磷脂酸含量测定法评价烟曲霉孢子对肺上皮细胞磷脂酶D活性的影响

目的 优化检测烟曲霉刺激A549细胞后磷脂酸（phosphotidic acid,PA）含量变化的方法,间接反应细胞内磷脂酶D（Phospholipase D,PLD）活性变化.方法 建立烟曲霉ATCC13073刺激肺上皮细胞模型;采用甲醇氯仿法提取胞内脂质;用改良的磷脂酸含量测定法测定PA标准品和细胞内PA水平变化规律.结果 PA标准品在5～ 250 μmol/L呈线性关系;经膨胀孢子刺激后,肺上皮细胞内PA含量显著升高,休眠孢子在这一过程中对肺上皮细胞内PA含量无明显作用.结论 改良的PA测量法能快速、稳定而有效地测定细胞内的PLD活性.烟曲霉膨胀孢子能显著激活肺上皮细胞内的PLD活性.     

Rapid detection of Phytophthora nicotianae by simple DNA extraction and real‐time loop‐mediated isothermal amplification assay

Phytophthora nicotianae is a phytopathogenic oomycete with a wide host range and worldwide distribution. Rapid detection and diagnosis at the early stages of disease development are important for the effective control of P. nicotianae. In this study, we designed a simple and rapid loop‐mediated isothermal amplification (LAMP)‐based detection method for P. nicotianae. We tested three DNA extraction methods and selected the Kaneka Easy DNA Extraction Kit version 2, which is rapid and robust for LAMP‐based detection. The designed primers were tested using mycelial DNA from 35 species (81 isolates) of Phytophthora, 12 species (12 isolates) of Pythium, one isolate of Phytopythium and one isolate each from seven other soil‐borne pathogens. All of the 42 P. nicotianae isolates were detected by these primers, and no other isolates gave positive results. Three isolates were tested for the sensitivity of the reaction, and the lowest amounts of template DNA that could be detected were 10 fg for two isolates and 1 fg for the third. The target was detected within 25 min in all tested samples, including DNA extracted from both inoculated and naturally infected plants. In contrast, PCR assays with P. nicotianae‐specific primers failed or showed weakened detection in several samples. Thus, we found that the rapid DNA extraction and LAMP assay methods developed in this study can be used to detect P. nicotianae with high sensitivity, specificity and stability.     

Error detection during VMAT delivery using EPID-based 3D transit dosimetry

PurposeTo investigate the effectiveness of an EPID-based 3D transit dosimetry system in detecting deliberately introduced errors during VMAT delivery.MethodsAn Alderson phantom was irradiated using four VMAT treatment plans (one prostate, two head-and-neck and one lung case) in which delivery, thickness and setup errors were introduced. EPID measurements were performed to reconstruct 3D dose distributions of “error” plans, which were compared with “no-error” plans using the mean gamma (γ_mean), near-maximum gamma (γ_1%) and the difference in isocenter dose (ΔD_isoc) as metrics.ResultsOut of a total of 42 serious errors, the number of errors detected was 33 (79%), and 27 out of 30 (90%) if setup errors are not included. The system was able to pick up errors of 5 mm movement of a leaf bank, a wrong collimator rotation angle and a wrong photon beam energy. A change in phantom thickness of 1 cm was detected for all cases, while only for the head-and-neck plans a 2 cm horizontal and vertical shift of the phantom were alerted. A single leaf error of 5 mm could be detected for the lung plan only.ConclusionAlthough performed for a limited number of cases and error types, this study shows that EPID-based 3D transit dosimetry is able to detect a number of serious errors in dose delivery, leaf bank position and patient thickness during VMAT delivery. Errors in patient setup and single leaf position can only be detected in specific cases.     

基于激光雷达数据的物种分布模拟: 以美国加州内华达山脉南部区域食鱼貂分布模拟为例

生态位模型通过拟合物种分布与环境变量之间的关系提供物种空间分布预测, 在生物多样性研究中有广泛应用。激光雷达(LiDAR)是一种新兴的主动遥感技术, 已被大量应用于森林三维结构信息的提取, 但其在物种分布模拟的应用研究比较缺乏。本研究以美国加州内华达山脉南部地区的食鱼貂(Martes pennanti)的分布模拟为例, 探索LiDAR技术在物种分布模拟中的有效性。生态位模型采用5种传统多类分类器, 包括神经网络、广义线性模型、广义可加模型、最大熵模型和多元自适应回归样条模型, 并使用正样本-背景学习(presence and background learning, PBL)算法进行模型校正; 同时对这5种模型使用加权平均进行模型集成, 作为第6个模型。此外, 一类最大熵模型也被用于模拟该物种的空间分布。模型的连续输出和二值输出分别使用AUC (area under the receiver operating characteristic curve)以及基于正样本-背景数据的评价指标F_pb进行评价。结果表明, 仅考虑气候因子(温度和降水)时, 7个模型的AUC和F_pb平均值分别为0.779和1.077; 当考虑LiDAR变量(冠层容重、枝下高、叶面积指数、高程、坡度等)后, AUC和F_pb分别为0.800和1.106。该研究表明, LiDAR数据能够提高食鱼貂空间分布的预测精度, 在物种分布模拟方面存在一定的应用价值。     

