在生物体外

Results as bellows: AtSIRT1 was located in Mitochondrial as hSIRT4 of human, and maybe take part in respiration and electron transformation chain, AtSIRT2 was located in nucleolus as hSIRT6 of human, maybe play important role in extend lifespan;mutation in AtSIRT1 leaded to cotyledon of plant turn to yellow and caused short life span. Mutation in AtSIRT2 could make the color of leaf turn to purple and accumulate a lot of anthocyanin;Sirtinol, a inhibitor of SIRT which did not cause the same model of the mutation of AtSIRT1 and AtSIRT2 indicated that the mechanism of Sirtinol was different from other organism;the structure of AtSIRT1 and AtSIRT2 were similar to other known Sir2, which indicated that they maybe have the same function;AtSIRT2 was overexpressed and its activity was detected.

结果表明，1，拟南芥AtSIRT1与人的同源蛋白hSIRT4相同，定位于线粒体，可能参与呼吸作用和电子传递，SIRT2与人的同源蛋白hSIRT6相同，定位于细胞核，可能同它的功能类似，在延缓衰老及调节细胞寿命方面起作用。2，AtSIRT1突变，可引起幼苗和植株的子叶变黄和早衰；AtSIRT2突变，可引起叶片发紫，沉积大量花青素。3，SIRT蛋白的抑制剂Sirtinol不能表型模写AtSIRT1和AtSIRT2突变体，说明Sirtinol在拟南芥中的作用机制不同于其他生物。4，AtSIRT1和AtSIRT2蛋白质结构预测表明与已知的Sir2蛋白相似，揭示其功能的相似性。5，在大肠杆菌中过量表达了其中一个基因(AtSIRT2)，可体外检测其酶学活性，进一步证明其功能。

BACKGROUND: Muscle-derived stem cells have some problems in purification, amplification and directional differentiation during in vitro culture. Basic fibroblast growth factor as a membrane of bioactive factor has been paid great attention, due to its biological activity.

背景：肌源性干细胞在体外培养过程中存在纯化、扩增、定向分化等难题，碱性成纤维细胞生长因子作为生物活性因子的一员，因其生物活性的多效性而备受关注。

At 10 weeks, smooth muscle bundles were observed and there were no significant difference between the two groups of the number of neogenesis capillary and positive VEGFR2 expression cells. To understand the release of exogenous growth factors from small intestinal submuscosa in bladder regeneration, the release of vascular endothelial growth factor and basic fibroblast growth factor from SIS in vitro were evaluated by ELISA and MTT method.

尽管文献报道制备的小肠粘膜下层中含有少量细胞生长因子，但尚不明确其在动物体内是否仍具有生物活性，为了解小肠粘膜下层在膀胱组织修复中外源性生长因子VEGF和bFGF释放，我们用ELISA法和MTT法体外测量冻干SIS在PBS孵育液中VEGF和bFGF的含量及对上皮细胞的增殖作用；用猪小肠粘膜下层行大鼠膀胱部分修复。

Transgenic tobacco plants were obtained through screening with kanamycin. The transgenic tobacco plants could delay TMV infection for about 25 days compared with non-transgenic tobacco plants. Pokeweed antiviral protein Ⅱ is expressed with high level in summer leaves. The expression of PAPⅡ is regulated by season. The total RNA was extracted from pokeweed (Phytolacca americana L.) leaves in summer using the method of TRIzol and used as template to amplify the PAPⅡ gene by RT-PCR and then the gene was cloned into E. coli expression vector and secreted expression pPIC9K vector. The two vectors with PAPⅡ gene were then transferred into E. coli strain BL21 (DE3)-plysS and Pachia pastoris GS115 strain respectively. The specific protein was produced induced by IPTG and methanol.

由于美洲商陆抗病毒蛋白Ⅱ是一个受季节调控表达的蛋白，本实验以美洲商陆夏季叶片为材料，通过对其总RNA的提取、反转录、并用PAPⅡ的特异引物进行PCR扩增，对PAPⅡ进行了基因克隆、序列分析，结果扩增出来的PAPⅡ基因与已经报道的序列同源性是99.9％，然后将该基因构建到原核、真核表达载体上，分别转化了大肠杆菌和毕赤酵母并对它们进行了诱导表达，在两个表达系统中均获得了有活性的PAPⅡ表达蛋白，体外生物测定表明表达的蛋白均具有抑制病毒的活性，PAPⅡ基因在酵母中还没有表达的报道，这为获得具活性PAPⅡ蛋白提供了一种简便可行的方法。

The transgenic tobacco plants could delay TMV infection for about 25 days compared with non-transgenic tobacco plants.Pokeweed antiviral protein II is expressed with high level in summer leaves. The expression of PAPII is regulated by season. The total RNA was extracted from pokeweed (Phytolacca americana L.) leaves in summer using the method of TRIzol and used as template to amplify the PAPII gene by RT-PCR and then the gene was cloned into E.coli expression vector and secreted expression pPIC9K vector. The two vectors with PAPII gene were then transferred into E.coli strain BL21 (DE3)-plysS and Pachia pastor is GS115 strain respectively. The specific protein was produced induced by IPTG and methanol.

