THE INVESTIGATION OF DIFFERENT POPULATION＇S TRANSFUSION-TRANSMITTED VIRUS-DNA IN SHAANXI PROVINCE

Objective To investigate transfusion transmitted virus (TTV) infection among population of different groups in Shaanxi Province. Methods A nested polymerase chain reaction (PCR) with primers from ORF1 of TTV genome was established to detect TTV-DNA in serum of the patients. Results TTV-DAN was detected in the sera of 3 of 50 cases of general population(6％), 2 of 30 cases of vocational blood donors(6.7%),21 of 97 cases with Type B hepatitis (21. 6％), 9 of 35 cases of Type C hepatitis (25. 7％),and 23 of 40 cases with non-A～ non-G hepatitis (57.5 ％ ). Conclusion There is TTV infection among general population in Shaanxi Province. TTV may be an impor- tant agent to cause non-A～non-G hepatitis. And the patients with HBV or HCV can have overlapping TTV infection.     

Effect of carbon nanotube concentration on cooling behaviors of oil-based nanofluids during the immersion quenching

Oil-based nanofluids including 0.75%—1.75%(mass fraction) carbon nanotubes(CNTs) without any surfactants have been synthesized by a two-step process. The probes machined from 45~# steel with 22 mm diameter and 50 mm length are quenched in the as-synthesized CNT nanofluids for testing the cooling behaviors of the nanofluids. The laser diffraction particle size analyzer, scanning electronic microscope(SEM), X-ray diffractometer and transmission electron microscope(TEM) are used to characterize the quality and distribution of CNTs in the nanofluids. The wettability and viscosity of 30~# oil and oil-based CNT nanofluids are measured by a goniometer at 15℃ and a rotational type viscometer at 40℃, respectively. The results show that the cooling efficiency of the oil-based CNT nanofluids is better than that of 30~# oil. Moreover, the cooling rate of the naonofluids increases with the further increase of the CNT concentration. When the mass fraction of CNTs increases to 1.75%, the cooling rate of the naonofluids reaches a maximum at 760℃ and it is increased by 77.8% as compared to that of base oil. The improved cooling rate of oil by CNTs is mainly due to the uniform distribution and excellent thermal conductivity of CNTs.     

Parallel Distributed Implementation of Truss Optimization on a Local Area Network

A two-level optimization method for the design of complex truss and parallel distributed implementation on a LAN is presented using parallel virtual machine (PVM) for Win 32 as message passing between PCs. The volumes of truss are minimized by decomposing the original optimization problem into a number of bar optimization problems executed concurrently and a coordinate optimization problem, subject to constraints on nodal displacements, and stresses, buckling and crippling of bars, etc. The system sensitivity analysis that derives the partial derivatives of displacements and stresses with respect to areas are also performed in parallel so as to shorten the analysis time. The convergence and the speedup performances as well as parallel computing efficiency of the method are investigated by the optimization examples of a 52-bar planar truss and a 3 126-bar three-diraensional truss. The results show that the ideal speedup is obtained in the cases of 2 PCs for the 3 126-bar space truss optimization, while no speedup is observed for the 52-bar truss. It is concluded that (1) the parallel distributed algorithm proposed is efficient on the PC-based LAN for the coarsegrained large optimization problem; (2) to get a high speedup, the problem granularity should match with the network granularity;and (3) the larger the problem size is, the higher the parallel efficiency is.     

Electron spin resonance analyse on the production of hydroxyl radicals by ozone photolysis

AbstractIn order to explore the production of hydroxyl radical(·OH)in a confined space,a novel ozone-light irradiation system is constructed in this study,and the·OH radical is measured by spin-trapping electron spin resonance(ESR)method in which 5,5-dimethyl-1-pyrroline-N-oxide(DMPO)is selected as the spin-trap.Several influence factors including the light intensity,the irradiation time and DMPO mass concentration are discussed. The results show that in this experimental system,with DMPO mass concentration of 1g/L and the irradiation time of 30 min,the·OH radical can be best captured.Besides,both wavelength and intensity of the irradiation light could effect the generation of·OH radical.These results are of great importance to further study the sterilization effect of·OH radical in confined space.     

