以β-半乳糖苷酶为标记选出高效表达乙型肝炎病毒表面抗原的天坛株重组痘苗病毒

我们从天坛株痘苗病毒的基因组中,分离编码11K及25K蛋白的双向转录启动子,以胸苦激酶(TK)基因为旁侧序列,插入到质粒pAT153中,构建成可以同时表达两个外源基因的载体质粒,并将乙型肝炎病毒表面抗原(HBsAg)基因及大肠杆菌的β-半乳糖苦酶(LacZ)基因,插入该载体质粒,使它们分别在11K及25K启动子钠控制下,构建成共表达质粒。用钙沉淀法将此质粒DNA同天坛株痘苗病毒在细胞内进行同源重组,在含X-gal的培养基挑蓝色蚀斑,简便、准确地选出重组痘苗病毒。所选到的重组痘苗病毒高效表达了HBsAg。我们还把上述重组痘苗病毒接种动物,观察了它们的毒力及免疫原性。     

带有乙型肝炎表面抗原基因的重组痘苗病毒——一种可能的乙型肝炎活疫苗

我们组建了带有痘苗病毒胸腺嘧啶核苷激酶(TK)基因的痘苗病毒载体,并将乙型肝炎(以下简称乙肝)表面抗原(HBsAg)基因(adr亚型)插入载体质粒,置于痘苗病毒早期启动基因的调控下。通过感染与转染相结合的方法,得到了带有乙型肝炎表面抗原基因的重组痘苗病毒。它保持了原有的感染性,并能有效地表达乙肝表面抗原。当病毒接种量为每个细胞1PFU(空斑形成单位)时,每1×10~6细胞感染后24h可产生2—3μg HBsAg,其中大部分释放于细胞培养液中。每升培养液可以产生0.5—1mg HBsAg。表达产物HBsAg能形成颗粒,其大小、浮力密度,多肽组份等都与病人血清中的HBsAg颗粒相同。用重组痘苗病毒免疫家兔,可以产生抗乙肝表面抗原的专一性抗体。实验结果表明,利用本文组建的重组痘苗病毒不仅可以提供一个生产乙肝表面抗原的系统,而且这种重组痘苗病毒就可作为活疫苗使用,来预防乙肝病毒感染。     

极谱法快速同时测定锌电解液中微量铜、镉、钴

铜、镉、钻在了丁二酮肟－氯化铵－柠檬酸钠－ＨＡｃ－ＮａＡｃ体系中有灵敏的极谱波。其峰电位分别为－０．２４Ｖ、－ ０．７９Ｖ和－０．９３Ｖ,线性范围分别为４ × １０~（－６）～ ４ × １０~（－３）、５ × １０~（－６）～ ５×１０~（－３）、１×１０~（－７）～１×１０~（－４）ｇ／Ｌ,检测限分别为２×１０~（－６）、４ × １０~（－６）和５×１０~（－８）ｇ／Ｌ。方法无需预处理直接用于锌电解液中微量铜、镉、钴的测定,结果令人满意。     

阿霉素在钴离子注入修饰电极上的电化学行为及其应用

阿霉素在０．１ｍｏｌ／Ｌ ＨＡｃ－ＮａＡｃ（ｐＨ４．６２）缓冲溶液中用注入钴的修饰玻碳电极为工作电极进行伏安测定,得到一良好的还原峰;Ｅｐ＝－０．５２２Ｖ（ｖｓ．ＳＣＥ）。峰电流与阿霉素浓度在１．４×１０＾－６－１．４×１０＾－６ｍｏｌ／Ｌ和１．４×１０＾－６－５．６×１０＾－５ｍｏｌ／Ｌ范围内呈线性关系,。     

铟(Ⅲ)-7-(1-苯偶氮)-8-羟基喹啉-5-磺酸钠的配位吸附波及其应用

在ｐＨ４．５的 ０．０１５ ｍｏｌ／Ｌ ＨＡｃ－ＮａＡｃ介质中，Ｉｎ铟（Ⅲ）－７－（１－苯偶氮）－８－羟基喹啉－５－磺酸钠（ＢＱ）在滴汞电极上于－０．６６５－０．７０ Ｖ（ｖｓ．ＳＣＥ）电位处得到络合物的吸附还原波，其一阶导数峰电流Ｉ＇ｐ与Ｉｎ（Ⅲ）浓度在４．０× １０~（－８）～３．５ × １０~（－６）ｍｏｌ／Ｌ范围内呈良好的线性关系，检测限为１．６ × １０~（－８）ｍｏｌ／Ｌ Ｉｎ（Ⅲ）．应用该法测定了铅粉和氢氧化铝样品中的微量铟，结果令人满意。对电极反应机理进行了探讨。     

