铜绿假单胞菌中群体感应系统研究进展

群体感应系统(Quorum-sensing system,QS)是一个依赖于细胞数量的基因调控系统。系统中的自诱导物(Autoinducer或AI)随细胞的数量增加而变化,当细胞数达到一定数量时,系统中的自诱导物达到一定的域值时可以与一类转录调节蛋白结合,开始诱导或抑制数量众多的基因表达,使细菌表现多细胞特性的群体行为。同时,群体感应系统受到许多外界环境因素的影响,其调节途径是一个极其复杂的级联过程。此外,以群体感应系统为药物靶点来筛选新型抗菌药物越来越受到人们的重视。结合作者本人的工作及铜绿假单胞菌中群体感应系统的最新研究进展,对该系统在铜绿假单胞菌中的作用及其调控途径进行分析、探讨和总结。     

Effect of warming and grazing on litter mass loss and temperature sensitivity of litter and dung mass loss on the Tibetan plateau

Knowledge about the role of litter and dung decomposition in nutrient cycling and response to climate change and grazing in alpine ecosystems is still rudimentary. We conducted two separate studies to assess the relative role of warming and grazing on litter mass loss and on the temperature sensitivity of litter and dung mass loss. Experiments were conducted for 1–2 years under a controlled warming–grazing system and along an elevation gradient from 3200 to 3800 m. A free‐air temperature enhancement system (FATE) using infrared heaters and grazing significantly increased soil temperatures (average 0.5–1.6 °C) from 0 to 40 cm depth, but neither warming nor grazing affected soil moisture except early in the growing seasons at 30 cm soil depth. Heaters caused greater soil warming at night‐time compared with daytime, but grazing resulted in greater soil warming during daytime compared with night‐time. Annual average values of the soil temperature at 5 cm were 3.2, 2.4 and 0.3 °C at 3200, 3600 and 3800 m, respectively. Neither warming nor grazing caused changes of litter quality for the first year of the controlled warming–grazing experiment. The effects of warming and grazing on litter mass losses were additive, increasing litter mass losses by about 19.3% and 8.3%, respectively, for the 2‐year decomposition periods. The temperature sensitivity of litter mass losses was approximately 11% °C^?1 based on the controlled warming–grazing experiment. The annual cumulative litter mass loss was approximately 2.5 times that of dung along the elevation gradient. However, the temperature sensitivity (about 18% °C^?1) of the dung mass loss was about three times that of the litter mass loss. These results suggest greater warming at night‐time compared with daytime may accelerate litter mass loss, and grazing will enhance carbon loss to atmosphere in the region through a decrease of litter biomass and an increase of dung production with an increase of stocking rate in future warmer conditions.     

新型重组人内毒素结合肽突变体的构建及原核表达纯化

目的构建新型人内毒素结合肽（a new endotoxin binding peptide consisting of 25 amino acid residues,EBP25）及其突变体（mutant of EBP25,mEBP25）的原核表达重组质粒,并在大肠埃希菌中诱导表达。方法采用PCR法,扩增EBP25基因,构建pET-30-EBP25.融合表达载体并转化Ecoli DH5α扩增。重组质粒经酶切和测序鉴定后,应用快速定点突变法将EBP25第2位缬氨酸和第5位谷氨酰胺所对应碱基均替换成赖氨酸所对应的碱基,突变后重组质粒再经测序鉴定后,将二者转化至E．coli BL21（DE3）PlysS后经IPTG诱导表达,表达产物采用Western印迹进行鉴定后,用His—Tag亲和层析对融合蛋白进行纯化。结果两次测序结果显示人EBP25,和mEBP25重组序列和理论设计序列完全一致后,经IPTG诱导表达获得目的融合蛋白,通过SDS—PAGE电泳、Western印迹证实蛋白表达的特异性,并对蛋白进行纯化,获得EBP25和mEBP25融合蛋白。结论构建、表达纯化了EBP25和mEBP25融合蛋白,为进一步研究其中和内毒素／月旨多糖活性奠定了基础。     

