应用细胞感染技术检测水中活性隐孢子虫

根据活性隐孢子虫感染人结肠癌细胞（HCT-8）产生感染斑（族）的特点,利用体外细胞培养技术和免疫荧光分析技术建立了活性隐孢子虫的检测方法,并利用最大可能数统计原理对样品中的活性隐孢子虫进行了定量.     

Comparison of cytopathogenicity, immunofluorescence and In situ DNA hybridization as methods for the detection of adenoviruses

Three different methods were compared for their efficiency at detection of adenoviruses. The samples examined for viral analysis consisted of concentrates prepared from raw sewage, chosen as providing a representation of the spectrum of viruses being intestinally shed from a large population at any given time. When using one single cell line, HEp-2, the overall numbers of adenoviruses detected using cytopathogenicity and immunofluorescence were roughly equal. In situ hybridization was approx. 40% more sensitive than either of these other methods as determined by average virus titers for the different samples, and also proved to be better by means of a nonparametric comparison. The 293 cell line was approx. 5 times more sensitive for detecting adenoviruses by cytopathogenicity as compared with the HEp-2 cell line, but proved unsuitable in our hands for quantitatively detecting indigenous adenoviruses by immunofluorescence. The relative number of indigenous adenoviruses present in the sewage concentrates we examined was, on average, 94-fold greater than that of enteroviruses. Assay of enteroviruses was performed by plaque assay in the BGM cell line.     

用PCR技术检测水中隐孢子虫

介绍了一种检测水中隐孢子虫的巢式PCR方法：根据Cryptosporidium parvum 18s rRNA基因序列选用了两对引物，特异性扩增了该基因可变区中1056bp和435bp目标片段。加标试验表明，该方法的检测限为10个卵囊。     

Functional display of amylase on yeast surface from Rhizopus oryzae as a novel enzyme delivery method

Fungal amylase has great importance in fermentation industry such as brewing, food fermentation, starch hydrolysis and for improving microbial populations in chicken intestine through feed applications. In the present investigation, alpha amylase cDNA from Rhizopus oryzae was cloned, sequenced, and successfully surface anchored in functional form using Saccharomyces cerevisiae EBY100 as host, yielding enzyme activity of 4.35 (±0.5) U/ml. The surface displayed yeast expressed amylase activity using plate assay, produced glucose and maltose as hydrolysis products using starch as substrate. The targeted and armed yeast with displayed enzyme was evaluated for its characterization at various pH and temperatures. The engineered yeast showed optimal activity at neutral pH and at 50°C incubation temperature. Reducing sugars produced by displayed enzyme were visualized by paper chromatography. The data suggested successful heterologous expression and display of amylase enzyme on yeast cell surface. The displayed system could further be utilized in fermentation industry for improving cost-effectiveness of the process.     

Determination of surface-induced platelet activation by applying time-dependency dissipation factor versus frequency using quartz crystal microbalance with dissipation

Platelet adhesion and activation rates are frequently used to assess the thrombogenicity of biomaterials, which is a crucial step for the development of blood-contacting devices. Until now, electron and confocal microscopes have been used to investigate platelet activation but they failed to characterize this activation quantitatively and in real time. In order to overcome these limitations, quartz crystal microbalance with dissipation (QCM-D) was employed and an explicit time scale introduced in the dissipation versus frequency plots (Df–t) provided us with quantitative data at different stages of platelet activation. The QCM-D chips were coated with thrombogenic and non-thrombogenic model proteins to develop the methodology, further extended to investigate polymer thrombogenicity. Electron microscopy and immunofluorescence labelling were used to validate the QCM-D data and confirmed the relevance of Df–t plots to discriminate the activation rate among protein-modified surfaces. The responses showed the predominant role of surface hydrophobicity and roughness towards platelet activation and thereby towards polymer thrombogenicity. Modelling experimental data obtained with QCM-D with a Matlab code allowed us to define the rate at which mass change occurs (A/B), to obtain an A/B value for each polymer and correlate this value with polymer thrombogenicity.     

