太子参内生细菌RPB-32的分类鉴定及其代谢物对小鼠肠道微生物群落的影响

【背景】肠道微生物在宿主肠道微环境稳态中起着至关重要的作用。众多因子,如抗生素、饮食和年龄等会干扰这种微平衡,引起菌群平衡发生改变,进而影响到机体健康状况。太子参具有心肌保护、增加免疫、抗氧化、抗糖尿病、抗应激、抗疲劳等药理活性,而太子参内生菌代谢物对肠道微生物的调控作用以及可能对机体健康的影响目前还未见报道。【目的】研究太子参内生菌RPB-32分类地位及其代谢提取物对小鼠肠道微生物的调节作用,以探讨其对机体健康的影响。【方法】对RPB-32进行传统方法和分子鉴定。将120只KM小鼠随机分为溶剂对照组(3%乙醇)及石油醚提取物、正丁醇提取物和乙酸乙酯提取物低、中、高剂量组(n=12)。小鼠灌胃处理14 d后采集粪便,通过选择性培养基培养及宏基因组测序[溶剂对照(S.F.1为3%乙醇溶液)、乙酸乙酯提取物高剂量组(S.F.2)]检测灌胃前后小鼠肠道微生物的变化。【结果】常规传统方法与分子鉴定结果表明RPB-32为芽孢杆菌(Bacillus sp.)。与空白对照组相比,给予内生菌代谢提取物的乙酸乙酯中剂量组、乙酸乙酯高剂量组、正丁醇低剂量组、正丁醇中剂量组及正丁醇高剂量组乳酸菌数量明显增加...     

Effect of normoxia and hypoxia on K(+) current and resting membrane potential of fetal rabbit pulmonary artery smooth muscle

At birth, the increase in O(2) tension (pO(2)) is an important cause of the decrease in pulmonary vascular resistance. In adult animals there are impressive interspecies differences in the level of hypoxia required to elicit a pulmonary vasoconstrictor response and in the amplitude of the response. Hypoxic inhibition of some potassium (K(+)) channels in the membrane of pulmonary arterial smooth muscle cells (PASMCs) helps to initiate hypoxic pulmonary vasoconstriction. To determine the effect of the change in pO(2) on fetal rabbit PASMCs and to investigate possible species-dependent differences, we measured the current-voltage relationship and the resting membrane potential, in PASMCs from fetal resistance arteries using the amphotericin-perforated patch-clamp technique under hypoxic and normoxic conditions. Under hypoxic conditions, the K(+) current in PASMCs was small, and could be inhibited by 4-aminopyridine, iberiotoxin and glibenclamide, reflecting contributions by Kv, K(Ca) and K(ATP) channels. The average resting membrane potential was -44.3+/-1.3 mV (n=29) and could be depolarized by 4-AP (5 mM) and ITX (100 nM) but not by glibenclamide (10 microM). Changing from hypoxia, that mimicked fetal life, to normoxia dramatically increased the K(Ca) and consequently hyperpolarized (-9.3+/-1.7 mV; n=8) fetal rabbit PASMCs. Under normoxic conditions K(+) current was reduced by 4-AP with a significant change in resting membrane potential (11.1+/-1.7 mV; n=8). We conclude that resting membrane potential in fetal rabbit PASMCs under both hypoxic and normoxic conditions depends on both Kv and K(Ca) channels, in contrast to fetal lamb or porcine PASMCs. Potential species differences in the K(+) channels that control resting membrane potential must be taken into consideration in the interpretation of studies of neonatal pulmonary vascular reactivity to changes in O(2) tension.     

模拟氮沉降对中亚热带森林土壤Ni、Cu、Zn含量的影响

通过野外模拟实验,研究3个氮沉降水平,CK(对照,0 kg·hm^-2·a^-1)、LN(低氮,30 kg·hm^-2·a^-1)和HN(高氮,100 kg·hm^-2·a^-1)处理对亚热带针叶(杉木)和阔叶(罗浮栲、浙江桂)森林土壤中微量元素Ni、Cu、Zn含量的影响。结果表明:就不同的林分来看,3种微量元素的含量大致呈杉木林>罗浮栲林>浙江桂林,施氮3 d后,浙江桂林和罗浮栲林土壤中3种微量元素在各处理之间差异不显著,仅发现杉木林土壤中的Ni含量在CK处理与LN及HN处理之间和无凋落物土壤中Cu含量在HN与CK及LN之间的差异显著; 3片林分中土壤表面有无凋落物处理总体对3种微量元素含量的影响不大。与施氮前相比,3片林分土壤中的Ni、Cu、Zn含量均有所下降,且浙江桂林在LN处理的降幅最大。     

