三唑醇分子烙印聚合物的吸附特性

实验采用分子烙印技术合成了对农药三唑醇有特异性作用的分子烙印聚合物(molecularly im-printed polymer,MIP).通过平衡吸附实验,评价了其对三唑醇的亲和力和选择性.与非烙印聚合物相比,分子烙印聚合物对三唑醇表现了较强的亲和力;Scatchard分析表明,以甲基丙烯酸为功能单体,以三唑醇为模板的分子烙印聚合物,通过离子作用和氢键作用可形成两类结合位点.用多点结合模型计算两类不同结合位点的离解常数为Kd1=2.80×10-4mol/L,Kd2=9.71×10-3 mol/L.实验表明,该聚合物对三唑醇有高亲和力和选择性,为在生物样品中选择富集三唑醇提供了可能性.     

安徽省自然科学基金结题项目简介(14)

1.主要研究结果和创新之处 (1)利用牛肾上腺皮质束、网状带细胞(BAC细胞)培养,放射性同位素受体结合标记,首次证明本地黄牛BAC细胞表面分布有机能性碱性成纤维细胞生长因子(FGF2)的高、低亲和力受体。它们高亲和力受体(具有酪氨酸激酶活性)的数目为4500个/细胞,低亲和力受体(硫酸乙酰肝素蛋白多糖HSPG)数目为28000个/细胞。两种受体的平衡解离常数(Kd)分别为23.5和381.9pm;转化生长因子β1(TGFβ1)和促肾上腺皮质激素(ACTH)刺激均可增加BAC细胞表面的FGF2受体数目。前者增加的高、低亲和力受体数目分别为6500个/细胞和140000个/细胞(Kd分别为3.151×10~(-11)mol/l     

利福平分子印迹聚合物微球的制备及性能研究

以聚乙烯醇1735为分散剂、利福平为模板分子、甲基丙烯酸为功能单体、乙二醇二甲基丙烯酸酯为交联剂,用水溶液微悬浮聚合法制备了利福平分子印迹聚合物微球;在水溶液中,利用静态平衡结合法和Scatchard分析法研究了印迹聚合物的结合能力和选择性.结果表明,利福平溶液的浓度范围在5×10-5~1×10-4mol/L内,所制得的印迹聚合物内存在两类不等价的结合位点,根据实验数据计算得到高亲和力结合位点的离解常数和最大表观结合量分别为Kd1=1.08×10-2mg/mL和Qmax1=14.40 mg/g;低亲和力结合位点的离解常数和最大表观结合量分别为Kd2=7.18×10-3mg/mL和Qmax2=12.51 mg/g.结合底物的实验结果表明,利福平分子印迹聚合物微球对利福平呈现出了较高的吸附性能和选择识别特性,能较简便地用于利福平的分离和富集.     

血糖控制良好的2型糖尿病患者血清VEGF—A及其受体VEGFR1和VEGFR2含量检测

研究目的：探讨血糖控制良好的2型糖尿病患者血管内皮生长因子-A（VEGF—A）及其受体VEGFR1和VEGFR2含量及其临床意义。研究方法：本文以31例血糖得到良好控制且没有明显大血管或微血管病变的2型糖尿病患者作为研究对象（实验组）,30位健康志愿者为对照组,同期检测患者及健康志愿者的空腹血糖、血脂、糖化血红蛋白水平,及血清VEGF—A、VEGFR1和VEGFR2含量,并加以比较和统计学分析。重要结论：研究结果表明,血糖控制良好的2型糖尿病患者血清中的VEGF-A及其受体VEGFR1和VEGFR2的含量和健康志愿者基本一致,两者无统计学意义,这可能显示血糖水平的合理控制能延缓血管并发症的产生。同时,2型糖尿病患者的血清VEGFR2含量和高密度脂蛋白胆固醇水平之间存在负相关,而血清VEGF-A、VEGFR2含量和甘油三脂水平之间存在正相关,这也表明糖尿病患者的血脂紊乱可能参与了血管生成的调节。     

