细胞源与鸡胚源传染性支气管炎病毒激活JAK-STAT信号通路的差异性研究

鸡传染性支气管炎病毒(Infectious Bronchitis Virus,IBV)对鸡的呼吸道、肾脏和输卵管等器官造成严重损伤,主要引起鸡产蛋率下降和雏鸡死亡.目前通过鸡胚传代获得IBV疫苗株和流行毒株.IBV Beaudette株是目前实验室研究的经典毒株,已经适应人源和猴源细胞,可利用Vero细胞进行制备.有研究表明,IBV感染延迟干扰素的表达,并对JAK-STAT信号通路具有拮抗作用.本研究中发现,通过Vero细胞制备的IBV Beaudette株,在感染早期激活STAT1,通过鸡胚制备的IBV Beaudette,则不能有效激活STAT1.进一步研究发现,鸡胚传代的IBV QX株和新城疫病毒(Newcastle Disease Virus,NDV)亦不能有效刺激JAK-STAT信号通路.两种制备病毒的方式分别得到不同的试验结果,推测是由于在病毒感染条件下,Vero细胞分泌到培养液中的细胞因子所致.进一步研究揭示,病毒感染条件下,细胞分泌的因子,瞬时激活了 STAT1.本研究对于病毒的制备方式对天然免疫信号通路的影响,具有一定的参考和借鉴作用.     

四株单纯疱疹病毒(HSV)感染 四种细胞的SDS—PAGE分析

我们用 SDS—PAGE 测定四株 HSV 感染 HEp—2细胞、HeLa 细胞、人胚肺细胞、鸡胚细胞多肽与未感染细胞多肽的电泳图结果表明:四小时感染细胞多肽与未感染细胞多肽相比未见有区别,二十四小时的感染细胞多肽与未感染细胞多肽的电泳带显示有所区别。其中感染 HSV—1和 HSV—2型的鸡胚细胞多肽电泳带亦显示有所区别。说明鸡胚细胞对 HSV—1型和 HSV—2型的敏感性差异,亦可以从电泳带反映出来,说明电泳带可作为 HSV 分型的又一方法。经蔗糖梯度超离心纯化的未标记同位素的地方株 HSV—1CC21株和 HSV—2W 株病毒颗粒用 SDS 裂解,并进行 SDS—PAGE 表明这两株病毒的结构多肽分别为12和15种,显示型的区别。本文还就关于在制备疫苗时以选择何种细胞为宜的问题进行了讨论。     

腮腺炎病毒抗血清的制备

以制备适用于疫苗生产检定中病毒鉴别试验和外源因子检查的高效价腮腺炎病毒抗血清为目的。用腮腺炎病毒接种SPF鸡胚尿囊腔,培养收取病毒尿液免疫SPF鸡,采集抗血清。腮腺炎病毒接种Vero细胞,培养病毒抗原经PEG沉淀,超速离心法纯化后免疫家兔采集抗血清。比较两种免疫方法所得病毒抗血清效价。结果显示SPF鸡抗腮腺炎病毒血清中和抗体GMT为1:1716,兔抗腮腺炎病毒血清中和抗体GMT为1:732。两种动物抗血清均适用于疫苗生产相关检定。免疫SPF鸡制备的病毒抗血清无特定病原及抗体污染,是毒种外源因子检测和疫苗鉴别试验的理想试剂。免疫SPF鸡制备病毒抗血清的程序简单,结果易于验证,有利于生物试剂标准化。     

MDCK单克隆细胞库的建立及检定

目的:建立甲型H1N1流感病毒疫苗株高适应MDCK单克隆细胞库,用于培养生产流感病毒疫苗,为细胞代替鸡胚制备流感病毒疫苗提供保证。方法:用有限稀释法将MDCK细胞进行单克隆化,通过血凝和TCID50筛选甲型H1N1流感病毒疫苗株的高适应性单克隆化细胞株,扩大培养建立细胞库并进行检定。结果:共制备了97株单克隆化MDCK细胞,筛选到甲型H1N1流感病毒疫苗株高适应性MDCK单克隆细胞株,扩大培养建立了细胞库,经检定符合《中华人民共和国药典.三部》(2010版)对细胞库的要求。结论:建立了甲型H1N1流感病毒疫苗株高适应性MDCK单克隆细胞库,为细胞培养生产流感病毒疫苗奠定了基础。     

