重组牛皮蝇素A蛋白的免疫试验

用由毕赤酵母表达的皮蝇素A（Hypdermin A，HA）蛋白分别免疫新西兰大白兔和牦牛，了解该蛋白免疫效果。用两种不同分子量的重组HA蛋白分别免疫新西兰大白兔1次和2次，ELISA测其抗体消长规律。结果表明，3周后抗体水平达到峰值，后缓慢下降。50KD蛋白免疫效果较30KD蛋白好，差异非常显著（P〈0．01）。用其中50KD的重组HA蛋白分别在7月和9月免疫自然感染牛皮蝇蛆病牦牛，观察感染率和感染强度，ELISA测其抗体消长规律。结果表明，7月免疫组与9月免疫组感染率分别为83．3％和36．3％（对照组为55．5％）；平均感染强度分别为6．5和4．2（对照组为13．0），差异不显著（P〉0．05）。免疫后，抗体水平上升，3个月后缓慢下降。9月免疫组免疫效果较7月免疫组好，三组抗体水平差异非常显著（P〈0．01）。上述研究结果为防治牛皮蝇蛆病疫苗的深入研究和实践运用提供了参考依据。     

玉米灰斑病菌生理分化的鉴别寄主体系

玉米灰斑病现已成为北方玉米产区的主要病害之一，但国内外关于玉米灰斑病菌Cercospora zeaemaydis Tehon ＆ Daniels致病力分化等方面的研究较少，尚未建立该病菌生理分化的鉴别寄主系统，无法明确我国是否存在玉米灰斑病菌致病性分化类群或生理小种及优势菌群和分布，限制了有针对性地进行品种合理布局和抗病育种工作，因此研究玉米灰斑病菌致病性分化鉴别寄主体系是一项重要的基础性工作。作者采用田间成株期鉴定技术，对不同的玉米灰斑病菌菌株进行致病力划分，研究其鉴别寄主体系。     

Yield-height correlation and QTL localization for plant height in two lowland switchgrass populations

Switchgrass (Panicum virgatum L.), as a model herbaceous crop species for bioenergy production, is targeted to improve biomass yield and feedstock quality. Plant height is a major component contributing to biomass yield. Accordingly, the objectives of this research were to analyze phenotypic variation for biomass and plant height and the association between them and to localize associated plant height QTLs. Two lowland switchgrass mapping populations, one selfed and another hybrid population established in the field at Perkins and Stillwater, Oklahoma, were deployed in the experiment for two years post establishment. Large genetic variation existed for plant biomass and height within the two populations. Plant height was positively correlated with biomass yield in the selfed population (r = 0.39, P<0.0001) and the hybrid population (r = 0.41, P<0.0001). In the selfed population, a joint analysis across all environments revealed 10 QTLs and separate analysis for each environment, collectively revealed 39 QTLs related to plant height. In the hybrid population, the joint analysis across overall environments revealed 35 QTLs and the separate analysis for each environment revealed 38 QTLs. The findings of this research contribute new information about the genetic control for plant height and will be useful for future plant breeding and genetic improvement programs in lowland switchgrass.     

鸡胚外泌体miRNAs功能分析

旨在基于鸡胚外泌体及所含有的miRNAs解析其产生生理功效的活性物质基础及作用机制。本研究收集13胚龄惠阳胡须鸡胚蛋18枚（3组,每组6枚）,采用酶消化、蔗糖密度梯度和差速超速离心等方法分离鸡胚组织来源的外泌体,经透射电镜、纳米流式和标志蛋白流式检测等方法鉴定其特征。利用高通量测序方法研究外泌体样本中miRNAs表达谱,对各样本top 10且共表达的miRNAs进行靶基因预测与功能分析。结果表明,分别获得了3个浮力密度范围（S1：1.13 g·mL^-1、S2：1.21 g·mL^-1和S3：1.32 g·mL^-1）具有外泌体典型特征的胞外囊泡;与miRBase V21进行比对发现,S1、S2和S3样品中分别含有675、661和845个已知的miRNAs,这些miRNAs中有488个（52.6%）三者共有,包括了广泛参与细胞增殖、分化和免疫应答等过程的功能性miRNAs。TargetScan 7.2和miRDB预测得到3个样本中top 10且共表达的56个miRNAs的靶基因,获得的3 650个靶基因经DAVID分析,显著富集了24个与机体生长、发育、免疫等有关的生物学通路（P<0.05）,包括MAPK信号通路、mTOR信号通路、Notch信号通路、Wnt信号通路和GnRH信号通路等。本研究建立了鸡胚组织中有效分离外泌体的方法,发现鸡胚组织来源的外泌体miRNAs会影响机体生长发育和免疫调节有关的生物学通路。     