An Effective Hybrid Optimization Algorithm for Capacitated Vehicle Routing Problem

IntroductionVehicle routing problem(VRP)is a well-known combinatorial optimization problem and isalso a focal problem of distribution managementwithin the area of service operations managementand logistics.Capacitated vehicle routing problem(CVRP)is a very hard vehicle routing problem.The problem is to simultaneously determine theroutes for several vehicles from a central depot to aset of customers,and then return to the depotwithout exceeding the capacity constraints of eachvehicle.In prac…     

Investigation of springback behavior and process control for stamping of ring-shaped workpiece

Due to the elastic deformation of metal stamping, springback is a serious problem to cause shape deviation and thus reduce precision. The springback behavior of ring-shaped workpiece is investigated in this paper and an auxiliary boss device is proposed to control the springback deformation. A finite element model is developed.The characteristic parameters including boss height, boss angle and boss gap are discussed systematically. It can be concluded that the gap has a positive linear correlation with springback percentage, and the boss angle has an optimal interval of 30°—60° to reduce springback. When the boss height increases, the springback decreases,but the effect weakens when the boss height is larger than 1.0 mm. Then, a response surface analysis is conducted with Design Expert 8.0.5 software and an optimal parameter process window(i.e., boss height from 2.1 to 3.0 mm,boss angle from 53° to 60° and the gap from 0 to 1.2 mm) is obtained. Finally, a set of molds are fabricated and the stamping experiments are conducted. The springback behavior is well controlled. The springback percentage can be decreased from 33% to 6% by using the auxiliary boss device of ring-shaped workpiece.     

High-Accuracy Pneumatic Position Control by Applying Nonlinear Control and Arranging Transient Process

By applying a nonlinear control and arranging a transient process, the initiative error of the pneumatic servo positioning system is reduced largely, and a larger gain of the controller is used to improve the responding speed of the system at the same damping ratio. Therefore, a compromise is made among the responding speed, overshoot, robustness, adaptability and stability. In addition, a dynamic output feedback controller, including position velocity and acceleration (PVA) feedback, is designed to improve the performance of the system. And a nonlinear controller is reconstructed based on the linear output feedback controller to decrease noises and disturbances. The dynamic responses of the system are simulated and tested. Results show that the error is kept within 0.02 mm under different mass loads and the positioning transient process is smooth, without overshoot and speedy.     

Fine Mapping of a Deafness Mutation hml on Mouse Chromosome 10

Objective To map a mouse deafness gene.identify the underlying mutation and develop a mouse model for human deafncss.Methods Genetic lindage cross and genome scan wer used to map a novel mutation named hypoplasia of the membranous labyrinth (hml),which causes hearing loss in mutant mice.Results ① hml was mapped on mouse Chr 10 (～43 cM from the centromere) suggests that the homologous human gene is on 12q22--q24, which was defined on the basis of known mouse-human homologies (OMIM,2004). ② This study has generated 25 polymorphic microsateUite markers, placed 3 known human genes in the correct order in a high-resolution mouse map and narrowed the hml candidate gene region to a 500 kb area.     

Distributed Broker-Agent Architecture for Multimedia Communications Traversing NAT／Firewall in NGN

The forthcoming Next Generation Network (NGN) is an all IP network. Multimedia communications over IP networks are a type of bundled session communications, which cannot directly traverse Network Address Translations (NATs) and firewalls even in NGN. To solve the problem that the existing traversal methods are not suitable for service providers to set up a real system in NGN,a Distributed Broker-agent Architecture (DBA) is addressed. DBA is secure and realizable for service providers and enterprises because it is easy to set up and does not need to upgrade the existing devices like Firewalls, NATs or endpoint devices of subscribers.DBA is composed of two-layer distributed agents, the server proxies and the client agents, in which all multimedia communications use shared tunnels to carry signaling messages and media data between broker-agents, and the call signaling is encrypted over Security Socket Layer (SSL) to guarantee the security of calling. Moreover, the function model and multiplexed connection messages format of DBA are designed, which lays a basis for the protocol in the future NGN. In addition, a simple implementation based on H.323 verifyies the main function of traversing firewalls and NATs.     

