醇水混合体系中藏花素褪色的动力学反应及机理研究

从成熟的植物栀子果实中提取分离得到藏花素(1),采用半合成法制得藏红花酸二乙酯(2).利用紫外光谱研究了醇水混合体系中藏花素在紫外光照和自然光照下褪色的动力学反应,利用电子自旋共振技术研究了藏花素褪色过程中的自由基种类,利用气相色谱/质谱联用对藏红花酸二乙酯的褪色产物进行了结构推断.结果表明:藏花素的褪色反应遵循动力学一级反应,反应速率常数随醇/水混合体系中乙醇含量的增加而减少,且紫外光照下比自然光照下减小的更快;单线态氧和超氧阴离子参与了藏花素的褪色反应.其褪色原因主要是共轭双键被氧化加成和酯键水解光催化重排所致.最后,提出了藏花素褪色反应的可能途径.     

固液两步法的藏红花素生物合成

为提高现代生物技术生产藏红花素的产率和效率,该研究采取固液两步培养法将在固体培养基中获得的藏红花愈伤组织转移到液体培养基中进行生物合成,结果显示生物反应器比摇瓶培养更有利于生物量的积累,也更有利于2号藏红花素的合成。碳源、氮源和磷酸的消耗与藏红花素的合成密切相关。采取固液两步培养法可以有效解决藏红花素合成与藏红花素抑制细胞快速增殖的相互矛盾。该结果对具有抗癌作用的藏红花素产业化有重要意义。     

用扣除杂交法分离藏红花苷合成相关基因的克隆

对藏红花苷合成的相关基因进行了初步分离与克隆实验研究,旨在分离与藏红花苷合成相关的特异性表达的cDNA克隆,为进一步从分子水平上研究藏红花素的合成机理奠定基础。实验采用经添加诱导子和合成前体、处于藏红花苷合成状态下的藏红花细胞和未添加诱导子和合成前体、处于生长状态的对照藏红花细胞为材料,采用扣除杂交法成功地分离到6条与藏红花苷合成相关的特异性表达的cDNA克隆。结果表明:其中一条与胡杨中由高盐环境引起的差异表达的一条mRNA的序列有较高的同源性。     

对马鞭草中环烯醚萜苷类生物合成路径的探讨

目的 :探讨马鞭草中环烯醚萜苷类的生物合成路径。方法 :分析马鞭草中环烯醚萜苷类化合物结构并与其生物合成路径对照。结果 :新化合物 3 ,4 二氢马鞭草苷的结构不符合这一生物合成路径。结论 :马鞭草中环烯醚萜苷类的生物合成可能存在其它路径。     

Cryptic chlorination by a non-haem iron enzyme during cyclopropyl amino acid biosynthesis

Enzymatic incorporation of chlorine, bromine or iodine atoms occurs during the biosynthesis of more than 4,000 natural products. Halogenation can have significant consequences for the bioactivity of these products so there is great interest in understanding the biological catalysts that perform these reactions. Enzymes that halogenate unactivated aliphatic groups have not previously been characterized. Here we report the activity of five proteins-CmaA, CmaB, CmaC, CmaD and CmaE-in the construction of coronamic acid (CMA; 1-amino-1-carboxy-2-ethylcyclopropane), a constituent of the phytotoxin coronatine synthesized by the phytopathogenic bacterium Pseudomonas syringae. CMA derives from l-allo-isoleucine, which is covalently attached to CmaD through the actions of CmaA, a non-ribosomal peptide synthetase module, and CmaE, an unusual acyltransferase. We show that CmaB, a member of the non-haem Fe(2+), alpha-ketoglutarate-dependent enzyme superfamily, is the first of its class to show halogenase activity, chlorinating the gamma-position of l-allo-isoleucine. Another previously undescribed enzyme, CmaC, catalyses the formation of the cyclopropyl ring from the gamma-Cl-l-allo-isoleucine product of the CmaB reaction. Together, CmaB and CmaC execute gamma-halogenation followed by intramolecular gamma-elimination, in which biological chlorination is a cryptic strategy for cyclopropyl ring formation.     