燕山板栗种质资源遗传多样性的RAPD分析

为研究燕山板栗的遗传多样性,采用随机扩增多态DNA(randomamplifiedpolymorphicDNA,RAPD)技术对36份燕山板栗种质进行了分析。分析了燕山板栗的遗传丰富度,并对包括36个燕山板栗品种和8份外来板栗品种在内的44份板栗种质进行聚类分析。结果表明,RAPD能有效地区分品种间的差异,用16个随机引物经PCR扩增共得到132个片段,其中有83个多态性片段,占62.9%;不同遗传位点之间遗传多样度最大可达0.444,最小值为0.096,平均多样度为0.187;UPGMA法聚类,将44份板栗种质聚成4个大的类群,36份燕山板栗可分为3个大的类群,外来种质聚为一类。燕山板栗明显不同于外来品种。在RAPD图谱中,找到了19个品种(类型)的特异性标记和标记组合,可作为品种(类型)分子鉴别的依据。     

菜豆种质资源对枯萎病菌的抗性研究

对供试的21份菜豆种质资源材料的幼苗采用下胚轴双孔注射法进行接种菜豆枯萎病菌(Fusarium oxyxporum f.sp.phaseoli),接种7 d后调查下胚轴维管束发病情况,计算病情指数,其中感病材料2份;中抗枯萎病材料11份;抗枯萎病材料7份;高抗枯萎病材料1份.     

团头鲂幼鱼饲料中α-亚麻酸、亚油酸的适宜含量

在半纯化饲料配方的基础上,分别设计6个α-亚麻酸含量(0.02%、0.55%、1.08%、1.60%、2.13%、2.65%)、6个亚油酸含量(0.86%、1.29%、1.73%、2.16%、2.59%、3.03%),以亚麻籽油、玉米油、棕榈油调节饲料中α-亚麻酸、亚油酸的含量,配制等氮等能(粗蛋白质含量为30.09%,粗脂肪含量为6.87%)的12种半纯化试验饲料,探讨团头鲂幼鱼[初始均重为(59.5±0.5)g]饲料中α-亚麻酸、亚油酸的适宜含量。养殖试验分为α-亚麻酸和亚油酸试验2部分,均设6组,每组4个重复,每个重复20尾,养殖周期为85 d。结果表明:在α-亚麻酸试验中,依据回归方程计算得到,在饲料α-亚麻酸含量分别为1.32%、1.33%时,团头鲂幼鱼具有最大的特定生长率和最小的饲料系数;0.02%组的脏体指数显著低于除1.08%组外的其他各组(P0.05),肥满度、肝体指数各组间无显著差异(P0.05);0.02%组血清总胆固醇、甘油三酯含量显著高于0.55%、1.08%组(P0.05),1.60%组血清高密度脂蛋白含量显著高于0.55%、2.13%组(P0.05),0.02%组血清低密度脂蛋白含量显著高于其他各组(P0.05)。在亚油酸试验中,依据回归方程计算得到,在饲料亚油酸含量分别为2.02%、2.03%时,团头鲂幼鱼具有最大的特定生长率和最小的饲料系数;3.03%组的肝体指数显著低于其他各组(P0.05),肥满度、脏体指数各组间无显著差异(P0.05);1.29%组血清总胆固醇含量显著高于1.73%组(P0.05),0.86%组血清甘油三酯含量显著高于1.73%组(P0.05),1.29%组血清高密度脂蛋白含量显著高于0.86%、3.03%组(P0.05),各组血清低密度脂蛋白含量无显著差异(P0.05)。以特定生长率、饲料系数作为主要评价指标,结合部分血清生化指标和形体指标,得到适合团头鲂幼鱼快速生长、维持鱼体正常健康的饲料中适宜的亚麻酸、亚油酸含量分别为1.32%~1.33%、2.02%~2.03%。     