血清中锂的原子吸收法测定

对血清中锂的火焰原子吸收分光光度分析法（ＦＡＡＳ）进行了研究，发现血清在０．２４ｍｏｌ／ＬＨＮＯ３介质中，用３０００×１０－６Ａｌ３＋作为碱土金属干扰的抑制剂，用ＦＡＡＳ法测定血清理，其回收率和精密度都很好，分别为１０４．７％和２．８％．本法１％吸收的灵敏度为０．０３３μｇ／ｍＬ。采用本法对服用碳酸锂治疗苯中毒的患者血清中锂进行了测定，其含量在０．７５～４．６４μｇ／ｍＬ之间，结果令人满意。     

CHO-EPO-EGFR细胞株的构建及在光固定EGF材料上的生长

采用人工细胞（ｌｉｐｏｓｏｍｅ）法将两种质粒ｐＣＥＰＯ及ｐＲＣ／ＣＭＶ／ＥＧＦＲ转染宿主细胞－中国仓鼠卵巢细胞（ＣＨＯ）,得到表达水平稳定,形态良好的细胞株ＣＨＯ－ＥＰＯ－ＥＧＦＲ,研究该细胞株在光固定ＥＧＦ的聚苯乙烯材料上生长情况。     

几个分子和负离子的QCI势能曲线

用ＱＣＩＳＤ（Ｔ）／６３１１＋Ｇ（３ｄｆ，２ｐ）（简称ＱＣＩ）计算了ＢＨ－（２Π），ＢＦ－（２Π），ＢＨ（１∑），ＢｅＨ－（１∑），ＢｅＦ－（１∑）和ＢＦ（１∑）的较完整的势能曲线，探讨了这些体系的稳定性和解离过程．同时用Ｍｏｒｓｅ函数和ＥＲ函数拟合得到势能曲线，用拟合结果计算谐性频率和非谐性频率，并与已有的实验数据（ＢＨ和ＢＦ分子）进行比较，由此考察了ＱＣＩ理论方法在远离平衡构型时的可靠性．对于因单参考态近似导致的反常势能曲线进行了ＣＡＳＳＣＦ计算．还比较了ＱＣＩ和Ｇ２理论中标准的ＭＰ２（ｆｕｌ）／６３１平衡几何构型及其对ＱＣＩ总能量的影响．     

聚乙烯醇缩对甲酰基偶氮磺的合成和液—固萃取分离镍,铜,锌的研究

报道了２－（４－磺酸苯偶氮）－７－（４－甲酰基苯偶氮）－１,８－二羟基－３,６－萘二磺酸以及在偶氮磺与聚乙烯醇（ＰＶＡ）的缩合产物的合成及其性质。并从试剂的结构和反应机理方面对ＳＡｐＦ和ＰＶ·ＳＡｐＦ的分析性能进行了对比研究。实验结果表明,采用高分子显色剂做萃取剂,易获得萃取率和好的分离效果,且萃取不受被萃物结构和电性的影响。     

电位型砷传感器的研究及分析应用

以ＣＴＭＡＢ修饰碳糊电极将砷钼杂多酸固定于碳糊电极表面,使其表面形成了杂多酸薄膜。该电极对４．００×１０＾－６～１．００×１０＾－３ｍｏｌ／Ｌ范围的砷钼杂多酸有能斯特响应,斜率为４７．５ｍＶ／ｐＡｓ（ｖ）,从而可间接测定砷。     

二氧化碳和丙烯直接合成甲基丙烯酸的NiPMo/TiO2催化剂的研究

用溶胶-凝胶法以磷钼酸(MPA)的镍盐溶液水解钛酸四丁酯制备了NiPMo/TiO2催化剂.使用ICP、 XRD、 TG-DTA、 IR、 TPD-MS和微反应技术研究了催化剂的化学组成、热稳定性、化学吸附性质和催化反应性能.杂多钼酸盐与TiO2通过O2-在TiO2表面发生了键合.在623 K下,杂多阴离子仍保持原有的Keggin结构.CO2在Lewis酸位Ni(Ⅱ)和Lewis碱位Ni-O-Mo的桥氧协同作用下生成CO2卧式吸附态Ni(Ⅱ)←O-(CO)←(O--Ni).丙烯有多种吸附态在催化剂上吸附.在563 K、 1 MPa和空速1500 h-1的反应条件下,丙烯的摩尔转化率为3.2%,产物MAA选择性为95%.     

CoFe2O4自形成磁性液体场致结构化对磁化的影响

The Langevin paramagnetic theory can’t describe the relation between magnetization of ferrofluids and applied magnetic field. The structuralization of ferrofluids, which is considered the main influence factor of the magnetization, is regarded. The part of magnetization works is deposited when the structure is forming. This action influences the magnetization of ferrofluids directly or indirectly. On the base of the “compressing” model, the Langevin function that usually describes the magnetization of ferrofluid is modified, and a well-fitted curve is obtained. An equation of the relation between the equivalent volume fraction after being “compressed” and the intensity of magnetic field is discovered, which approximately describes the process of magnetization. The relation between the approximate initial susceptibility and the volume fraction can be obtained from modified formula.     

