血清10型鸭疫里默氏杆菌灭活油乳剂疫苗的研制

鸭疫里默氏杆菌(Riemerella anatipestifer,RA)感染是主要危害2~8周龄雏鸭的高致病性细菌性传染病。为研制血清10型RA灭活油乳剂疫苗,选取10株血清10型RA分离株进行动物体复壮、分离和鉴定后,分别对部分毒株进行了毒力测定和灭活油乳剂疫苗的研制。结果表明,所有试验用分离株对7日龄雏鸭均具有较强的致病力。分离株YXL1和HXb2的半数致死量分别为2.8×108 cfu和82 cfu,不同菌株之间的毒力差异很大。用研制的灭活油乳剂疫苗对雏鸭进行2次免疫后,免疫鸭可获得高水平的特异性抗体,并可抵抗强毒RA的攻击,以HXb2免疫后的攻毒保护性最好。抗体水平检测和攻毒保护试验结果表明,用血清10型RA分离株HXb2制备的油乳剂灭活疫苗具有良好的免疫保护作用。     

RGO/Nylon-6 composite mat with unique structural features and electrical properties obtained from electrospinning and hydrothermal process

In this work, the reduced graphene oxide (RGO) sheets were effectively uploaded through nylon-6 fibers using combined process of electrospinning and hydrothermal treatment. Good dispersion of graphene oxide (GO) with nylon-6 solution could allow to upload GO sheets through nylon-6 fibers and facilitate the formation of spider-wave-like nano-nets during electrospinning. GO sheets present on/into nylon-6 spider-wave-like nano-nets were further reduced to RGO using hydrothermal treatment. The impregnated GO sheets into nylon-6 nanofibers and their reduction during hydrothermal treatment were confirmed by FE-SEM, TEM, FT-IR and Raman spectra. The electrical characteristics of pristine nylon-6, GO/nylon-6 and RGO/nylon-6 nanofibers were investigated and it was found that RGO/nylon-6 composite mat had better electrical conductivity than others. The formation of spider-wave-like nano-nets as well as indirect route of incorporation of RGO sheets on electrospun nylon-6 mat may open a new direction for future graphene/polymer electronics.     

Expression of CD4 T cell cytokine genes in the colorectal mucosa of inflammatory colorectal polyps in miniature dachshunds

Inflammatory colorectal polyps (ICRPs) in miniature dachshunds are recently recognized as a major cause of large bowel diarrhea in this dog breed in Japan. ICRPs are characterized by the formation of multiple small polyps and a space-occupying large polyp in the colorectal area, and are thought to be a novel form of inflammatory bowel disease (IBD). In humans, specific cytokine patterns attributed to T helper (Th)1, Th17 and regulatory T cells have important roles in the pathogenesis of IBD. Thus, the aim of the present study was to assess the gene expression of cytokines of T cell subsets in the colorectal mucosa from dogs with ICRPs. Colorectal mucosal specimens from 10 dogs with ICRPs and 14 control dogs were used in this study. Interferon (IFN)-γ, interleukin (IL)-4, IL-17A and IL-10 mRNA expression was assessed using quantitative real-time PCR. IL-17A mRNA expression was significantly increased in large polyps compared to small polyps and controls. IFN-γ and IL-10 mRNA expression in large polyps were significantly higher than in controls. There was no significant difference in IL-4 mRNA expression among the three groups. IL-17A is thought to play important roles in the pathogenesis of ICRPs. IL-10 up-regulation could oppose the proinflammatory function of IL-17A.     

新城疫病毒ClassⅠ强毒株磷蛋白在细胞中的表达与定位

根据Class Ⅰ新城疫病毒分离株9a5b编码序列设计特异性引物扩增P蛋白基因,原核表达后利用纯化的蛋白免疫BALB/c小鼠制备单克隆抗体,经二次亚克隆后筛选到4株针对P蛋白的单克隆抗体,利用所得单抗进行P蛋白在细胞内定位分析,NDV 9a5b株按5 M.O.I感染量接种DF1单层细胞,当PI=4 h时,P蛋白呈点状弥散分布于细胞浆内;PI=6~8 h时,P蛋白聚集于细胞核周围;PI=12 h时,P蛋白呈单极化聚集于细胞核一侧;而PI=16 h后开始形成合胞体,P蛋白在单个细胞中仍呈现单极分布;当PI=48 h时,细胞核已发生碎裂,荧光强度有所减弱。本研究为深入开展P蛋白在病毒增殖以及细胞周期中的作用机制研究奠定了基础。     