葡萄霜霉菌候选效应子RXLR5信号肽的鉴定

利用Getorf、SignalP 3.0、BLASTP等生物信息学软件和PERL语言从葡萄霜霉菌基因组中预测到一条编码RXLR-EER结构域的效应蛋白候选基因,将该基因编码的蛋白命名为RXLR5。将RXLR5信号肽SP5编码序列连接到pSUC2T7M13ORI载体并转化到酵母蔗糖酶缺陷菌株YTK12后,重组菌株能成功分泌蔗糖酶,促使棉籽糖分解成单糖,因此,能在YPRAA培养基上正常生长;同时生成的单糖能与TTC反应产生红色不溶于水的氯化三苯基四氮唑。由此推测,SP5具有信号肽活性,可能在RXLR5从葡萄霜霉菌细胞内泌到胞外的过程中起重要作用。     

调亏灌溉和氮处理对小桐子生长及水分利用的影响

为了研究阶段水分亏缺(亏水+不亏水W_LW_H、不亏水+亏水W_HW_L、亏水+亏水W_LW_L、不亏水+不亏水W_HW_H)和不同施氮量(N_Z:0,N_L:0.2 g/kg,N_H:0.4 g/kg)对小桐子生长、生理指标和灌溉水利用效率的影响,通过盆栽试验,发现W_LW_H的生长和灌溉水利用效率均显著高于W_HW_L;作物的灌溉水利用效率随施氮量的增大而呈现先增大后减小的趋势,在N_L水平下达到最大值;与高水高氮的处理N_HW_HW_H相比,中水低氮处理N_LW_LW_H节约灌溉水27%,节约氮肥使用量50%,小桐子株高减少31%、总干物质量减少35%,灌溉水利用效率减少13%,但茎粗增加13%,根冠比增加20%.可见小桐子在第一阶段处理(40~90 d)幼树期对水分的需求量较小,适度的亏缺灌溉可提高灌溉水利用效率;小桐子在第二阶段处理(90~140 d)处于旺长期,对水分的需求量较大,增大灌水量可大幅度促进小桐子生长及其干物质量的积累.全生育期实施亏缺灌溉,可提高小桐子自身适应外界环境能力,抗干旱胁迫能力也逐渐增强,但W_LW_L水平下的小桐子生长缓慢.经综合分析,认为处理N_LW_LW_H可作为干旱地区条件下的小桐子灌溉和施氮制度.     

Association of tumor budding with tumor-infiltrating lymphocytes and prognosis in breast cancer patients

AIM: To investigate the relationship of tumor budding with clinicopathologic parameters, tumor-infiltrating lymphocytes (TILs) of tumor microenvironment and the prognosis in breast cancer patients.METHODS: A total of 178 HE section samples were collected from the breast cancer patients treated with surgery in the First Affilated Hospital of Jinan University during Jan. 2012 to Dec. 2016. The tumor budding and stromal tumor-infiltrating lymphocytes were observed under light microscope. The correlation of tumor budding with the clinicopathologic status and TILs were analyzed by χ² test. Kaplan-Meier survival analysis and Log-rank test were used to estimate the disease-free survival (DFS) and overall survival (OS).RESULTS: High tumor budding level was associated with more positive lymph nodes, higher grade, and more lymphovascular invasion. In addition, the patients with higher tumor budding level showed fewer TILs, while the patients with lower tumor budding level had more TILs. Furthermore, the patients with higher tumor budding level had a worse disease-free survival and overall survival than those with lower tumor budding level.CONCLUSION: Tumor budding is significantly associated with adverse clinicopathological characteristics of breast cancer and negatively correlated with TILs. Therefore, tumor budding may serve as a potential biomarker to predict the prognosis of breast cancer.