纤维素多孔微载体的制备及其用于动物细胞培养

将纤维素铜氨溶液喷洒至-40℃的硅油：正己烷=1：4的冷冻液中形成含冰晶的微球,用-30℃、40%的H₂SO₄再生纤维素,并用EDAE盐酸盐修饰其表面,制成适合动物细胞培养的纤维素多孔微载体。利用该微载体培养能分泌尿激酶原(Pro-UK)的重组CHO细胞,在100mL搅拌瓶中换液培养25d,细胞最高密度为6.3×10⁶/mL,尿激酶原最高活性为2325IU/mL,共获28.7mg产品。之后转入1000mL搅拌瓶中培养,可观察到细胞可从种子微载体中自动转移到新微载体中生长繁殖直至所有微载体中都长有细胞。在25d二级培养中,细胞最高密度为7.3×10⁶/mL,尿激酶原最高活性为3108IU/mL,共获含353mg尿激酶原的上清13.7L。在培养后期换用无血清培养基培养,细胞生长及蛋白表达水平正常。     

新疆分离细菌的提取物对λ噬菌体紫外诱导的抑制作用

本研究从我国夏季高温、强辐射的新疆塔里木地区采集样品,进行了细菌的分离、纯化及分类鉴定。用丙酮萃取法抽提39个分离菌株的培养产物,通过高效液相色谱和紫外-可见吸收光谱分析其主要组分;利用λ噬菌体紫外诱导系统研究抽提物清除自由基的能力。高效液相色谱和紫外-可见吸收光谱分析的结果表明:多数分离菌株丙酮提取物的主要组分为类胡萝卜素物质。λ噬菌体紫外诱导系统抑制实验结果表明:29个菌株的丙酮提取物对λ噬菌体的紫外诱导具有不同程度的抑制作用,抑制率最高达81%。不同菌株之间的比较试验显示不同属细菌的丙酮提取物对λ噬菌体紫外诱导系统的抑制效果存在很大差异,其中以Methylobacterium的抑制作用最强;不同颜色菌株提取物的抑制效果也出现较大差异;Koanria中不同颜色菌株的抑制效果也有类似结果。本文为类胡萝卜素可能是该类细菌具有抗辐射能力的内因之一提供了直接证据。同时,也证实了λ噬菌体紫外诱导系统是一种很好的探讨细菌抗辐射机理的研究模型。     

荆条叶性状对野外不同光环境的表型可塑性

光照是影响植物生长和分布的重要环境因子。对生长在野外5种不同光环境下(林外、阔叶林林缘、阔叶林林下、针叶林林窗和针叶林林下)的荆条的叶片进行取样研究,通过对光合作用光响应曲线、叶绿素荧光、叶绿素含量、叶片氮磷含量以及叶片形态的测量,来反映荆条对不同光环境的表型可塑性。研究结果表明,荆条叶片对于野外不同的光环境具有很好的适应机制,叶片功能性状受到结构性状的调节。低光下通过高的比叶面积(SLA)、单位质量叶绿素含量、光系统II最大量子产量,低的暗呼吸速率、光饱和点、光补偿点、叶绿素a,b的比值来提高对光能的利用效率,维持生长;高光下则通过与SLA有关的叶片结构的变化对光合作用进行调节。大多数的叶性状只受到日光照总量的影响,SLA的大小与日最高光强有关,可以对不同日变化模式的光照做出迅速的响应,是适应不同光照的敏感指标。尽管光照是不同光环境下影响荆条叶性状的主要环境因子,土壤养分含量同样会对叶性状产生影响,高土壤养分下的高叶长与叶柄长的比值体现了植物对资源获取和支撑结构之间分配的权衡。     

