产气荚膜梭菌α毒素基因工程菌表达产物的免疫原性研究

已构建的能表达产气荚膜梭菌α毒素保护性抗原基因工程菌株Ｅ．ｃｏｌｉＢＬ２１（ＤＥ３）（ｐＸＥＴＡ１）经动物试验证实没有毒性。从ＩＰＴＧ诱导后的工程菌中提取包涵体,再辅以氢氧化铝胶制成抗原,免疫小鼠３０ｄ后,用产气荚膜梭菌强毒株培养上清及培养菌体攻击,结果免疫鼠能抵抗至少２ＬＤ１００的攻击,证明Ｅ．ｃｏｌｉＢＬ２１（ＤＥ３）（ｐＸＥＴＡ１）工程菌株表达产物具有良好的免疫原性。     

基于突变分级模型的淤地坝坝系规划方案优选

淤地坝坝系规划方案优选需要考虑水沙控制能力、拦泥能力、淤地能力与投资效益等多方面的因素,是一个典型的复杂系统多准则决策问题,复杂系统多准则决策通常具有模糊性与不确定性.利用突变理论处理模糊与不确定问题的优势,提出了淤地坝坝系规划方案评价指标体系,建立了基于突变分级评价法的淤地坝坝系规划方案优选模型,给出了应用该模型的具体步骤.将该模型应用于偃师市浏涧河流域坝系方案优选中,对底层指标量纲一化处理并进行归一运算,利用归一公式的聚集机理对各指标的重要性进行量化,比较各方案的总突变隶属函数值,从而确定淤地坝坝系规划方案优选结果.结果表明:方案Ⅰ,Ⅱ,Ⅲ总突变隶属函数值分别为0.959 2,0.918 1,0.948 4,方案Ⅰ最优,其评价结果与传统的人工对比分析方法评价结果一致,从而验证了突变分级评价模型用于淤地坝坝系规划方案优选的合理性和可行性.     

硫酸化黄芪多糖对雏鸡十二指肠黏膜组织形态和免疫细胞的影响

【目的】制备硫酸化黄芪多糖(Sulfated astragalus polysaccharide,sAPS),研究其对雏鸡十二指肠组织发育及黏膜免疫相关细胞分布的影响。【方法】采用氯磺酸-吡啶法对黄芪多糖(Astragalus polysaccharid,APS)进行硫酸化修饰,制备sAPS,氯化钡-明胶浊度法测定sAPS中硫含量,并计算sAPS的硫酸基取代度。运用组织学制片技术和染色方法分别观察APS及sAPS干预后7d和14d鸡十二指肠的组织学特点,对肠壁黏膜层上皮内淋巴细胞(Intraepithelial lymphocytes,IEL)、柱状上皮细胞间杯状细胞(Goblet cells,GC)和肠壁内肥大细胞(Mast cells,MC)数量进行统计分析。【结果】制得的sAPS中硫含量为14.3%,计算得到sAPS的硫酸基取代度为1.325。给予APS和sAPS干预后,sAPS组雏鸡十二指肠的绒毛高度及绒毛高度/隐窝深度比值均明显高于对照组和APS组,隐窝深度明显低于对照组和APS组,sAPS组黏膜上皮内淋巴细胞、杯状细胞及肠壁肥大细胞的数量也明显高于对照组和APS组。【结论】sAPS对雏鸡十二指肠组织发育及肠道黏膜免疫的促进作用较APS强。     

冬小麦叶面积测算方法的再探讨

为使小麦叶面积测算的方法更为精确,以田间种植的冬小麦植株为材料,测算了冬小麦叶面积的校正值K。在小麦每个主茎叶片展开时和主要生育时期取样,用直尺测定小麦叶片的长度（L）和宽度（W）,并用Li-3000便携式叶面积仪测定对应的叶面积（A）,用K=A /(L·W)进行计算。结果表明,冬小麦冬前主茎叶的平均K值为0.748 8≈0.75,春生主茎叶的平均K值为0.780 6≈0.78,全株主茎叶的平均K值为0.766 2≈0.77。按生育时期测定,越冬前全部叶片的平均K值为0.729 8≈0.73,春季各生育时期平均K值为0.768 0≈0.77,全部生育时期的平均K值为0.760 4≈0.76。通过与叶面积仪实测的值比较,用全部生育时期的平均值K=0.76,依据公式A=K∑（L·W）计算的小麦各生育时期的单株叶面积和叶面积指数,比用常用的K=0.83得到的结果更准确,因而建议在利用系数法计算叶面积时采用K=0.76。     