苦参碱对小麦旗叶中蔗糖磷酸合成酶活性的调节

小麦旗叶中碳水化合物水平将影响穗的生长和物质的积累 .本文实验结果表明苦参碱对小麦旗叶蔗糖生物合成有明显的影响 ,并与叶片中蔗糖磷酸合成酶和酸性转化酶活性变化有关     

二十五味珊瑚丸中西红花苷Ⅰ&Ⅱ的含量测定研究

目的:建立高效液相色谱法测定二十五味珊瑚丸中西红花苷含量的方法。方法:采用HPLC法,色谱柱:Dionex C18柱(250mm×4.60mm,5um);流动相为甲醇-水(52:48),流速为0.8ml/min,检测波长为440nm,柱温为25℃;进样量10μl;用标准曲线法定量,测定二十五味珊瑚丸中西红花苷含量。结果:西红花苷Ⅰ在33μg～330μg范围内与峰面积呈良好的线性关系(r=0.9999),西红花苷Ⅱ在62μg～310μg范围内与峰面积呈良好的线性关系(r=0.9998),平均回收率为103.73%,RSD为1.68%(n=6)。结论:本方法操作简单、重现性好,可用于测定二十五味珊瑚丸中西红花苷Ⅰ&Ⅱ含量。     

Protein biosynthesis in organelles requires misaminoacylation of tRNA

In the course of our studies on transfer RNA involvement in chlorophyll biosynthesis, we have determined the structure of chloroplast glutamate tRNA species. Barley chloroplasts contain in addition to a tRNA(Glu) species at least two other glutamate-accepting tRNAs. We now show that the sequences of these tRNAs differ significantly: they are differentially modified forms of tRNA(Gln) (as judged by their UUG anticodon). These mischarged Glu-tRNA(Gln) species can be converted in crude chloroplast extracts to Gln-tRNA(Gln). This reaction requires a specific amidotransferase and glutamine or asparagine as amide donors. Aminoacylation studies show that chloroplasts, plant and animal mitochondria, as well as cyanobacteria, lack any detectable glutaminyl-tRNA synthetase activity. Therefore, the requirement for glutamine in protein synthesis in these cells and organelles is provided by the conversion of glutamate attached to an 'incorrectly' charged tRNA. A similar situation has been described for several species of Gram-positive bacteria. Thus, it appears that the occurrence of this pathway of Gln-tRNA(Gln) formation is widespread among organisms and is a function conserved during evolution. These findings raise questions about the origin of organelles and about the evolution of the mechanisms maintaining accuracy in protein biosynthesis.     

含不同官能团聚羧酸减水剂的吸附分散性能

通过水溶液自由基聚合法合成了含有不同官能团的聚羧酸减水剂,并研究了不同结构聚羧酸减水剂的吸附-分散性能,以及其对水泥水化性能的影响.结果表明:含酰胺基的聚羧酸减水剂对水泥浆体流动度的削弱程度最大,含酯基官能团的聚羧酸减水剂对水泥浆体流动度的影响程度较小.含磺酸基团的聚羧酸减水剂吸附性能增强;而含酰胺基及酯基的聚羧酸减水剂的吸附性能削弱.含酯基官能团的聚羧酸减水剂显著延缓了水泥水化诱导期,相比之下,含磺酸基官能团的聚羧酸减水剂提高了水泥水化加速期的最大水化放热速率.     

邛海鱼类区系的形成及其演变

本文作者根据50-60年代的调查,分析和探讨了邛海的自然鱼类区系形成同演变的规律.并对其形成提出了应从四个方面去认识:(1)雅砻江原来向南的流向;(2)长江上游的袭夺;(3)喜马拉雅山的造山运动.(4)高原地势和平原气候的渗合.对其演变分析了四种趋向:(1)流水性种类消失,(2)鱼类食性中植物性比重加大;(3)肉食性鱼类比重加大;(4)出现了种一级的分化.提出邛海与程海在鱼类区系上可列为同一类型,可与除程海以外的其它云南高原湖泊所属的另一类型相对比.并对Worthington(1940)提出的关于种的分化论点作了补充.     

表面活性剂和促进剂对重组里氏木霉生物合成t-PA的影响

重组里氏木霉(Trichoderma reesei)是染色体上整合有组织型纤溶酶原激活剂(t-PA)cDNA的基因工程菌株.对高产t-PA的重组里氏木霉菌株进行了筛选,并研究了乙酸钠和抗坏血酸(Vc)等促进剂及曲拉通(Triton100)、吐温80(Tween80)、十二烷基磺酸钠(SDS)和蔗糖酯等表面活性剂对菌株生物合成t-PA的影响.结果表明:乙酸钠能促进t-PA的生物合成,在发酵培养基中,其浓度为0.1%时,酶活可提高36.42%;吐温80对t-PA的合成有促进作用,其浓度为0.1%时,酶活可提高82.04%;抗坏血酸也可促进t-PA的合成,其浓度为0.05%时,酶活可提高124.82%;曲拉通和十二烷基磺酸钠加入时完全抑制t-PA的合成;蔗糖酯对t-PA的合成有部分抑制作用.     

