结核分枝杆菌特异性IFNγ体外释放试验全血检测试剂盒的质量评价

目的评价结核分枝杆菌特异性IFNγ体外释放试验(interferonγrelease assay,IGRA)全血检测试剂盒的质量。方法采用10个结核分枝杆菌特异性IGRA全血检测试剂盒检测参考样品,以参考样品检测值和样品浓度进行线性回归,获得线性回归方程及相关系数(R2)。按各试剂盒说明书判定参考样品,以阳性的最低参考样品浓度作为相应试剂盒的最低检测浓度。结果 10个结核分枝杆菌特异性IGRA全血检测试剂盒的R2均高于0. 9,最低检测浓度分布在0. 4～2 IU/mL之间。结论结核分枝杆菌特异性IGRA全血检测试剂盒的质量稳定,线性分析和最低检测浓度检测可用于该试剂盒IFNγ测定部分的质量控制。     

IFNγ释放试验评价水痘减毒活疫苗的细胞免疫效果

目的应用IFNγ释放试验评价水痘减毒活疫苗的细胞免疫效果。方法利用IFNγELISA定量试剂盒检测不同灭活抗原(热灭活、紫外灭活、甲醛灭活)刺激、不同孵育时间(24、48、72 h)、不同个体(12人接种水痘减毒活疫苗前后)对IFNγ释放水平的影响。结果紫外灭活方式制备的刺激抗原及新鲜全血与刺激抗原共孵育48 h,IFNγ释放水平最高;12人接种疫苗后全血中IFNγ释放水平均有不同程度提高,接种前后几何平均值(GMT)分别为6.13和234.75 pg/ml,增长倍数和GMT值在个体间均存在较大差异。结论 IFNγ释放试验操作简便、快速,可定量,适用于水痘疫苗细胞免疫效果的评价。     

控制释放技术及其在农药中的应用

综述了控制释放技术的类型、特点及最新的研究进展,介绍了控制释放技术在农药领域的应用及其进展,讨论了控制释放技术存在的问题和发展趋势。     

结核分枝杆菌肝素结合血凝黏附素的原核表达及其在结核病血清学诊断中的应用

目的原核表达与纯化结核分枝杆菌(Mycobacterium tuberculosis,M.tb)肝素结合血凝黏附素(heparin-binding haemagglutinin adhesion,HBHA),并初步评估其在结核病血清学诊断中的价值。方法采用PCR法从M.tb H37Rv基因组中扩增hbha基因,克隆至原核表达载体p ET-28a(+),构建重组表达质粒p ET-28ah,转化大肠埃希菌BL21(DE3),IPTG诱导表达。表达的重组HBHA(r HBHA)蛋白经Ni2+亲和层析柱纯化后,Western blot法分析其抗原特异性。收集71例确诊的结核病患者和30名健康体检者血清,采用ELISA法检测血清中的抗HBHA Ig A、Ig G抗体水平,并以ESAT-6、Ag85A作为对照抗原,评估r HBHA在结核病血清学诊断中的价值。结果重组表达质粒p ET-28ah经酶切及测序证实构建正确;表达的重组蛋白主要以包涵体形式存在,经Ni2+亲和层析柱纯化,可得到高纯度的r HBHA蛋白;纯化的r HBHA蛋白可与小鼠抗His-tag单克隆抗体特异性结合;结核病患者(TB)组血清中抗r HBHA、ESAT-6、Ag85A抗原特异性Ig G水平均显著高于健康对照(HC)组(P0.05),且TB组的r HBHA抗原特异性Ig A水平也显著高于HC组(P0.05),而两组间ESAT-6、Ag85A抗原特异性Ig A水平差异无统计学意义(P0.05);ROC曲线分析显示,r HBHA抗原检测Ig A的诊断效能较好,曲线下面积为0.711,其他两种对照抗原ESAT-6、Ag85A的Ig A诊断效能一般,曲线下面积分别为0.512和0.531。结论 r HBHA抗原用于结核病的诊断效能较好,可作为结核病血清学诊断的备选抗原之一。     

