Development of a stable attenuated double-mutant of tobacco mosaic virus for cross-protection

Tobacco(Nicotiana tabacum)and tomato(Solanum lycopersicum)are two major economic crops in China.Tobacco mosaic virus(TMV;genus Tobamovirus)is the most prevalent virus infecting both crops.Currently,some widely cultivated tobacco and tomato cultivars are susceptible to TMV and there is no effective strategy to control this virus.Cross-protection can be a safe and environmentally friendly strategy to prevent viral diseases.However,stable attenuated TMV mutants are scarce.In this study,we found that the substitutions in the replicase p126,arginine at position 196(R^(196))with aspartic acid(D),glutamic acid at position 614(E^(614))with glycine(G),serine at position 643(S^(643))with phenylalanine(F),or D at position 730(D^(730))with S,significantly reduced the virulence and replication of TMV.However,only the mutation of S^(643) to F reduced the RNA silencing suppression activity of TMV p126.A double-mutant TMV-E614G-S643F induced no visible symptom and was genetically stable through six successive passages in tobacco plants.Furthermore,our results showed that TMV-E614G-S643F double-mutant could provide effective protection against the wild-type TMV infection in tobacco and tomato plants.This study reports a promising mild mutant for cross-protection to control TMV in tobacco and tomato plants.

Improving Cross-Protection against Influenza Virus Using Recombinant Vaccinia Vaccine Expressing NP and M2 Ectodomain Tandem Repeats

Conventional influenza vaccines need to be designed and manufactured yearly.However,they occasionally provide poor protection owing to antigenic mismatch.Hence,there is an urgent need to develop universal vaccines against influenza virus.Using nucleoprotein(NP)and extracellular domain of matrix protein 2(M2e)genes from the influenza A virus A/Beijing/30/95(H3N2),we constructed four recombinant vaccinia virus-based influenza vaccines carrying NP fused with one or four copies of M2e genes in different orders.The recombinant vaccinia viruses were used to immunize BALB/C mice.Humoral and cellular responses were measured,and then the immunized mice were challenged with the influenza A virus A/Puerto Rico/8/34(PR8).NP-specific humoral response was elicited in mice immunized with recombinant vaccinia viruses carrying full-length NP,while robust M2e-specific humoral response was elicited only in the mice immunized with recombinant vaccinia viruses carrying multiple copies of M2e.All recombinant viruses elicited NP-and M2e-specific cellular immune responses in mice.Only immunization with RVJ-4M2eNP induced remarkably higher levels of IL-2 and IL-10 cytokines specific to M2e.Furthermore,RVJ-4M2eNP immunization provided the highest cross-protection in mice challenged with 20 MLD5〇of PR8.Therefore,the cross-protection potentially correlates with both NP and M2e-specific humoral and cellular immune responses induced by RVJ-4M2eNP,which expresses a fusion antigen of full-length NP preceded by four M2e repeats.These results suggest that the rational fusion of NP and multiple M2e antigens is critical toward inducing protective immune responses,and the 4M2eNP fusion antigen may be employed to develop a universal influenza vaccine.

盐业法律的跨区域实施困境与出路——以宜宾丰源盐业公司案为例

地方保护主义是构建国内统一大市场的重要障碍,同时也是法律跨区域实施的一大难题。通过宜宾丰源盐业公司延边分公司诉D市盐务管理局行政强制案,可以看出地方保护主义在盐业法律跨区域实施上的阻碍。按照《盐业体制改革方案》的基本精神和新修订实施的《食盐专营办法》相关规定,国家已经取消了食盐跨区域经营的禁止性规定,明确支持食盐经营企业可以开展跨区域经营。但是一些地方盐务管理部门仍然以修订前的行政法规作为执法依据,这是地方保护主义在盐务执法上的体现。为此,需要建立公正独立的司法机制,以克服地方保护主义,保障法律的跨区域顺利实施,促进全国统一大市场建设。

美国跨境数据流动的国际法规制路径与中国的因应

作为互联网大国,美国在规制跨境数据流动方面从一开始就确立了重数据自由流动而轻政府监管的路径,并将之贯穿于历届政府的对外贸易政策,以及世界贸易组织框架下的相关议题谈判和自由贸易协定谈判。这样的路径背后体现出美国国内法与国际法的紧密联系、更加关注消费者权利保护等特点。对其他国家所采取的限制数据流动的措施,美国常将之认定为'数据保护主义',但这样的认定带有明显的'双重标准'。对于我国而言,除了完善有关个人隐私保护的国内法,也应当通过政府间的磋商机制与美国展开双边层面的谈判,厘清数据保护主义与数据保护之间的界限。从宏观层面上看,我国也应当推动国际社会针对跨境数据流动形成多边国际法规范。

An Attenuated Strain of Cucumber Green Mottle Mosaic Virus as a Biological Control Agent against Pathogenic Viral Strains

