A/California/7/2009与A/California/4/2009病毒感染力比较

目的甲型H1N1流感病毒A/California/7/2009与A/California/4/2009病毒序列比较同源性在99%以上,本实验旨在比较两株病毒感染BALB/c小鼠研究感染力强弱。方法分别将A/California/7/2009（CA7）与A/California/4/2009（CA4）两株病毒分别连续10倍稀释后,对4~6周龄雌性BALB/c小鼠经乙醚麻醉后进行滴鼻攻毒,每个稀释度接种10只实验小鼠,测定CA7 MLD50为101.24/0.05 mL,检测小鼠感染、致病的多项指标,观察期为14 d。结果相同TCID50的CA7和CA4病毒感染小鼠,CA4感染小鼠后14 d内死亡率为20%,而CA7感染小鼠后8 d内死亡率为100%。CA7 106TCID50感染的小鼠病理表现为重度弥漫性间质性肺炎,CA4 106TCID50感染的小鼠病理表现为中度-重度间质性肺炎。结论在相同条件下,CA7感染力明显强于CA4。     

YidC/Oxa/Alb3蛋白家族

YidC/Oxa/Alb3蛋白家族是进化上保守的蛋白质转运酶,分别负责将一些与能量合成有关的膜蛋白转运到细菌内膜、线粒体内膜和叶绿体类囊体膜。它们具有保守的跨膜结构域,广泛存在于三界生物中。本文主要综述近年来对YidC/Oxa/Alb3家族成员的分布、结构、功能及进化的研究进展。     

NO3-/NO2-对渤海湾油藏中硫酸盐还原菌SO42-还原活性的抑制效应

长期注水开发促进了渤海湾海域油藏中硫酸盐还原菌(SRP)的生长繁殖,产生了大量H₂S,引起油藏酸化(souring)等问题. 本文首先以改进的API RP 38培养基富集了渤海湾海域某油藏采出井井口采出液中的SRP,再通过批次试验研究了不同浓度NO₃^-和NO₂^-对SRP富集培养物SO₄^2-还原活性的抑制效应. 结果表明： 渤海湾海域油藏中的SRP富集培养物SO₄^2-还原活性较强,SO₄^2-还原速率为10.4 mmol SO₄^2-·d^-1·g^-1 dry cell;加入浓度为0.4、0.8、1.8、4.2 mmol·L^-1NO₃^-时,SRP富集培养物的SO₄^2-还原活性均可被抑制,维持时间分别为5、9、20和大于35 d;加入浓度为0.6、0.9、1.4、2.6或4.6 mmol·L^-1的NO₂^-时,SO₄^2-还原活性也被抑制,维持时间分别为3、12、22和大于39 d. SRP富集培养物具有异化NO₃^-还原成NH₄⁺的代谢途径.当环境中同时存在SO₄^2-、NO₃^-、NO₂^-时,SRP富集培养物优先利用NO₃^-和NO₂^-. SRP富集培养物对电子受体的优先利用及NO₂^-的毒性效应是NO₃^-/NO₂^-抑制渤海湾海域油藏中SO₄^2-还原活性的主要原因.     

结肠癌中NGX6抑制EGFR/K-ras/JNK/c-Jun/cyclin D1信号通路的研究

候选抑瘤基因NGX6具有抑制结肠癌增殖和转移的作用,研究表明其为表皮生长因子受体(epidermal growth factor receptor, EGFR)的负性调控因子,并可下调JNK通路中重要分子MADD (MAP-kinase activating death domain)的表达,其抑瘤机制是否与抑制EGFR介导的JNK信号通路的活性有关?在已建立的转染NGX6的细胞模型基础上,借助蛋白质印迹(Western blot)和免疫组化方法在细胞和组织水平检测NGX6转染前后EGFR/K-ras/JNK/c-Jun/cyclin D1信号通路中重要蛋白质的表达.结果表明,NGX6转染后结肠癌细胞HT-29在裸鼠体内成瘤明显受抑,差异有统计学意义.Western blot结果显示,在结肠癌细胞中NGX6可明显下调EGFR、K-ras、p-JNK、c-Jun和cyclin D1的表达;进一步采用Western blot和免疫组化法验证NGX6在体内对上述关键分子表达的影响,发现NGX6可抑制裸鼠移植瘤组织中EGFR、K-ras、p-JNK、c-Jun和cyclin D1的表达,与体外结果一致.上述研究表明,NGX6在结肠癌中主要通过抑制EGFR介导的JNK通路的活性而发挥其抑瘤作用,该研究为深入探讨NGX6的机制提供了重要的实验依据.     

