4种植物源性成分多重real-time PCR检测方法的建立及其在食用淀粉中的应用

建立一种可同时快速检测红薯、木薯、马铃薯、玉米源性成分的多重实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)方法。分别以红薯g3pdh基因、木薯g3pdh基因、马铃薯UGPase基因、玉米zSSIIb基因为靶基因设计特异性引物和TaqMan探针,以18S rRNA基因为内参基因,建立多重real-time PCR方法,开展方法学验证,并对不同掺入比例模拟样品和实际淀粉样品进行检测。结果显示,该方法具有高通量、特异性强、灵敏度高等优点。与15种非目标源性均无交叉反应;对目标DNA的检测灵敏度可达到3×10^(-3) ng/μL,且具有良好的线性关系和扩增效率;对淀粉样品的检出限可达0.1%,对50份实际样品进行检测,结果与参比方法一致,说明建立的多重real-time PCR法可用于食用淀粉种类掺假鉴别检测。

血-迷路屏障通透性与突聋发病机制及预后的关系:在3D real IR的发现

目的分析突聋患者的内耳钆造影MRI三维真实重建反转恢复(three dimensional real inversion recovery,3D real IR)成像上的表现,探讨血-迷路屏障的通透性与突聋发病机制及其预后的关系。方法对41例单侧突聋患者行内耳钆造影MRI,测量患耳和健耳的耳蜗信号强度,并测量延髓信号强度,分别计算出耳蜗/延髓比值(cochlear/medulla ratio,CM ratio),以CM比值作为血-迷路屏障通透性的标志物,分析突聋患者患耳、健耳CM比值的不对称程度与疗效之间的关系。结果41例患者中,33例(80.48%)患耳的CM比值高于健耳,差异有统计学意义(P<0.05);患耳CM比值为健耳的1.5倍以下者18例,治疗有效率为77.78%(14/18);患侧CM比值不高于健侧者8例,治疗有效率为100%;达到健耳的1.5倍至1.75倍之间者7例,治疗有效率为100%(7/7);达到健耳的1.75倍至2.0倍之间者2例,治疗有效率为50%(1/2);达到健耳的2.0倍以上者14例,治疗有效率为14.28%(12/14);差异有统计学意义(P<0.05)。结论内耳3D Real IR可显示突聋患者血-迷路屏障通透性的改变,80.48%的突聋患者患侧耳蜗出现高信号,患耳CM比值达健耳的1.75倍以上者多数预后不良。

基于病菌孢子捕捉和real-time PCR技术的田间空气中小麦白粉病菌孢子动态监测及病情估计模型研究

利用Burkard定容式孢子捕捉器结合real-time PCR定量技术,分别对种植高抗、中感和高感白粉病小麦品种的田间空气中白粉病菌分生孢子浓度进行监测,结果表明,real-time PCR定量与传统的显微观察计数两种方法测得的孢子浓度呈显著正相关(P≤0.01),且两种病菌孢子计数方法在同一抗性品种上监测到的孢子浓度动态相近。此外,两种方法测得的孢子浓度与各气象因子的相关性分析结果一致,空气中的白粉病菌孢子浓度主要与空气相对湿度显著正相关。在此基础上,利用两种方法测定的田间空气中白粉病菌孢子浓度分别建立了基于累积孢子浓度的田间病情估计模型。分析发现,基于两种孢子浓度测定方法建立的病情估计模型间无显著性差异,表明real-time PCR定量技术测定的孢子浓度在构建白粉病病情估计模型上具有一定可行性。该结果为real-time PCR定量技术与病菌孢子捕捉技术相结合用于小麦白粉病的监测和预测提供理论依据。

马乳酒样乳杆菌马乳酒样亚种real-time PCR检测方法的建立与应用

本研究建立了一种特异性实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)检测方法,根据模式菌株马乳酒样乳杆菌马乳酒样亚种ZW3的16S rDNA序列和全基因组序列设计筛选特异性引物,采用SYBR Green I荧光染料建立real-time PCR方法,并对方法的特异性、灵敏度、重复性和混合体系等进行检测。结果表明,本研究所建立的方法特异性强、灵敏度高、重复性好,建立real-time PCR的标准曲线,其决定系数R2为0.965,具有良好的线性关系,且在马乳酒样乳杆菌马乳酒样亚种及混合体系中可以特异性检出。综上,本研究建立的real-time PCR法可以快速、准确地检测马乳酒样乳杆菌马乳酒样亚种,为马乳酒样乳杆菌的特异性定性定量检测提供了一种新的方法。

