基于高通量测序技术的微生物检测数据分析方法

高通量测序技术的发展正在逐渐改变诸多生物学领域的研究方法.为应对突发疫情以及新发未知微生物威胁的需求,微生物鉴定技术逐渐从传统的物理化学方法及核酸杂交等分子水平方法进一步走向利用无需培养的测序数据进行快速分析检测.随之而来的是对高通量数据分析在精度及速度的要求.基于高通量测序数据的微生物检测数据分析方法在近些年得到了快速的发展.本文分析了目前基于高通量测序数据的微生物检测数据分析方法,对其数据分析的处理流程和计算方法进行了研究,比较了各个微生物检测数据分析方法的特点及适用场景.最后结合本实验室工作总结微生物检测数据分析方法在实际应用中可能遇到的问题,希望对该应用领域的研究有一定的参考意义.     

胡杨无性系幼苗响应盐胁迫的miRNA表达差异研究

植物miRNA在调控基因表达、细胞周期、生物体发育、抗逆等方面起重要作用。为研究胡杨(Populus euphratica Oliv.)的耐盐机制,以1年生胡杨无性系幼苗为材料,构建具有空间代表性的盐胁迫胡杨cDNA文库,利用二代测序技术测定NaCl胁迫下和正常培养条件下胡杨叶和根miRNA表达情况。结果表明,不同的miRNA之间表达量存在明显差异,表达丰度最高的miRNA有miR156、miR157、miR165、miR166和miR167等,合计占总表达量的90%以上。胡杨根部存在特异表达的miRNA,在整个耐盐调控机制中发挥着生理调节、分子调控和信号传导等极为重要的作用。盐处理样品中发现大量响应盐胁迫的miRNA,对这些转录因子进行靶基因预测和注释后,发现很多盐胁迫响应的miRNA与NAC和SPL等重要转录因子家族相关,与前人的结论一致,另外还发现许多miRNA的调控对象是ATP酶和激素响应因子。     

Tonsilar NK Cells Restrict B Cell Transformation by the Epstein-Barr Virus via IFN-��

Cells of the innate immune system act in synergy to provide a first line of defense against pathogens. Here we describe that dendritic cells (DCs), matured with viral products or mimics thereof, including Epstein-Barr virus (EBV), activated natural killer (NK) cells more efficiently than other mature DC preparations. CD56^brightCD16⁻ NK cells, which are enriched in human secondary lymphoid tissues, responded primarily to this DC activation. DCs elicited 50-fold stronger interferon-γ (IFN-γ) secretion from tonsilar NK cells than from peripheral blood NK cells, reaching levels that inhibited B cell transformation by EBV. In fact, 100- to 1,000-fold less tonsilar than peripheral blood NK cells were required to achieve the same protection in vitro, indicating that innate immune control of EBV by NK cells is most efficient at this primary site of EBV infection. The high IFN-γ concentrations, produced by tonsilar NK cells, delayed latent EBV antigen expression, resulting in decreased B cell proliferation during the first week after EBV infection in vitro. These results suggest that NK cell activation by DCs can limit primary EBV infection in tonsils until adaptive immunity establishes immune control of this persistent and oncogenic human pathogen.     

Tonsilar NK cells restrict B cell transformation by the Epstein-Barr virus via IFN-gamma

Cells of the innate immune system act in synergy to provide a first line of defense against pathogens. Here we describe that dendritic cells (DCs), matured with viral products or mimics thereof, including Epstein-Barr virus (EBV), activated natural killer (NK) cells more efficiently than other mature DC preparations. CD56(bright)CD16(-) NK cells, which are enriched in human secondary lymphoid tissues, responded primarily to this DC activation. DCs elicited 50-fold stronger interferon-gamma (IFN-gamma) secretion from tonsilar NK cells than from peripheral blood NK cells, reaching levels that inhibited B cell transformation by EBV. In fact, 100- to 1,000-fold less tonsilar than peripheral blood NK cells were required to achieve the same protection in vitro, indicating that innate immune control of EBV by NK cells is most efficient at this primary site of EBV infection. The high IFN-gamma concentrations, produced by tonsilar NK cells, delayed latent EBV antigen expression, resulting in decreased B cell proliferation during the first week after EBV infection in vitro. These results suggest that NK cell activation by DCs can limit primary EBV infection in tonsils until adaptive immunity establishes immune control of this persistent and oncogenic human pathogen.     

