自然感染鼠痘的小鼠肝脾内病毒超微结构的观察

应用自然感染鼠痘存档5年蜡块转制超薄切片，在电镜下观察发现鼠痘病毒的复制部位是在肝脾细胞浆的基质区和包涵体的周围。此部位有大量不同发育阶段的病毒颗粒。未成熟病毒颗粒可分为球状颗粒和含有拟核的球状颗粒。成熟病毒颗粒的典型形态呈年椭圆形，外被双层囊膜，中心含有哑铃形核心，核心两侧各有一个测体。本实验结果证实应用存档蜡块转制超薄切片，可成功地应用于鼠痘感染及追溯性鼠痘病原和传染源的电镜诊断。并在电镜下证实光镜下A、B型包涵体及免疫酶组化病毒抗原的阳性部位与鼠痘病毒生物合成和装配的部位是一致的。     

免疫组织化学抗原修复微波方法的一点改进

本文主要探讨了免疫组织化学抗原修复微波的一点改进方法。     

表达HIV-1 Gag-Pol抗原的重组痘病毒的构建及免疫原性

目的构建稳定高效表达HIV-1Gag-Pol蛋白的重组痘病毒载体疫苗,并检测其体液免疫效果。方法将修饰型HIV-1Gag-Pol基因插入到痘病毒表达载体pSC11m2启动子P7·5下游,经与野生型痘病毒同源重组后,进行蓝白斑筛选。用筛选得到的重组病毒rM-GP转染HEK293细胞,经SDS-PAGE和Westernblot检测表达蛋白。用该重组病毒免疫BALB/c小鼠,并检测体液免疫应答。结果已筛选到稳定高效表达HIV-1Gag-Pol蛋白的重组痘病毒载体rM-GP,经Westernblot,在免疫小鼠血清中检测到抗HIV-1p24蛋白的特异性抗体。结论重组痘病毒载体rM-GP能稳定高效表达HIV-1Gag-Pol蛋白,并能引起有效的体液免疫应答。     

病理诊断中免疫组化技术的应用研究

在临床病理诊断和形态学研究中,免疫组化染色是应用抗体结合组织中特异性抗原的特殊染色。目前其多应用于肿瘤的病理诊断,提高恶性肿瘤诊断和鉴别诊断准确性。但是,随着免疫组化的广泛应用,发现免疫组化染色过程中存在一些问题,分析问题的原因,找到解决问题的办法。本文就此问题作如下综述。     

草原兔尾鼠卵透明带3融合蛋白抗原的制备及免疫反应性

目的大量制备重组的草原兔尾鼠卵透明带3(Recom binant Laguruslagurus zona pellucida 3,r-LZP3)融合蛋白,并检测其免疫反应性。方法经密码子优化的Lzp3基因在E.coli BL21(DE3)中以包涵体形式表达融合蛋白,表达产物经洗涤、纯化、溶解、复性后,用Western blot、ELISA和抗生育试验鉴定免疫反应性。结果得到纯度达93%的r-LZP3蛋白。Western blot显示表达产物与LZP3小鼠抗血清出现特异性反应条带。ELISA表明免疫小鼠血清中LZP3抗体滴度达1∶45000以上。抗生育试验表明r-LZP3蛋白免疫小鼠的避孕率为25%,平均每胎产仔数减少4~5只。结论已获得了大量具有显著免疫活性的r-LZP3蛋白,为控制草原兔尾鼠鼠害的研究提供了重要的抗原。     

骨样骨瘤的影像学诊断与病理分析

目的探讨骨样骨瘤的影像学表现和病理变化。方法对29例骨样骨瘤的影像学资料和病理进行回顾性分析。结果本组病例29例骨样骨瘤均为单发,其中皮质型12例;中心型5例;松质骨型8例;骨膜型4例。结论瘤巢是骨样骨瘤的特征性表现,了解骨样骨瘤的病理变化和影像学表现,诊断就比较明确,可减少误诊发生。     