Kv1.3 钾离子通道在卵巢癌细胞株中的表达及活性

目的:研究Kv1.3钾离子通道在SKOV3卵巢癌细胞中的表达及其在细胞增殖和细胞周期中的作用。方法:应用RT-PCR和免疫细胞化学鉴别Kv1.3钾离子通道在SKOV3卵巢癌细胞中的表达。应用MTT和流式细胞技术观察KV1.3钾离子通道对SKOV3卵巢癌细胞增殖及细胞周期的影响。结果:4-氨基吡啶是Kv1.3钾离子通道特异性阻滞剂。不同浓度的4-氨基吡啶可以明显抑制SKOV3细胞的增殖,并且细胞周期也受到影响。G0/G1细胞比例增加,S期和G2/M期细胞比例下降。结论:Kv1.3钾离子通道在SKOV3卵巢癌细胞中表达,并且在细胞增殖及细胞周期变换中扮演着重要的角色。     

Fucoidin enhances dendritic cell-mediated T-cell cytotoxicity against NY-ESO-1 expressing human cancer cells

Scavenger receptor A (SR-A) plays a crucial role in affecting the dendritic cell-mediated presentation of cancer testis antigens to T cells against human cancer cells. Here we use a dendritic cell-mediated model to verify that a sulphated polysaccharide, fucoidin, can regulate the adverse regulatory function of SR-A, and lead to the up-regulation of the anti-tumor immunological response. SR-A is a receptor of calreticulin (CRT) existing on the surface of dendritic cells (DCs). CRT is a specific receptor for a NY-ESO-1 cancer testis antigen, and CRT itself is responsible for the cross-presentation of NY-ESO-1 to CD8+ cells and the induction of anti-tumor immunity. Flow cytometrical analysis (FACS) showed that fucoidin was able to significantly enhance the binding ratio of NY-ESO-1 to human DCs in a concentration dependent manner, and that the addition of fucoidin promoted the DC maturation upon stimulation of NY-ESO-1. Results from a cytotoxicity assay indicated that fucoidin-treated DCs stimulated the CD8+ T cells more effectively than non-treated DCs via a cross-presentation pathway. Furthermore, it was found that after stimulated by fucoidin-treated DCs, the CD8+ T cells can release more IFN-γ than non-fucoidin-treated cells as detected by intracellular IFN-γ staining. We conclude that fucoidin enhances the cross-presentation of NY-ESO-1 to T cells leading to an increase of T-cell cytotoxicity against NY-ESO-1 expressing human cancer cells.     

Important role for the CA-NC spacer region in the assembly of bovine immunodeficiency virus Gag protein

Lentiviral Gag proteins contain a short spacer sequence that separates the capsid (CA) from the downstream nucleocapsid (NC) domain. This short spacer has been shown to play an important role in the assembly of human immunodeficiency virus type 1 (HIV-1). We have now extended this finding to the CA-NC spacer motif within the Gag protein of bovine immunodeficiency virus (BIV). Mutation of this latter spacer sequence led to dramatic reductions in virus production, which was mainly attributed to the severely disrupted association of the mutated Gag with the plasma membrane, as shown by the results of membrane flotation assays and confocal microscopy. Detailed mutagenesis analysis of the BIV CA-NC spacer region for virus assembly determinants led to the identification of two key residues, L368 and M372, which are separated by three amino acids, 369-VAA-371. Incidentally, the same two residues are present within the HIV-1 CA-NC spacer region at positions 364 and 368 and have also been shown to be crucial for HIV-1 assembly. Regardless of this conservation between these two viruses, the BIV CA-NC spacer could not be replaced by its HIV-1 counterpart without decreasing virus production, as opposed to its successful replacement by the CA-NC spacer sequences from the nonprimate lentiviruses such as feline immunodeficiency virus (FIV), equine infectious anemia virus and visna virus, with the sequence from FIV showing the highest effectiveness in this regard. Taken together, these data suggest a pivotal role for the CA-NC spacer region in the assembly of BIV Gag; however, the mechanism involved therein may differ from that for the HIV-1 CA-NC spacer.     

重庆市26个南亚果实蝇种群mtDNA 16S rRNA基因部分序列及其系统进化

对重庆市26个南亚果实蝇Bactrocera(Zeugodacus)tau(Walker)种群线粒体16S rRNA基因进行测序,获得长约350bp片段的序列。对获得的序列分析表明,A,T,C,G平均含量分别为35·0%,41·3%,7·2%,16·5%,其中保守位点数342个,变异位点数5个,简约信息位点2个,自裔位点2个,所有碱基转换总数为136,替换总数为50。利用MEGA2·1软件重建系统发生树,发现其中21个南亚果实蝇种群未出现分化,另外有5个南亚果实蝇种群出现了分化,但遗传分化程度小。