JURASSIC PALAEONTINIDAE FROM CHINA AND THE HIGHER SYSTEMATICS OF PALAEONTINOIDEA (INSECTA: HEMIPTERA: CICADOMORPHA)

Abstract:  Genera of Palaeontinidae (Insecta, Hemiptera: Cicadomorpha) are reviewed based on some newly-discovered material. An updated list of palaeontinid genera is provided. The genus Palaeontinopsis Martynov is assigned to Palaeontinidae incertae sedis . A new genus and species, Ningchengia aspera gen. et sp. nov., is described based on forewings and hindwings from the Middle Jurassic of Daohugou, China. Fletcheriana colorata Wang, Zhang and Fang and F . minuta Wang, Zhang and Fang are transferred to Suljuktocossus and Ningchengia respectively, resulting in Suljuktocossus coloratus comb. nov. and Ningchengia minuta comb. nov. Papilioncossus Wang, Ren and Shih, 2007 syn. nov. is considered to be a junior synonym of Eoiocossus Wang and Zhang, 2006 and Quadraticossus Wang and Ren 2007 syn. nov. is a junior synonym of Sinopalaeocossus   Hong, 1983 . The genus Fletcheriana is believed to represent a transition between Dunstaniidae and Palaeontinidae. This result is consistent with the previous view that Dunstaniidae is ancestral to Palaeontinidae. Furthermore, Suljuktocossus is a transitional genus between Fletcheriana and Cretaceous palaeontinids. The fusion of veins RP and M₁ in the palaeontinid hindwings implies a change of the mechanism of flight and an improvement in flight ability. The presence of a reduced costal area and the rigid basal leading edge of the palaeontinid forewings suggest gain of lift.     

刺五加提取物对断奶仔猪血常规和抗氧化功能的影响

为研究日粮添加刺五加提取物对断奶仔猪血常规和抗氧化功能的影响,选择21日龄断奶仔猪60头,随机分为3个处理,每个处理20个重复,单栏饲养,分别饲喂基础日粮+0.1%刺五加提取物、基础日粮+0.02%粘杆菌素和基础日粮.于试验开始后第7、14和28 d每个处理随机选取5头试猪,前腔静脉采血,测定血常规和抗氧化指标.结果表明:刺五加提取物能显著改善断奶仔猪的抗病能力,减轻应激反应,机体抗氧化酶活性明显升高;第7 d时提取物组MID、PLT、MDA和NO含量显著小于基础日粮组(P＜0.05);第14 d时提取物组WBC显著小于、CAT活性显著大于基础日粮组(P＜0.05);第28 d时提取物组WBC、MID和GRA显著小于(P＜0.05),SOD活性大于基础日粮组(P＞0.05).     

花生种子含油量的遗传分析

花生是重要的油料作物,对花生种子含油量进行遗传分析具有重要意义.本研究以高油花生新品种SPI056(P1)和低油花生新品种花育17(P2)及其杂交组合的F1、F2群体为材料,应用P1、P2、F1和F2 4个世代的数量性状主基因+多基因混合遗传模型联合分离分析方法,对种子含油量进行了遗传分析.结果表明:花生种子含油量由两对加性-显性-上位性主基因+加性-显性多基因共同控制,主基因遗传率为72 55%.     

高灵敏焦测序结合生物发光分析仪检测甲型H1N1流感病毒

目的:建立基于核酸序列分析的快速、准确、低成本的甲型H1N1流感病毒检测方法。方法:通过优化焦测序反应体系中ATP硫酸化酶和荧光素酶的浓度,建立高灵敏的焦测序反应体系;将该体系应用于低成本、小型化的便携式生物发光分析仪,焦测序分析流感病毒M、NP、HA基因片段的核酸序列。结果:优化后的焦测序反应体系可检测低至10 fmol的DNA样本,检测灵敏度较传统焦测序提高了10倍以上。对两例样本进行检测,根据所测得的M、NP、HA基因特异性片段序列,可以确认其均为甲型H1N1感染;另外,对M2蛋白阻断剂耐药性标志位点(S31N突变)的测定结果显示该病毒存在S31N突变,为M2蛋白阻断剂耐药型。结论:高灵敏焦测序体系结合便携式生物发光分析仪成功实现了对甲型H1N1流感病毒快速、准确的低成本检测。     

G蛋白偶联受体与G蛋白相互作用的最新研究进展

传统观念认为,在激动剂作用下,G蛋白偶联受体(GPCRs)能够激活G蛋白的α亚基,从而使Gα亚基与Gβγ亚基分离,被激活的Gα亚基通过信号转导进一步参与细胞的生理过程。但是,最新研究发现GPCRs和G蛋白存在多种偶联关系,GPCRs不仅能够激活Gα亚基,还可以与Gβγ亚基相互靠近,甚至会使G蛋白亚基构象发生重排而不分离,这对于疾病发病机制的研究及新的药物靶点的发现具有重要意义。就GPCRs与G蛋白之间的相互作用以及最新研究技术作一简要综述。