食品、药品微生物检测实验室质量控制方法分析

系统分析食品、药品微生物检测实验室的质量控制方法,从总体环境条件控制、微生物室管理、样品管理、仪器设备管理、人员管理、培养基质量控制、标准菌(毒)种管理、操作过程控制、记录管理等9个方面进行系统分析,对微生物实验室的质量控制工作提供系统指导,提高微生物检测结果的准确性.     

DNA methylation based biomarkers: Practical considerations and applications

A biomarker is a molecular target analyzed in a qualitative or quantitative manner to detect and diagnose the presence of a disease, to predict the outcome and the response to a specific treatment allowing personalized tailoring of patient management. Biomarkers can belong to different types of biochemical molecules such as proteins, DNA, RNA or lipids, whereby protein biomarkers have been the most extensively studied and used, notably in blood-based protein quantification tests or immunohistochemistry. The rise of interest in epigenetic mechanisms has allowed the identification of a new type of biomarker, DNA methylation, which is of great potential for many applications. This stable and heritable covalent modification mostly affects cytosines in the context of a CpG dinucleotide in humans. It can be detected and quantified by a number of technologies including genome-wide screening methods as well as locus- or gene-specific high-resolution analysis in different types of samples such as frozen tissues and FFPE samples, but also in body fluids such as urine, plasma, and serum obtained through non-invasive procedures. In some cases, DNA methylation based biomarkers have proven to be more specific and sensitive than commonly used protein biomarkers, which could clearly justify their use in clinics. However, very few of them are at the moment used in clinics and even less commercial tests are currently available. The objective of this review is to discuss the advantages of DNA methylation as a biomarker, the practical considerations for their development, and their use in disease detection, prediction of outcome or treatment response, through multiple examples mainly focusing on cancer, but also to evoke their potential for complex diseases and prenatal diagnostics.     

Outlier detection methods are still effective even using virtual species created with the probabilistic approach

Liu et al. (Journal of Biogeography, 2018, 45 :164–176) presented an approach to detect outliers in species distribution data by developing virtual species created using the threshold approach. Meynard et al. (Journal of biogeography, 2019, 46 :2141–2144) raised concerns about this approach stating that ‘using a probabilistic approach … may significantly change results’. Here we provide a new series of simulations using the two approaches and demonstrate that the outlier detection approach based on pseudo species distribution models was still effective when using the probabilistic approach, although the detection rate was lower than when using the threshold approach.     

Rapid determination of plasmid-carrying yeast cells by using an imaging sensor system

A novel imaging sensor system for the determination of plasmid carrying yeast cells was developed. The sensor system consisted of an Silicon Intensifier Target (SIT) video camera, a fluorescent microscope, and a personal computer system equipped with an image memory board. This system was based on the fact that the membrane integrity of only plasmid-carrying cells is lost following cell growth in 5-fluoro-orotic acid (5-FOA) containing medium, and consequently these target cell can be stained with fluorescent probes and detected. In this study, plasmid-carrying cells were detected and their fraction determined in a mixture of both plasmid-carring and plasmid-free cells. A good correlation was observed between the values determined by this sensor system and the conventional method in the 30%-80% range, and one assay was possible within 4 h. This sensor system could be used for the monitoring of plasmid-carrying fraction in recombinant yeast cells during cultivation.     

西加毒素的危害及其检测技术

西加毒素是由少数几种海洋底栖微藻产生,具有极强毒性的生物毒素,它能够通过食物链传递而累计在多种珊瑚鱼体内,继而造成人类因食用鱼类而中毒。近年来,随着珊瑚鱼类在世界范围内的广泛贸易,西加毒素中毒已经成为全球性的健康问题。为给预防西加毒素引起的食物中毒提供借鉴,对西加毒素的分子结构、化学性质、生物来源、制毒机理、中毒症状和对人类的危害进行了综述。就目前主要的检测方法进行了技术特性的介绍,并对常用检测方法的优缺点进行了比较分析。