4种适生植物对陇东地区油污土壤场地生态修复的响应

[目的]选取陇东黄土高原地区4种本地植物作为试验材料,进行了为期3个月的场地生态修复工程,旨在为该地区开展油污土壤植物场地生态修复技术提供新的植物品种和基础数据参考。[方法]采用常规方法测定了不同油污浓度胁迫下受试植物根际土壤总石油烃(TPH)降解率、降解菌数量及部分生长指标在不同油污浓度胁迫时响应情况。[结果](1)紫花苜蓿和金盏菊根际TPH降解率在轻度污染时相对较高,而重度污染时菊科植物显著高于豆科植物(p0.05);(2)油污浓度增加可显著抑制菊科植物根长以及豆科植物根际石油降解菌数量(p0.05),亦对紫花苜蓿地上部干重及金盏菊地下部干重和株高则有显著促进作用(p0.05);(3)方差分解结果显示:石油降解菌数量×TPH转移效率以及植物生长指标×石油降解菌数量的交互作用是决定两种菊科植物TPH降解率的关键因素。[结论]4种适生植物各项指标对陇东地区油污土壤场地生态修复的响应方式因种而异,而两种菊科植物油污耐受性和修复效果相对较好,因此在该地区具备一定的应用潜力。     

Isolation and sequence analysis of a Dof protein gene (PtDof1) in Populus

Both cDNA and DNA clones of PtDof1 (GenBank Accession No. FJ402844 and FJ402845) were isolated from plants grown in tissue culture of Populus tomentosa. The DNA sequence is 1597 bp including two exons and one intron. The cDNA is 969 bp in length with a 765 bp open reading frame which is capable of encoding 255 amino acids. The deduced amino acids sequence of the PtDof1 protein shares 65%, 56% and 55% identity with Vitis vinifera (CAO48618), Nicotiana tabacum (CAA08755) and Glycine max (ABI16022) Dof protein by blast analysis in GenBank. Phylogenic analysis suggests PtDof1 gene could belong to the Dof gene family. PtDof1 protein contains an unusual conserved single zinc finger with the pattern of C-X2-C-X21-C-X2-C, which may play a functional role in tissue-specific expression and possibly the auxin response of endogenous plant genes.     

冰冻保存鱿鱼体内氧化三甲胺脱甲基酶活性测定方法的建立

建立了冰冻保存鱿鱼体内氧化三甲胺脱甲基酶(TMAOase)活性的测定方法,初步探索了鱿鱼体内甲醛的产生机理。根据TMAOase可在体外催化TMAO生成甲醛,后者可用乙酰丙酮法定量,并结合酶促反应动力学的原理,建立针对冰冻保存鱿鱼体内TMAOase活性的检测方法。建立的活性检测方法可用于检测冰冻保存的鱿鱼体内TMAOase的活性;冰冻鱿鱼体内TMAOase的活性随保存时间延长而增高。该方法简便易行,重复性较好,对深入研究TMAOase生物学特性具有一定的应用价值;在冰冻保存过程中,鱿鱼体内TMAOase活性是造成甲醛含量增高的重要原因。     