鸭胚多肽对小鼠抗疲劳、耐缺氧能力的影响

实验研究鸭胚多肽对小鼠机体抗疲劳、耐缺氧能力的影响。实验设对照组和实验组(鸭胚多肽低剂量、中剂量和高剂量组),连续灌胃小鼠30 d后,测定小鼠负重游泳力竭至死时间,并测定小鼠游泳后血清中乳酸(LD)含量、尿素氮(BUN)含量、葡萄糖(LG)含量、乳酸脱氢酶(LDH)活性,小鼠常压耐缺氧存活时间、亚硝酸钠中毒存活时间、急性脑缺血性存活时间。结果表明:鸭胚多肽能显著增加小鼠负重游泳时间、血清中LDH活性、小鼠亚硝酸钠中毒存活时间及急性脑缺血性存活时间,并能显著降低LD、BUN含量。说明鸭胚多肽能有效提高小鼠的抗疲劳、耐缺氧能力。     

大鼠心肌缺血再灌注模型构建的改良

目的改良大鼠心肌缺血再灌注模型的构建方法,增强实用性。方法32只SD雄性大鼠随机分为四组（A组：伪手术组;B组：缺血组;C组：缺血30 min再灌注组;D组：缺血60 min再灌注组）。缺血过程在人工机械通气条件下完成,术前术中监测心电图变化,模型成功72 h后抽血进行心肌酶谱分析,并观察左室压变化。结果造模成功率为91%。B、C、D组各心肌酶含量明显高于A组（P〈0.05）,C、D组低于B组（P〈0.05）。B、C、D组左室压值低于A组（P〈0.05）,C、D组高于B组（P〈0.05）。结论改进后的模型制备方法简便易行,成功率高,评价指标符合临床应用。     

重庆酸雨区马尾松香樟混交林的土壤化学性质和林木生长特征

为给遭受酸化危害的重庆马尾松(Pinus massoniana)纯林的恢复与管理提供科学依据,在属于酸雨区的重庆市江北区铁山坪林场,调查了马尾松香樟(Cinnamomum camphora)混交林的土壤化学性质和林木生长状况,并与毗邻的马尾松纯林和香樟纯林进行了比较。结果表明,与马尾松纯林相比,混交林腐殖质层和土壤酸度降低,交换性盐基离子Ca2+和Mg2+含量增加,酸性阳离子Al3+和H+含量明显减少(p0.05)。混交林马尾松树冠下腐殖质层和0–60cm土层马尾松细根的干重、长度、表面积、体积和根尖数的总密度都显著低于马尾松纯林(p0.05),比值分别为0.44、0.71、0.59、0.46和0.71,马尾松树高显著低于马尾松纯林(p0.05),其针叶变色率为15%,显著高于马尾松纯林(p0.05),香樟的生长特征与香樟纯林相似。结果说明,尽管混交林的土壤化学性质优于马尾松纯林,但林中马尾松的生长状况表现出了明显的恶化迹象,而香樟依然比较健康,这可能与香樟具有明显的竞争优势有关。因此,在利用香樟营造与马尾松的混交林来改善遭受酸化危害的马尾松纯林的生长状况时,应注意调控香樟的竞争优势。     

数据点选择与参数估计方法对杉木人工林自疏边界线的影响

自疏边界线是指植物种群发生密度依赖死亡时种群最大收获量的上边界线。已有研究由于在拟合自疏边界线的过程中对数据点的选择和参数估计的方法存在诸多的差异,进而导致产生对自疏法则的争议。该研究采用26年生杉木(Cunninghamia lanceolata)人工林的定位观测数据,对视觉法、死亡率法、等距区间法和相对密度法等4种数据点选择方法以及最小二乘法、降维分析法、分位数回归法和随机边界方程等4种参数拟合方法进行对比分析,以探寻客观选择自疏拟合数据和正确拟合方法的途径。比较4种不同的数据选择方法得出:视觉法具有较强的主观性;对于没有发生非密度依赖死亡的林分,死亡率法可以准确地确定林分自疏的起始点;等距区间法可以减少非密度依赖死亡的影响,得到的数据点能充分反映林分的自疏过程;相对密度法可以保证临界密度阈值以上的数据点拟合林分自疏边界线的有效性,并能排除非密度依赖死亡的影响。比较分析4种不同的拟合方法发现:最小二乘法和降维分析法拟合的林分自疏边界线均从实测数据"中心"穿过,与林分自疏边界线为林分收获量上边界线的涵义不相符合,无法真实反映林分的自疏进程;分位数回归和随机边界方程的拟合结果均与实测数据一致,能够较为准确地反映林分自疏的真实过程,但二者的统计推断要求都比较严格。分位数值的正确选取和残差足够小且趋于0,是分位数回归法和随机边界方程能否正确反映林分自疏动态的前提。     