基于Real-time PCR法检测乳粉中牛源性成分定量研究

基于Real-timePCR建立了乳粉中牛源性成分相对定量检测方法,并对牛的特异性引物与探针进行了特异性、灵敏度和稳定性测试。通过模拟不同浓度牛乳粉与马乳粉混合样本,根据其△Ct值的函数关系进行线性拟合进而绘制标准曲线,建立乳粉中牛源性成分的相对定量检测。结果显示,该方法的最低检测限为0.00001 mg/mL,回收率为91.11%~119.2%,组间变异系数≤0.58%、组内变异系数≤1.44%。说明该方法在特异性与稳定性上适用于乳粉中牛源性成分及含量的掺假检测。

一种基于real-time PCR技术的TTV检测方法的建立及应用

目的:本研究旨在开发一种具有更高灵敏度和特异性的TTV检测技术,为揭示TTV在多种疾病过程中的作用提供重要的技术支持。方法:为了更精确、灵敏的检测TTV,本研究分析了目前公布的所有亚型的TTV基因序列,在此基础上建立了一种基于UTR区域的real-time PCR检测方法,并与文献报道应用较为广泛的PCR检测方法进行了对比。结果:本研究建立的方法在1×10^(7)~1×10^(1) copies/μL标准品浓度范围内具有良好的线性关系,相关系数为1.000,斜率为-3.446,检测下限为1×10^(1) copies/μL。重复性试验结果显示,组内变异系数为7.22%,表明本方法重复性、稳定性较强。针对30份临床样本,使用本研究建立的real-time PCR检测方法及目前被多个研究所使用的4套引物进行对比。结果表明,本研究所建立的方法灵敏度显著高于文献中报道的4种方法(P<0.01);Sanger测序结果表明,本方法检测出的30份阳性样本均为TTV,检测特异性为100%。结论:本研究采用基于TaqMan探针的real-time PCR检测方法,检测灵敏性高、覆盖基因型范围广,尤其对于TTV病毒载量较低的情况下能够进行定量检测,对于TTV病毒的致病性及作为免疫标志物的应用提供重要的技术支持。

基于γ-Clahe和Real-esrgan的红外图像增强方法

针对红外图像对比度低和清晰度差的问题,提出一种基于γ-Clahe和Real-esrgan的红外图像增强方法。通过Haar小波变换将红外图像分解为低频和高频分量,对低频和高频分量分别进行γ-Clahe变换和高斯滤波,将处理后低频和高频分量进行重构得到重构的红外图像,采用Real-esrgan算法对重构的红外图像进行超分辨率重建。结果表明,所提出的红外图像增强算法的主观和客观指标均优于HE、Clahe和Gamma算法,相较于上述三种传统算法PSNR平均提高了3.525、9.141和9.631,SSIM平均提高了0.085、0.295和0.162,使重建后的红外图像对比度和清晰度得到了增强。

基于Real ESRGAN的视频修复系统研究

图像修复和视频修复是计算机视觉的一项重要任务,其中图像修复又是视频修复的基础。为此,如何有效提升图像质量是实现视频质量提升的关键。传统的图像修复算法主要以样本信息为基础,通过对样本内容的扩撒来实现对破损区域的修复;由于这种方式对于图像样本有一定要求,从而制约传统图像修复技术的发展。为此,以生成新图像内容为基础的神经网络如GAN的出现,为图像修复技术转向深度学习提供方向。该课题主要以Real ESRGAN网络的图像修复技术为基础,通过对音频视频数据的隔离处理以及相同帧数据的优化和标记,构建视频修复处理流程。通过对随机视频样本的测试,并通过对单帧图片质量和视频数据流畅性与协调性的评估,该视频处理方法表现出较好的系统性能。

Real-World Evidence in Localized Pancreatic: Coping with Uncertainty in Unselected Populations