靶向重组Aif-T18融合蛋白对TEM1阳性肿瘤细胞的杀伤作用的研究

验证Aif-T18融合蛋白对肿瘤内皮细胞标志分子1(TEM1)阳性肿瘤细胞的杀伤作用。采用基因工程技术方法构建重组质粒pIRES-TEM1-EGFP,转染TEM1阴性表达细胞MS1,经G418筛选得到TEM1阳性细胞株(MS1-TEM1);通过构建重组质粒pET302-Aif和pET302-Aif-T18,转化至表达宿主菌BL21中,经异丙基-β-D-硫代吡喃半乳糖苷(IPTG)诱导菌体得到所需的目的蛋白,采用流式细胞仪和MTT技术检测融合蛋白对MS1和MS1-TEM1细胞的亲和力和特异性杀伤的效果。本研究中,主要利用单链抗体sc FvT18选择性携带凋亡诱导因子AIF,靶向抑杀TEM1阳性细胞,从而为Aif-T18融合蛋白在肿瘤治疗中的开发应用提供实验依据。     

融合聚精氨酸九肽的小热休克蛋白 原核表达及蛋白纯化

目的构建小热休克蛋白-聚精氨酸九肽融合蛋白的原核表达载体,表达纯化融合蛋白.方法在小热休克蛋白表达载体的基础上利用点突变方法引入聚精氨酸九肽对应序列,转入BL21大肠杆菌感受态细胞进行原核表达,利用亲和层析方法对表达蛋白进行纯化.结果成功构建了融合聚精氨酸九肽的HSP16. 5融合蛋白表达载体,并对其在大肠杆菌细胞中的原核表达进行条件优化,研究显示:在诱导剂IPTG浓度为0. 5 mmol、37℃条件下诱导4 h目的蛋白产量较高.结论此实验成功构建了小热休克蛋白-聚精氨酸九肽融合蛋白的原核表达载体,得到了高纯度的小热休克蛋白-聚精氨酸九肽融合蛋白,为进一步研究其功能奠定基础.     

血管内皮生长因子基因反义核酸设计及其对HL60细胞生长的影响

目的:用计算机设计筛选出血管内皮生长因子(VEGF)的高效反义核酸;用实验方法研究VEGF反义核酸对HL60细胞生长的影响。方法:用RNAstructure(version3 7)软件,选择总自由能(Overall△G37)的相对低的反义核酸,共计7条,长度18～20核苷酸,全硫代修饰;细胞培养72h,采用胎盼蓝拒染法观察存活细胞,用ELISA法检测培养液中VEGF蛋白水平,分析反义核酸对HL60细胞的作用。结果:筛选出6条反义药物对HL60细胞生长有明显抑制作用,其中4条优于阳性对照组(A2);A7组细胞生长抑制率达41 74%。培养液中VEGF蛋白表达抑制率达47 81%。Overall△G37与反义核酸活性密切相关(r=-0 842,P<0 01)。结论:计算机辅助设计有助于获得更好的反义药物,VEGF反义核酸可抑制HL60细胞生长及VEGF蛋白表达,VEGF可能成为白血病治疗的新靶点。     

表达EGFR与HER2双特异性抗体融合蛋白腺病毒载体的构建及表达

目的：构建携带Cetuximab全长抗体与Herstatin C末端79个氨基酸（Herin）融合蛋白的腺病毒,探讨其在体外表达情况,及其与EGFR和HER2的结合能力。方法：通过合拉PCR,将抗体cetuximab基因与编码Herstatin C末端79个氨基酸（Herin）的序列融合,构建携带西妥昔单抗全长基因与Herin融合基因的腺病毒穿梭载体pDC339-AT2-Herin,并在293细胞内包装成重组病毒Ad5-AT2-Herin。通过酶联免疫吸附试验（ELISA）检测Ad5-AT2-Herin表达全长抗体融合蛋白的体外表达量;应用Western印迹法检测所表达全长抗体轻重链的完整性及均衡性;应用间接免疫荧光法（IFA）检测所表达的融合蛋白与EGFR( )Her2(-)的A431细胞,以及EGFR(-)Her2( )的SK-OV-3细胞膜结合的特异性。结果：ELISA检测分析表明,Ad5-AT2-Herin在293细胞中的表达量为209.3 ng/ml-317.3 ng/ml;Western blot显示轻重链表达平衡,大小符合预期;间接免疫荧光(IFA)显示腺病毒表达的融合蛋白与A431细胞表面受体EGFR及SK-OV-3细胞表面受体Her2均能特异性结合。 结论：成功构建携带EGFR和HER2双特异性抗体基因的腺病毒载体Ad5-AT2-Herin,该载体在体外能高效表达双特异性抗体蛋白AT2-HERIN,而且AT2-HERIN蛋白能在体外与EGFR和HER2特异结合。     