气候试验及其在粉末涂料涂装中的应用

本文介绍了气候试验的种类、测试原理、工作流程及其在粉末涂料涂装研发中的应用,说明气候试验对粉末涂料与涂装产品老化性能的评价以及原材料的选择都有着重要的指导作用。     

ZIF-8的制备及其在药物特征响应释放中的应用

以硝酸锌和2-甲基咪唑为原料,采用溶剂法合成了ZIF-8纳米微粒,考察了反应溶剂、反应温度和反应时间等因素对其产率、结构和性能的影响。并以合成的ZIF-8纳米微粒作为药物载体,研究了其对抗癌药物盐酸阿霉素(DOX)的载药及酸响应释放性能。结果表明,溶剂的质子化能力对ZIF-8微粒的结构和形貌有明显影响,以质子化能力适中的甲醇作反应溶剂可获得粒径大小均一、形貌规整的微粒;以甲醇为溶剂,在硝酸锌和2-甲基咪唑的物质的量比为1∶4、25℃下反应24h时,ZIF-8产率可达36%;浊度检测表明,以反应48h的浊度为基准,反应3 h时,反应进程达到80%。合成的ZIF-8微粒均具有良好的热稳定性,但呈现明显酸不稳定性。制备的ZIF-8纳米微粒载药效率最高可达19.73%,体外溶出实验表明,DOX@ZIF-8在酸性环境中释放效率明显快于在中性环境中的释放效率,呈现出优异的酸响应释放性能。     

结核分枝杆菌Rv3873抗原的同源表达及其在结核病血清学诊断中的应用

目的 在耻垢分枝杆菌（Mycobacterium smegmatis,Msm）中克隆并表达结核分枝杆菌（M. tuberculosis,Mtb）RD1区蛋白Rv3873,并评价其在结核病（tuberculosis,TB）血清学诊断中的价值。方法 以Mtb H37Rv标准株全基因组DNA为模板,扩增得到rv3873基因完整序列,克隆至分枝杆菌表达载体pMF406,构建重组质粒pMF406-rv3873,电转化至Msm mc2155,加入乙酰胺诱导表达Rv3873同源重组蛋白（mRv3873）,利用亲和层析进行纯化,Western blot分析抗原特异性;ELISA法检测27份TB患者和31份健康者血清IgG抗体,以大肠埃希菌表达的Rv3873(r Rv3873)和免疫优势抗原Ag85A作为对照抗原,评价mRv3873在TB血清学诊断中的作用。结果 在Msm中成功表达了重组Mtb蛋白mRv3873,主要以可溶形式存在,经Ni2+亲和层析柱纯化可获得高纯度（95%）的可溶性重组蛋白,蛋白浓度约为2.0 mg/mL;TB患者组血清中抗mRv3873和Ag85A抗原特异性IgG水平均显著高于健康...     

结核分枝杆菌ESAT-6重组二聚体的表达、纯化及其在血清学诊断中的初步应用

目的表达和纯化结核分枝杆菌早期分泌蛋白ESAT-6重组二聚体(rdESAT-6),并探讨其在结核病血清学诊断中的价值。方法以HG856A核酸疫苗质粒为模板,经PCR扩增获得2×esat-6基因,克隆至pET-28a质粒,构建原核表达质粒pET2E6;转化大肠杆菌BL21(DE3),IPTG诱导表达;用Ni2+柱亲和层析纯化重组蛋白,并用Western blot鉴定其反应原性,ELISA检测其敏感性和特异性。结果ESAT-6重组二聚体以包涵体形式表达,表达量约占菌体总蛋白的35%;纯化的rdESAT-6纯度可达95%,可与ESAT-6小鼠抗血清发生特异性反应;用于结核病血清学诊断的敏感性为30%,特异性为95.8%。结论已成功表达并纯化了rdESAT-6,可作为结核病血清学诊断或皮试检测用的抗原之一。     

护理干预在慢性阻塞性肺疾病患者中的应用研究

目的探讨护理干预在慢性阻塞性肺疾病患者中的护理效果,改善患者的肺功能,提高患者的生存质量。方法随机将80例慢性阻塞性肺疾病患者分为观察组和对照组各40例,观察组实施为期6个月的护理干预,对照组仅在就诊时进行宣教,比较2组患者的肺功能和生活质量。结果观察组护理效果明显好于对照组,如肺功能改善情况和生活质量均高于对照组(P<0.05)。结论护理干预可以提高慢性阻塞性肺疾病患者的肺功能和生活质量,促进患者健康。     