Visual learning and classification of human epithelial type 2 cell images through spontaneous activity patterns

Identifying the presence of anti-nuclear antibody (ANA) in human epithelial type 2 (HEp-2) cells via the indirect immunofluorescence (IIF) protocol is commonly used to diagnose various connective tissue diseases in clinical pathology tests. As it is a labour and time intensive diagnostic process, several computer aided diagnostic (CAD) systems have been proposed. However, the existing CAD systems suffer from numerous shortcomings due to the selection of features, which is commonly based on expert experience. Such a choice of features may not work well when the CAD systems are retasked to another dataset. To address this, in our previous work, we proposed a novel approach that learns a set of filters from HEp-2 cell images. It is inspired by the receptive fields in the mammalian's vision system, since the receptive fields can be thought as a set of filters for similar shapes. We obtain robust filters for HEp-2 cell classification by employing the independent component analysis (ICA) framework. Although, this approach may be held back due to one particular problem; ICA learning requires a sufficiently large volume of training data which is not always available. In this paper, we demonstrate a biologically inspired solution to address this issue via the use of spontaneous activity patterns (SAP). The spontaneous activity patterns, which are related to the spontaneous neural activities initialised by the chemical release in the brain, are found as the typical stimuli for the visual cell development of newborn animals. In the classification system for HEp-2 cells, we propose to model SAP as a set of small image patches containing randomly positioned Gaussian spots. The SAP image patches are generated and mixed with the training images in order to learn filters via the ICA framework. The obtained filters are adopted to extract the set of responses from a HEp-2 cell image. We then employ regions from this set of responses and stack them into “cubic regions”, and apply a classification based on the correlation information of the features. We show that applying the additional SAP leads to a better classification performance on HEp-2 cell images compared to using only the existing patterns for training ICA filters. The improvement on classification is particularly significant when there are not enough specimen images available in the training set, as SAP adds more variations to the existing data that makes the learned ICA model more robust. We show that the proposed approach consistently outperforms three recently proposed CAD systems on two publicly available datasets: ICPR HEp-2 contest and SNPHEp-2.     

免疫荧光定量分析仪设计

设计了免疫荧光定量分析仪,用以对人体血液和尿液中的各种分析物(CRP、PCT、NTpro BNP、c Tn I等)含量进行快速准确的定量分析。光源采用大功率LED灯珠,采用窄带干涉滤光片对激发光和荧光进行滤光,采用内置运放的光电转换芯片OPT101进行荧光强度的检测。采用步进电机驱动,皮带传动方式带动双排滚珠宽体滑块在单根精密直线导轨上滑动,实现对检测样品的扫描式检测。通过与标准仪器进行对比试验,结果表明样机在小型化、快速性以及低成本的基础上,测量结果准确,测量精度高,稳定性好,能满足临床应用要求。     

藻胆蛋白免疫荧光法的初步探索

采用CNBr活化的琼脂糖凝胶小珠作为固相载体,应用藻胆蛋白荧光标记物对血清可溶性抗原检测作了初步的探索. 结果表明, 藻胆蛋白免疫荧光法用于可溶性抗原检测不但可行,而且具有荧光强度高、可长期保存且无明显衰减等特点.     

几种常见抗体介导的肾小球肾炎免疫荧光和超微结构的形态对比观察

本文通过比较观察几种常见抗体介导的肾小球肾炎的免疫荧光和超微结构的特征，以进一步提高对其识别能力和诊断水平。结果显示，一些常见抗体介导的肾小球病，如IgA肾病（包括紫癜性肾炎）、膜性肾病、急性弥漫增生性肾小球肾炎、膜性增生性肾小球肾炎（Ⅰ型）、狼疮性肾炎（Ⅳ型）的免疫荧光和电镜检查所显示的抗体或免疫反应产物有较好的可比性和一致性。提高对抗体介导的肾小球肾炎典型肾小球超微结构形态特征的认识和鉴别能力将十分有助于对肾小球疾病作出正确的病理学诊断。     

鼻咽癌增殖与转移机制荧光检测的研究进展

鼻咽癌是一种高转移性的头颈部肿瘤,EB病毒编 码潜伏膜蛋白-1(LMP1)被认为与鼻咽癌的发展、侵 袭、转移关系密切,但是LMP1诱导鼻咽癌增殖、转移的机制尚不明确。本文对近年鼻咽癌增 殖、转移机制 方面的研究进展作简要介绍;荧光检测技术作为一种有效技术手段被广泛应用于生命科学, 对应用于鼻咽 癌增殖、转移机制研究中的荧光检测技术(免疫荧光、双分子荧光互补、荧光共振能量转移 )进行详细总 结,并分析了各自的优缺点。针对鼻咽癌增殖、转移与肿瘤细胞质膜关系密切的特点,展望 了利用全内反 射荧光显微镜(TIRF)结合荧光共振能量转移(FRET)技术获取鼻咽癌细胞质膜上LMP1蛋白高分 辨率成像,破解鼻咽癌增殖、转移机制的前景。