非洲猪瘟研究进展

非洲猪瘟是由非洲猪瘟病毒感染家猪或野猪后引发的一种急性、烈性传染病,主要通过病猪及其周围环境传播,蜱是中间宿主。1921年该病首次暴发于非洲肯尼亚,2018年8月传入我国,目前已有24个省级行政区发生疫情。非洲猪瘟病毒主要经呼吸道和消化道进入猪体内,感染靶细胞主要是单核-巨噬细胞,目前受体还不明确。非洲猪瘟病毒是单分子双链DNA病毒,长度为170~190kb,编码150~200种蛋白,包括多种免疫调控蛋白,可以抵抗机体免疫。非洲猪瘟病毒疫苗研究较多,包括灭活疫苗、减毒疫苗、亚单位疫苗和基因疫苗等,但迄今这些疫苗都不能保护家猪免受非洲猪瘟病毒感染。今后需要对非洲猪瘟病毒及其发病机制做详细系统的研究,为开发有效防治方案提供资料。     

非洲猪瘟病毒微滴式数字PCR检测方法的建立与应用

为建立一种特异性强、灵敏度高、重复性好的非洲猪瘟病毒微滴数字PCR检测方法,本研究根据非洲猪瘟病毒核酸的2段保守序列(VP72和K205基因)设计合成了3对引物和探针,摸索其反应条件,优化方法,比较方法的线性、特异性、灵敏性和重复性。结果发现,这3对引物和探针的非洲猪瘟病毒方法检出限均小于6拷贝数/反应,在10～106拷贝数/反应之间均呈良好的线性关系,定量范围内的检测变异系数均低于15%,且不与猪常见的2种病毒发生交叉反应,适用于猪肉及其制品中非洲猪瘟病毒的检测,有利于在源头、食品加工、食品销售等各个环节加强对生鲜猪肉及各类猪肉制品中非洲猪瘟病毒的检测,切实降低潜在风险。     

非洲猪瘟抗体胶体金免疫层析检测方法的建立及评价

为建立一种快速、高效、便捷的非洲猪瘟病毒抗体检测方法，本试验采用生物信息学方法筛选得到了性能优异的非洲猪瘟病毒抗原表位多肽P54t，利用P54t多肽作为检测抗体的特异性抗原，建立了一种特异性、敏感性好的胶体金免疫层析抗体检测方法，用于检测待测样品中是否含有非洲猪瘟病毒抗体。结果显示，该方法的敏感性为100%，特异性为100%，最低检测限度可达1:12800，与商品化试剂盒比较，符合率为94.3%。结果表明，该方法敏感性好、特异性强，检测结果准确，可用于非洲猪瘟病毒特异性抗体检测。     

非洲猪瘟防控及疫苗研发：挑战与对策

非洲猪瘟是由非洲猪瘟病毒引起的一种接触传染性、广泛出血性猪烈性传染病,最急性和急性感染死亡率高达100%。自2018年8月我国发生首起非洲猪瘟疫情后,3个多月内,已有18个省份累计暴发69起,给我国养猪业造成了沉重打击。从目前非洲猪瘟全球流行态势及世界各国防控经验来看,我国非洲猪瘟防控和根除面临的形势不容乐观,亟需安全有效的疫苗用于该病的防控。文中结合当前非洲猪瘟病原学最新研究成果,系统总结了非洲猪瘟防控策略、疫苗研究进展及其面临的挑战,重点分析了疫苗研发历程、存在的问题、未来发展方向以及商业化应用所面临的关键科学问题,以期为我国非洲猪瘟防控及病原和疫苗研究提供借鉴。     