Background: Localized pancreatic cancer, including resectable (R), borderline resectable (BR) and locally advanced unresectable disease (LAU), is considered in clinical guidelines for diverse treatment options based on clinical trials in selected populations. Hence, exploring with real world evidence (RWE) clinicians’ preferences for treatment options and their results seems pertinent. Methods: In a set of consecutive patients with localized pancreatic cancer assisted in a third level hospital from January 2013 to December 2022, medical records, symptoms, diagnostic process, distribution between subtypes, and treatment plans, with safety and efficacy results, were assessed. Results: A total of 152 patients with localized disease were included (43.4% R, 21.0% BR, 33.6% LAU). The population characteristics exemplified differences between daily practice and clinical trials. Tumor location and symptoms were as expected. Treatment plan was conditioned by PS or comorbidities in 23.0% of patients. In patients with R disease, surgery followed by different adjuvant chemotherapy (CT) regimes was the antineoplastic treatment of choice (64.8%) with efficacy results (OS 37.5 months;95% CI 18.4 - 56.7), in the range of contemporary standards. The common use of neoadjuvant CT for BR disease (94.4%), with surgery in 50% of them, and its results (OS 30.8 months;95% CI 10.5 - 51.2) reflected current controversies of treatment recommendations and evolution in this scenario. Paliative CT with or without radiotherapy was the standard specific treatment in LAU disease (95.1%) with survival results (PFS: 10.8 months;95% CI 8.8 - 12.7. OS: 20.3 months;95% CI 13.5 - 27.2) that justify the distinct character and the specific study of this entity. Conclusion: RWE for localized pancreatic cancer aroused from the analysis of this population confirms the distinct nature of patients assisted in daily practice, as well as mirrors the complexity of decision making in clinical assumptions in which achieving stronger evidence should be paramount.

Prevalence and Antibiotic Resistance of Urinary Tract Pathogens, with Molecular Identification of Klebsiella pneumoniae, Klebsiella oxytoca, and Acinetobacter spp., Using Multiplex Real-Time PCR

Urinary tract infections (UTIs) caused by uropathogens are a significant public health problem, and their treatment primarily relies on antibiotic therapy. However, the increasing global development of antibiotic resistance necessitates updating diagnostic techniques to ensure higher sensitivity and specificity, especially with advancements in science and medicine. This study aimed to evaluate the prevalence of UTIs and antibiotic resistance profiles through urine culture, as well as to identify Klebsiella pneumoniae, Klebsiella oxytoca, and Acinetobacter spp. in urine samples using a molecular approach with multiplex real-time PCR. From May 3 to July 25, 2023, at the Pietro Annigoni Biomolecular Research Center (CERBA) and Saint Camille Hospital of Ouagadougou (HOSCO), 209 urine samples collected from patients with suspected UTIs were analyzed using both urine culture and multiplex real-time PCR. Among the 209 patients, 52.15% were male and 47.85% female, with an average age of 46.87 ± 21.33 years. Urine cultures revealed an overall UTI prevalence of 23.44%, with a prevalence of 8.13% in men versus 15.31% in women (P = 0.023). The bacterial prevalence rates were as follows: Escherichia coli (12.92%), Klebsiella spp. (7.18%), Enterobacter cloacae (1.44%), Staphylococcus aureus (0.96%), and other bacteria. Klebsiella spp. demonstrated 100% resistance to Amoxicillin and Amoxicillin/Clavulanic Acid, while Escherichia coli showed 96.2% and 65.4% resistance to Amoxicillin and Amoxicillin/Clavulanic Acid, respectively. PCR analysis of the target bacteria revealed mono-infection prevalence rates of Klebsiella pneumoniae (10.39%), Klebsiella oxytoca (7.79%), and Acinetobacter spp. (7.79%), along with a co-infection prevalence rate of Klebsiella pneumoniae/Acinetobacter spp. (1.30%). This study demonstrated that PCR, with its high sensitivity and specificity, could effectively distinguish Klebsiella pneumoniae from Klebsiella oxytoca and detect Acinetobacter spp. in less than 24 hours—something urine culture alone could not achieve. The relative ease of automating urine PCR testing, combined with its diagnostic accuracy and rapid turnaround time, makes it a valuable addition to modern medical practice for the laboratory diagnosis of UTIs.

24重荧光real-time PCR技术在食物中毒快速检测中的应用

目的:应用24重荧光real-time PCR检测技术快速筛检食物中毒病原菌,结合国家标准中的培养法探讨24重荧光real-time PCR检测技术符合性和应用价值。方法:采用高灵敏度、高特异性的24重荧光real-time PCR检测技术作为中毒病原菌的初筛方法,国标方法进行细菌分离培养,并对分离出的病原菌进行生化鉴定。结果:5份食物中毒患者肛拭子在增菌前检出4份霍乱弧菌核酸(非O1/非O139群,24重荧光real-time PCR),9份患者肛拭子在增菌后检出5株霍乱弧菌(非O1/非O139群,国标培养法),其中包含4份PCR技术初筛阳性样品,两个方法的符合率为80%。所有样本均未检出沙门氏菌、志贺菌、副溶血性弧菌、致泻性大肠埃希菌以及金黄色葡萄球菌。结论:应用24重荧光real-time PCR检测技术同时检测24种常见致病病原菌,能高效锁定中毒病原菌。将其与国标培养法相结合,对临床治疗和食物中毒快速处置能起到积极作用,值得应用和推广。