白介素24融合蛋白的表达、纯化和促伤口愈合功能的鉴定

本研究通过优化小鼠白介素24(mIL-24)基因序列,构建以Fc-tag为标签的mIL-24-Fc融合蛋白.对CHO进行MTX分级加压筛选,获得了稳定、高表达mIL-24-Fc蛋白的阳性克隆,同时优化了CHO细胞无血清驯化与稳定表达蛋白体系.经protein A亲和层析纯化mIL-24-Fc蛋白并利用AP binding实验检测生物学活性后,小鼠皮内注射mIL-24-Fc蛋白和伤口愈合实验证明:注射mIL-24-Fc蛋白后,小鼠表皮产生2~3层细胞增生,伤口愈合速度加快.     

基于小热休克蛋白-细胞穿膜肽蛋白纳米粒的siRNA递送系统

利用大肠杆菌表达系统原核表达并纯化小热休克蛋白-细胞穿膜肽融合蛋白（Hsp-Tat）; 用动态光散射和透射电镜成像分析Hsp-Tat纳米粒的粒径和形貌; 用噻唑蓝（MTT）法和活/死细胞染色法研究蛋白纳米粒的细胞相容性; 用凝胶阻滞实验检测纳米粒的小干扰RNA（siRNA）复合特性; 用激光共聚焦荧光显微镜成像研究蛋白纳米粒的siRNA胞内递送能力. 实验结果表明： 亲和层析得到高纯度的Hsp-Tat融合蛋白, 融合蛋白可自组装成40.2 nm空心球形结构; 蛋白纳米粒在HEK-293T细胞上显示出良好的细胞相容性; Hsp-Tat NP与siRNA形成的复合物可阻止siRNA在琼脂糖凝胶中迁移; Hsp-Tat 纳米粒能将Cy3-siRNA转运至细胞内, 可作为一种安全高效的非病毒核酸递送载体.        

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.     

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.     

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.     

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.     

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.     

Quantitative trait loci （QTLs） for quality traits related to protein and starch in wheat

Quality traits in wheat （Triticum aestirum L.） were studied by quantitative trait locus （QTL） analysis in a recombinant inbred line （RIL） population, a set of 131 lines derived from Chuan 35050 × Shannong 483 cross （ChSh）. Grains from RILs were assayed for 21 quality traits related to protein and starch. A total of 35 putative QTLs for 19 traits with a single QTL explaining 7.99-40.52% of phenotypic variations were detected on 10 chromosomes, 1D, 2A, 2D, 3B, 3D, 5A, 6A, 6B, 6D, and 7B. The additive effects of 30 QTLs were positive, contributed by Chuan 35050, the remaining 5 QTLs were negative with the additive effect contributed by Shannong 483. For protein traits, 15 QTLs were obtained and most of them were located on chromosomes 1 D, 3B and 6D, while 20 QTLs for starch traits were detected and most of them were located on chromosomes 3D, 6B and 7B. Only 7 QTLs for protein and starch traits were co-located in three regions on chromosomes 1D, 2A and 2D. These protein and starch trait QTLs showed a distinct distribution pattern in certain regions and chromosomes. Twenty-two QTLs were clustered in 6 regions of 5 chromosomes. Two QTL clusters for protein traits were located on chromosomes 1D and 3B, respectively, three clusters for starch traits on chromosomes 3D, 6B and 7B, and one cluster including protein and starch traits on chromosome 1D.