鸡胚细胞的传代培养和麻疹病毒的增殖

为了建立原代鸡胚细胞的传代培养工艺,探究传代鸡胚细胞对麻疹病毒的敏感性和适应性,本研究将原代鸡胚细胞进行传代培养,分别采用原代鸡胚细胞和传代鸡胚细胞培养麻疹病毒沪-191(Shanghai-191,S-191)株毒种,并对病毒收获液进行滴度检测和基因序列测定。结果显示,原代鸡胚细胞可稳定传代培养至第10代,各代次细胞生长趋势相似;第5代鸡胚细胞染色体检查为正常染色体核型;第8代鸡胚细胞成瘤性检查未见成瘤;采用第3、5代鸡胚细胞制备的麻疹病毒滴度水平高于原代鸡胚细胞,但无显著性差异(n＝3,P＞0.05),编码病毒核蛋白(nucleoprotein,N)和血凝素蛋白(hemagglutinin,H)的基因序列与S-191株完全一致,未发生变异。本研究证实,原代鸡胚细胞可进行传代培养,各代次鸡胚细胞对麻疹病毒的敏感性不变,产毒水平无显著差异,可用于培养麻疹病毒。     

H3N2v疫苗病毒的鸡胚适应性突变能提高病毒收量,而又不失去抗原性和免疫原性

<正>最近发现的人畜共患H3N2型变异甲型流感病毒(H3N2v)已导致有记录的343例与猪接触相关的人类感染。需要研制针对病毒的有效疫苗,这些病毒可能获得了在人群中的遗传稳定性。然而,从人体中分离的病毒在鸡胚中不能良好复制,这构成了基于鸡胚的疫苗生产的障碍。为了解决这个问题,我们想得到表面蛋白的鸡胚适应性突变以在保留免     

抗体选择压作用下H9N2亚型禽流感病毒HA基因的变异

摘要：【目的】了解H9N2亚型禽流感病毒(AIV)在抗体选择压作用下的遗传变异。【方法】将制备疫苗用的LG1株H9N2亚型AIV分别接种含有母源抗体鸡胚(A组)和不含有母源抗体的SPF鸡胚(B组),并连续传代。其中A组再分为4 个独立的传代系列A1-4,B组再分为2 个独立传代系列B1-2。在每个传代系列,分别对第10,20,30,40,50 代病毒的HA基因进行扩增克隆测序,并与原始病毒的序列比较。【结果】LG1株H9N2在没有抗体的鸡胚的传代过程中,仅发生少数碱基的不稳定随机变异,且多为无义突变。在2 个传代系列的10 个代次病毒,共出现了29 个位点变异,有义突变(NS)与无义突变(S)比值NS/S为1.42 。但在有抗体的鸡胚的传代过程中,发生了多个呈现稳定遗传的有义突变。在4 个传代系列的20 个代次病毒,共出现了45 个位点变异,有义突变(NS)与无义突变(S)比值NS/S为3.46。【结论】在鸡胚传代过程中母源抗体提供的免疫选择压确实能影响H9N2的HA基因的变异。同时表明,带有母源抗体的鸡胚是实验室条件下研究免疫选择压对病毒抗原性变异影响的一种有效的实验模型。     

疫苗

<正> 考虑了用鸡细胞生产人病毒疫苗问题。使用30个11日龄Spafas Cofal种的无马立克病毒的有胚鸡蛋的鸡胚为原始细胞悬液,并使用Cutodex3微载体。培基中含胎牛血清,或胎牛血清、鸡血清和tryptose pho-sphate肉汤,以及组织培养瓶或     