ZIF-8的制备及其在药物特征响应释放中的应用研究（0892退后重投）

以硝酸锌和2-甲基咪唑为原料,采用溶剂法合成了ZIF-8纳米微粒,考察了反应溶剂、反应温度和反应时间等因素对其产率、结构和性能的影响。并以合成的ZIF-8纳米微粒作为药物载体,研究了其对抗癌药物盐酸阿霉素(DOX)的载药及酸响应释放性能。结果表明,溶剂的质子化能力对ZIF-8微粒的结构和形貌有明显影响,以质子化能力适中的甲醇作反应溶剂可获得粒径大小均一、形貌规整的微粒;以甲醇为溶剂,在硝酸锌和2-甲基咪唑的物质的量比为1∶4、25℃下反应24h时,ZIF-8产率可达36%;浊度检测表明,以反应48h的浊度为基准,反应3 h时,反应进程达到80%。合成的ZIF-8微粒均具有良好的热稳定性,但呈现明显酸不稳定性。制备的ZIF-8纳米微粒载药效率最高可达19.73%,体外溶出实验表明,DOX@ZIF-8在酸性环境中释放效率明显快于在中性环境中的释放效率,呈现出优异的酸响应释放性能。     

The carboxyl-terminal region of human interferon {gamma} is important for biological activity: mutagenic and NMR analysis

Deletion of nine amino acids from the carboxyl terminus of humanIFN (residues 138–146; LFRGRRASQ) resulted in a 7-foldincrease in specific antiviral activity. Similar increases inreceptor binding affinity were seen. Deletion of residues 136and 137 (QM) had little additional effect, but removal of Ser135resulted in a sharp drop in antiviral activity. Further removalof residues 133 and 134 (KR) lowered antiviral activity to 1%of the peak value. Comparison of the proton NMR spectra of selecteddeletions down to residue 132 showed that there was no significantchange in the core protein structure. Deletions down to residue125 had the same antiviral activity as those to 132, but changescould now be seen in the aromatic proton NMR spectrum of thisshorter derivative. Substitution of the homologous murine sequencebetween residues 124 and 130 (human SPAAKTG; murine LPESSLR)resulted in only a small decrease in antiviral activity, furthersuggesting that the precise sequence in this region was notcritical for activity. Ser135 was substituted with a numberof other amino acids with little or no change in activity. Theimportance of the residues between 131 and 134 for biologicalactivity was corroborated by mutagenesis, although some substitutionsin this region were tolerated.     

Production and characterization of anti-human interferon {gamma} receptor antibody fragments that inhibit cytokine binding to the receptor

Three single-chain antibody fragments that recognize the extracellularhuman interferon receptor -chain (IFNR), and inhibit the bindingof human IFN, have been produced in Escherichia coli. Thesefragments are derived from murine anti-receptor monoclonal antibodies,and comprise the variable heavy (V_H) domain linked to the variablelight (V_L) chain through a 15 amino acid linker [(GGGGS)₃].Using surface plasmon resonance technology (BIAcore), the solubleproteins were shown to retain a high affinity for recombinantIFNR, and by radioimmunoassay to possess high inhibitory activitytowards IFN-binding to human Raji cells. The antibody fragmentsmost likely recognize epitopes that overlap the cytokine bindingsite on the receptor surface. Attempts to dissect further theantibodies to isolated V_H- and V_L-chains and to synthetic linearand cyclic peptides derived from the individual complementaritydetermining regions failed to afford fragments with significantIFNR binding affinity. Nevertheless, these native-like variableregion fragments and petidomimetics derived from them are ofinterest in the design of novel IFNR antagonists.     

Development and validation of fluorescence spectroscopic assays to evaluate antioxidant efficacy. Application to metal chelators

Two fluorescence-based assays were developed for rapid evaluation of compounds for antioxidant activity. These assays were based on the quenching of intensity of the fluorescent probe and an increase in its fluorescence anisotropy due to the free radicals generated during lipid peroxidation. A large unilamellar vesicle system, containing the fluorescence probe diphenylhexatriene-propionic acid, was used to study the effects of chelators on metal-ion-induced lipid peroxidation. In this paper, the actions of the chelating agents ethylenediaminetetraacetic acid disodium salt (EDTA), nitrilotriacetic acid trisodium salt (NTA), adenosine-5′-diphosphate disodium salt (ADP), and sodium citrate on Fe(II)- and Fe(III)-induced peroxidation were compared. The effects of chelators on metal-ion-induced peroxidation depended on the type of metal used to initiate peroxidation and, for citrate, also on the concentration of chelator used. EDTA strongly suppressed both Fe(II)- and Fe(III)-induced peroxidation in this system. NTA and ADP inhibited Fe(III)-induced peroxidation but enhanced Fe(II)-induced peroxidation at all concentrations tested. Citrate promoted both Fe(II)- and Fe(III)-induced peroxidations at lower chelator-to-metal ratios; however, at higher ratios, it inhibited both peroxidations. The results of the two fluorescence-based assays agreed well with the quantitation of conjugated dienes and hydroperoxides by high-performance liquid chromatography. The combination of sensitivity, speed, and general utility associated with these methods suggests that these methods will be useful in rapid screening of extracts and purified compounds for antioxidant activity.     

