Regulation of avoidant behaviors and pain by the anti-inflammatory tyrosine phosphatase SHP-1

The protein tyrosine phosphatase SHP-1 is a critical regulator of cytokine signaling and inflammation. Mice homozygous for a null allele at the SHP-1 locus have a phenotype of severe inflammation and are hyper-responsive to the TLR4 ligand LPS. TLR4 stimulation in the CNS has been linked to both neuropathic pain and sickness behaviors. To determine if reduction in SHP-1 expression affects LPS-induced behaviors, responses of heterozygous SHP-1-deficient (me/+) and wild-type (+/+) mice to LPS were measured. Chronic (4-week) treatment with LPS induced avoidant behaviors indicative of fear/anxiety in me/+, but not +/+, mice. These behaviors were correlated with a LPS-induced type 2 cytokine, cytokine receptor, and immune effector arginase profile in the brains of me/+ mice not found in +/+ mice. Me/+ mice also had a constitutively greater level of TLR4 in the CNS than +/+ mice. Additionally, me/+ mice displayed constitutively increased thermal sensitivity compared to +/+ mice, measured by the tail-flick test. Moreover, me/+ glial cultures were more responsive to LPS than +/+ glia. Therefore, the reduced expression of SHP-1 in me/+ imparts haploinsufficiency with respect to the control of CNS TLR4 and pain signaling. Furthermore, type 2 cytokines become prevalent during chronic TLR4 hyperstimulation in the CNS and are associated positively with behaviors that are usually linked to type 1 pro-inflammatory cytokines. These findings question the notion that type 2 immunity is solely anti-inflammatory in the CNS and indicate that type 2 immunity induces/potentiates CNS inflammatory processes.     

尼古丁在帕金森病中的神经保护作用

帕金森病(Parkinson’s disease,PD)是常见的中老年神经退行性疾病。研究表明,尼古丁具有抵抗黑质多巴胺神经元损伤的作用,其通过烟碱型乙酰胆碱受体(nicotinic acetylcholine receptor,nAChR)途径与非受体途径抑制帕金森病的发生与发展。本文就尼古丁在帕金森病中的神经保护作用以及保护机制的相关研究进行综述。     

盐生植物角果碱蓬种子二型性对环境的适应策略

角果碱蓬(Suaeda corniculata)是藜科一年生盐生植物, 在我国分布于北方盐碱滩涂和盐碱荒漠地区。角果碱蓬具有棕色和黑色两种异型体种子(简称棕色和黑色种子)。对采自内蒙古鄂托克前旗盐渍化生境的角果碱蓬二型种子的形态、休眠和萌发特性开展对比研究, 测定了二型种子休眠和萌发行为对温度、光照和盐分(NaCl)的响应, 以揭示盐生植物异型种子对温带盐漠生境的适应对策。结果表明: (1)二型性种子在大小、种皮特性和结实比例方面有显著差异。与黑色种子相比, 棕色种子个体较大, 种皮透水性强。黑色种子与棕色种子的结实比例约为5.6 : 1。(2)新成熟的棕色种子的萌发对各温度梯度和光照条件不敏感, 萌发率较高(84%-100%); 而新成熟的黑色种子萌发率较低(8%-78%), 萌发对光照敏感。(3)黑色种子具有浅度生理休眠, 种皮划破、赤霉素处理和低温层积均可有效地提高种子的萌发率。(4)二型种子萌发对土壤盐分的胁迫具有不同的响应。与黑色种子相比, 棕色种子对盐分胁迫不敏感, 在较高的盐分浓度下仍有较高的萌发率, 低温层积处理能够降低黑色种子对盐胁迫的敏感性, 有效地提高种子的初始萌发率、萌发恢复率和最终萌发率。角果碱蓬二型种子不同的形态、休眠和萌发特性, 提高了该物种在高度异质性生境中的适合度, 对种群成功地适应温带盐漠环境具有重要的意义。     