非洲猪瘟病毒编码蛋白功能研究进展

非洲猪瘟(African swine fever,ASF)是非洲猪瘟病毒(African swine fever virus,ASFV)感染家猪或野猪引起的一种急性、出血性、高度接触性传染病,其特征是病程短、高热和出血性病变,急性感染死亡率高达100%,严重威胁全球养猪业但目前尚未开发出有效的疫苗和治疗方法。ASFV是非洲猪瘟病毒科非洲猪瘟病毒属的唯一成员,为大型双链DNA病毒,主要在巨噬细胞胞质中复制,其基因组约170?193 kb,含有150?167个开放阅读框,编码150?200种蛋白质。目前已知功能的病毒编码蛋白约有50个,大部分为病毒的结构蛋白,仍有一半以上的ASFV编码蛋白功能尚不清楚。除结构蛋白以外,病毒含有完整的酶和与病毒转录有关的因子,编码调节宿主细胞功能及与病毒免疫逃逸相关的蛋白等。本文综述了ASFV的结构蛋白、非结构蛋白以及参与免疫逃逸等相关蛋白功能的研究进展,以期为ASFV病毒蛋白研究及疫苗研发提供相关借鉴。     

一种针对非洲猪瘟病毒的具有交叉保护功能的新型重组减毒活疫苗

正非洲猪瘟是世界动物卫生组织(OIE)强制声明的一种具有高度传染性的病毒性疾病。缺乏可用的疫苗使这种疾病很难控制,因此,非洲猪瘟病毒(ASFV)成为养猪业的一大主要威胁。灭活疫苗并不会对ASFV提供坚实有力的保护。相反,减毒活病毒(LAV),无论是自然分离的还是通过基因操作获得的,都证明了具有对同源ASFV病毒株的可靠保护力,尽管对异种病毒的保护力很低或没有保护力。     

非洲猪瘟病毒多基因家族研究进展

非洲猪瘟(African swine fever,ASF)是由非洲猪瘟病毒(African swine fever virus,ASFV)感染引起的一种急性、热性、高度接触性、致死性动物传染病.由于ASFV基因组庞大,变异能力强,免疫逃逸机制复杂,至今无有效药物和疫苗.近年来,对多基因家族的研究取得了很大的进展,可变区多基因家族拷贝数变化是导致ASFV变异的主要原因,且陆续发现多基因家族在决定细胞宿主范围、影响病毒毒力强弱、抑制Ⅰ型干扰素信号通路、抑制干扰素抗病毒效应和促进病毒蜱源感染中具有重要作用.本文对当前的ASFV多基因家族研究进展进行总结,阐述其在基因变异和病毒感染中的作用,以期为非洲猪瘟免疫逃逸机制的探索和疫苗的研发提供理论依据.     

管式磁微粒非洲猪瘟病毒pp62蛋白化学发光抗体检测方法的建立

【背景】非洲猪瘟(African swine fever, ASF)作为高致病性传染病,给我国生猪养殖业造成了严重的经济损失,因此建立快捷、灵敏的诊断方法至关重要。【目的】建立一种管式非洲猪瘟病毒pp62蛋白化学发光抗体检测方法。【方法】以重组pp62蛋白作为包被抗原,羧基磁珠(carboxylic magnetic beads)作为固相载体,碱性磷酸酶(alkaline phosphatase, AP)标记的兔抗猪IgG作为酶标二抗,通过对反应条件进行优化,采用非洲猪瘟国家参考品作为溯源血清绘制出标准曲线,建立基于ASFV pp62蛋白的化学发光抗体检测方法。【结果】用建立的化学发光方法检测277份临床样品血清,通过受试者工作特征(receiver operating characteristic, ROC)曲线分析确定阴性、阳性临界值,并确定判定标准：浓度值>140.02 U时为抗体阳性,浓度值<140.02 U时为抗体阴性。此方法对6种不同的病原血清抗体进行检测均无交叉反应,且批内和批间变异系数均在10%以内,与商品化非洲猪瘟抗体检测试剂盒的符合率达96.7%。【结论】本研究建立的管式非洲猪瘟病毒化学发光抗体检测方法具有良好的特异性、敏感性和重复性,可为非洲猪瘟早期监测及试剂盒研发提供参考。     

非洲猪瘟病毒结构蛋白在病毒感染过程中的作用

非洲猪瘟(African swine fever,ASF)是由非洲猪瘟病毒(African swine fever virus,ASFV)引起的一种急性、烈性、出血性的猪传染性疫病,给疫情发生国家(地区)的养猪业带来严重的经济损失。ASFV作为双股DNA病毒,含有150-167个开放阅读框(ORFs),编码200余种蛋白质,其中结构蛋白约50种。结构蛋白作为病毒颗粒的主要组分,在病毒吸附、侵入和复制等感染过程中起着重要作用。综述了ASFV结构蛋白在病毒感染中的作用,以期为ASFV结构蛋白的进一步研究提供参考。     