以结核分枝杆菌38kD重组蛋白为抗原的血清学诊断

目的克隆表达结核分枝杆菌38kD抗原基因,并以此蛋白为抗原,进行分枝杆菌的血清学诊断。方法采用PCR自结核分枝杆菌基因组DNA中扩增38kD抗原基因,经测序鉴定正确后,克隆于pGEX-4T-2表达载体,转化大肠杆菌BL21进行诱导表达,利用GSTrapFF蛋白柱纯化表达产物,经ELISA检测其抗原的灵敏度与特异性。结果目的蛋白表达量约占菌体总蛋白量的28%,ELISA检测灵敏度为79.8%,特异性为99.0%。结论重组38kD抗原可用于结核分枝杆菌的诊断。     

关于HCV不同区段抗原的反应性研究

目的　探讨各种丙型肝炎病毒 (HCV)抗原在诊断中的作用及其与HCVRNA的关系。方法　利用HCV不同区段抗原抗 -HCV酶联免疫试验 (EIA)和逆转录聚合酶链反应法 (RT PCR) ,对不同临床型HCV患者进行检测。结果　各组患者均以抗 NS3和抗 C阳性率最高。但均有一定比例患者抗 NS4和抗 NS5阳性。与各单个片段抗 HCV阳性率相比 ,抗 HCV总抗体阳性率最高。结论　NS3和C抗原在检测抗 HCV中起很重要作用 ,但NS4和NS5抗原在抗 HCVEIA中的作用也不容忽视。联合应用不同区段HCV抗原将大大提高抗 HCVEIA试剂的灵敏度。     

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...     

Interplay of Cellular mRNA,miRNA and Viral miRNA during Infection of a Cell

The understanding of the kinetics of gene expression in cells infected by viruses is currently limited. As a rule, the corresponding models do not take viral microRNAs (miRNAs) into account. Such RNAs are, however, operative during the replication of some viruses, including, e.g., herpesvirus. To clarify the kinetics of this category (with emphasis on the information available for herpesvirus), I introduce a generic model describing the transient interplay of cellular mRNA, protein, miRNA and viral miRNA. In the absence of viral miRNA, the cellular miRNA is considered to suppress the populations of mRNA and protein due to association with mRNA and subsequent degradation. During infection, the viral miRNA suppresses the population of cellular miRNA and via this pathway makes the mRNA and protein populations larger. This effect becomes appreciable with the progress of intracellular viral replication. Using biologically reasonable parameters, I investigate the corresponding mean-field kinetics and show the scale of the effect of viral miRNAs on cellular miRNA and mRNA. The scale of fluctuations of the populations of these species is illustrated as well by employing Monte Carlo simulations.     

高效液相色谱与质谱联用法分离鉴定己烯雌酚合成抗原

采用L ichrospher C-18柱,甲醇和水为流动相,流速0.3 mL/m in,甲醇从60%~100%(体积分数)梯度洗脱25 m in,液质联用,电喷雾(ESI),对己烯雌酚(DES)及其衍生物己烯雌酚-单(双)-丁酸乙酯基-醚进行了分离和结构鉴定。以混合酸酐法将己烯雌酚-单-羧丙基-醚与载体蛋白偶联形成完全抗原,并以紫外扫描分析了完全抗原的偶联比率。结果表明:在极性溶剂中,己烯雌酚和其衍生物存在着顺反异构转化现象,极性有机溶剂的溶剂化作用促使顺反异构体的转化。计算了DES-MCPE-BSA的偶联比率为10。     

飞机发动机油液故障诊断技术的现状与发展趋势1

油液分析技术是飞机发动机故障诊断中一种非常重要的手段。综述了常规理化分析、光谱分析、铁谱分析、颗粒计数器和磁塞监测等几种常用的油液分析技术,并进行了对比分析。最后指出,综合应用多种油液分析技术才能实现飞机发动机的全面、准确、快速的故障诊断。     

挤压法重质纯碱技术的完善与装置国产化

介绍鸿化第二套挤压法重质纯碱装置工艺、设备进一步改进完善及大部分引进设备国产化的情况。     

Cellular Redox Metabolism Is Modulated by the Distinct Localization of Cyclic Nucleotide Phosphodiesterase 5A Isoforms