青杄功能性状及其在各器官间分配格局的时空变化

为研究青杄在不同环境和发育阶段的适应性和生存策略,以关帝山典型森林分布区的青杄(Piceawilsonii)为对象,分析其叶片鲜干比、叶面积、叶绿素和C、N、P含量等重要功能性状在不同年龄(5、15、25、35、45a)和不同海拔梯度(1760、1820、1880、1940、2000m)上的变化,以及C、N、P在各器官间分配格局随年龄和海拔的时空变化。结果表明:1)随树龄的增加,青杄叶面积、叶鲜干比呈增大趋势,叶P和根P含量呈下降趋势,叶有机碳含量下调,但根有机碳在上调。2)随海拔增加,叶绿素、叶面积显著增加,叶有机碳呈下降趋势,而根有机碳在增加。叶、枝、根中的N、P含量均呈先增加后减小趋势,相比之下,N、P随海拔的变化较有机碳更为显著。3)随树龄增加,C、N、P在各器官间分配格局表现为:P在幼年时于叶、根中分配比例相对较大,成年时主要集中在枝中。在研究的5~45a范围内,N分配呈叶>枝>根的稳定格局,未随年龄而显著变化。有机碳则在各器官呈近乎均匀分配的基础上随年龄增加根中比例略有增大,相反叶中比例略有下降。C、N、P并未随海拔增加呈现各器官间分配格局上的显著变化。青杄各功能性状随年龄和海拔变化并通过在各器官间相互关联和协调分配来增强其生态适应性。     

An artificial inoculation protocol for Uromycladium acaciae,cause of a serious disease of Acacia mearnsii in southern Africa

Uromycladium acaciae is the cause of a severe wattle rust epidemic in plantations of Acacia mearnsii (black wattle) in southern Africa. Research on the biology of this damaging rust is assisting in the development of control strategies. One strategy under investigation is the identification and deployment of resistant lines of A. mearnsii. Selection of resistant families currently relies on large-scale, time-consuming and expensive field trials. In this study, we present a detailed artificial inoculation protocol for U. acaciae, which can be used to screen for resistance. The results of an experiment that used the protocol to screen the relative resistance of 12 families of A. mearnsii to U. acaciae are also presented. The developed artificial inoculation protocol can also be used to investigate several other aspects of this host–pathogen system.     

鸡TIGAR基因真核表达质粒的构建及其抗凋亡功能评价

【背景】 TP53诱导的糖酵解和凋亡调节因子（TP53-induced glycolysis and apoptosis regulator,TIGAR）是p53下游的靶基因,具有调节糖酵解水平和去除活性氧（Reactive oxygen species,ROS）并降低由活性氧诱发的细胞凋亡水平。【目的】 构建鸡源TIGAR基因的真核表达质粒,并检测TIGAR基因在DF1细胞中的抗凋亡作用,为建立稳定表达鸡TIGAR基因的细胞系做准备。【方法】根据GenBank（登录号：XM_417232.6）中预测基因设计引物,利用RT-PCR的方法从SPF鸡脾脏中扩增鸡TIGAR基因,将扩增产物克隆至载体（Flag-CMV14）后送公司测序验证;随后构建进化树对鸡TIGAR基因与其他哺乳动物及水生动物的TIGAR基因进行同源性分析。将重组质粒（Flag-TIGAR）转染入DF1细胞24 h后,使用新城疫病毒诱导细胞凋亡,利用Western Blot检测重组质粒表达情况以及Poly ADP-Ribose Polymerase（PARP）裂解情况。此外还将重组质粒（Flag-TIGAR）转染入DF1细胞,并于收样前2 h使用Staurosporine刺激细胞发生凋亡,分别在转染后24、48 h收集样品,并用流式细胞仪检测细胞凋亡情况。【结果】 RT-PCR扩增TIGAR基因,在843 bp处出现目的条带与预测相符,构建的TIGAR真核表达质粒（Flag-TIGAR）经测序,结果显示其序列与GenBank上预测的基本一致。Western Blot结果显示在30 h、36 h收集的样品中PARP均被裂解且转染重组质粒（Flag-TIGAR）的实验组与未转染质粒（MOCK）组或转染空载体（Flag-CMV14）组的样品相比,裂解的PARP表达量明显降低,且差异极显著（P<0.01）。流式结果显示：24 h 检测转染Flag-CMV14后细胞总凋亡率为11%（早期凋亡7.8%,晚期凋亡3.2%）,而转染Flag-TIGAR后细胞总凋亡率仅为4%（早期凋亡3.7%,晚期凋亡0.3%）,转染Flag-CMV14的早期凋亡率和晚期凋亡率均高于转染Flag-TIGAR组,且差异显著（P<0.05）。48 h转染Flag-CMV14后细胞总凋亡率为20.3%（早期凋亡14.3%,晚期凋亡6.0%）,而转染Flag-TIGAR后细胞总凋亡率为6.4%（早期凋亡4.8%,晚期凋亡1.6%）,转染Flag-CMV14组的细胞早期凋亡和晚期凋亡率均高于转染Flag-TIGAR的组,且差异极显著（P<0.01）。【结论】成功扩增出鸡TIGAR基因,并构建了其真核表达质粒,通过Western Blot和流式实验均证实过表达TIGAR后可降低细胞的凋亡程度并有利于细胞存活。     

