一种制备鱼类染色体的新方法

自1966年Ojima建立鱼类染色体空气干燥法以来,鱼类染色体研究技术有了很大进展.目前有肾细胞培养法、上皮细胞培养法和外周血培养法等。细胞培养需要在有一定条件的实验室进行(如无菌操作),因而简便快速的制备鱼类染色体方法便不断产生,有PHA+秋水仙碱注射法、秋水仙碱体外注射法和CoCl+秋水仙碱体外注射等方法.本文介绍一种适合于在野外条件下进行的小型鱼类和鱼苗染色体制片方法。     

Identification of two critical amino acid residues of the severe acute respiratory syndrome coronavirus spike protein for its variation in zoonotic tropism transition via a double substitution strategy

Severe acute respiratory syndrome coronavirus (SARS-CoV) is a recently identified human coronavirus. The extremely high homology of the viral genomic sequences between the viruses isolated from human (huSARS-CoV) and those of palm civet origin (pcSARS-CoV) suggested possible palm civet-to-human transmission. Genetic analysis revealed that the spike (S) protein of pcSARS-CoV and huSARS-CoV was subjected to the strongest positive selection pressure during transmission, and there were six amino acid residues within the receptor-binding domain of the S protein being potentially important for SARS progression and tropism. Using the single-round infection assay, we found that a two-amino acid substitution (N479K/T487S) of a huSARS-CoV for those of pcSARS-CoV almost abolished its infection of human cells expressing the SARS-CoV receptor ACE2 but no effect upon the infection of mouse ACE2 cells. Although single substitution of these two residues had no effects on the infectivity of huSARS-CoV, these recombinant S proteins bound to human ACE2 with different levels of reduced affinity, and the two-amino acid-substituted S protein showed extremely low affinity. On the contrary, substitution of these two amino acid residues of pcSARS-CoV for those of huSRAS-CoV made pcSARS-CoV capable of infecting human ACE2-expressing cells. These results suggest that amino acid residues at position 479 and 487 of the S protein are important determinants for SARS-CoV tropism and animal-to-human transmission.     

蚂蚁光顾云南紫胶虫对其天敌紫胶黑虫种群的影响

为了弄清蚂蚁光顾云南紫胶虫Kerria yunnanensis Ouet Hong对其天敌紫胶黑虫Holcocera pulverea Meyr种群的影响,于云南紫胶虫成虫期,对有无蚂蚁光顾的胶被抽样,调查胶被上紫胶黑虫的为害率及种群数量变化。结果显示,紫胶黑虫对有无蚂蚁光顾的胶被的为害率均很高,并逐月增加;其中有蚂蚁光顾的胶被紫胶黑虫的为害率略小于无蚂蚁光顾的胶被。蚂蚁光顾能明显减少紫胶黑虫的种群数量(︱t︱=2.764,df=356,P<0.01),其原因可能是蚂蚁光顾能干扰紫胶黑虫的产卵行为、破坏卵、取食卵和幼虫并且这种保护主要发生在云南紫胶虫幼虫期;云南紫胶虫进入成虫期后,由于紫胶黑虫生活于胶被内,并且产卵于胶被的凹陷处或雄虫胶壳内或雌虫肛突孔处,蚂蚁很少与紫胶黑虫相遇,故蚂蚁光顾对紫胶黑虫每月增长量没有显著影响(︱t︱=0.970,df=161,P>0.05)。紫胶黑虫和蚂蚁相遇的行为反应存在显著差异(χ2=4.781,df=1,P<0.05),蚂蚁对紫胶黑虫有捕食作用。蚂蚁与云南紫胶虫之间存在互利关系。蚂蚁取食云南紫胶虫的蜜露,能降低紫胶黑虫的为害率,并减少紫胶黑虫的种群数量,从而保护云南紫胶虫。     

Expression profiling and binding properties of fibrinogen-related proteins (FREPs), plasma proteins from the schistosome snail host Biomphalaria glabrata

A growing body of evidence suggests an important role for fibrinogen-like proteins in innate immunity in both vertebrates and invertebrates. It has been shown that fibrinogen-related proteins (FREPs), plasma proteins present in the freshwater snail Biomphalaria glabrata, the intermediate host for the human blood fluke Schistosoma mansoni, are diverse and involved in snail innate defense responses. To gain further insight into the functions of FREPs, recombinant FREP proteins (rFREPs) were produced in Escherichia coli and antibodies (Abs) were raised against the corresponding rFREPs. We first show that most FREP proteins exist in their native conformation in snail hemolymph as multimeric proteins. Western blot analyses reveal that expression of multiple FREPs including FREP4 in plasma from M line and BS-90 snails, which are susceptible and resistant to S. mansoni infection, respectively, is up-regulated significantly after infection with the trematode Echinostoma paraensei. Moreover, our assays demonstrate that FREPs are able to bind E. paraensei sporocysts and their secretory/excretory products (SEPs), and a variety of microbes (Gram-positive and Gram-negative bacteria and yeast). Furthermore, this binding capability shows evidence of specificity with respect to pathogen type; for example, 65-75-kDa FREPs (mainly FREP4) bind to E. paraensei sporocysts and their SEPs whereas 95-kDa and 125-kDa FREPs bind the microbes assayed. Our results suggest that FREPs can recognize a wide range of pathogens, from prokaryotes to eukaryotes, and different categories of FREPs seem to exhibit functional specialization with respect to the pathogen encountered.     