医用多孔支架材料的制备及缓释性能分析

采用溶液浇铸法制备了聚己内酯⁃姜黄素（PCL⁃CUR）多孔支架,通过冷场发射扫描电子显微镜（SEM）和热重分析仪（TG）等对支架材料的孔隙率、载药量及缓释性等进行了表征,并分析了其体外释药模型。结果表明,CUR在支架材料中含量为2 %（质量分数,下同）,PCL在醋酸中浓度为10 %,壳聚糖在支架材料中含量为2.86 %时,其释药模型符合1级方程,支架材料的孔隙率达95 %以上,载药量达到1.63 %,在PBS缓冲液中90 h内CUR累计释放率为76.2 %;其他配方的支架材料孔隙率均为95 %以上,90 h内CUR累计缓释率在60 %~87 %之间,表明制得的支架材料具有较理想的孔隙率和明显的CUR药物缓释作用,在组织工程领域有较好的应用前景。     

Application of graph theory to process design and analysis

Graph Theory and related computational techniques have been applied to a wide range of problems in process design and analysis to facilitate visualization, formulation, computation and interpretation. These applications are illustrated with selected examples. Most problems of practical importance are of such size and complexity that their solution requires using algorithms on a computer. Many problems we encountered in the last 20 years were solved using pollynomial time algorithms. We are now likely to have to conend with some harder and some ill-defined problems, which call for different approaches. Recent development in computational complexity theory provides us with a very useful perspective and some valuable tools, which we did not have 10 years ago. Alternatives for dealing with these hard but practically important problems are outlined and discussed in this paper.     

HAZOP及LOPA分析在氧化反应器的应用

详细阐述了HAZOP分析方法及步骤,并将其应用到PTA装置中的氧化反应器,对反应器进行系统的分析,找出反应器压力高和尾氧高的原因,针对后果等级较高的采用LOPA分析,通过对保护措施进行定量评估,从而确认风险等级以及风险是否可接受。     

生物传感器在水质分析监测中的应用

简要介绍了生物传感器的基本原理。重点介绍了生物传感器在水质分析监测中的应用,主要有生化需氧量的测定、细菌总数、硫化物、有机农药、酚的测定等。同时探讨了生物传感器存在的不足,并展望了生物传感技术的发展和应用前景。     

Application of digital image processing to the analysis of two-phase materials with spherical inclusions

Digital image processing was used to experimentally determine the two-dimensional pair distribution function of suspensions of solid spheres (cross-linked polystyrene spheres in silicone oil) and of composite materials with spherical inclusions (surface-treated glass spheres in unsaturated polyester resin). An assembly of optical devices and computers was used to obtain and store images of two-phase materials. Software was developed to analyze the information in the images in order to determine the pair distribution function. The technique developed is simple yet powerful, and with some modification could easily be adapted for the analysis of other multiphase systems.     

Application of ultraviolet spectrophotometry to the analysis of fats and derivatives

Summary There are several spectrophotometric methods available which allow a high degree of accuracy in the analysis of the common fats and oils. Further there are methods available for those fats and oils that contain unsaturated acids up to and including hexaenoic acid which allow good estimations of the composition. Several sources of error are discussed in an effort to point the way to their elimination. Recently published spectrophotometric data on certain polyunsaturated acids isolated from marine oils are discussed in relation to the possibility of extending the method to include these oils. More accurate spectrophotometric constants to be employed in the analysis of certain drying oils are suggested. Presented at the American Oil Chemists' Society, Minneapolis, Minn., October 11–13, 1954. A laboratory of the Eastern Utilization Research Branch, Agricultural Research Service, United States Department of Agriculture.     

重力式纤维束滤池在给水中的应用

重力式纤维束滤池是一种新的重力式滤池,它以新型的纤维束软填料作为滤元,取代了传统滤池中的石英砂,充分发挥了纤维滤料比表面积大、孔隙率高、截污能力强的特长,在给水领域中应用取得了一定效果.