A Comparative Study between Landmark Based and Real Time Ultrasound Guided Sub Arachnoid Block

Background: Sub arachnoid block (SAB) performed by traditional landmark palpation technique can be inaccurate. This problem is exacerbated by altered patient anatomy due to obesity and age-related changes. A pre-procedural ultrasound scan of the lumbar spine has been shown to be of benefit in guiding lumbar epidural insertion in obstetric patients. Information on the use of real-time ultrasound (RUS) guided SAB, to date, been limited. This study compared RUS guided SAB to traditional landmark guided technique in patients undergoing spinal anesthesia for different surgical procedures. Methods: This was a prospective, single center, comparative observational study conducted in the department of anesthesiology at our center. 560 patients who underwent spinal anesthesia either by landmark based technique or real-time ultrasound-guided methods. The primary outcome was the first attempt success rate of dural puncture when employing the two methods. Results: Baseline characteristics were similar in the two study groups. The first attempt success rate of dural puncture in landmark guided group was 64.3% compared to 72.6% in the ultrasound guided group. This difference was not statistically significant. The procedure performance time was significantly shorter with landmark palpation compared to use of real-time ultrasound guided method. Conclusion: Use of RUS-guided technique does not significantly improve the first attempt success rate of SAB dural puncture during spinal anesthesia compared to the traditional landmark-guided technique.

An Effective Prediction Method for Supporting Decision Making in Real Estate Area Selection

Real estate has been a dominant industry in many countries. One problem for real estate companies is determining the most valuable area before starting a new project. Previous studies on this issue mainly focused on market needs and economic prospects, ignoring the impact of natural disasters. We observe that natural disasters are important for real estate area selection because they will introduce considerable losses to real estate enterprises. Following this observation, we first develop a self-defined new indicator named Average Loss Ratio to predict the losses caused by natural disasters in an area. Then, we adopt the existing ARIMA model to predict the Average Loss Ratio of an area. After that, we propose to integrate the TOPSIS model and the Grey Prediction Model to rank the recommendation levels for candidate areas, thereby assisting real estate companies in their decision-making process. We conduct experiments on real datasets to validate our proposal, and the results suggest the effectiveness of the proposed method.

Real-Time Monitoring of Meteorological Data at In-Situ GCW Remediation Sites

To optimize the self-organization network, self-adaptation, real-time monitoring, remote management capability, and equipment reuse level of the meteorological station supporting the portable groundwater circulation wells, and to provide real-time and effective technical services and environmental data support for groundwater remediation, a real-time monitoring system design of the meteorological station supporting the portable groundwater circulation wells based on the existing equipment is proposed. A variety of environmental element information is collected and transmitted to the embedded web server by the intelligent weather transmitter, and then processed by the algorithm and stored internally, displayed locally, and published on the web. The system monitoring algorithm and user interface are designed in the CNWSCADA development environment to realize real-time processing and analysis of environmental data and monitoring, control, management, and maintenance of the system status. The PLC-controlled photovoltaic power generating panels and lithium battery packs are in line with the concept of energy saving and emission reduction, and at the same time, as an emergency power supply to guarantee the safety of equipment and data when the utility power fails to meet the requirements. The experiment proves that the system has the characteristics of remote control, real-time interaction, simple station deployment, reliable operation, convenient maintenance, and green environment protection, which is conducive to improving the comprehensive utilization efficiency of various types of environmental information and providing reliable data support, theoretical basis and guidance suggestions for the research of groundwater remediation technology and its disciplines, and the research and development of the movable groundwater cycling well monitoring system.