生物活性多甲氧基黄酮羧酸和氨基酸衍生物的合成及其结构表征

多甲氧基黄酮(PMFs)是一类存在于柑橘属植物中具有显著抗癌、抗炎和抗氧化等生物活性的天然产物。为了提高多甲氧基黄酮类化合物的水溶性和药用价值,以2种来源丰富且抗癌活性高的多甲氧基黄酮橘皮素和川陈皮素为底物,分别经过氧丙酮氧化得到多甲氧基黄酮醇[3-羟基橘皮素(7)和3-羟基川陈皮素(8)],化合物7和化合物8分别在K₂CO₃和N,N-二甲基甲酰胺条件下与氯乙酸乙酯进行Williamson反应,生成了多甲氧基黄酮羧酸酯类衍生物,再经碱性水解合成了2种未见文献报道的多甲氧基黄酮羧酸衍生物[橘皮素-3-O-乙酸(1)和川陈皮素-3-O-乙酸(2)]。化合物1和化合物2在CH₂Cl₂作溶剂、1-(3-二甲基丙胺)-3-乙基碳二亚胺盐酸盐为缩合剂和4-N,N-二甲氨基吡啶为辅助剂的条件下,分别与2种不同的氨基酸甲酯盐酸盐发生缩合反应,得橘皮素和川陈皮素的氨基酸甲酯衍生物,然后经水解反应得到了4种新型多甲氧基黄酮氨基酸衍生物3～6。对所合成的化合物用¹H NMR、¹³C NMR和MS等方法进行了结构表征。     

肿瘤磁感应热疗用115kHz中频交变磁场对小鼠生殖系统的影响

 为了研究肿瘤磁感应热疗用115kHz中频交变磁场对精子密度、活动率、精子运动系数和睾丸重量的影响,以及观察睾丸组织病理变化,探讨中频交变磁场下生殖系统的生物学现象,采用115kHz中频磁场对Balb/C小鼠进行全身短期连续辐照（7d）和长期连续辐照（28d）,辐照强度为12mT,时间为1h/d,分为对照组（0T）和辐照组（12mT）,每组8只小鼠。辐照后取小鼠双侧睾丸称重、固定,并取附睾精子进行分析。结果表明,短期辐照可使睾丸重量增加,长期辐照可使精子密度、活动率增加,短期与长期辐照中频交变磁场对小鼠雄性生殖系统无损伤效应。由此可得,肿瘤磁感应热疗用115kHz中频交变磁场具有促进睾丸组织细胞增殖的作用,且无损伤效应。     

Immobilization of biomacromolecules on poly-L-lactide surface via a layer-by-layer method for the improving of its cytocompatibility to bone marrow stromal cells

Hyaluronic acid （HA） and chitosan （CS） were immobilized on the surface of poly-L-lactide （PLLA） by the following procedure： Firstly, PLLA was aminolyzed with 1, 6-hexanediamine, and part of the PLLA surface ester groups were converted to free amino groups. Then negatively charged hyaluronic acid and positively charged chitosan were deposited onto the surface of aminolyzed PLLA film in a layer-by-layer assembly manner. The effect of the layer-by- layer deposition was evaluated by ATRoFTIR spectroscopy, Raman spectroscopy and static contact angle measurements. The cytocompatibility of PLLA sample to bone marrow stromal cells （BMSCs） was improved after modification with chitosan and HA. The cell attachment, activity, and proliferation on CS/HA modified PLLA films were enhanced comparing with the control. The cells cultured on the modified PLLA samples excreted abundant cytoplasm and can differentiate to vascular smooth muscle （SM）-like （SM-like） cells. A macroporous three-dimensional PLLA scaffold was prepared by integrating both the technique of freeze-drying and particle leaching. Layer-by-layer modification by HA/CS and cell culture was also applied on this scaffold. The scaffold cultured with BMSCs for 2 weeks has been tested successfully in vivo as a patch for repairing the artificial incision on canine pulmonary artery.     

八种天然黄酮类化合物的抗氧化构效关系

采用H2O2－CTMAB－Luminol化学发光体系和Fe^2＋诱发脂蛋白PUFA过氧化比色体系，研究了八种高纯度的天然黄酮类化合物清除H2O2、LO和LOO的构效关系。结果表明，这八种天然黄酮类化合物都能 有效地清除H2O2、LO和LOO。根据其清除作用和化学结构分析，得出如下结论：B环上羟基是清除H2O2、LO和LOO的主要活性基团，A环上羟基是清除H2O2、LO和LOO的重要基团，并且B环上羟基相邻清除作用就大大增强；糖甙对清除H2O2、LO和LOO也有贡献。     