3′-5′ cyclic nucleotide phosphodiesterases (PDEs) are a family of evolutionarily conserved cAMP and/or cGMP hydrolyzing enzymes, components of transduction pathways regulating crucial aspects of cell life. Among them, cGMP-specific PDE5—being a regulator of vascular smooth muscle contraction—is the molecular target of several drugs used to treat erectile dysfunction and pulmonary hypertension. Production of full-length murine PDE5A isoforms in the milk-yeast Kluyveromyces lactis showed that the quaternary assembly of MmPDE5A1 is a mixture of dimers and tetramers, while MmPDE5A2 and MmPDE5A3 only assembled as dimers. We showed that the N-terminal peptide is responsible for the tetramer assembly of MmPDE5A1, while that of the MmPDE5A2 is responsible for its mitochondrial localization. Overexpression of the three isoforms alters at different levels the cAMP/cGMP equilibrium as well as the NAD(P)⁺/NAD(P)H balance and induces a metabolic switch from oxidative to fermentative. In particular, the mitochondrial localization of MmPDE5A2 unveiled the existence of a cAMP-cGMP signaling cascade in this organelle, for which we propose a metabolic model that could explain the role of PDE5 in some cardiomyopathies and some of the side effects of its inhibitors.     

The Lack of STING Impairs the MHC-I Dependent Antigen Presentation and JAK/STAT Signaling in Murine Macrophages

STING is a transmembrane ER resident protein that was initially described as a regulator of innate immune response triggered by viral DNA and later found to be involved in a broader range of immune processes. Here, we assessed its role in the antigen presentation by generating a STING KO macrophage cell line. In the absence of STING, we observed an impaired OVA-derived SIINFEKL peptide presentation together with a decreased level of MHC-I complex on the plasma membrane, likely due to a decreased mRNA expression of β2 m light chain as no relevant alterations of the peptide-loading complex (TAPs) were found. Moreover, JAK-STAT signaling resulted in impaired STING KO cells following OVA and LPS treatments, suggesting a dampened activation of immune response. Our data revealed a new molecular role of STING in immune mechanisms that could elucidate its role in the pathogenesis of autoimmune disorders and cancer.     

Human Milk Oligosaccharide 2′-Fucosyllactose Modulates Local Viral Immune Defense by Supporting the Regulatory Functions of Intestinal Epithelial and Immune Cells

Human milk contains bioactive components that provide protection against viral infections in early life. In particular, intestinal epithelial cells (IEC) have key regulatory roles in the prevention of enteric viral infections. Here we established an in vitro model to study the modulation of host responses against enteric viruses mimicked by poly I:C (pIC). The effects of 2′-fucosyllactose (2′FL), abundantly present in human milk, were studied on IEC and/or innate immune cells, and the subsequent functional response of the adaptive immune cells. IEC were pre-incubated with 2′FL and stimulated with naked or Lyovec™-complexed pIC (LV-pIC). Additionally, monocyte-derived dendritic cells (moDC) alone or in co-culture with IEC were stimulated with LV-pIC. Then, conditioned-moDC were co-cultured with naïve CD4⁺ T helper (Th)-cells. IEC stimulation with naked or LV-pIC promoted pro-inflammatory IL-8, CCL20, GROα and CXCL10 cytokine secretion. However, only exposure to LV-pIC additionally induced IFNβ, IFNλ1 and CCL5 secretion. Pre-incubation with 2′FL further increased pIC induced CCL20 secretion and LV-pIC induced CXCL10 secretion. LV-pIC-exposed IEC/moDC and moDC cultures showed increased secretion of IL-8, GROα, IFNλ1 and CXCL10, and in the presence of 2′FL galectin-4 and -9 were increased. The LV-pIC-exposed moDC showed a more pronounced secretion of CCL20, CXCL10 and CCL5. The moDC from IEC/moDC cultures did not drive T-cell development in moDC/T-cell cultures, while moDC directly exposed to LV-pIC secreted Th1 driving IL-12p70 and promoted IFNγ secretion by Th-cells. Hereby, a novel intestinal model was established to study mucosal host-defense upon a viral trigger. IEC may support intestinal homeostasis, regulating local viral defense which may be modulated by 2′FL. These results provide insights regarding the protective capacity of human milk components in early life.     

往复压缩机监测与诊断技术研究现状与展望

分析了往复压缩机的常见故障模式与机理,综述了往复压缩机故障诊断方法和监测装置的研究现状,指出了往复压缩机在线监测与诊断技术的发展方向。