重组人截短型IL-6在大肠杆菌DH5α中的高效表达与纯化

为研究重组人截短型白细胞介素6(rhIL-6)工程菌高效表达的影响因素及纯化方法的优化.观察不同培养温度对重组人截短型IL-6工程菌生长密度和IL-6表达的影响;复性蛋白终浓度对复性效率的影响;细菌诱导培养后,经破菌-洗包-溶包初步纯化后,再经柱层析纯化IL-6.其结果为30℃培养后42℃诱导培养5h的IL-6表达效率最高,达32.8%;复性蛋白终浓度在0.5 g/L以下时,蛋白复性效率最佳,比活性为4.42×108 μmol/min*mg-1;进一步柱层析后,IL-6的纯度达96.5%,得率可达46.5%.最后得出培养和纯化工艺简便易行,可获得高纯度、高比活性的截短型rhIL-6的结论.     

重组人血管内皮细胞生长因子的表达及活性研究

目的 :用基因工程的方法在大肠杆菌中诱导表达人血管内皮生长因子 (VEGF) ,分离纯化并检测其生物学活性 ,以研究其在药学领域潜在的药用价值。方法 :利用PCR技术扩增VEGF基因片段 ,克隆到pQE30表达载体中 ,转化E .coliM15菌株后用IPTG进行诱导表达。经裂解细胞、变性、复性和Ni-NTAagarose金属螯合柱层析等方法纯化得到VEGF。用鸡胚绒毛尿囊膜 (CAM)血管生成实验检测VEGF的生物活性。结果 :重组表达质粒在大肠杆菌中成功地表达了相对分子质量为 2 0 6 0 0的融合蛋白 ,它以不溶性的包涵体形式存在 ,占菌体总蛋白的 30 %左右。经分离纯化融合蛋白SDS -PAGE显示为单一区带。CAM结果表明给药组血管生成数 (2 1 7± 3 1、39 3± 2 8)与对照组 (15 4± 1 9、2 9 2± 4 2 )相比有明显增加 (P <0 0 5 )。结论 :利用原核表达系统得到血管内皮生长因子具有天然VEGF生物学活性 ,为进一步的应用研究奠定了基础。     

重组人内皮抑素层析复性的相关研究

研究了由大肠杆菌表达的重组人内皮抑素(Recombinant human endostatin,rhES)的复性、纯化过程.比较先层析复性后纯化(工艺1)和先纯化后层析复性(工艺2)两种工艺的结果表明:工艺1所得rhES的收率和活性回收率分别为64.44%和213.19%,与工艺2相比分别提高8.70%和33.04%.工艺1中,研究了3种凝胶过滤柱复性及复性pH和上样蛋白浓度对复性的影响,并优化相关工艺.结果表明:在pH9.0,rhES浓度3.91 mg/mL的条件下,采用双梯度凝胶过滤Sephacryl S-100 HR进行层析复性,Sephadex G-25脱盐后,用SP Sepharose FF阳离子交换吸附纯化,获得活性回收率为213.19%,HPLC纯度为97.07%的rhES,其IC50为1.58 ug/mL.     

Na_2WO_4溶液结晶过程中除砷、硅等杂质

以机械活化碱分解黑白钨混合中矿及常压碱煮钨渣所得高杂粗钨酸钠溶液 (m(As) /m(WO3) =(0 .32～ 2 .94)× 10 -3,m(Si) /m(WO3) =(1.2 6～ 5 .84)× 10 -3)为原料进行钨酸钠结晶过程除砷、硅等杂质的研究 ,分析了结晶率及氧化铝的添加量对除杂效果的影响 .试验结果表明 :除杂效果与结晶率有关 ,当结晶率为 80 %～ 90 %时 ,除砷率和除硅率均可达 90 %以上 ,对m(As) /m(WO3)和m(Si) /m(WO3)分别为 3 .2 1× 10 -4 和 2 .41× 10 -3 的料液而言 ,除杂后的精液中m(As) /m(WO3)和m(Si) /m(WO3)可分别降至 3 .5 0× 10 -5和 3 .5 6× 10 -4 以下 .添加Al2 O3 可明显改善结晶过程中的除杂效果 ,且Al2 O3 经化学改性处理后 ,活性增强 ,除杂效果更好 ;在相同条件下 ,与不加铝盐相比 ,添加活性Al2 O3 可使精液中的杂质含量明显降低 ,除砷率由 88.48%增加到 94.6 5 % ,除硅率由 89.77%提高至97.74% .     

Long R^3-IGF-Ⅰ的克隆表达

在重组质粒pExSecI-IGF-Ⅰ的基础上,采用基因克隆方法合成了长链人胰岛素样生长因子(Long R3-IGF-Ⅰ)基因序列,构建基于该基因的重组原核表达质粒.将重组质粒转入E.coli,并对其表达条件进行了优化.实验结果表明,在37 ℃,IPTG终浓度0.6 mmol/L,诱导3 h条件下,Long R3-IGF-Ⅰ以包涵体形式可得到高效表达.将包涵体变性溶解后通过分离纯化,目的蛋白纯度达到95%以上.通过尿素梯度透析复性法对变性包涵体复性,复性率达到66%.     