黄鳝肠道蛋白酶的应用研究

将黄鳝内脏混合物提取液作同工酶电泳,得到3条蛋白酶同工酶条带,其中ProteaseⅠ条带与从黄鳝肠道中提取的蛋白酶纯品SDS-PAGE条带重合,表明从黄鳝内脏混合物中可以提取到黄鳝肠道蛋白酶纯品.金属离子Cu2 和Ba2 对酶有激活作用,EDTA、Mn2 、Pb2 对酶有抑制作用,而Ca2 、Mg2 、K 、Na 对酶活性无明显影响.通过酶与化学试剂的配伍试验测得:CO23-、SO32-、硼砂对酶活性无影响,甘露醇、橄榄油、聚乙烯醇对酶活性有轻微抑制作用,SO42-、H2PO4-、BO32-和海藻酸钠对酶有轻微激活作用,而洗涤剂中常用的表面活性剂SDS、LAS、CHAPS和TritonX-100在低浓度时对酶活性无明显影响,酶在0·02%的SDS、0·05%的LAS和0·1%的CHAPS中保持相对稳定,40min酶活保持在50%左右.酶专一底物测试结果表明:黄鳝肠道蛋白酶对弹性蛋白酶的专一底物刚果红弹性蛋白(elastin-CongoRed)有水解活性,而对胰蛋白酶和胰凝乳蛋白酶的专一底物N苯甲酰L精氨酸乙酯(BAEE)和N乙酰L酪氨酸乙酯(ATEE)无水解活性,表明黄鳝肠道蛋白酶为弹性蛋白酶.     

汽爆玉米秸秆渣诱导产纤维素酶及其水解特性

为提高玉米秸秆的生物降解效率,研究了汽爆处理玉米秸秆渣对纤维素酶的诱导作用及所产纤维素酶的水解特性。结果表明：玉米秸秆采用汽爆处理可以去除大量的半纤维素成分,获得的汽爆渣对里氏木霉NL02合成纤维素酶的诱导作用良好,以10 g/L的汽爆渣为碳源发酵7 d滤纸酶活达到1.90 U /mL。发酵5 d获得的汽爆酶水解效果最佳,当酶用量25 U/g,底物质量浓度50 g/L条件下,水解24 h酶解得率达到95.9%,水解48 h葡萄糖产率为76.4%。当汽爆渣质量浓度为80 g/L,酶用量为15 U/g时,48 h酶解效率达到90.1%,葡萄糖产率为62.9%。     

Disruption of retinogeniculate afferent segregation by antagonists to NMDA receptors.

Afferent activity has an important role in the formation of connections in the developing mammalian visual system. But the extent to which the activity of target neurons shapes patterns of afferent termination and synaptic contact is not known. In the ferret's visual pathway, retinal ganglion cell axons from each eye segregate early in development into eye-specific laminae in the lateral geniculate nucleus (LGN). The dorsal laminae (termed laminae A and A1) then segregate further into inner and outer sublaminae that retain input from on-centre and off-centre retinal axons, respectively. Thus, individual retinogeniculate axons form terminal arbors within laminae A and A1 that are restricted to one inner or outer sublamina. We report here that blockade of N-methyl-D-aspartate (NMDA) receptors on LGN cells with specific antagonists during the period of sublamina formation prevents retinal afferents from segregating into 'On' and 'Off' sublaminae. Retinogeniculate axons have arbors that are not restricted appropriately, or are restricted in size but inappropriately positioned within the eye-specific laminae. NMDA receptor antagonists may specifically disrupt a mechanism by which LGN neurons detect correlated afferent and target activity, and have been shown to reduce retinogeniculate transmission more generally, causing LGN cells to have markedly reduced levels of activity. These results therefore indicate that the activity of postsynaptic cells can significantly influence the patterning of inputs and the structure of presynaptic afferents during development.     

半育耳蕨化学成分及其抑制α-葡萄糖苷酶活性研究

采用MCI柱色谱、硅胶柱色谱、Sephadex LH-20凝胶柱色谱和半制备HPLC等色谱分离手段对半育耳蕨的化学成分进行研究.利用理化性质及波谱数据(MS、NMR)对其结构进行鉴定,从半育耳蕨体积分数95%的乙醇提取物中分离并鉴定了10个化合物,分别是2R, 3R-3, 5, 6, 7, 8,4′-hexahydroxyflavane(1)、(-)-表儿茶素(2)、(+)-epicatechin(3)、arachniodesin A(4)、4β-carboxymethyl-(-)-epicatechin methyl ester(5)、eriodictyol(6)、daucosterol(7)、3, 4-二羟基苯甲酸乙酯(8)、圣草酚7-O-β-D-(6′-甲酯基)-吡喃葡萄糖醛酸苷(9)、圣草酚7-O-β-D-(6′-乙酯基)-吡喃葡萄糖醛酸苷(10).化合物1～10均为首次从耳蕨属中分离得到.活性测试结果表明,化合物4、5和6对α-葡萄糖苷酶具有较好的抑制活性,其IC_(50)分别为(26.2±1.9)、(32.6±2.5)和(51.5±3.5)μmol·L~(-1).