Tempo and mode of concerted evolution in the L1 repeat family of mice

A 300-bp DNA sequence has been determined for 30 (10 from each of three species of mice) random isolates of a subset of the long interspersed repeat family L1. From these data we conclude that members of the L1 family are evolving in concert at the DNA sequence level in Mus domesticus, Mus caroli, and Mus platythrix. The mechanism responsible for this phenomenon may be either duplicative transposition, gene conversion, or a combination of the two. The amount of intraspecies divergence averages 4.4%, although between species base substitutions accumulate at the rate of approximately 0.85%/Myr to a maximum divergence of 9.1% between M. platythrix and both M. domesticus and M. caroli. Parsimony analysis reveals that the M. platythrix L1 family has evolved into a distinct clade in the 10-12 Myr since M. platythrix last shared a common ancestor with M. domesticus and M. caroli. The parsimony tree also provides a means to derive the average half-life of L1 sequences in the genome. The rates of gain and loss of individual copies of L1 were estimated to be approximately equal, such that approximately one-half of them turn over every 3.3 Myr.      

核心组蛋白的融合表达和纯化

目的:克隆核心组蛋白H2A、H2B、H3和H4的基因,表达并纯化组蛋白与谷胱甘肽S-转移酶(GST)的融合蛋白。方法:用PCR方法从乳腺文库中扩增核心组蛋白H2A、H2B、H3和H4的编码序列,分别将其以正确相位与pGEX-KG载体中的GST编码序列融合,得到重组质粒pGST-H2A、pGST-H2B、pGST-H3和pGST-H4,分别转化大肠杆菌BL21,表达融合蛋白GST-H2A、GST-H2B、GST-H3和GST-H4;用谷胱甘肽-Sepharose 4B亲和纯化融合蛋白;用Western印迹检测融合蛋白的表达及纯化。结果:分别构建了核心组蛋白H2A、H2B、H3和H4的融合表达载体;Western印迹检测表明,融合蛋白GST-H2A、GST-H2B、GST-H3和GST-H4获得表达及纯化。结论:表达并纯化了H2A、H2B、H3和H4的融合蛋白,为进一步研究核心组蛋白的功能奠定了基础。     

单链胰岛素前体在酵母中的分泌表达及其转变成人胰岛素

将化学合成的单链猪胰岛素前体(PIP)基因和用聚合酶链反应得到的α交配因子前导顺序(αMFL)基因插入质粒pVT102-U的醇脱氢酶基因ADH1的启动子和3’终止顺序之间而生成质粒pVT102-U/αMFL-PIP.被pVT102-U/αMFL-PIP转化的酵母(Saccharomyces cerevistae)可表达单链前体并分泌到培养基中.前体在纯化后可通过胰蛋白酶的转肽作用转变成人胰岛素.纯化的人胰岛素具有全部活力并可结晶.人胰岛素的总收率为每升培养液25mg.     

苎麻光合生理生态特性研究

以大田栽培的苎麻植株为材料,用TPS-2便携式光合作用测定系统测定自然条件下生长的苎麻叶片的光合气体交换参数,以及光响应曲线和CO2响应曲线,并通过回归和相关法分析探讨净光合速率与主要生理、生态因子间的关系.结果表明:(1)苎麻叶片的净光合速率(Pn)日变化曲线呈现双峰型,2个光合峰高度接近,其净光合速率具有典型的午休"现象;蒸腾速率(Tr)日变化曲线呈现单峰型,其走势与气孔导度(Gs)日变化一致.(2)苎麻叶片光合作用的光饱和点(LSP)为1 568.5μmol?m-2?s-1,光补偿点(LCP)为54.18μmol?m-2?s-1,表观量子效率(AQY)为0.025 8 mol?mol-1;而其CO2补偿点(CCP)、饱和点(CSP)和羧化效率(CE)分别为49.25、1 746.9μmol?mol-1和0.045;因此,苎麻属于喜光性阳生植物,且对强光有一定的耐受能力.(3)苎麻叶片Pn日变化的主要限制因子是胞间CO2浓度(Ci),主要决定生理因子是气孔导度(Gs).