Preparation of copoly (γ-benzyl-l-glutamyl-N5-β-d-glucopyranosyl-l-glutamine) and its interaction with fibroblast cells

Copoly(α-amino acid)s consisting of γ-benzyl-l-glutamate and $\text{N}{}^5\text{-β-}\text{d}\text{-glucopyranosyl-}\text{l}\text{-glutamine}$ were prepared by the reaction of copoly(l-glutamate) containing succinimide ester, which served as active site for the coupling reaction with β-d-glucopyranosylamine. The α-helical conformation of these copolymers became unstable in DMF as the content of glutamine derivative increased. A dry film made from this copolymer could take a full α-helical conformation even at such a high content as 80% of the glutamine derivative, but in a wet film this ordered structure was partially disrupted by hydration. The hydraulic permeability of this copoly(α-amino acid) was clearly dependent on the molar content of glucopyranosyl groups. The attachment of fibroblast cells to these hydrated copolymer films was effectively depressed in the presence of a serum-free medium. The cells attached to the substrate were spherical in shape.     

Metabolism of Δ4-cis-PGF1α in the monkey

This isomer of PGF_2α is relatively resistant to metabolic degradation in the Cynomolgus monkey. Thus, 16–20 per cent of the amount injected was excreted unchanged in the urine. Five metabolites with 20, 18, 16 and 14 carbon atoms in the skeleton were identified. The data are similar to those earlier seen in the rat and further support the idea that this analogue of PGF_2α could have a long half-life time in the mammalian body and thus a long duration of its pharmacological actions.     

原始森林土壤NH4+/NO3-生境特征与某些针叶树种的适应性

在陆地生态系统中,生存地段的土壤养分环境构成了植物的“营养生境”。植物在长期进化过程中往往产生对原生营养生境的生态适应,其中对NH4 和NO3-两种无机氮源的吸收、利用特性便可能是这种适应的一个重要方面。由于硝化抑制(限制)或微生物对NO3-的强烈吸收、固持作用,酸性、弱酸性的原始森林土壤中NH4 含量大都远高于NO3-,从而形成了以NH4 占绝对优势的“氮营养生境”。很多针叶树种(尤其是演替晚期阶段占优势者)对其长期所处的NH4 优势生境产生了充分适应,以致对非还原态氮(NO3-)的吸收、利用能力严重下降。这些针叶树往往表现出典型的“喜铵性”,而在NO3-优势环境中则会引起氮代谢失调和生长下降。从氮同化酶、高耐铵性、根对NH4 和NO3-的相对吸收能力及NO3-吸收的反馈控制、养分关系与养分平衡、根部碳流失、光合作用及耐荫性等多方面阐述了喜铵针叶树适应的生理生化机制。这种生态适应可能是顶极森林群落维持长期稳定的重要机制之一,而采伐干扰后NO3-明显增加的立地条件则可能会导致喜铵的“原优势针叶树种”更新困难。在温带退化森林生态系统恢复与重建过程中,顶极针叶树种对NH4 营养生境的固有适应性是必须充分考虑的问题。     

转录因子cpcR同源基因cpcR-c1和cpcR-t的功能鉴定

苏云金芽胞杆菌（Bacillus thuringiensis,Bt） LM1212菌株与典型的Bt菌株表型不同,可分化形成芽胞、形成细胞和晶体产生细胞。在LM1212菌株中,转录因子CpcR不仅参与了细胞分化过程,而且能够激活晶体蛋白基因cry35-like的启动子(P₃₅)。【目的】筛选cpcR同源基因,验证其生物学功能。【方法】本研究克隆了2个cpcR同源基因,来源于蜡样芽胞杆菌的cpcR-c1和来源于东洋芽胞杆菌的cpcR-t,将cpcR及其同源基因分别构建在pHT304-P₃₅-gfp、pHT304-P₃₅-lacZ报告载体上,获得的重组质粒转入无cpcR基因且无晶体蛋白基因的Bt HD73^–菌株中。利用激光共聚焦显微镜观察重组菌HD^–(cpcR-c1-P₃₅-gfp)和HD^–(cpcR-t-P₃₅-gfp)的细胞表型并进行芽胞计数实验。测定HD^–(cpcR-c1-P_35<...     