Fairness-embedded governmental coordination by dominated retailer in heterogeneous circumstance

Market demand is positively affected by the investments in marketing and quality improvement. In a retailer dominated supply chain, the manufacturer takes charge of product quality improvement and the retailer focuses on marketing. The investments on cooperative marketing and product quality improvement in supply chains are known to be heterogeneous investment, where the manufacturer(he) is fairness sensitive and the retailer(she) is fairness insensitive. Due to the manufacturer's conservative investment resulting from this sensitivity of fairness, the retailer shares a proportion of the manufacturer's investment to increase his investment. It is found that the manufacturer's fairness negatively affects his investment and the manufacturer sometimes tends to be a ‘free rider' under decentralized decision patterns, which lead to a poor supply chain performance. This study also proves that both the manufacturer and the retailer are motivated to cooperate under centralized patterns,where the retailer's objective is additional expected profit and the manufacturer's objective is enlarging his utility.Based on the comparison of equilibrium solutions in decentralized and centralized patterns, this study suggests the range of cost-sharing proportion for supply chain coordination. The example shows the effectiveness of the provided supply chain coordination.     

丙型肝炎病毒NS3-NS4基因的表达及鉴定

目的　构建丙型肝炎病毒 (HCV)非结构蛋白NS3 NS4全长基因重组质粒并诱导表达 ,鉴定NS3 NS4蛋白的抗原性。方法　应用PCR技术从pUC19/HCV中扩增目的基因 ,构建重组表达质粒 ,进行原核表达 ,用SDS PAGE、ELISA技术对表达产物的抗原性进行鉴定。结果　成功构建、表达了重组质粒pBV2 2 0 /NS3 NS4 ,用 5 0份质控标准血清对表达蛋白质进行抗原性检测 ,与第二代诊断试剂相比 ,总符合率为 96 %。结论　全长NS3 NS4蛋白是一个优势免疫原性区 ,应该是HCVEIA诊断试剂的有效成分。     

丙型肝炎病毒非结构蛋白3全长基因的克隆和表达

目的克隆和表达丙型肝炎病毒(HCV)非结构基因NS3,为进一步研究其基因功能奠定基础。方法设计特异性引物,以HCV全长cDNA质粒为模板进行PCR扩增,获得全长NS3基因。将其插入克隆载体pMD18-Tvector中,经酶切、PCR及测序鉴定后,与表达载体pET32a重组,后转入BL21菌株并诱导表达。采用SDS-PAGE电泳及Western-blot检测NS3基因的蛋白表达水平。结果含有NS3的重组体构建成功并在大肠杆菌中表达。结论运用原核细胞基因工程技术成功构建和表达了HCV非结构基因NS3蛋白,为尽一步研究HCVNS3基因功能奠定了基础。     

人神经生长因子成熟蛋白基因片段在大肠杆菌中的表达和生物活性鉴定

目的　在大肠杆菌中表达人神经生长因子成熟蛋白基因片段(hNGFβ)并检测其生物活性。方法　将 NGFβ目的基因亚克隆入表达载体pBV220的BamHI- BamHI位点,连接产物转化大肠杆菌DH5α菌株通过热诱导表达NGFβ蛋白,利用溶解包涵体和重折叠使表达产物复性,将复性后的蛋白质加入体外培养的鸡胚背根神经节及 PC12 细胞BrdU掺入实验,观察表达蛋白的生物学活性。结果　成功构建了重组质粒 pBV220/NGFβ。使用该质粒转化 DH5α宿主菌,使用热诱导方法表达了NGFβ蛋白,SDS PAGE结果提示表达蛋白主要存在于包涵体中。通过肝素亲和层析可以获得纯度大于90%的 NGFβ。每升表达菌可以获得 1.8~2.0 mg NGFβ。鸡胚背根神经节突起生长实验和PC12细胞培养生长刺激BrdU掺入实验表明原核表达的 NGFβ是具有生物活性的。它的生物活性按生物制品鉴定章程判断为1×105 BU/g。结论　在大肠杆菌中可以表达出有活性的NGFβ蛋白。     