表达绿色荧光蛋白重组新城疫病毒 LaSota疫苗株的构建

新城疫病毒是理想的新型活病毒疫苗载体,具有巨大的优势和应用前景。采用生产实践中广泛应用、免疫效果良好的NDV LaSota弱毒疫苗株,建立了反向遗传操作系统。在此基础上,进一步构建了表达绿色荧光蛋白(GFP)的重组NDV基因组cDNA克隆,成功救获了重组病毒rLaSota-EGFP,病毒F1代尿囊病毒液按1×104EID50接种9~10日龄SPF鸡胚尿囊腔,接种后分别于24h、48h、72h及96h收获尿囊液,检测平均HA滴度分别为28、210.3、211.3和211,每mL尿囊液病毒量EID50分别为108.64、109.22、109.21和109.64,重组病毒与亲本株生长滴度在相近时间达到峰值,生长动力学特性与亲本株无明显差异。各代次重组病毒按1×106EID50病毒量接种9~10日龄SPF鸡胚,96h内完全不致死鸡胚。救获重组病毒保持了LaSota弱毒疫苗亲本毒株对鸡胚良好的高滴度生长适应和低致病特性,并且鸡胚连续传9代次仍保持GFP的稳定表达及生物学特性不变。重组病毒rLaSota-EGFP的成功救获为开展新城疫病毒活载体疫苗研制提供了可行的技术平台。     

共表达HIV-1中国流行株gp120与IL-18重组鸡痘病毒的构建及其免疫原性观察

为筛选我国的HIV-1疫苗候选株,以鸡痘病毒282E4中国疫苗株为载体,构建了共表达中国流行株HIV-1外膜蛋白gp120和IL-18的重组鸡痘病毒,并将该重组鸡痘病毒免疫BALB/c小鼠,检测免疫小鼠脾特异性CTL杀伤活性和血清抗体水平。结果显示,HIV_1外膜蛋白gp120和IL-18不但可在重组鸡痘病毒感染的鸡胚成纤维细胞中表达,而且可在重组鸡痘病毒感染的哺乳动物细胞中表达。重组病毒具有良好的免疫原性,可诱导小鼠产生特异性抗体和脾特异性CTL反应,且IL_18发挥了免疫佐剂的作用。本研究结果为制备安全、有效的HIV-1基因工程活载体疫苗奠定了基础。     

Neutrino intergalactic communication,metal life,and viruses: Part 1 quo vadis ex machina

At one spectrum extreme, Astrobiology conjectures that for exoplanets with Goldilocks conditions, terrestrial-like life is inevitable. Moreover, it is envisaged that via panspermia, terrestrial-like life and its precursors are transferred among galaxies, stars, and within solar systems via transiting comets, asteroids, and planetoids. In addition, expelled stars, which have solar systems, it is inferred, transfer life as well. However, at the other extreme, we propose a paradigm shift that on some planets, subject to non- Goldilocks conditions, metal machine life could arise, ab initio, and evolve viruses, intelligence, and civilizations, conjointly. Accordingly, intelligent mechanized civilizations could readily and efficiently commence space exploration. Furthermore, as a counter paradigm shift, such civilizations could experiment and produce non-metallic life, based on carbon and other non-metal elements, under suitable conditions, related to Goldilocks life. Even a single example of validated interstellar or intergalactic communication received on the Earth would support the existence of life elsewhere. However, the communication platform should not be restricted to electromagnetic radiation. Other platforms should be included as well - one such example, which would require sophisticated technology, is neutrino communication. This is the case for any advanced civilization, be it metal-machine based, biological-based, and carbon-based. In sum, civilizations based on machine life, would be highly productive due to the longevity and hardiness of machine life. However, significant caveats are raised in this brief report, because possibly dissimilar psychologies and intelligence may lead to conflicts between metal machine life and biological life, inter-paradigm conflict.     

Molecular phylogeny of the gobioid fishes (Teleostei: Perciformes: Gobioidei)

The phylogeny of groups within Gobioidei is examined with molecular sequence data. Gobioidei is a speciose, morphologically diverse group of teleost fishes, most of which are small, benthic, and marine. Efforts to hypothesize relationships among the gobioid groups have been hampered by the prevalence of reductive evolution among goby species; such reduction can make identification of informative morphological characters particularly difficult. Gobies have been variously grouped into two to nine families, several with included subfamilies, but most existing taxonomies are not phylogenetic and few cladistic hypotheses of relationships among goby groups have been advanced. In this study, representatives of eight of the nine gobioid familes (Eleotridae, Odontobutidae, Xenisthmidae, Gobiidae, Kraemeriidae, Schindleriidae, Microdesmidae, and Ptereleotridae), selected to sample broadly from the range of goby diversity, were examined. Complete sequence from the mitochondrial ND1, ND2, and COI genes (3573 bp) was used in a cladistic parsimony analysis to hypothesize relationships among the gobioid groups. A single most parsimonious topology was obtained, with decay indices indicating strong support for most nodes. Major phylogenetic conclusions include that Xenisthmidae is part of Eleotridae, and Eleotridae is paraphyletic with respect to a clade composed of Gobiidae, Microdesmidae, Ptereleotridae, Kraemeriidae, and Schindleriidae. Within this five-family clade, two clades are recovered. One includes Gobionellinae, which is paraphyletic with respect to Kraemeriidae, Sicydiinae, Oxudercinae, and Amblyopinae. The other contains Gobiinae, also paraphyletic, and including Microdesmidae, Ptereleotridae, and Schindleriidae. Previous morphological evidence for goby groupings is discussed; the phylogenetic hypothesis indicates that the morphological reduction observed in many goby species has been derived several times independently.     

