共表达H5、H7亚型AIV HA基因与鸡IL-18基因的重组鸡痘病毒构建

以鸡痘病毒(FPV)疫苗株为载体,将H5和H7亚型禽流感病毒血凝素(AIV HA)基因串联后(拥有同一个阅读框)和鸡白细胞介素-18(IL-18)基因分别插入到鸡痘病毒表达载体pUTA-16-LacZ复合启动子(ATI-P7.5×20)和单一启动子1(P7.5)下游,构建了携带AIV HA基因和鸡白细胞介素-18基因的重组鸡痘病毒转移载体质粒pUTAL-H5-H7-IL18;用相同的方法构建重组鸡痘病毒转移载体质粒pUTAL-H5-IL18;将H5亚型AIV HA基因插入到鸡痘病毒表达载体pUTA2复合启动子(ATI-P7.5×20)下游,构建了携带H5亚型AIV HA基因的重组鸡痘病毒转移载体质粒pUTA2-H5.应用脂质体转染法,将重组鸡痘病毒转移载体质粒与282E4株鸡痘病毒共转染鸡胚成纤维细胞(CEF),经BrdU进行三次加压蚀斑筛选后,以不同代次的细胞mRNA为模板,利用H5亚型AIV HA基因、H7亚型AIV HA基因和鸡IL-18基因特异引物进行RT-PCR和蛋白印迹检测,筛选出H5HA-H7HA融合蛋白基因和鸡IL-18基因共表达的重组鸡痘病毒rFPV-H5HA-H7HA-IL18,H5亚型AIV HA基因和鸡IL-18基因共表达的重组鸡痘病毒rFPV-H5HA-IL18以及单独表达H5亚型AIV HA基因的重组鸡痘病毒rFPV-H5HA.这些重组鸡痘病毒的构建为AIV活载体疫苗的研制奠定了基础.     

共表达FMDV P1-2A和3C重组鸡痘病毒疫苗与核酸疫苗的实验免疫研究

将口蹄疫病毒(FMDV)衣壳蛋白前体P1-2A基因和蛋白酶3C基因分别插入到鸡痘病毒表达载体质粒pUTA-16Lac Z的复合启动子和单一启动子的下游,构建共表达重组鸡痘病毒转移载体质粒pUTAL3CP1,与282E4株FPV共转染鸡胚成纤维细胞,经RT-PCR、IFA及Western blotting检测,成功获得能正确表达目的蛋白的重组鸡痘病毒株vU-TAL3CP1。并与FMDV重组核酸疫苗免疫小鼠实验表明,各实验免疫组均能诱导小鼠产生特异性的细胞免疫和体液免疫反应,其中以重组鸡痘病毒与核酸疫苗联合免疫效果好于其它实验各组。     

HIV-1核心蛋白Gag与IL-6共表达重组鸡痘病毒的构建及其免疫原性研究

分别将IL-6和HIV-1gag基因插入到鸡痘病毒表达载体质粒pUTAL串联启动子和复合启动子(ATI-P7．5)下游，构建了重组鸡痘病毒表达质粒pUTA-GAG-IL6。经同源重组和Western blot检测，获得重组鸡痘病毒，并将该重组病毒免疫BALB／c小鼠，检测免疫小鼠血清抗体水平和特异性CTL杀伤活性。结果获得的重组鸡痘病毒可同时表达HIV-1核心蛋白和IL-6，并在感染细胞内形成病毒样粒子。重组病毒具有良好的免疫原性，可诱导小鼠产生特异性体液免疫和细胞免疫。     

口蹄疫病毒二价DNA疫苗的构建及实验免疫研究

通过PCR方法从本室已构建的克隆载体pGEMT-P1-2A获得O型口蹄疫病毒主要保护性抗原VP1基因,以基因突变获得A型VP1基因。将这两种血清型的VP1基因串联后连接到真核表达载体PVAX1 PCMV启动子下游,构建成口蹄疫二价核酸疫苗pVAX1-OA。经Western blot和IFA检测,目的蛋白在HeLa细胞中获得正确表达。动物实验表明,免疫小鼠T淋巴细胞明显增殖,特异性CTL杀伤活性较对照组显著提高;血清抗体能分别与O型和A型抗原反应,抗体效价均高于空白对照组,但较灭活苗低。     