Highly regioselective Buchwald–Hartwig amination at C-2 of 2,4-dichloropyridine enabling a novel approach to 2,4-bisanilinopyridine (BAPyd) libraries

The highly regioselective Buchwald–Hartwig amination at C-2 of the cheap and readily accessible reagent, 2,4-dichloropyridine with a range of anilines and heterocyclic amines is described. This new methodology is robust and provides a facile access to 4-chloro-N-phenylpyridin-2-amines on 0.25 mol scale. These intermediates undergo a further Buchwald–Hartwig amination at higher temperature to enable rapid exploration of the chemical space at C-4 and to provide a library of 2,4-bisaminopyridines.     

Sulfur-directed metal-free and regiospecific methyl C(sp3)–H imidation of thioanisoles

Regiospecific and direct imidation of the methyl C(sp³)–H bond of thioanisoles is realized under mild and metal-free conditions with N-fluorobis(benzenesulfonyl)imide as an oxidant and nitrogen source. Proposed mechanism suggests that thionium ion intermediates and a Pummerer-type reaction are involved. The imidation has advantages such as high step-economy, excellent functionality tolerance, and regiospecificity, giving structurally diverse imidation products.     

Selective syntheses of 2H-1,3-oxazines and 1H-pyrrol-3(2H)-ones via temperature-dependent Rh(II)-carbenoid-mediated 2H-azirine-ring expansion

The Rh(II)-catalyzed reaction of 2-carbonyl-substituted 2H-azirines with ethyl 2-cyano-2-diazoacetate or 2-diazo-3,3,3-trifluoropropionate provides an easy access to 2H-1,3-oxazines and 1H-pyrrol-3(2H)-ones. These compounds can be selectively prepared from the same starting material using temperature as the only varied parameter. The 2-azabuta-1,3-diene intermediate, a common precursor for both heterocyclic products, isomerizes into 2H-1,3-oxazine under kinetic control, while 1H-pyrrol-3(2H)-one is the sole product of the reaction at elevated temperatures. According to DFT-calculations a one-atom oxazine ring contraction involving ring-opening to a 2-azabuta-1,3-diene intermediate, followed by a 1,5- and 1,2-prototropic shift leads to the consecutive formation of imidoylketene and azomethine ylide, which then further undergo cyclization to the pyrrole derivative.     

Synthesis of 1-benzyloxypyrazin-2(1H)-one derivatives

Different approaches for the synthesis of 1-benzyloxypyrazin-2(1H)-one derivatives from simple amino acids have been investigated. A library of 33 precursors for the preparation of N-hydroxy pyrazinones was obtained in moderate to good yields.     

Copper catalyzed/mediated synthetic methodology for ebselen and related isoselenazolones

Scope of the copper catalyzed/mediated selenium-nitrogen coupling reaction has been studied for the synthesis of isoselenazolones. It is noticed that the 2-chloro, 2-bromo-, and 2-iodo-aryl amides substrates can be exploited in the selenium-nitrogen coupling reaction by employing 25-100 mol % of CuI/1,10-phenanthroline (L) and potassium carbonate as a base in DMF. Furthermore, electron rich 2-chloro-arylamides also underwent selenium-nitrogen coupling reaction to give biologically important selenium-nitrogen heterocycles. Also, copper-catalyzed selenium-nitrogen coupling reaction has been meticulously applied for the synthesis of diaryl diselenides having methoxy, amine, and amide functionality from respective aryl iodides in the presence of stoichiometric amount of succinimide as an external Se-N coupling partner.     

Knoevenagel condensation catalyzed by novel Nmm-based ionic liquids in water

A series of novel N-methyl morpholine (Nmm) based ionic liquids with 1,2-propanediol group were synthesized and used as catalysts for Knoevenagel condensation at room temperature in water. Under the effect of the catalyst, various aldehydes or aliphatic ketones could react with a wide range of activated methylene compounds well, including malononitrile, alkyl cyanoacetate, cyanoacetamide, β-diketone, barbituric acid, 2-arylacetonitrile and thiazolidinedione. Furthermore, most of the products could be separated just by filtrating and washing with water. Additionally, the catalyst is recyclable and applicable for the large-scale synthesis.     

Construction of polyheterocyclic spirotetrahydrothiophene derivatives via sulfa-Michael/aldol cascade reaction

A series of polyheterocyclic spirotetrahydrothiophene derivatives were obtained in moderate to excellent yields via a catalyst-free sulfa-Michael/aldol cascade reaction of chalcones 1 and commercially available 1,4-dithiane-2,5-diol 2 under mild conditions. We also present the first asymmetric sulfa-Michael/aldol cascade reaction of chalcones 1 and commercially available 1,4-dithiane-2,5-diol 2 with moderate to good enantioselectivities catalyzed by readily available chiral phase-transfer catalysts (PTCs).