丙型肝炎病毒C区截短型基因原核表达载体的构建及在大肠杆菌中的表达

目的建立稳定表达丙型肝炎病毒(HCV)核心蛋白的原核表达系统,获得高产量的纯化核心蛋白。方法应用多聚酶链反应(PCR),以HCV-H株全长cDNA序列为模板,扩增获得C区羧基端部分缺失的基因片段,克隆入原核表达载体pBVIL1,构建原核表达载体pBVIL1-Ct,转化HB101宿主菌,通过温度诱导表达截短型C蛋白。结果扩增得到目的基因长度为462bp,构建pBVIL1-Ct表达载体,在HB101宿主菌中通过温度诱导获得稳定表达,表达蛋白占菌体总蛋白含量的24%。Western-Blot及ELISA检测证实表达产物可与HCV患者阳性血清发生特异性结合反应。结论羧基端部分缺失的HCV C区基因片段可在大肠杆菌中稳定表达并具有良好的反应原性。     

丙型肝炎病毒1b DY株ns5a基因的克隆及其在大肠杆菌中的表达

目的克隆和表达丙型肝炎病毒（HCV）1b型地方株DY株ns5a基因。方法运用原核细胞基因工程技术。设计目的基因的特异引物，采用巢式PCR法，从含HCV1bDY株全长eDNA的质粒HCV17中扩增出约480bp的目的片段，将其插入克隆载体pMD18-Tvector中，再亚克隆到原核表达载体pET-28a中；经酶切、PCR及测序鉴定后转化入BL21菌株，在IPTG诱导下进行融合蛋白的表达；采用SDS-PAGE电泳及Western-blot检测NS5A蛋白的表达水平。结果成功构建了含有HCV1b DY株ns5a基因的重组体，并得以表达。结论成功构建和表达了HCV1b DY株ns5a基因，为进一步研究HCVns5a的基因型及探讨该基因编码的Ns5A蛋白的性质和生物学活性创造了条件。     

融合基因Aβ-HBcAg的原核表达及表达蛋白的免疫反应性和免疫原性分析

目的　研究 β 淀粉样肽 (β amyloidpeptide,Aβ)与乙型肝炎核心抗原 (HBcAg)融合基因Aβ HBcAg的原核表达 ,检测融合蛋白的免疫反应性及免疫原性。方法　将重组质粒pBV2 2 0 /Aβ HBcAg转化大肠杆菌DH5α,温控表达 ,超声破碎细菌 ,通过SDS PAGE、考马斯亮蓝染色和ELISA等方法 ,观察Aβ HBcAg的表达及融合蛋白的免疫反应性 ;用融合蛋白免疫Balb/c小鼠 ,ELISA法检测血清中的抗 Aβ抗体和抗 HBc抗体滴度。结果　融合蛋白以包涵体的形式存在于沉淀中 ,表达量为菌体沉淀的 5 % ,融合蛋白既有Aβ的免疫反应性 ,又有HBcAg的免疫反应性。Balb/c小鼠经过 3次免疫后 ,血清中的抗体滴度很低 ,继续强化免疫 2次后 ,血清中抗 Aβ抗体滴度、抗 HBc抗体滴度分别上升为 1∶80 0和1∶32 0 0。结论　融合基因Aβ HBcAg可在大肠杆菌DH5α中表达 ,表达蛋白有一定的免疫反应性和免疫原性 ,但表达量较低 ,免疫反应性和免疫原性较弱。提示融合基因Aβ HBcAg的表达量和融合蛋白免疫原性的提高尚需进一步研究。     