联用抗口蹄疫病毒重组多肽及DNA疫苗诱发豚鼠产生特异免疫应答

选择一株O型口蹄病毒（FMDV）外壳蛋白VPI中21－40位及141－160位氨基酸残基两个抗原表位,组成串联结构141－160（20AA）－21－40（20AA）－141－160（20AA）,并与大肠杆菌β－半乳糖苷酶相连,在大肠杆菌中表达了此融合蛋白。同时将编码此融合蛋白的基因克隆进真核表达载体中,构建成重组表达质粒pCDM8FZ1。将pCDM8FZ1DNA与融合蛋白混合,免疫豚鼠一次,观察豚鼠产生免疫应答情况。结果显示,豚鼠可产生抗FMDV中和抗体及特异性T细胞增殖反应,部分豚鼠能抵抗FMDV的攻击,保护率为50％,证明将DNA疫苗与重组多肽疫苗结合起来作一次免疫,是可望用于预防FMDV感染的新途径,值得进一步探索。     

人工设计的HIV-1重组鸡痘病毒诱导恒河猴产生免疫应答的研究

将人工设计的HIV-1多表位-p24嵌合基因重组鸡痘病毒rFPV-MEGp24大量制备并纯化后,分别在0周、8周、18周肌肉注射至实验恒河猴中,在第2次免疫后2周和第3次免疫后2周采集血液,分离血清和外周血淋巴细胞(PBMC),ELISA法检测血清HIV-1抗体和PBMC培养上清Th1类细胞因子的含量,MTT法测定PBMC的增殖能力,乳酸脱氢酶(LDH)释放法检测PBMC特异性CTL杀伤活性.结果表明:HIV-1多表位-p24嵌合基因重组鸡痘病毒可刺激恒河猴产生HIV-1特异性抗体;免疫猴的PBMC在HIV-1抗原肽刺激下细胞增殖能力加强,针对HIV-1靶细胞的特异性CTL杀伤活性产生,分泌Th1类细胞因子IFN-γ、IL-2的水平升高.研究提示,HIV-1多表位-p24嵌合基因重组鸡痘病毒能诱导恒河猴产生特异性细胞和体液免疫应答.     

猪瘟病毒E2蛋白4重复抗原表位的构建及抗原活性研究

利用PCR方法获得4次重复的猪瘟病毒E2蛋白中和性抗原表位基因，将其克隆到pGEM-5ZF( )载体，测序正确后亚克隆到pGEX-3X载体构建得到重组质粒pGEX-3X-4P。重组质粒在大肠杆菌中诱导表达了含4重复抗原表位的融合蛋白。该蛋白经纯化后，利用间接ELISA检测其与血清的反应性，结果表明，纯化的融合蛋白与兔抗CSFV E2血清有很强的反应性，与兔抗BVDV E2血清不反应，说明该重复抗原表位在鉴别诊断CSFV与猪的BVDV感染方面具有潜在的应用价值。     

共表达VP3与hIL-18基因真核重组质粒的构建及对人结肠癌细胞的影响

先将鸡贫血病毒VP3基因插入真核表达载体pVAXl的多克隆位点，再将pVAX1载体上的IRES启动子及新霉素基因一同切下插入pVAXl的VP3基因下游，然后切下新霉素基因，将pIRES1基因插入其中，构建成pVVP3IL-18，将pVVP3IL-18转染Heda细胞。间接免疫荧光表明，在细胞膜和细胞浆观察到了特异的黄绿色荧光，证明表达了hIL-18。pVVP3IL-18转染人结肠癌细胞HCY-116后，经AO／EB染色及电镜观察，可见大量细胞凋亡。说明真核重组质粒pVVP3IL-18可在HeLa细胞中表达，并可在体外诱导人结肠癌细胞HCT-116凋亡。     

小鼠对HIV-2嵌合结构基因重组鸡痘病毒的免疫反应

将HIV-2嵌合结构基因gag-gp105插入到转移载体pUTA2复合启动子ATI-P7.5×20下游,构建出鸡痘病毒重组转移质粒pA-gg;经转染和BrdU加压筛选,以基因组PCR和Western blot法鉴定重组病毒;将获得的重组病毒大量制备并纯化后,肌肉注射免疫BALB/c小鼠,ELISA法检测小鼠血清HIV-2抗体,流式细胞仪测定CD4 、CD8 T淋巴细胞亚类数量,乳酸脱氢酶(LDH)释放法检测脾特异性CTL杀伤活性.结果表明,成功筛选出一株可稳定表达HIV-2嵌合蛋白Gag-gp105的重组鸡痘病毒rFPV-gg;该重组病毒免疫组小鼠的血清出现HIV-2抗体,脾T细胞亚类数量增加,并产生针对HIV-2靶细胞的特异性CTL杀伤活性.本研究提示,HIV-2 gag-gp105重组病毒能诱导小鼠产生特异性细胞和体液免疫应答.     