伪狂犬病病毒上海株gE-/gI-/GFP+缺失株的生物学特性初步研究

在构建了伪狂犬病病毒上海株的缺失载体pgEI-GFP基础上,将pgEI-GFP转染感染了PRV-SH株的BHK-21细胞,待出现80%以上的细胞病变时收获病毒,并以绿色荧光蛋白为标志,通过蚀斑法得到纯化重组病毒gE-/gI-/GFP+缺失株.研究了该缺失株的的安全性、在细胞上的生长特性以及对断奶仔猪的安全性、免疫原性等生物学特性.试验结果显示,缺失了gE-和gI-后,不影响其在RK细胞上的生长状况和病毒的滴度.该疫苗株对小鼠的半数致死量比亲本毒低且对家兔的致死性的时间延长了,这表明该疫苗的毒力比亲本毒有所下降.该缺失株对断奶仔猪安全,无不良接种反应,接种断奶仔猪能抵御高剂量PRV-SH株强毒的感染,攻毒后试验猪的发热期、散毒天数均低于对照组.该缺失株接种仔猪后在试验期间一直维持较高水平的中和抗体.     

长江三角洲地区雨水中NH4+-N/NO3--N和δ15NH4+值的变化

2003年6月至2005年7月,利用自行设计的雨水收集器对位于长江三角洲地区的常熟、南京和杭州3个观测点进行了全年性雨水观测,分析了雨水中NH₄⁺-N/NO₃^--N和铵态氮自然丰度(δ¹⁵NH₄⁺)值的变化.结果表明：研究区3个观测点雨水中NH₄⁺-N/NO₃^--N和δ¹⁵NH₄⁺值均呈相似的季节性变化规律,两者的规律性变化在以田间农事耕作为主的常熟观测点尤其明显,而位于市区的南京观测点和位于城乡结合部的杭州观测点的规律性次之;雨水中NH₄⁺-N/NO₃^--N的峰值出现在6月下旬到8月上旬,然后逐渐下降,冬季降到最低;雨水中δ¹⁵NH₄⁺值在6月下旬到8月中旬为负值,在8月下旬到11月中下旬为正值,12月至翌年3月又变为负值,5至6月中旬又转变为正值.雨水中NH₄⁺-N/NO₃^--N和δ¹⁵NH₄⁺值的季节变化与不同作物生育期间氮肥的施用、当地气候的季节性变化以及其他NH3释放源的NH3挥发有关（人和动物排泄物、氮污染水体及有机氮源中的氨挥发）,其对大气湿沉降中NH₄⁺的来源、形态组成及陆地不同NH₃排放源的强度具有明显的指示意义.     

血清7型胸膜肺炎放线杆菌apxⅡC-/apxⅠA+基因缺失重组菌株的构建与鉴定

通过接合转移和SacB负向筛选方法,成功构建了一株apxⅡC缺失的血清7型胸膜肺炎放线杆菌重组菌株。首先构建重组转移质粒pEHA1。将pEHA1转化供体菌大肠杆菌(E.coliβ2155),并将其与野生型APP血清7型亲本菌混合培养约5h,然后涂到含氯霉素抗性的培养基培养,挑取阳性克隆,接种到无抗性液体培养基,培养后涂于含有蔗糖的的固体培养基,培养一定时间后挑取蔗糖抗性的克隆,即可得到目的突变株。通过PCR、遗传稳定性、外毒素分泌、重组位点序列分析证明重组菌构建成功。通过对重组菌生物学特性进行初步研究,表明突变株生长能力未受影响,对小鼠毒力显著降低。该突变株构建体系的建立为猪传染性胸膜肺炎减毒活疫苗的开发及对胸膜肺炎放线杆菌新基因的功能研究奠定了良好基础。     