基因工程人表皮生长因子（rhEGF）单克隆抗体的制备与初步鉴定

以基因工程人表皮生长因子(rhEGF)为抗原,免疫BALB/c小鼠,将免疫的小鼠脾细胞与小鼠Sp2/0-Ag14骨髓瘤细胞进行融合,经多次筛选和再克隆化,获得3株能稳定分泌抗rhEGF单克隆抗体(McAbs)的杂交瘤细胞株.对该3株杂交瘤细胞小鼠腹水进行了抗体效价测定,结果显示在1×10-6～2×10-7之间.对上述3种杂交瘤单抗(分别命名为:AE-2A4,AE-2G5和AE-3E11)进行了特异性、抗原决定簇及相对亲和力测定.结果表明,它们都具有抗rhEGF高特异性;并分别抗两个不同抗原决定簇;相对亲和力大小次序为:AE-3E11>AE-2A4>AE-2G5.Ig亚类鉴定结果显示:AE-2A4,AE-3E11同为IgG1亚类,而AE-2G5为IgG2b亚类.确定AE-2A4单克隆抗体可用于制备高纯rhEGF.     

超级电容器用活性炭的纯化与表征

主要对超级电容器用活性炭的纯化工艺进行了研究.采用不同的实验条件,对实验结果进行了对比,通过正交实验得到了最优的工艺条件.本研究将活性炭中的钾离子含量由3000×10-6以上降到100×10-6以下,铬离子含量由600×10-6以上降到了10×10-6以下,并对处理后活性炭的孔貌、孔径、比表面积进行了表征,结果表明,制备的超级电容器用活性炭具有纯度高、钾离子和铬离子含量低、比表面积大等优点.     

石龙子rADVR突触结构的定量分析

用实物及计算机投影系统对石龙子前背侧室嵴嘴外侧区突触的电镜图像进行放大处理和统计分析,测量结果:突触前膜平均厚度为70.3×10-10m,后膜平均厚度为117.7×10-10m,突触间隙平均宽度为147×10-10m,突触界面曲率为:平均值±标准差=(1.123±0.389)×10-10m,突触活性带长度为:平均值±标准差=(3 514.93±982.75)×10-10m,空心小泡直径为:平均值±标准差=(477.3±38.45)×10-10m(n=58),实心小泡直径为:平均值±标准差=(892±55.83)×10-10m(n=28).     

双氯醇胺对大鼠肝脏氮和葡萄糖代谢

利用离体肝脏灌流技术测定了CL(二个实验组,CL给药量分别为1×10-8 mol和1×10-6 mol)对大鼠肝脏物质代谢及相关酶活性的影响.结果表明CL可使大鼠离体肝脏灌流液中尿素氮浓度下降,并有一定的剂量效应和时间效应.在给药后灌流的1、2、3、4 h内,与对照组相比,1×10-6 mol的CL使大鼠离体灌流的肝脏产生的尿素氮分别下降了14.79%(P>0.05),17.88%(P>0.05),26.79%(P<0.05)和29.95%(P<0.01). 1×10-8 mol的CL具有类似的效应.而CL对大鼠离体肝脏灌流液中葡萄糖的水平影响不大.CL可抑制大鼠肝组织中GPT的活性,灌流4 h后1×10-6 mol的CL使得肝组织中GPT活性下降了24.65%(P<0.05),但1×10-8 mol的CL仅使GPT活性下降7.50%(P>0.05).CL还抑制大鼠肝组织中G6PDH的活性,1×10-6 mol的CL可使肝脏中G6PDH活性下降36.04%(P<0.05),同样1×10-8 mol的CL对G6PDH活性影响不显著.提示CL可直接通过影响肝脏的氮代谢及相关酶的活性继而影响机体的物质代谢.     

单克隆抗体亲和层析纯化基因工程人γ-干扰素研究

用抗基因工程人γ－干扰素单克隆抗体（２Ａ１２）细胞株亲和层析从表达人γ－干扰素的大肠杆菌抽提液中纯化经稀释复性后的γ－干扰素。一步纯化后的γ－干扰素含量达９５％以上，蛋白质的比活性达 １．２×１０ｕ／ｇ，收率为７８％。洗脱液用 ０．５ｍｏｌ／ＬＮａＣＩ的 ＰＢＳ溶液，洗脱率达９２．８％。     

Markedness and its Interpretation in Language Use

Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.     