由于美洲商陆抗病毒蛋白Ⅱ是一个受季节调控表达的蛋白，本实验以美洲商陆夏季叶片为材料，通过对其总RNA的提取、反转录、并用PAPⅡ的特异引物进行PCR扩增，对PAPⅡ进行了基因克隆、序列分析，结果扩增出来的PAPⅡ基因与已经报道的序列同源性是99.9％，然后将该基因构建到原核、真核表达载体上，分别转化了大肠杆菌和毕赤酵母并对它们进行了诱导表达，在两个表达系统中均获得了有活性的PAPⅡ表达蛋白，体外生物测定表明表达的蛋白均具有抑制病毒的活性，PAPⅡ基因在酵母中还没有表达的报道，这为获得具活性PAPⅡ蛋白提供了一种简便可行的方法。

Methods SH-SY5Y cells, a human neuroblastoma cell line, were incubated with different concentrations of fluoride for 48 hr and somle of them were treated with vitamin E precedently. The functional situation of cells was measured by MTT method; lipid peroxidation was detected by High-Performance Liquid Chromatography; phospholipid was separated by a Silica SepPak cartridge and neutral lipids by HPLC.

体外培养SH-SY5Y人脑神经母细胞瘤细胞，在培养液中加入不同浓度的氟化物或加入抗氧化剂，培养48h后用测定细胞MTT的方法来了解细胞的损伤程度，用高效液相色谱法分离和测定培养液中脂质过氧化物水平，用过柱和比色法测定细胞生物膜磷脂含量，用高效液相色谱法测定细胞生物膜辅酶Q和胆固醇含量。

Since Anfinsen and his co-workers brought forward the plus of the stability of thermodynamics is the right folding drive in the protein folding process basis on the study of the reversible ribotide-enzyme stretch and refold in 1960s, the investigation on the study of kinetics and thermodynamics of protein folding and binding have been greatly promoted.

自从上世纪60年代Anfinsen和他的合作者根据核糖核酸酶可逆伸展与重折叠的研究提出蛋白质折叠过程在热力学稳定方面获得的增益是控制蛋白质在体外正确折叠的驱动力以来，关于蛋白质折叠的动力学和热力学研究取得了很大的进展，特别是Wolynes所建立的能量形貌图理论对生物物理学的蛋白质折叠问题提供了强有力的理论模型。

By developing the optional measurement condition for ruminal fermentation products and comparing the effects of different levels of tannic acid on fermentation and methane production, the proper adding levels of tannic acid in vitro was established. We also examined the characteristics of rumen fluid fermentation and methane production of tannic acid unsupplemented or supplemented with different forage : concentrate ratios by using gas production method.

通过对瘤胃体外发酵系统中单宁酸的生物降解以及对瘤胃内环境影响的研究，初步确立了单宁酸改善瘤胃发酵的适宜添加量及其效果；并探讨了在不同精粗比底物条件下单宁酸对瘤胃体外发酵和甲烷产量的作用。

Methods:(1) Dissoluble PGN and CpG DNA were immobilized onto the surface of biotin cuvette for establishing target. Another effective tracking approach was established by immobilizing Escherichia lipid A F583 onto the surface of Non-derivatised cuvett. The biosensor technology was applied to screen anti-inflammatory TCM targeting on three key molecules.(2) The active compositions were isolated by AB-8 macroreticular resin from lycium bark. After the activities of compositions were evaluated, the most effective compositions was confirmed. In vitro, the affinities of different concentrations composition E binding with PGN, CpG DNA and lipid A were measured separately. The effect of composition E on vigor of RAW264.7 cells were tested by MTT and CCK-8, and its inhibition on TNF-α, which was released from RAW264.7 cells induced by PGN, CpG DNA and LPS, was also tested by ELISA. In vivo, murine sepsis models were made by intravenously heat-killed E.coli and heat-killed S.aureus, then protection of composition E on mice sepsis model were observed.

(1)将PGN及CpG DNA包被于生物素样品池，将lipid A包被于非衍生样品池，分别建立以PGN、CpG DNA及lipid A为靶点的技术平台，对114种抗炎中药水提物进行筛选、评价其活性物质含量，并评估出针对上述三种病原分子均具有较高结合活性的中药；(2)利用生物传感器跟踪检测技术、大孔吸附树脂分离技术，从地骨皮中定向分离与PGN、CpG DNA及lipid A均具有较高亲和力的活性组分；在体外实验中，测定不同浓度活性组分与PGN、CpG DNA及lipid A亲和力；MTT法及CCK-8法检测活性组分对RAW264.7细胞活力的影响；ELISA法检测活性组分对PGN(2μg/ml)、CpG DNA(10μg/ml)及LPS(100ng/ml)刺激小鼠RAW264.7细胞分泌TNF-α的抑制作用；在体内实验中，采用尾静脉注射致死剂量热灭活大肠杆菌和热灭活金黄色葡萄球菌，建立细菌脓毒症小鼠模型，观察活性组分对脓毒症模型小鼠的保护作用。

RDSSF could increase the incorporation of 〓H-Thymidine into DNA of Africa green monkey kidney cells (Vero E6) within the range of 1ug/ml～6ug/ml, but beyond the concentration of 2ug/ml, there is no increase of biological activity. RDSSF is stable within a range of pH 5.0～9.0 and sensitive to proteinase K and temperature.

RDSSF在体外可显著提高非洲绿猴肾细胞DNA中〓H-TdR掺入量，RDSSF浓度在1ug/ml～6ug/ml时活性明显，浓度达到2ug/ml后，增加RDSSF含量其活性不再增加；该物质在pH5～9范围内生物活性稳定；对蛋白酶k敏感；不耐热。

Fancy gold-plated dangling earrings with facetted White Opal crystals.

花式镀金悬垂耳环与facetted白欧泊水晶。

This essay chooses the study aim from biology teachers in middle school in Shi Jiazhuang which tells us that most of the middle school biology teachers in Shi Jiazhuang have the"burnout", lower successfulness, individualize.

本文选取石家庄市初中生物教师作为研究对象，运用问卷调查的方法对石家庄市初中生物教师职业倦怠的现状进行调查，调查结果发现，石家庄市初中生物教师这一群体普遍存在职业倦怠，情感枯竭程度偏高，成就感偏低，去个性化程度最为严重。