“稻—小龙虾”连作生态种养技术

稻虾连作作为一种绿色高效的农业生产模式,提高了土地资源利用率,产出的小龙虾品质更优,同时生产出绿色稻米,也解决了低湖田因惧怕洪涝威胁而长时间闲置的问题,技术简单可行,能达到稻丰虾肥的效果。介绍了稻虾田标准池田建设、小龙虾高产养殖技术和水稻高产栽培技术。     

Residual patterns of alkyl polyoxyethylene surfactant droplets after water evaporation

Using a nonionic, alkyl polyoxyethylene surfactant (X-77) in aqueous solutions, sessile droplet spreading, pinning, evaporation, contraction, and post-evaporation deposits are characterized. X-77 is widely used in the agricultural field as a spreader/adherent, intended to optimize pathenogenic agent coverage. Using a single droplet size under monitored temperature conditions, we control humidity, substrate hydrophobicity, and surfactant concentration to mimic varying agricultural conditions. For hydrophilic surfaces, the droplet spreads, reaching and retaining a maximum, stationary size. At this stage, a ring accretion occurs at the maximum spread diameter. During the final stage, the water film retracts, resulting in deposition of small islands of surfactant residue inside the ring. At lower concentrations of surfactant, we discover ring formations that break-up into "ring islands" at late-stage evaporation, accompanied by a distribution of the smaller islands in the interior portion of the substrate contact area. These are promoted by higher relative humidity. At higher concentrations, only a solid ring of surfactant remains, post-evaporation. Increasing surfactant concentration tends to increase the mean of the interior island size and to broaden the overall island size distribution. On sufficiently hydrophobic surfaces, surfactant-laden droplets do not evidence pinning, ring formations, or post-evaporation interior islands. Interestingly, lower humidity increases spreading at higher surfactant concentrations. Such pattern formations of surfactant deposit are reported for the first time and are of significance in projecting how surfactants such as X-77 distribute pesticides or other chemicals on leaf surfaces.     

Worldwide genotyping of castor bean germplasm (<Emphasis Type="Italic">Ricinus communis</Emphasis> L.) using AFLPs and SSRs

Worldwide genetic diversity in 200 individuals comprising 41 castor bean accessions was assessed using amplified fragment polymorphisms (AFLPs) and simple sequence repeats (SSRs). We found that, despite surveying five continents and 35 countries, genetic diversity in castor bean germplasm is relatively low (overall H _e = 0.126 for AFLPs and 0.188 for SSRs) compared to estimates of genetic diversity in other plant species. Our data also show no geographic structuring of genotypes across continents or countries within continents. An assessment of the congruence between AFLP and SSRs indicates a low correlation (R ² = 0.19) between the two data sets, but each marker class nonetheless shows similar patterns of low-genetic diversity and a lack of geographic structure. Our data do suggest that SSRs yield a higher percentage of polymorphic loci, higher heterozyosity and a greater range of genetic distances, and are therefore more informative than are AFLPs on a locus-by-locus basis. Based on comparisons with numerous other plant species, we suggest that the lower genetic variation in this worldwide collection may be due to one or more factors including: sampling strategies that have not captured the full extent of genetic variation in the species; artifactual variation due to long-term germplasm storage and seed regeneration; or intense selection followed by domestic cultivation of a limited number of castor bean genotypes, which are widely propagated for their horticultural and agro-economic value.