Lower Paleozoic oil relationships within Williston Basin, Canada

The Williston Basin is a significant petroleum province, containing oil production zones that include the Middle Cambrian to Lower Ordovician, Upper Ordovician, Middle Devonian, Upper Devonian and Mississippian and within the Jurassic and Cretaceous. The oils of the Williston Basin exhibit a wide range of geochemical characteristics defined as "oil families", although the geochemical signature of the Cambrian Deadwood Formation and Lower Ordovician Winnipeg reservoired oils does not match any "oil family". Despite their close stratigraphic proximity, it is evident that the oils of the Lower Palaeozoic within the Williston Basin are distinct. This suggests the presence of a new "oil family" within the Williston Basin. Diagnostic geochemical signatures occur in the gasoline range chromatograms, within saturate fraction gas chromatograms and biomarker fingerprints. However, some of the established criteria and cross-plots that are currently used to segregate oils into distinct genetic families within the basin do not always meet with success, particularly when applied to the Lower Palaeozoic oils of the Deadwood and Winnipeg Formation.     

An Analysis of Catherine＇s Character——Wuthering Heights

Wuthering Heights, Emily Bronte＇s only novel, was published in December of 1847 under the pseudonym Ellis Bell. The book did not gain immediate success, but it is now thought one of the finest novels in the English language. Catherine is the key character of this masterpiece, because everybody and everything center on her though she had a short life. We can understand this masterpiece better if we know Catherine well.     

Characterization of Ordovician carbonate reservoirs, southeastern Saskatchewan, Canada

The discovery of the prolific Ordovician Red River reservoirs in 1995 in southeastern Saskatchewan was the catalyst for extensive exploration activity which resulted in the discovery of more than 15 new Red River pools. The best yields of Red River production to date have been from dolomite reservoirs. Understanding the processes of dolomitization is, therefore, crucial for the prediction of the connectivity, spatial distribution and heterogeneity of dolomite reservoirs.The Red River reservoirs in the Midale area consist of 3～4 thin dolomitized zones, with a total thickness of about 20 m, which occur at the top of the Yeoman Formation. Two types of replacement dolomite were recognized in the Red River reservoir: dolomitized burrow infills and dolomitized host matrix. The spatial distribution of dolomite suggests that burrowing organisms played an important role in facilitating the fluid flow in the backfilled sediments. This resulted in penecontemporaneous dolomitization of burrow infills by normal seawater. The dolomite in the host matrix is interpreted as having occurred at shallow burial by evaporitic seawater during precipitation of Lake Almar anhydrite that immediately overlies the Yeoman Formation. However, the low δ18O values of dolomited burrow infills (-5.9‰～ -7.8‰, PDB) and matrix dolomites (-6.6‰～ -8.1‰, avg. -7.4‰ PDB) compared to the estimated values for the late Ordovician marine dolomite could be attributed to modification and alteration of dolomite at higher temperatures during deeper burial, which could also be responsible for its 87Sr/86Sr ratios (0.7084～0.7088) that are higher than suggested for the late Ordovician seawaters (0.7078～0.7080). The trace amounts of saddle dolomite cement in the Red River carbonates are probably related to "cannibalization" of earlier replacement dolomite during the chemical compaction.     

Features of Women's language

Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women＇s language from these perspectives： pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women＇s language, more attention should be paid to language use in social context. What＇s more, the linguistic phenomena in a speaking community can be understood more thoroughly.     

Design and analysis of generic LBS application developing platform

Location based services is promising due to its novel working style and contents.A software platform is proposed to provide application programs of typical location based services and support new applications developing efficiently. The analysis shows that this scheme is easy implemented, low cost and adapt to all kinds of mobile nework system.     

橡胶颗粒沥青混合料低温抗裂性试验研究

以AC-13级配为基础,将橡胶颗粒代替部分集料掺入混合料中,以低温弯曲试验为评价方法对不同橡胶颗粒掺量下沥青混合料的低温抗裂性进行研究,并引入应变能密度值对混合料的低温抗裂性进行综合评价.试验结果表明:橡胶颗粒沥青混合料试件的破坏微应变均超过2 300,满足冬寒区的技术指标;无论是否掺加橡胶颗粒,随着温度的下降,沥青混合料破坏时的最大弯拉强度增大,弯拉应变降低,劲度模量增大;弯曲应变能密度在胶粒掺量为1%左右时具有较大的弯曲应变能密度值,此时橡胶颗粒沥青混合料具有较好的低温抗裂性.     

A computer generator for randomly layered structures

AcomputergeneratorforrandomlylayeredstructuresYUJia shun1,2,HEZhen hua2(1.TheInstituteofGeologicalandNuclearSciences,NewZealand;2.StateKeyLaboratoryofOilandGasReservoirGeologyandExploitation,ChengduUniversityofTechnology,China)Abstract:Analgorithmisintrod…     

低渗透非均质砂岩油藏启动压力梯度研究

理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。     

On Tragic Consciousness In The Works By Ernest Hemingway

As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.