Cu(C6H9N3O2)2Cl2对小麦的生态毒理效应

以冬小麦为实验材料,比较研究了(1)不同浓度配合物对小麦生长的影响;(2)相同浓度的CuC l2、配体C6H9N3O2和配合物Cu(C6H9N3O2)2C l2对冬小麦种子萌发、苗期生长及其保护酶活性的影响。结果表明:与对照相比,(1)不同浓度新配合物对小麦生长具有不同程度的抑制作用,随着浓度的增高抑制作用逐渐增大;(2)CuC l2、配合物Cu(C6H9N3O2)2C l2对小麦种子总淀粉酶活性、蛋白酶活性、萌发率、生长势、根长、株高、总生物量均具有显著的抑制作用,配合物Cu(C6H9N3O2)2C l2的抑制作用小于CuC l2,而配体C6H9N3O2对上述生物学参数具有促进作用;(3)CuC l2、配合物Cu(C6H9N3O2)2C l2处理引起膜脂过氧化,显著的提高了幼苗的M DA浓度,导致SOD、POD、CAT活性降低,CuC l2的抑制作用大于配合物Cu(C6H9N3O2)2C l2,而配体C6H9N3O2处理对SOD、POD、CAT活性的提高有促进作用。上述结果说明C6H9N3O2对CuC l2生理胁迫具有保护作用,结合态的Cu2 (配合物Cu(C6H9N3O2)2C l2)的毒性显著的降低。在此基础上探讨了配合物抑制小麦生长发育的生物学机制。     

Conformational studies on poly(N-δ-trimethyl-l-ornithine)

$\text{Poly}\text{(N-δ-}\text{trimethyl-}\text{l}\text{-ornithine}$), (Me₃Orn)_n, is usually not able to attain the α-helical conformation in aqueous solution independent of its pH value; however, it becomes α-helical at low concentrations of sodium perchlorate over a wide pH range according to the circular dichorism (c.d.) spectra. Cl^?, SO₄^2? and H₂PO₄^? do not induce α-helix formation. One can conclude that a distinct topology of the anions bound by the side chains is responsible for the α-helix-inducing effect of some water-structure-breaking anions such as perchlorate. This means that the anions are inserted between the ?N⁺ of the side groups shielding the positive charges repelling one another. The insertion of the anions requires that the water molecules surrounding the ions can be stripped off, which is easily possible if they are water-structure-breaking ones. At higher perchlorate concentrations, the c.d. spectrum changes. It is characterized by a negative shoulder near 208 nm and a pronounced minimum at ≈ 226 nm. With increasing temperature, the c.d. spectrum of the α-helix occurs. Finally the α-helix undergoes a conformational change to the random coil. The apparent transition enthalpy ΔH_vH is remarkably lower than that of the homologue (Me₃Lys)_n, obviously due to a lower cooperativity of the transition. In contrast to poly(l-ornithine), (Orn)_n, the c.d. spectrum of (Me₃Orn)_n remains almost unchanged after adding anionic surfactants such as sodium octyl sulphate (SOS) or sodium dodecyl sulphate (SDS). In organic solvents like methanol or isopropanol, in contrast to (Orn)_a and (Lys)_n, no α-helix formation occurs. However, in mixtures of these alcohols or dioxane with water, α-helix formation is induced by perchlorate, as in pure water. The thermal stability of the α-helix in these systems is increased.     

禽流感病毒A/Chicken/Guangdong/SS/94(H9N2)HA基因的克隆及序列分析

禽流感是由A型流感病毒引起的禽的一种疾病综合症.1878年首次在意大利爆发流行,当时称该病为"鸡瘟".之后,许多国家和地区都有该病的报道,包括美国、英国、澳大利亚、爱尔兰、比利时、荷兰、法国、俄罗斯、加拿大、以色列、匈牙利、日本、中国(包括香港)等[1-3].     

Dissimilatory nitrate uptake in Paracoccus denitrificans via a Δ\̃gmH+-dependent system and a nitrate-nitrite antiport system

Respiration-driven proton translocation has been studied with the oxidant pulse method for cells of denitrifying Paracoccus denitrificans oxidizing H₂ during reduction of O₂, NO^?₃, NO^?₂ or N₂O. A simplified scheme of anaerobic electron transport and associated proton translocation is shown that is consistent with the measured $\text{H}{}^{\mathrm{+}}\text{oxidant ratios}$. Furthermore, the kinetics and energetics of NO^?₃ uptake in whole cells of P. denitrificans were studied. For this purpose, we measured H₂ consumption or N₂O production after addition of NO^?₃ to a cell suspension, which indirectly gave information about uptake (and reduction) of NO^?₃. It was found that a lag phase in H₂ consumption or N₂O production appeared whenever the membrane potential was dissipated by addition of thiocyanate, carbonyl cyanide m-chlorophenylhydrazone or triphenyl-methylphosphonium bromide. However, these lag phases were not observed when NO^?₂ was present at the moment of introduction of NO^?₃. On the basis of these findings we conclude that there are two uptake systems for NO^?₃. One system is dependent on the proton-motive force and is probably used for initiation of NO^?₃ uptake. The other is an $\text{NO}{}^{\mathrm{?}}{}_3\text{NO}{}^{\mathrm{?}}{}_2$ antiport and its function is to take over NO^?₃ uptake from the first system.     