联合应用凋亡素基因、新城疫病毒HN基因及IL-18基因对黑色素瘤的抑制效应研究

以前的研究结果表明,来自于鸡贫血病毒的凋亡素基因、来自于鸡新城疫病毒的HN基因和IL-18基因具有不同的抗肿瘤效应,但上述基因抗肿瘤的机制不同,本研究在双启动子真核表达质粒pIRES中的CMV启动子下游插入凋亡素基因,IRES启动子下游插入IL-18HN嵌合基因,构建能同时表达三种基因的真核表达质粒pIRVP3IL-18HN。复制C57BL／6小鼠荷黑色素瘤模型,并将质脂体包裹的重组质粒进行瘤内注射,共注射3次,每次100μg质粒,拟观察凋亡素、新城疫病毒HN基因和IL-18基因以核酸疫苗的形式联合应用对小鼠黑色素瘤的抑制效应。结果显示pVIL-18HN、pVlL-18和pVVP3IL-18均有抑制肿瘤作用,但pIRVP3IL-18HN组抑瘤率高于pVIL-18HN组、pVIL-18VP3组、pVIL-18组和Hanks液对照组。研究结果提示,上述3种基因联合应用具有协同抑瘤效应。     

The Research of Anti-Wear Properties of Organomolybdenum Compounds in the Engine Oils

A four-ball tester was used to evaluate the anti-wear performance of three kinds of organomolybdemun compounds in the engine oils, i. e., molybdenum dialkyldithiophosphate (MoDDP), molybdenum dialkyldithiocarbamate ( MoDTC), and sulphur and phosphorus freeorganomolybdeum (Molybdate). The results indicate that a low concentration of MoDDP doesn' t improve the anti-wear properties of the commercial engine oils, but a high concentration of MoDDP can obviously improve the anti-wear properties and the load-carrying capacity of the engine oils. MoDTC doesn' t improve the antiwear properties of the engine oils, but worsens the anti-wear properties of the oils. Signifi can timprove ment of frictional and wear characteristics is obtained with Molybdate added in the commercial engine oils and the formulated oils.     

Certification Becomes Important for the I Quality; and Safety Supervision of the Chinese Liquor Sector

The end of 2007, over 200 unit products of more than 80 Chinese firms have passed the quality grade certification for liquor products. These products involve distilled spirits, beer, wine, yellow wine, fruit syrup wine and others, and cover over 80% of the national top-branded liquor products.……     

Deposition and Coating Properties on CVD Tungsten

Surface characterization and microstructure studies are performed on chemical vapor deposited (CVD) tungsten coating. There is about 2 μm thickness diffusion layer of tungsten in the molybdenum substrate. The thermal shock test shows tungsten coating has good adhesion with molybdenum substrate, but the elements of oxygen and carbon in the tungsten coating have the bad affection to the adhesion. The result of high-temperature diffusion experiment is the diffusion rate from molybdenum substrate to tungsten coating is faster.     

Analysis of marine hydrobiont lipid extracts as possible cryoprotective agents

We studied a combined effect of the cryoprotectants both of lipid and carbohydrate origin at the presence of DMSO or without it on cell viability and the RNA synthesis in the embryonic mollusc and echinoderm cell cultures obtained at different developmental stages. Cryoprotective properties of exogenous lipids correlated with their thermotropic behavior. Lipid extracts from marine hydrobiontes, which thermal transitions are characterized by relatively small part of calorimetric thermogram area lying at temperatures above 0 °C (6–18%), were found to possess a considerable cryoprotective activity. The results are discussed in relation to the prediction of the cryoprotective activity of lipid extracts for marine animal cells. In addition, it was found that marine invertebrate cells can be cryopreserved without DMSO at the presence of trehalose and lipid extracts of some marine hydrobiontes.     

