科学院水生生物研究所武汉430072

鱼腥藻粉作为肉鸡、全雌鲤、青虾和对虾的饲料研究结果表明：2％的鱼腥藻粉替代鱼粉作为肉鸡饲料试验，肉鸡的个体平均增重效果与对照相当，屠宰率、半净膛率和全净膛率都有所提高；同时还具有改善肉质色泽的效果．对重金属的含量分析... 详细信息

鱼腥藻粉作为肉鸡、全雌鲤、青虾和对虾的饲料研究结果表明：2％的鱼腥藻粉替代鱼粉作为肉鸡饲料试验，肉鸡的个体平均增重效果与对照相当，屠宰率、半净膛率和全净膛率都有所提高；同时还具有改善肉质色泽的效果．对重金属的含量分析发现，食鱼腥藻的鸡除锌的含量略高外，铜、铁、锰、镉及铝的含量都低于对照组．添加2％的鱼腥藻粉在网箱中进行小试和大面积试验，结果都表明鱼腥藻粉对全雌鲤的生长有明显的效果，并能提高全雌鲤鱼种的成活率和降低饵料系数．利用鱼腥藻粉配合饲料在网箱和大面积养殖青虾试验表明，具有促进生长，提高产量的作用．对虾试验结果表明，在饵料中添加2％的鱼腥藻粉，饲喂45d后，对虾的体长增加了14．49％；体重增加了10．43％这些研究结果表明利用鱼腥藻作为家禽及水产养殖动物的饲料具有相当大的应用前景．

关键词：鱼腥藻粉饲料

清瘟败毒饮颗粒结合双嘧达莫治疗儿童川崎病临床疗效及安全性观察

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中华中医药学刊 2024年第5期42卷 170-173页

作者：曹小彩宋文涛胡云芝路楠楠焦作市人民医院河南焦作454000

目的分析清瘟败毒饮颗粒结合双嘧达莫治疗儿童川崎病临床疗效及安全性。方法研究纳入2019年1月—2022年10月小儿内科收治的82例儿童川崎病进行分组研究,分组方法为随机数字表法,将其分为西医组和中医组,各41例,西医组给予双嘧达莫治疗,... 详细信息

目的分析清瘟败毒饮颗粒结合双嘧达莫治疗儿童川崎病临床疗效及安全性。方法研究纳入2019年1月—2022年10月小儿内科收治的82例儿童川崎病进行分组研究,分组方法为随机数字表法,将其分为西医组和中医组,各41例,西医组给予双嘧达莫治疗,中医组治疗方法同西医组,并结合清瘟败毒饮颗粒治疗,各组数据观察:临床治疗效果、治疗前后患儿中医证候(壮热不退、肌肤斑疹、唇赤干裂、咽红目赤、关节疼痛、手足硬肿等)积分变化、患儿冠脉损伤发生率、发热持续时间、淋巴结肿大消退时间及四肢肿胀消退时间、黏膜充血消退时间、住院时间、治疗前后患儿降钙素原(procalcitonin,PCT)、C反应蛋白(C-reactive protein,CRP)、红细胞沉降率(erythrocyte sedimentation rate,ESR)等水平炎症因子水平变化、免疫球蛋白G(immunoglobulin G,IgG)及免疫球蛋白M(immunoglobulin M,IgM)、补体C_(3)(complement 3,C_(3))等免疫指标水平变化、不良反应发生情况。结果中医组患儿治疗总有效率比西医组高,P0.05,治疗后各组患儿中医证候(壮热不退、肌肤斑疹、唇赤干裂、咽红目赤、关节疼痛、手足硬肿等)积分、PCT及CRP、ESR、IgG、IgM、C_(3)等指标均改善,中医组患儿治疗后中医证候(壮热不退、肌肤斑疹、唇赤干裂、咽红目赤、关节疼痛、手足硬肿等)积分、PCT及CRP、ESR、IgG、IgM、C_(3)等指标均优于西医组,均P0.05。结论清瘟败毒饮颗粒结合双嘧达莫治疗儿童川崎病临床疗效显著,患儿症状改善,免疫功能提升,冠脉损伤发生率低,恢复快,治疗安全可靠,值得应用。

关键词：儿童川崎病清瘟败毒饮颗粒双嘧达莫治疗效果免疫功能不良反应

Recent advances of microneedles for biomedical applications: drug delivery and beyond

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Acta Pharmaceutica Sinica B 2019年第3期9卷 469-483页

作者： Jian Yang Xinli Liu yunzhi Fu Yujun song College of Materials Science and Chemical Engineering Hainan UniversityHaikou570228China College of Engineering and Applied Sciences Nanjing UniversityNanjing210093China

The microneedle(MN), a highly efficient and versatile device, has attracted extensive scientific and industrial interests in the past decades due to prominent properties including painless penetration, low cost, excellent therapeutic efficacy, and relative safety. The robust microneedle enabling transdermal delivery has a paramount potential to create advanced functional devices with superior nature for biomedical applications. In this review, a great effort has been made to summarize the advance of microneedles including their materials and latest fabrication method, such as three-dimensional printing(3DP). Importantly, a variety of representative biomedical applications of microneedles such as disease treatment, immunobiological administration, disease diagnosis and cosmetic field, are highlighted in detail. At last, conclusions and future perspectives for development of advanced microneedles in biomedical fields have been discussed systematically. Taken together, as an emerging tool, microneedles have showed profound promise for biomedical applications.

关键词： Microneedle patches Biomedical applications Microfabricated device Drug delivery Disease treatment Disease diagnosis Immunobiological administration 3D printing