野猪在非洲猪瘟流行中的重要作用

非洲猪瘟因其高死亡率、没有疫苗防疫、影响国际贸易而备受广泛关注。1921年首次确认非洲猪瘟疫情以来,先后在非洲、欧洲、美洲等多个国家和地区发病造成重大经济损失。野猪作为该病传播的重要生物媒介在俄罗斯等多个国家非洲猪瘟疫情散播中发挥了重要作用。充分了解全球非洲猪瘟疫情状况和野猪在俄罗斯非洲猪瘟疫情中的影响,分析我国野猪分布、疫病监测和管理现状,将为我国非洲猪瘟外来疫情防控策略制定提供参考。     

房室海绵化石的微机研究:性状演化

利用微机对房室海绵进行了１）属的清理工作，２）标准化术语的建立与属的标准化术语描述，３）建数据库，４）性状的统计分析。并发现了房室海绵出水管类型、房室排列方式、房室充填物类型、孔和管、体型和房室形态方面的六大演化趋向。     

Super Hybrid Rice Breeding in China：Achievements and Prospects

Hybrid rice has contributed greatly to the self-sufficiency of food supply In China. To meet the future demand for rice production, a national program on super rice breeding was established In China In 1996. The corresponding targets, breeding strategies and most significant advances are reviewed In this paper. New plant type models have been modified to adjust to various rice growing regions. In recognition of the Importance of applying parents with Intermediate subspecies differentiation In Increasing F1 yield, medium type parental lines were selected from populations derived from Inter-subspecies crosses with the assistance of DNA markers for subspecies differentiation. Results also indicate that a substantial increase of blomass Is the basis for further enhancement of the grain yield potential, and amelioration of leaf characteristics Is helpful In Increasing the photosynthetic rate. Thirty-four super hybrid rice varieties have been released commercially, growing In a total area of 13.5 million hm2 and producing 6.7 thousand million kg more rice In 1998-2005. Although remarkable progress has been made In super hybrid rice breeding In China, selections on the root system and Integration of more blotechnologlcal tools remain a great challenge.     

Integrons found in different locations have identical 5' ends but variable 3' ends.

The positions of the outer boundaries of the 5'- and 3'-conserved segment sequences of integrons found at several different locations have been determined. The position of the 5' end of the 5'-conserved segment is the same for six independently located integrons, In1 (R46), In2 (Tn21), In3 (R388), In4 (Tn1696), In5 (pSCH884), and In0 (pVS1). However, the extent of the 3'-conserved segment differs in each integron. The sequences of In2 and In0 diverge first from the conserved sequence, and their divergence point corresponds to the 3'-conserved segment endpoint defined previously (H.W. Stokes and R.M. Hall, Mol. Microbiol. 3:1669-1683, 1989), which now represents the endpoint of a 359-base deletion in In0 and In2. The sequence identity in In3, In1, In4, and In5 extends beyond this point, but each sequence diverges from the conserved sequence at a different point within a short region. Insertions of IS6100 were identified adjacent to the end of the conserved region in In1 and 123 bases beyond the divergence point of In4. These 123 bases are identical to the sequence found at the mer end of the 11.2-kb insertion in Tn21 but are inverted. In5 and In0 are bounded by the same 25-base inverted repeat that bounds the 11.2-kb insert in Tn21, and this insert now corresponds to In2. However, while In0, In2, and In5 have features characteristic of transposable elements, differences in the structures of these three integrons and the absence of evidence of mobility currently preclude the identification of all of the sequences associated with a functional transposon of this type.     