人杀菌渗透增强性蛋白N末端片段在大肠杆菌中的表达

目的构建表达人杀菌渗透增强性蛋白(BPI)N-端193个氨基酸的重组表达质粒,诱导表达BPI193重组蛋白。方法应用RT-PCR的方法从HL-60细胞内扩增出BPI氨基端1-193个氨基酸的基因序列,克隆入T载体。将BPI193基因片段定向克隆入原核表达载体PET-28a中,构建重组的原核表达质粒PET-BPI193,转化大肠杆菌BL21菌株,用IPTG诱导表达蛋白。应用SDS-PAGE检测蛋白表达情况,计算机薄层扫描分析蛋白占菌体总蛋白百分比;Western-blot技术检测表达蛋白的特异性。结果从HL-60细胞中扩增得到579bp的BPI193基因片段,构建了T-BPI193亚克隆和PET-BPI193重组表达质粒。转化大肠杆菌BL21,通过IPTG诱导,经SDS-PAGE检测表明,成功表达了6His-BPI193融合蛋白。计算机薄层扫描分析表达量占菌体总蛋白的12%。Western-blot检测显示表达产物具有特异性。结论成功构建了PET-BPI193重组表达质粒。在大肠杆菌BL21内,诱导获得了BPI193与组氨酸融合表达的重组蛋白。     

丙型肝炎病毒核心蛋白酵母双杂交诱饵载体的构建及人胎肝cDNA文库的扩增、纯化和鉴定

目的用含丙型肝炎病毒核心(HCV core)蛋白的cDNA片段构建酵母双杂交诱饵载体,并进行人胎肝cDNA文库的扩增、纯化和鉴定。方法PCR扩增含HCV core蛋白不同大小的cDNA片段,分别克隆入pUC19质粒,经测序正确后,再亚克隆入酵母双杂交诱饵载体pGBKT7中。扩增人胎肝cDNA文库并纯化、鉴定。结果获得含HCVcore蛋白的cDNA片段,并成功克隆入pGBKT7中。待转化的人肝cDNA文库滴度在5×108左右,纯化后的质粒DNA质量浓度约1 g/L。用EcoRⅠ、XhoⅠ双酶切显示插入片段大小不一。结论成功构建了核心蛋白的酵母双杂交诱饵载体;扩增、纯化的人胎肝cDNA文库的多样性很好,适合于筛选。为用酵母双杂交技术研究与HCV core蛋白相互作用的蛋白打下坚实基础。     

抗人γ-精浆蛋白V_H单域抗体基因的构建、表达及空间构象分析

目的　扩增出抗人γ 精浆蛋白 (γ Sm)单克隆抗体 (mAb)的重链可变区 (VH)单域抗体基因 ,并进行原核表达及空间构象分析。方法　从分泌抗γ SmmAb的杂交瘤细胞系E4B7中提取总RNA ,经RT PCR扩增VH 单域抗体基因 ,将其克隆到融合蛋白表达载体 pGEX 4T 1中进行表达 ,并利用计算机软件对表达产物进行空间构象分析。结果　VH 单域抗体基因全长 363bp ,含起始码和终止码。将其克隆到 pGEX 4T 1内 ,转化大肠杆菌DH5α ,获得高效表达 ,表达量占全菌总蛋白质的 38% ,以包涵体形式存在。经初步纯化和复性后 ,用谷胱甘肽 (GST)亲和色谱纯化 ,再经凝血酶水解获得抗γ SmVH 单域抗体。竞争结合抑制实验证明 ,该表达产物具有前列腺癌细胞亲和活性。空间构象分析结果显示 ,该抗体具有完整的抗原结合位点。结论　构建成功抗γ Sm的VH 单域抗体 ,为进一步临床应用奠定基础。     

饰胶蛋白(Decorin)基因的cDNA克隆与表达

目的　通过基因克隆技术获得饰胶蛋白基因cDNA克隆并表达。为研究饰胶蛋白的生物学功能及应用打基础。方法　应用PCR技术从T细胞cDNA文库获得饰胶蛋白全长基因cDNA片段 ,并克隆到pUC1 9载体中 ,采用Sanger双脱氧末端终止法测定全部cDNA序列 ,将测序正确的饰胶蛋白cDNA序列插入pGEX 4T 1表达载体中 ,经BamHI和EcoRI双酶切后 1 2 %凝胶电泳鉴定 ,IPTG诱导表达产物经SDS PAGE分析。结果　克隆的饰胶蛋白cDNA序列与GenBank中AF1 3830 0记载的序列完全一致 ,所表达的融合蛋白质分子量与理论计算值(65 6KD)也一致并为可溶性蛋白。结论　本研究成功地克隆了饰胶蛋白基因和表达了GST Decorin融合蛋白