研究: 奶牛与水牛初乳中乳寡糖组分比较研究

乳寡糖是由乳汁中含量丰富的固体物质组成．研究结果表明,乳寡糖有提高免疫、益生元及抗感染等作用,已发现与婴儿肠道发育、神经智力发育等多方面关系密切．水牛奶是除牛奶外的第二大奶源,国际上公认其为营养含量高、口感好的优质乳制品,但目前针对水牛乳寡糖的研究多以美洲水牛为研究对象,尚无中国水牛的相关研究．本研究利用固相萃取对已脱脂和除去蛋白质的广西水牛初乳乳汁样品进行纯化,并采用苯胺 (aniline,Bn)衍生化试剂对其进行衍生化处理,通过UPLC-ESI-Q-TOF-MS液相质谱进行优化后,对水牛初乳中的寡糖组分进行测定并与牛乳进行了对比,最终测得奶牛初乳中19种及水牛初乳中的9种乳寡糖组分,并对二者的种类及含量进行比较,发现在两种初乳的乳寡糖中,中性糖二糖m/z 385.15和中性糖三糖m/z 547.21以及酸性糖m/z 635.23均为其主要寡糖成分,与其他乳寡糖相比含量相对较高．总体而言水牛初乳中的中性寡糖占比比奶牛初乳高,二者中性糖占乳寡糖总量的比例分别为88.88%和63.16%．     

紫胶林-农田复合生态系统地表甲虫多样性——以云南绿春为例

为揭示紫胶林-农田复合生态系统地表甲虫多样性, 于2006-2007年在云南省绿春县牛孔乡采用陷阱法调查了天然紫胶林、人工紫胶林和旱地的地表甲虫群落。共采集标本1 678头, 分别隶属于24科120种, 其中步甲科(Carabidae)和金龟科(Scarabaeidae)种类最丰富, 均占全部种类的12.50%。拟步甲科(Tenebrionidae)个体数量最丰富, 占个体总数的64.48%; 金龟科次之, 占个体总数的17.58%。大多数科的物种数和个体数在不同土地利用生境中的分布没有显著差异, 而步甲科、隐翅甲科(Staphylinidae)、叩甲科(Elateridae)、拟步甲科、瓢虫科(Coccinellidae)和小蠧科(Scolytidae)在不同土地利用生境中的分布有显著差异。天然紫胶林地表甲虫个体数量最少, 物种较丰富, 优势度最低, 多样性最高; 人工紫胶林个体数和物种数均丰富, 优势度和多样性居中; 旱地个体数量最多, 物种最贫乏, 优势度最高, 多样性最低。种级水平的聚类分析体现出人工紫胶林与旱地之间在种类组成上距离较近; 而科级水平的聚类反映出人工紫胶林和天然紫胶林更接近。结果提示, 紫胶林-农田复合生态系统具有区域内土地利用方式多样化的特点; 天然紫胶林在维持地表甲虫多样性水平上具有重要作用, 而人工紫胶林虽具有积极作用, 但仍需进一步恢复。     

Acupuncture Prevents the Impairment of Hippocampal LTP Through β1-AR in Vascular Dementia Rats

It is widely accepted that the synaptic dysfunction and synapse loss contribute to the cognitive deficits of vascular dementia (VD) patients. We have previously reported that acupuncture improved cognitive function in rats with VD. However, the mechanisms involved in acupuncture improving cognitive ability remain to be elucidated. The present study aims to investigate the pathways and molecules involved in the neuroprotective effect of acupuncture. We assessed the effects of acupuncture on hippocampal long-term potentiation (LTP), the most prominent cellular model of memory formation. Acupuncture enhanced LTP and norepinephrine (NE) levels in the hippocampus. Inhibition of the β-adrenergic receptor (AR), but not the α-AR, was able to block the effects of acupuncture on hippocampal LTP. Furthermore, inhibition of β1-AR, not β2-AR, abolished the enhanced LTP induced by acupuncture. The expression analysis revealed a significant upregulation of β1-AR and unchanged β2-AR with acupuncture, which supported the above findings. Specifically, increased β1-ARs in the dentate gyrus were expressed on neurons exclusively. Taken together, the present data supports a beneficial role of acupuncture in synaptic plasticity challenged with VD. A likely mechanism is the increase of NE and activation of β1-AR in the hippocampus.     

应用PCR重组技术构建大肠杆菌分泌型表达载体

应用ＰＣＲ重组技术,对大肠杆菌分泌型表达载体ｐＩＮ－ⅢｏｍｐＡ进行改建,除去原载体的ＨｉｎｄⅢ位点,将信号肽序列末端两个密码子ＣＡＧＧＣＣ改为ＣＡＡＧＣＴ,从而引入一个新的ＨｉｎｄⅢ位点,并在其下游接上一段多克隆位点序列。改建后的载体可用于直接插入外源ＤＮＡ编码序列,表达产物在分泌过程中被切除信号肽而成为天然有活性的蛋白质,操作十分简便。     

林木基因组学研究进展

林木基因组学研究进展迅速。结构基因组学方面,已构建了近40个主要造林树种的遗传连锁图谱,在不同树种中定位了30余个重要的数量性状位点,在部分树种中开展了基因组比较和综合图谱构建研究,杨树的全基因组测序已经完成,桉树的全基因组测序正在进行。功能基因组学方面,已分析了主要造林树种多种组织的转录组EST序列,对林木次生生长与木材形成、开花和抗寒性的形成等过程开展了功能基因组学研究。另外,探讨了林木基因组学研究的发展趋势,以期为我国林木基因组学研究提供有益的参考。