"活的非可培养状态"细菌荧光观察方法

目的：建立一套稳定、简便、快捷,可靠的细菌活的非可培养状态（VBNC）荧光显微镜观察方法。利用市售的黑色钢笔墨水着染微孔滤膜,然后采用手动压片方式,将经LIVE/DEAD BacLightTM Bacterial Viability Kit（13152）染色的细菌样本固定在滤膜上,用落射荧光显微镜直接观察。结果：该法所得的荧光图像背景无光亮且黑暗,菌体荧光、形态、排列等均清晰可见,而且呈现绿色荧光的活菌与呈现红色荧光的死菌较易分辨。与现行技术相比,还能缩短染色时间近12～16h。结论：建立的VBNC细菌荧光观察方法能有效地克服现有技术存在的适用范围狭窄、取材困难、观察结果不准确、操作时间长等缺点,可为细菌VBNC研究提供值得借鉴的方法。     

Heme-protein covalent bonds in peroxidases and resistance to heme modification during halide oxidation

Plant peroxidases, as typified by horseradish peroxidase (HRP), primarily catalyze the one-electron oxidation of phenols and other low oxidation potential substrates. In contrast, the mammalian homologues such as lactoperoxidase (LPO) and myeloperoxidase primarily oxidize halides and pseudohalides to the corresponding hypohalides (e.g., Br(-) to HOBr, Cl(-) to HOCl). A further feature that distinguishes the mammalian from the plant and fungal enzymes is the presence of two or more covalent bonds between the heme and the protein only in the mammalian enzymes. The functional roles of these covalent links in mammalian peroxidases remain uncertain. We have previously reported that HRP can oxidize chloride and bromide ions, but during oxidation of these ions undergoes autocatalytic modification of its heme vinyl groups that virtually inactivates the enzyme. We report here that autocatalytic heme modification during halide oxidation is not unique to HRP but is a general feature of the oxidation of halide ions by fungal and plant peroxidases, as illustrated by studies with Arthromyces ramosus and soybean peroxidases. In contrast, LPO, a prototypical mammalian peroxidase, is protected from heme modification and its heme remains intact during the oxidation of halide ions. These results support the hypothesis that the covalent heme-protein links in the mammalian peroxidases protect the heme from modification during the oxidation of halide ions.     

羚羊石斛的离体快速繁殖

1植物名称羚羊石斛(DendrobiumcochliodesSchltr.)。2材料类别种子。3培养条件萌发培养基:(1)VW;(2)VW+椰子乳;(3)1/2MS;(4)1/2MS+椰子乳;(5)N6;(6)N6+椰子乳;(7)N6+椰子乳+活性炭2g·L-1。继代增殖和分化成苗培养基:(8)N6+椰子乳+6-BA1.0mg·L-1(单位下同)+NAA0.2;(9)N6+椰子乳+6-BA0.2+NAA0.1;(10)N6+椰子乳+6-BA0.2+NAA0.1+活性炭2g·L-1;(11)花宝1号(美国Haponex公司产品,N∶P∶K=7∶6∶19)1.5g·L-1+花宝2号(美国Haponex公司产品,N∶P∶K=20∶20∶20)1.5g·L-1+椰子乳+6-BA0.2+NAA0.1。生根壮苗培养基:(12)N6+NA…