Hydrogen cyanide synthesis and antifungal activity of the biocontrol strain Pseudomonas fluorescens In5 from Greenland is highly dependent on growth medium

Hydrogen cyanide (HCN) is a secondary metabolite produced by many antagonistic Pseudomonas species. In the present study, the gene cluster encoding HCN synthesis in a newly isolated Pseudomonas fluorescens strain, In5, from South Greenland was investigated. Sequence analysis showed that the Greenlandic hcn gene cluster comprises a novel hcn cluster. Transposon mutagenesis of strain In5 resulted in mutants In5-2E1 and In5-1H7 with no production of HCN, and mutant In5-6B9 with reduced HCN synthesis. In mutant In5-2E1, the transposon was inserted into the hcnC gene; in mutant In5-1H7, the Tn5 insertion was found in a region upstream of a putative malate:quinone oxidoreductase gene (mqo); and in mutant In5-6B9, the transposon disrupted a probable enoyl-CoA hydratase/isomerase gene. In vitro inhibition experiments with In5 (wild type) and In5-2E1 (mutant) showed that in nitrogen-rich Luria-Bertani medium, strain In5 but not the hcn mutant In5-2E1 produced HCN and inhibited the growth of hyphae of Rhizoctonia solani and Pythium aphanidermatum . In contrast, when cultivating the strains in the carbohydrate-rich potato dextrose medium, neither of the strains produced any HCN, and thus, they were unable to inhibit hyphal growth of fungi. These experiments strongly indicate that the synthesis of HCN is highly dependent on the growth medium used.     

化学交联质谱技术的研究进展

化学交联质谱技术是解析蛋白质结构和研究蛋白质相互作用的重要工具。近5年以来,该技术在方法和应用上都取得了很大的进步。方法上,一方面可断裂交联剂与新型分离富集方法展现了较好的应用前景,另一方面更加高效的交联肽段搜索引擎和质量控制方法为交联质谱数据分析提供了有力的工具。应用上,一方面与冷冻电镜技术结合解析了大量蛋白质的结构,另一方面从研究蛋白质复合物的相互作用发展到研究全蛋白质组水平的相互作用网络。化学交联质谱技术在方法和应用上的蓬勃发展,体现了这一技术的重要作用。本文对化学交联质谱技术的各个环节进行了详细的综述,包括交联剂选择、交联反应、酶切、交联肽段富集、液质联用、交联肽段鉴定、质量控制和生物学应用,重点介绍了最近5年的研究进展。最后,讨论了化学交联质谱技术面临的挑战及未来的发展方向。     

番茄受精作用及其间隔期的研究

利用常规石蜡切片法研究了番茄受精作用的全过程,具体研究结果为:(1)授粉后2 h,花粉粒在柱头上萌发;约2～4 h,花粉管长入柱头,且末端膨大;约8 h后,生殖细胞进入分裂期;并于约两小时后,分裂为两个精细胞.(2)约14 h,花粉管进入子房腔;约18～24 h,花粉管进入胚囊,破坏一个助细胞,并在其珠孔端释放两个精子;随后被释放的精子移到卵细胞与次生核附近.(3)授粉后约30 h精核进入卵细胞;约34 h,精核与卵核融合,并在卵核内出现分散的雄性染色质,进而出现雄性核仁;44～50 h,雌、雄性核仁融合,形成合子;合子的休眠期为10 h左右.60 h之后,合子分裂形成二细胞原胚.(4)约26 h,另一个精子的精核与次生核核膜相贴伏,随后与之融合;约30～34 h,次生核内出现分散的雄性染色质,随之出现雄性核仁;约38～42 h,雌、雄性核仁融合,形成初生胚乳核.约44 h后,初生胚乳核进行有丝分裂,形成两个胚乳细胞.番茄胚乳发育属于细胞型.初生胚乳核无休眠期.(5)精子与次生核的融合比与卵核的融合快.(6)番茄的受精作用属于有丝分裂前配子融合类型.     