陇东黄土高原油污土壤环境因子对场地生态修复的季节响应

[目的]探讨陇东黄土高原地区油污土壤场地修复过程中土壤环境因子对季节变化的响应机制,为油污土壤植物微生物联合生态修复的推广应用提供参考。[方法]利用陇东适生植物"金盏菊"联合土著石油降解菌剂开展为期285d的场地原位修复试验,采用常规方法测定不同季节土壤理化特性、酶活性和微生物群落特性等环境指标。[结果](1)场地修复过程中土壤TPHs降解率在夏(24.62%±3.96%)秋(29.93%±1.94%)两季明显高于春(3.82%±0.91%)冬(9.24%±0.87%)两季;(2)季节变化和强化处理对油污土壤酶活性和理化特性存在显著交互效应(p0.05)。土壤pH值、含盐率和有机质含量在秋季明显降低(p0.05),而土壤速效养分均在夏秋含量最高(p0.05);(3)春季土壤微生物群落Evenness指数偏低(p0.05),夏季Richness指数明显增加(p0.05),Shannon-Wiener指数在夏秋两季明显高于其他季节(p0.05);(4)NMDS排序结果显示,夏秋两季分异于其他季节主要与土壤微生物群落Richness指数(r2=0.706 3,p=0.002)和速效磷(r2=0.615 7,p=0.005)有关;(5)低温季节影响土壤TPHs降解率的因素主要是土壤理化特性和土壤酶活性的叠加效应(54%),而夏秋两季土壤理化特性、土壤酶活性和土壤微生物特性三者的共同作用可解释土壤TPHs降解率变异的74%。[结论]土壤微生物群落Evenness指数和土壤酶活性偏低是造成春冬两季土壤TPHs降解率偏低,而土壤微生物群落ShannonWiener指数、Richness指数、速效磷含量和生物强化处理有效化水平是陇东地区夏秋两季土壤TPHs降解率明显增加的主要环境因素。     

Prokaryotic Expression of the Cysteine Proteinase Gene of Xinjiang Strain of Theileria equi/Babesia caballi and Its Bioinformatics Analysis

This study aimed to clone and express cysteine proteinase (CP) gene of Theileria equi and Babesia caballi. The CP proteins were analyzed using bioinformatics tools and online databases. The total DNA of T. equi and B. caballi as the template sequence for PCR. The CP genes were cloned, expressed using the cloning technique and prokaryotic expression system, respectively. Then the recombinant CP (rCP) proteins were purified by Urea dialysis. Finally the specific reaction of polyclonal horse anti-CP serum with rCPs was analyzed through Dot blot method. The CPs were predicted and analyzed by the software MAGA, Prot Param, TMHMM, SignalP-5.0, Antibody Epitope Prediction, SYFPEITHⅡ, ProPred and EzMol^2.1 respectively. The CP genes were successfully amplified from DNA of T. equi and B. caballi and expressed in the inclusion body fractions, Dot blot assay indicated that the recombinant protein could react with polyclonal horse anti - CP serum. Compared with T. equi-CP and B. caballi-CP, different protozoon CP genes were highly conserved in evolution, and phylogenetic analysis results were consistent with their protozoon taxonomy. The bioinformatics analysis revealed that two CPs are the hydrophilic protein instead of a transmembrane one, no transmembrane domain, no Th cell epitope was found, and more localized in the cytoplasm, mitochondria and nuclei. The relative molecular weight (MW) of T. equi-CP was 29 948.60, the theoretical isoelectric point (pI) was 5.53, stability coefficient was 22.50; B. caballi-CP’MW was 14 603.62, the theoretical pI was 8.54, stability coefficient was 47.69, but signal peptide was contained. The CPs have good reactionogenicity, as the hydrophilic extracellular proteins with no Th cell epitope, and more localized in the cytoplasm, mitochondria and nuclei, which helped T. equi and B. caballi avoid host immune response outside the cell membrane and the CPs involved in proliferation, differentiation and programmed cell death. In conclusion, a theoretical basis was provided for the study on CP’s functions and the pathogenesis of T. equi and B. caballi.     

铅污染对水葱生理特性的影响

为进一步明确铅对植物的伤害机理以及植物的抗性机制,采用营养液水培法,测定不同Pb2+处理浓度下,重金属铅对水葱的根系活力、叶片含水量(RWC)、叶绿素含量、光合参数、丙二醛(MDA)含量以及抗氧化物酶活力的影响。结果表明:当Pb浓度达到800mg/L时,与对照组相比水葱根系活力逐渐下降,减小78.5%;叶片含水量(RWC)逐渐减小,减小到50.4%;叶绿素含量减小,光合速率降低19.6%;丙二醛(MDA)含量增加,MDA最大值可达到空白对照组的7.8倍,膜脂的过氧化程度加大;过氧化物酶(POD)活性显著升高,超氧化物歧化酶(SOD)和过氧化氢酶(CAT)的活性下降。结论:较高浓度铅污染对水葱的生理特性有很大的影响,不利于水葱的正常生长。