Low-Temperature Properties of Silver

Pure silver is used extensively in the preparation of high-temperature superconductor wires, tapes, films, and other configurations in which the silver not only shields the superconducting material from the surrounding materials, but also provides a degree of flexibility and strain relief, as well as stabilization and low-resistance electrical contact. Silver is relatively expensive, but at this stage of superconductor development, its unique combination of properties seems to offer the only reasonable means of achieving usable lengths of conductor. In this role, the low-temperature physical (electrical, thermal, magnetic, optical) and mechanical properties of the silver all become important. Here we present a collection of properties data extracted from the cryogenic literature and, to the extent possible, selected for reliability.     

Early ironwork in Biscay: Survey,excavation, experimentation and materials characterization. An integral study of the mountainside ironworks (ferrerías de monte or “haizeolak”)

ABSTRACT

The production of iron in Biscay (Basque Country, Spain) has been a hallmark of this territory of the North of the Iberian Peninsula throughout its history. The Archaeology team of the Basque Country Mining Museum has spent 13 years, so far, studying the origins of this activity through the evidences left at the old pre-hydraulic iron production workshops known as mountainside ironworks (ferrerías de monte) or haizeolak. It has catalogued 170 archaeometallurgical sites of this type in Biscay, and the archeological excavation of some of them has led to the knowledge of the different stages of the production process that took place in these workshops. The team has also experimentally reproduced the work carried out in a mountainside ironworks from the High Middle Ages, and it has currently started the characterization process of this type of activity through the study of the collected materials.     

Mechanism Study on the Wear of CrAlTiN Coated High-Speed Steel Twist Drills Under Dry Cutting Conditions

As demands about environment protection are growing up, dry cutting technology is getting more and more concerns from all over the world. Main works performed here are study on dry cutting performances and wear mechanisms of M2high-speed steel (HSS) twist drills with CrAlTiN multicomponent coatings, which was deposited using magnetron sputter ion plating system, in drilling 45 and 30CrMnSiA steel, and their comparisons to those in drilling the same steel but using monolayer CrN and commercial TiN coated drills. Drilling performances of drills are evaluated mainly through the measurements of width on outer corner flank land and the cutting forces. Results show that performances of CrAlTiN coated drills are better than those of monolayer CrN and TiN coated drills. In drilling 45 and 30CrMnSiA steel, the average tool life of the CrAlTiN coated drills are 17.2 and 11.8 times higher than those of the uncoated drills. Observing wear of the drill with scanning electronic microscope, results show that there is no crack or spallation of the CrAlTiN coatings in wear zones. Main mechanism of the wear here is adhesion.     

Study on Bond Ability of Arc-Spraying Coatings with Different Surface Pretreatment on Cast-Iron

Arc spraying coatings are widely used in various applications, but uncommon in cast iron substrate. Different surface pretreatment technology is tested on substrates of gray cast iron. Surface roughness and residual stress were measured by TR200 and X-ray diffraction analyzer. Influence of different surface pretreatment methods ( dry blasting and fusebond) on roughness and residual stress was analyzed. The arc-sprayed coatings of wire 3Cr13 (φ2mm) on gray cast iron substrate is studied. The microstructure and interface of bonding layer were observed by SEM. The bond strength was taken by tensile test. Results show that bond strength with grit blasting is higher than fuse-bond; it is feasible to make wire 3Cr13 coating with arc spraying on cast iron substrate roughened by grit blasting.     

Structural Characteristics of TiO2 Ceramic Coating by Micro-Plasma Oxidation

TiO2 ceramic coatings with thickness of 20 μm were formed on the surface of pure titanium by micro-plasma oxidation. Their micro-structures were investigated by by using X-ray diffraction and their surface images were detected by using scan electronic microscope. There were three kinds of TiO2 coatings, pure anatase type TiO2 phase, mixed phases consisted of rutile type TiO2 phase and anatase type TiO2 phase, pure rutile type TiO2 phase. The coating surface with the pure anatase type TiO2 phase is rough, while the coating surface with the pure rutile type TiO2phase is smooth. The upper coating surface with the mixed type TiO2 phases is anatase type TiO2 structure and the subsurface of the TiO2 coating is rutile type TiO2structure.     

Evaluation of fatigue life of steel using steel grain size

The insightful discussion of the relationship between the construction of metallographic and fatigue life and proposition of mathematical relationships describing this relationship was conducted in the paper. This paper presents a method for estimating the fatigue life, based on the construction of the microstructural material taking into account the grain size and the participation of ferrite and pearlite phase.