Cucumber green mottle mosaic virus (CGMMV), a member of the Tobamovirus genus, causes a severe disease of cucurbits. In the Moscow region of Russian Federation, the incidence of infection on cucumber plants in greenhouses is high;however, the virus is poorly studied. In this work, the full-length genomes of two pathogenic MC-1 and MC-2 strains of CGMMV isolated from cucumber plants grown in greenhouses in the Moscow region and the attenuated VIROG-43M strain were sequenced. Comparison of VIROG-43M nucleotide sequence with those of the pathogenic strains revealed three missense mutations. Their role in attenuation is discussed. For the first time, in a number of trials conducted under laboratory conditions and in commercial greenhouses, the efficiency of the attenuated VIROG-43M strain as a biocontrol agent for cucumber plant protection resulting in significant yield gain was demonstrated. Phylogenetic analysis with 83 full-length CGMMV coat protein genes isolated in 16 different countries showed that Russian strains are related to isolates from Spain, Greece, USA and Israel.

全球数字保护主义的兴起、发展和应对

数字贸易快速发展,对世界经济的贡献日益显著,但随之而来的隐私和个人数据安全、限制跨境数据流动等敏感问题引起了国际社会的广泛关注。一些国家通过立法设定与数字贸易相关的一系列壁垒,产生了以这些壁垒为主要特征的数字保护主义,对数字贸易发展构成严重威胁。数字保护主义在当前国际贸易中备受关注,应对全球数字保护主义蔓延,各国迫切需要借鉴和吸收一些区域或多边贸易协定制定数字贸易规则的经验与方法,推进世贸组织建立数字贸易国际规则进程。中国应当在数字贸易国际规则构建中发挥重要作用。

In search of a pan-coronavirus vaccine:next-generation vaccine design and immune mechanisms

Members of the coronaviridae family are endemic to human populations and have caused several epidemics and pandemics in recent history.In this review,we will discuss the feasibility of and progress toward the ultimate goal of creating a pan-coronavirus vaccine that can protect against infection and disease by all members of the coronavirus family.We will detail the unmet clinical need associated with the continued transmission of SARS-CoV-2,MERS-CoV and the four seasonal coronaviruses(HCoV-OC43,NL63,HKU1 and 229E)in humans and the potential for future zoonotic coronaviruses.We will highlight how first-generation SARS-CoV-2 vaccines and natural history studies have greatly increased our understanding of effective antiviral immunity to coronaviruses and have informed next-generation vaccine design.We will then consider the ideal properties of a pan-coronavirus vaccine and propose a blueprint for the type of immunity that may offer cross-protection.Finally,we will describe a subset of the diverse technologies and novel approaches being pursued with the goal of developing broadly or universally protective vaccines for coronaviruses.

公平竞争审查制度能否促进企业跨地区投资?

打破地方保护与市场分割,促进各类生产要素在地区间自由流动,是构建全国统一大市场的关键。以公平竞争审查制度作为准自然实验,检验该制度对企业跨地区投资的影响。研究表明,公平竞争审查制度显著促进了企业跨地区投资。作用机制检验证实,该制度通过提升市场竞争的充分性和公平性以及降低交易费用,促进企业跨地区投资。进一步检验发现,公平竞争审查制度更有效地促进了将该制度的执行纳入政府绩效评价体系和将第三方评估结果向社会公开地区的企业以及环境不确定程度高的企业进行跨地区投资。经济后果检验发现,该制度通过促进企业跨地区投资提高了企业异地经营绩效。研究结论拓展了政府监管与制度环境影响企业投资决策的经验证据,为完善公平竞争审查制度,以更好地实现有效市场与有为政府的有机统一、推进市场一体化建设、构建新发展格局,提供了微观层面的经验启示。

司法“去地方化”改革反思

基于“司法权是中央事权”这一论断,2013年以来的司法改革力图通过司法权的中央化来祛除司法的地方保护主义。在中央确定的两项主要改革举措中,推进省以下地方法院、检察院人财物省级统管,因受地方各级人大产生同级法院、检察院的宪制约束,以及各地方财政资源不平衡现实的影响,只能实行“有限统管”;设立跨行政区划法院、检察院,因存在宪制空间上的争议无法形成统一的认识,一直停留在试点阶段而踟蹰不前,改革路径也在司法机构创制和管辖制度调整之间来回切换。实际上,地方法院、检察院不仅具有“国家性”,而且“地方性”也是其内在基因,司法地方保护主义的产生,并非根源于法院、检察院的地方性,而在于法院、检察院行使司法权时被属地体制性地行政化。只有按照法治的要求和司法的规律,合理确定司法权在地方权力结构中的地位,法院、检察院依法独立公正行使职权,才具有可依赖的前提和基础。

地方法院人财物省级统管促进了跨地区投资

在2014年后,我国分阶段推出了“省以下地方法院人财物省级统一管理”改革。本研究发现,改革后外地企业赴改革地区的异地投资显著增加。该效应对于金融发展水平更高的地区、非国有企业、具有法律背景的企业及省内异地投资更为明显。机制分析表明,改革通过改善地区产权保护和降低企业制度性交易成本来促进企业异地投资。本文表明破除司法地方保护有助于打破市场壁垒和构建全国统一大市场。