Oxypred： Prediction and Classification of Oxygen-Binding Proteins

This study describes a method for predicting and classifying oxygen-binding pro- teins. Firstly, support vector machine (SVM) modules were developed using amino acid composition and dipeptide composition for predicting oxygen-binding pro- teins, and achieved maximum accuracy of 85.5% and 87.8%, respectively. Sec- ondly, an SVM module was developed based on amino acid composition, classify- ing the predicted oxygen-binding proteins into six classes with accuracy of 95.8%, 97.5%, 97.5%, 96.9%, 99.4%, and 96.0% for erythrocruorin, hemerythrin, hemo- cyanin, hemoglobin, leghemoglobin, and myoglobin proteins, respectively. Finally, an SVM module was developed using dipeptide composition for classifying the oxygen-binding proteins, and achieved maximum accuracy of 96.1%, 98.7%, 98.7%, 85.6%, 99.6%, and 93.3% for the above six classes, respectively. All modules were trained and tested by five-fold cross validation. Based on the above approach, a web server Oxypred was developed for predicting and classifying oxygen-binding proteins(available from http://www.imtech.res.in/raghava/oxypred/).     

Endogenous opiates: 1984

This paper is the seventh in an annual series of reviews of research involving the endogenous opiate peptides, each installment being restricted to work published during the previous year. As in the past three years, the review this year is limited to non-analgesic and behavioral studies of the opiate peptides. The specific topics this year include: stress, tolerance and dependence, consummatory responses, gastric and renal activity, alcohol, mental illness, learning and memory, cardiovascular responses, respiratory effects, thermoregulation, seizures and neurological disorders, activity, and miscellaneous other topics.     

External exoskeletal cavities in Coleoptera and their possible mycangial functions

This paper reviews the occurrence of external exoskeletal cavities in beetles and provides critical reassessment of their possible mycangial function. In most reported cases, the decision to attribute mycangial function to exoskeletal cavities was based solely on the combination of two factors: (i) observation of these cavities on beetle's body; and (ii) knowledge that this particular beetle species uses fungi as a food source. Such reasoning resulted in the assumption, occasionally premature and not supported by other evidence, that exoskeletal pits in the following families may function as mycangia: Rhysodidae, Ptiliidae, Staphylinidae, Hybosoridae, Scarabaeidae, Derodontidae, Ptinidae, Jacobsoniidae, Boganiidae, Cryptophagidae, Endomychidae, Erotylidae, Latridiidae, Nitidulidae, Phloeostichidae, Silvanidae, Sphindidae, Pyrochroidae, Anthribidae, Attelabidae and Curculionidae. We conclude that only two beetle families include species with adequately documented cases of external exoskeletal mycangia: (i) Curculionidae (some Scolytinae and Platypodinae); and (ii) the structurally complex mycangia of Attelabidae ( Euops females). One or more species of Sphindidae, Erotylidae, Silvanidae, and Latridiidae have likely functional mycangia. Exoskeletal pits with uncertain function are additionally reported from the following families: Cupedidae, Ommatidae, Lepiceridae, Carabidae, Histeridae, Hydraenidae, Leiodidae, Elmidae, Artematopodidae, Throscidae, Elateridae, Rhinorhipidae, Biphyllidae, Cerylonidae, Cyclaxyridae, Monotomidae, Mycetophagidae and Zopheridae. We also discuss the transport of fungal spores and conidia by waxy exudates or debris build up on beetle exoskeleton, as well as their passive attachment to the body.     