植物脱落酸PYR/PYL/RCAR受体

脱落酸(abscisic acid,ABA)对植物生长发育、生物与非生物胁迫的应答具有重要作用。近年研究发现ABA在植物体中引起的信号通路源于其受体的参与,以ABA受体及ABA信号通路为基础的研究成为新的研究热点。本文简略介绍了ABA受体的研究进展概况,重点介绍细胞内ABA受体PYR/PYL/RCAR蛋白对ABA信号感知和下游转导的研究进展,最后总结了PYR/PYL/RCAR介导的ABA信号通路。     

小流域内植被类型对土壤NO3-/NH4+空间变化的影响

流域内植被类型、地形地貌特征对土壤氮循环过程有重要的作用,是影响下游水体无机氮素来源以及富营养化的关键因子。通过比较小流域内4种植被类型(落叶松人工林、油松人工林、天然阔叶次生林和农田(玉米))对土壤NO3--N和NH4+-N含量空间变化的影响,揭示流域内不同立地条件下水源涵养林与土壤无机氮变化特征之间的关系。结果表明:4种植被类型土壤NO3--N和NH4+-N含量差异显著(P<0.05);由坡上到坡下土壤NO3--N和NH4+-N含量显著降低;在土壤表层NO3--N和NH4+-N含量最高,随着土层深度增加无机氮含量减少;与水源涵养林天然植被和人工林植被相比,农田土壤NO3--N含量最高(11.86mg·kg-1),有较高的氮流失风险。     

Comparison of red cell and kidney (Na+ + K+)-ATPase at 0°C

Human red cell and guinea pig kidney $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ were phosphorylated at 0°C. Using concentrations of ATP ranging from 10^?6 to 10^?8 M, ATP-dependent regulation of reactivity is observed with red cell but not kidney $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ at 0°C. In particular, with the red cell enzyme only, the following are observed: (i) the ratio of enzyme-bound ATP (E·ATP, measured by the pulse-chase method of Post, R.L., Kume, S., Tobin, T., Orcutt, B. and Sen, A.K. (1969) J. Gen. Physiol. 54, 306s-326s) to steady-state level of total phosphoenzyme (EP) decreases with decrease in ATP concentration and (ii) the apparent turnover of phosphoenzyme (ratio of Na⁺-stimulated ATP hydrolysis to level of total EP at steady state) also varies as a function of ATP concentration. In addition, when EP is formed at very low ATP (0.02 μM), and then EDTA is added, rapid disappearance of a fraction of EP occurs, presumably due to ATP resynthesis, only with the red cell enzyme. These differences in behaviour of the red cell and kidney enzymes are explained on the basis of the observed predominance of K⁺-insensitive EP in red cell, but K⁺-sensitive EP in kidney $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ at 0°C.     

Determination of H+e− ratios in chloroplasts with flashing light

Using a rapid pH electrode, measurements were made of the flash-induced proton transport in isolated spinach chloroplasts. To calibrate the system, we assumed that in the presence of ferricyanide and in steady-state flashing light, each flash liberates from water one proton per reaction chain. We concluded that with both ferricyanide and methylviologen as acceptors two protons per electron are translocated by the electron transport chain connecting Photosystem II and I. With methyl viologen but not with ferricyanide as an acceptor, two additional protons per electron are taken up due to Photosystem I activity. One of these latter protons is translocated to the inside of the thylakoid while the other is taken up in H₂O₂ formation. Assuming that the proton released during water splitting remains inside the thylakoid, we compute $\text{H}{}^{\mathrm{+}}\text{e}{}^{\mathrm{?}}$ ratios of 3 and 4 for ferricyanide and methyl viologen, respectively.In continuous light of low intensity, we obtained the same $\text{H}{}^{\mathrm{+}}\text{e}{}^{\mathrm{?}}$ ratios. However, with higher intensities where electron transport becomes rate limited by the internal pH, the $\text{H}{}^{\mathrm{+}}\text{e}{}^{\mathrm{?}}$ ratio approached 2 as a limit for both acceptors.A working model is presented which includes two sites of proton translocation, one between the photoacts, the other connected to Photosystem I, each of which translocates two protons per electron. Each site presents a ≈ 30 ms diffusion barrier to proton passage which can be lowered by uncouplers to 6–10 ms.