Expression, post-translational modification and biochemical characterization of proteins encoded by subgenomic mRNA8 of the severe acute respiratory syndrome coronavirus

The most striking difference between the subgenomic mRNA8 of severe acute respiratory syndrome coronavirus isolated from human and some animal species is the deletion of 29 nucleotides, resulting in splitting of a single ORF (ORF8) into two ORFs (ORF8a and ORF8b). ORF8a and ORF8b are predicted to encode two small proteins, 8a and 8b, and ORF8 a single protein, 8ab (a fusion form of 8a and 8b). To understand the functions of these proteins, we cloned cDNA fragments covering these ORFs into expression plasmids, and expressed the constructs in both in vitro and in vivo systems. Expression of a construct containing ORF8a and ORF8b generated only a single protein, 8a; no 8b protein expression was obtained. Expression of a construct containing ORF8 generated the 8ab fusion protein. Site-directed mutagenesis and enzymatic treatment revealed that protein 8ab is modified by N-linked glycosylation on the N81 residue and by ubiquitination. In the absence of the 8a region, protein 8b undergoes rapid degradation by proteasomes, and addition of proteasome inhibitors inhibits the degradation of protein 8b as well as the protein 8b-induced rapid degradation of the severe acute respiratory syndrome coronavirus E protein. Glycosylation could also stabilize protein 8ab. More interestingly, the two proteins could bind to monoubiquitin and polyubiquitin, suggesting the potential involvement of these proteins in the pathogenesis of severe acute respiratory syndrome coronavirus.     

海盐对绿竹叶片反射光谱及叶绿素荧光参数的影响

沿海防护林是海岸生态系统中最基本的生物资源,由于沿海地区地下水位高,土壤含盐量高,肥力低,生态环境恶劣等特点,所以优良植物种选择是加快沿海优良防护林体系建设的关键。为了探讨绿竹耐盐性以及为沿海地区防护林选择优良植物种,以2年生绿竹(Bambusa oldhamii)为材料,采用水培法进行不同浓度的海盐处理,利用Unispec-SC型单通道光纤光谱仪和非调制式叶绿素荧光仪对绿竹叶片反射光谱和叶绿素荧光参数等进行测定。结果表明:当海盐浓度小于1.2%时,绿竹叶片中叶绿素a、叶绿素b和类胡萝卜素含量,以及"三边"参数与对照无显著差异,当海盐浓度升高到1.6%时,色素与对照相比分别降低了63.2%、62.8%和47.2%(P0.01),红边位置(λr ed)和红边面积(Sred)相比对照显著减小(P0.05)。1.2%浓度的海盐处理,红边归一化指数(rNDVI)、绿度归一化指数(gNDVI)、类胡萝卜素反射指数2(CRI700)和光化学反射指数(PRI)显著降低(P0.05),与对照相比分别降低了27.3%、23.3%、19.5%和43.9%;当海盐浓度增加到1.6%时,rNDVI、改良红边比值指数(mND)、gNDVI、改良归一化差值指数(mSR700)、类胡萝卜素反射指数1(CRI550)、CRI700和PRI等参数与对照相比分别下降了42.4%、43.9%、32.6%、21.5%、47.2%、49.9%和58.5%。绿竹叶片PSⅡ最大量子产率(Fv/Fm)、电子传递的量子产额(ΦEo)、单位反应中心捕获的用于电子传递的能量(ETo/RC)、单位面积反应中心数目(RC/CS)和叶片性能指数(PIABS)等参数,1.6%海盐处理与对照相比分别降低了50.8%、28.6%、21.7%、52.1%和92.3%,单位反应中心复合体吸收的能量(ABS/RC)比对照提高了96.9%。说明绿竹具有一定的耐盐性。     

高原湿地纳帕海水生植物群落分布格局及变化

采用3S技术与植物群落研究法,对高原湿地纳帕海24a来的湿地植物群落分布格局及变化的研究结果表明：与24a前水生植物群落相比较。纳帕海水生植物群落类型、数量改变,原生群落不断减少或消失,耐污、喜富营养类群如水葱群落（Com．Scirpus tabernaemontani）、茭草群落（Com．Zizania caduciflora）、穗状狐尾藻群落（Com．Myriophyllum spicatum）、满江红（Com．Azolla imbricata）群落等大量出现;群落总数由24a前的9个增至当前的12个,其中挺水植物群落增加2个,浮叶植物群落增加1个,挺水植物群落增幅最大。由东向西、由南向北,纳帕海水生植物群落分布大致呈现出浮叶群落、挺水群落、沉水群落斑块状依次配置的水平格局规律。挺水植物群落分布面积最大,达528．42hm^2,其次是沉水植物群落,分布面积为362．50hm^2,浮叶植物群落分布面积最小,为70．23hm^2。随沉水群落、浮叶群落向挺水群落的演替,群落伴生种数量增加、优势种优势度减小、层次类型改变,群落结构变得更为复杂。纳帕海湿地水生植物群落分布格局及变化是对湿地环境变化的响应,表明了在人为干扰作用影响下,纳帕海湖岸线内移、水量减少、水质恶化等湿地水文条件的改变,致使湿地生态系统功能不断退化。     

盐酸胍对口蹄疫病毒完整颗粒和空衣壳的影响

在口蹄疫病毒的复制过程中，会形成完整的病毒粒子（146S）和一定量的空衣壳（75S）。本试验研究了盐酸胍在不同时间和浓度下对口蹄疫病毒146S和75S的影响，并筛选出75S产量最高的条件。通过蔗糖密度梯度离心、透射电镜、特异性单域抗体包被的双抗夹心ELISA等方法对75S和146S进行定性和定量检测。结果表明在口蹄疫病毒（Foot-and-mouth disease virus,FMDV）感染BHK-21细胞2 h时，加入4 mmol/mL的盐酸胍，75S产量比同时间下加入盐酸胍浓度为2 mmol/mL和8 mmol/mL时得到的75S含量高两倍，约为146S的三倍。本研究为定量口蹄疫病毒75S形态的免疫学方法建立奠定了前期研究基础。     

Effect of non-symmetric waveform on conduction block induced by high-frequency (kHz) biphasic stimulation in unmyelinated axon

The effect of a non-symmetric waveform on nerve conduction block induced by high-frequency biphasic stimulation is investigated using a lumped circuit model of the unmyelinated axon based on Hodgkin-Huxley equations. The simulation results reveal that the block threshold monotonically increases with the stimulation frequency for the symmetric stimulation waveform. However, a non-monotonic relationship between block threshold and stimulation frequency is observed when the stimulation waveform is non-symmetric. Constant activation of potassium channels by the high-frequency stimulation results in the increase of block threshold with increasing frequency. The non-symmetric waveform with a positive pulse 0.4–0.8 μs longer than the negative pulse blocks axonal conduction by hyperpolarizing the membrane and causes a decrease in block threshold as the frequency increases above 12–16 kHz. On the other hand, the non-symmetric waveform with a negative pulse 0.4–0.8 μs longer than the positive pulse blocks axonal conduction by depolarizing the membrane and causes a decrease in block threshold as the frequency increases above 40–53 kHz. This simulation study is important for understanding the potential mechanisms underlying the nerve block observed in animal studies, and may also help to design new animal experiments to further improve the nerve block method for clinical applications.