国内8款常用植物识别软件的识别能力评价

随着智能手机和人工智能技术的发展, 以手机app为载体的植物识别软件慢慢走进公众生活、科普活动和科研活动的各个方面。植物识别app的识别正确率是决定其使用价值和用户体验的关键因素。目前, 国内应用市场上有许多植物识别app, 它们的开发目的和应用范围各异, 软件本身的关注点、数据库来源、算法、硬件要求也存在很大差异。对于不同人群, 植物识别app有不同的意义, 如对于科研人员来说, 识别能力强的app是提高效率的一大工具; 对植物爱好者来说, 具一定准确率的识别app可以作为入门的工具。因此, 对各app的识别能力进行分析与评价显得尤为重要。本文选取了8款常用的app, 分别对400张已准确鉴定的植物图片进行识别, 其中干旱半干旱区、温带、热带和亚热带4个区各选取100张。这些图片共计122科164属340种, 涵盖了乔木、灌木、草本、草质藤本和木质藤本5种生长型, 包含23种国家级保护植物。种、属、科准确识别正确分别计4分、2分、1分, 以此标准对软件识别能力按总得分进行排序, 正确率得分由高到低依次为花帮主、百度识图、花伴侣、形色、花卉识别、植物识别、发现识花、微软识花。     

海蛎壳生物质资源再利用的研究进展*

海蛎是一种营养和医药价值较高的咸水双壳类动物,在世界各地被广泛养殖。海蛎壳副产品是一种天然生物质资源,由95%的碳酸钙和5%的有机基质组成。海蛎壳的多尺度、多层次“砖-泥”独特结构,使其具有良好的机械稳定性、生物相容性、可降解性和优异的吸附特性。首先,介绍了海蛎壳生物质的理化性质和天然独特微纳米结构,总结了海蛎壳在农业、工业、生物医药领域的研究现状,详细阐述了其在污水治理、土壤改良、天然抗菌剂(食品工业和生物医药)、骨组织工程、医药原料、生物填料、工业催化剂及分散载体、建筑工业填料、功能化涂料等领域的研究现状。其次,概述了利用生物转化技术将海蛎壳转化为生物能源、新型生物质材料等方面的研究进展。最后,展望了海蛎壳生物质资源及其衍生物未来在工业、农业、医药领域的潜在应用。     

Materials for Chinese Phyllanthoideae

The present paper is part of taxonomic study on Chinese Phyllanthoideae. Included in it are two new varieties, Leptopus esquirolii var. villosus and Drypetes hainanensis var. longistipitata, one new combination, Glochidion triandrum var. siamense, and seven new records in China: Drypetes salicifolia, D. hoaensis. Actephila subsessilis, Glochidion khasicum, G. nubigennum, Bridelia spiosa and B. poilanei. In addition, seventeen taxon names are newly reduced: Liodendron formosanum = Drypetes formosana, Liodendron matsumurae = Drypetes matsumurae, D. longipes = D. indica, Antidesma paxii = A. acidum, A. hiiranense, A. filipes and A. pentandrum var. hiiranense = A. japonicum, A. calvescens = A. montanum, A. microphyllum = A. venosum, Breynia stipitata var. formosana and B. jormosana = B. vitis-idaea, Glochidion zeylanicum var. tomentosum = G. hirsutum, G. rubidulum = G. thomsonii, G. acuminatum = G. triandrum, G. fagifolium and Phyllanthus fagifolius = Glochidion sphaerogynum, Bridelia penangiena = B. insulana, B. henryana = B. tomentosa. All the types are kept in SCBl and PE.     

Allenic and cumulenic lipids

Nowadays, about 200 natural allenic metabolites, more than 2700 synthetic allenic compounds, and about 1300 cumulenic structures are known. The present review describes research on natural as well as some biological active allenic and cumulenic lipids and related compounds isolated from different sources. Intensive searches for new classes of pharmacologically potent agents produced by living organisms have resulted in the discovery of dozens of such compounds possessing high anticancer, cytotoxic, antibacterial, antiviral, and other activities. Known allenic and cumulenic compounds can be subdivided on several structural classes: fatty acids, hydrocarbons, terpenes, steroids, carotenoids, marine bromoallenes, peptides, aromatic, cumulenic, and miscellaneous compounds. This review emphasizes the role of natural and synthetic allenic and cumulenic lipids and other related compounds as an important source of leads for drug discovery.