An indium:calcium phosphate colloid that specifically targets fibrin

The ability of indium to target fibrin in vitro was evaluated. The radionuclide ^114mIndium (^114mIn) was prepared as a soluble and colloidal (In:In) form, as well as, a mixed indium:calcium phosphate (In:CaP) colloid. Soluble ^114mIn was prepared by maintaining acid pH (50 mM HCl). Colloidal ^114mIn (In:In) was prepared under slightly basic conditions (50 mM Tris-Cl, pH 7.6). The mixed In:CaP colloid was prepared by incubation of ^114mIn with calcium (10 mM) and phosphate (250 M) under slightly basic conditions (50 mM Tris-Cl, pH 7.6). To assess fibrin binding, the three ^114mIn preparations were mixed with diluted human plasma (source of fibrinogen). Fibrin polymerization was initiated by addition of calcium (5 mM) and thrombin (0.5 U/ml). Following incubation (15 min, 37 °C), the fibrin matrix was condensed, removed from the reaction mixture, and washed briefly. Fibrin uptake of ^114mIn (soluble, colloidal, or In:CaP) was determined by gamma counting. Results demonstrated that soluble ^114mIn exclusively bound a plasma protein electrophoretically and immunologically identified as transferrin. Although both colloidal ^114mIn and ^114mIn:CaP bound fibrin, the mixed ^114mIn:CaP colloid demonstrated substantially higher fibrin binding activity (about 2-fold). The target of indium binding was confirmed as fibrin due to the presence of characteristic cross-linked – dimers (100 kDa) and -monomers (58 kDa) by SDS-PAGE. ^114mIn colloid and the mixed ^114mIn:CaP colloid demonstrated no ability to bind fibrins precursor, fibrinogen. ^114mIn:CaP fibrin binding was associated with formation of CaP, as evidenced by its dependence on phosphate concentration. The biocompatibility of CaP including its ability to bind ^114mIn and specifically target fibrin may be of potential value for diagnostic imaging studies to identify regions of occult vascular stenosis (i.e., atherosclerotic plaques, deep vein thrombosis, pulmonary embolus).In memoriam.     

不同生境内濒危植物长喙毛茛泽泻种群数量动态比较

湖南茶陵自然种群动态在一年内表现为春季数量下降，以后振荡变化。在以莼菜为主的样方中，种群在夏季以后一直呈增加趋势，至冬季末数量下降；在杂草拥挤的样方内，种群基本呈下降趋势。湖北武汉，网室中迁地保护的种群生长在理想条件下，单一种群呈逻辑斯缔（Ｌｏｇｉｓｔｉｃ）增长，方程为Ｎ（株／ｍ２）＝８４０１＋ｅ４．４７７－０．２６４ｔ。江西东乡回归原产地的种群在回归当年经过精心照顾，种群呈增加趋势，次年随着杂草茂盛生长，种群数量迅速下降，种群具消失的危险。长喙毛茛泽泻种群数量的变动很大程度上受水位变化和杂草干扰的影响。实验结果表明，长喙毛茛泽泻种群能成功地实施迁地保护。     

The association between inversion In(3R)Payne and clinally varying traits in Drosophila melanogaster

In Drosophila melanogaster, inversion In(3R)Payne increases in frequency towards low latitudes and has been putatively associated with variation in size and thermal resistance, traits that also vary clinally. To assess the association between size and inversion, we obtained isofemale lines of inverted and standard karyotype of In(3R)Payne from the ends of the Australian D. melanogaster east coast cline. In the northern population, there was a significant association between In(3R)Payne and body size, with standard lines from this population being relatively larger than inverted lines. In contrast, the inversion had no influence on development time or cold resistance. We strengthened our findings further in a separate study with flies from populations from the middle of the cline as well as from the cline ends. These flies were scored for wing size and the presence of In(3R)Payne using a molecular marker. In females, the inversion accounted for around 30% of the size difference between cline ends, while in males the equivalent figure was 60%. Adaptive shifts in size but not in the other traits are therefore likely to have involved genes closely associated with In(3R)Payne. Because the size difference between karyotypes was similar in different populations, there was no evidence for coadaptation within populations.     

Distribution of phenylalanine ammonia-lyase in etiolated and far-red irradiated radish seedlings

Summary In dark-grown Raphanus seedlings, most of the PAL activity is found in roots where it increases sigmoidally during organ development. In hypocotyls, the dark increase of enzyme activity is linear with time. In cotyledons and hooks, dark activity is very low and remains constant. After onset of continuous far-red irradiation, an activity increase is observed in all parts of the seedling. In cotyledons and hooks, the increase is followed by a decrease. This is comparable to light-induced PAL activity described in other materials. In roots and hypocotyls, the initial increase is not followed by a decrease. In dark-grown roots and hypocotyls PAL activity is correlated to fresh weight augmentation. In no part of the seedling could a correlation be found between light-induced PAL activity and anthocyanin formation.