垂体后叶粉中3种动物源性成分多重real-time PCR检测方法的建立及应用

目的基于三重实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)技术同时快速检测垂体后叶粉中的猪、牛、羊性动物源性成分。方法采用CTAB法提取样品及对照品DNA,以线粒体基因为靶基因设计并合成三重real-time PCR的引物探针,并对建立方法进行验证,用于检测垂体后叶粉样品是否与标识一致。结果猪、牛、羊源性成分的最低检测浓度为10 pg/μL,采用建立的方法检测3批垂体后叶粉样品,均只检测出猪源性成分。结论建立的方法可用于同时检测垂体后叶粉中的猪、牛、羊源性成分,可为相关工作提供技术支持。

A CNN-Based Single-Stage Occlusion Real-Time Target Detection Method

Aiming at the problem of low accuracy of traditional target detection methods for target detection in endoscopes in substation environments, a CNN-based real-time detection method for masked targets is proposed. The method adopts the overall design of backbone network, detection network and algorithmic parameter optimisation method, completes the model training on the self-constructed occlusion target dataset, and adopts the multi-scale perception method for target detection. The HNM algorithm is used to screen positive and negative samples during the training process, and the NMS algorithm is used to post-process the prediction results during the detection process to improve the detection efficiency. After experimental validation, the obtained model has the multi-class average predicted value (mAP) of the dataset. It has general advantages over traditional target detection methods. The detection time of a single target on FDDB dataset is 39 ms, which can meet the need of real-time target detection. In addition, the project team has successfully deployed the method into substations and put it into use in many places in Beijing, which is important for achieving the anomaly of occlusion target detection.

Cyber Resilience through Real-Time Threat Analysis in Information Security

This paper examines how cybersecurity is developing and how it relates to more conventional information security. Although information security and cyber security are sometimes used synonymously, this study contends that they are not the same. The concept of cyber security is explored, which goes beyond protecting information resources to include a wider variety of assets, including people [1]. Protecting information assets is the main goal of traditional information security, with consideration to the human element and how people fit into the security process. On the other hand, cyber security adds a new level of complexity, as people might unintentionally contribute to or become targets of cyberattacks. This aspect presents moral questions since it is becoming more widely accepted that society has a duty to protect weaker members of society, including children [1]. The study emphasizes how important cyber security is on a larger scale, with many countries creating plans and laws to counteract cyberattacks. Nevertheless, a lot of these sources frequently neglect to define the differences or the relationship between information security and cyber security [1]. The paper focus on differentiating between cybersecurity and information security on a larger scale. The study also highlights other areas of cybersecurity which includes defending people, social norms, and vital infrastructure from threats that arise from online in addition to information and technology protection. It contends that ethical issues and the human factor are becoming more and more important in protecting assets in the digital age, and that cyber security is a paradigm shift in this regard [1].

4种动物源性成分多重real-time PCR检测方法的建立及其在驴肉制品检测中的应用

建立一种同时快速检测驴、马、猪及鸭源性成分的四重实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)方法。分别以4种源性成分的Nad5、ATpase6、ATP8、cytb基因为靶基因设计特异性引物和TaqMan探针,以18S rRNA基因为内参基因,建立多重real-time PCR方法,并对该方法进行方法学验证,同时对不同掺入比例模拟样品、不同加工工艺模拟样品和实际驴肉样品进行检测。结果显示,该方法具有高通量、特异性强、灵敏度高等优点。当Ct值≤35.0时,方法对16种非目标源性具有良好特异性;灵敏度可检测到质量浓度为2×10^(-4)ng/μL的模板DNA;对生肉的检出限为肉含量的0.001%,对熟肉制品的检出限为肉含量的0.01%;对100份实际样品进行检测,结果与标准方法一致,说明建立的多重real-time PCR法可用于肉及肉制品中常见掺假源性成分的检测。

肉中猪源性成分Real-time PCR定量检测技术

【目的】建立一种快速、准确的肉中猪源性成分定量检测方法。【方法】首先从GenBank数据库中筛选猪特异性的微卫星DNA,根据微卫星DNA核酸序列设计引物,对常见10种动物基因组DNA进行PCR扩增,通过有无扩增产物判断筛选的微卫星DNA对猪源性成分的特异性。然后根据微卫星DNA核酸序列,设计特异性引物和探针,建立猪源性成分Real-time PCR检测方法,采用双标准曲线分别对猪源性成分和总动物源性成分进行定量,计算猪源性成分的百分含量。【结果】筛选到猪特异性微卫星DNA(Accession EF172428),根据其序列设计的引物SEQ-sus2-F/R只能从猪基因组DNA中扩增出目的条带,其他动物的基因组均无目的条带扩增。建立的Real-time PCR检测方法灵敏度为0.02 ng/25μL反应体系。该方法能够准确检测出混合DNA样品中猪源性成分和混合肉样品中猪源性成分,百分误差分别约为1.32%和1.06%-7.12%。【结论】本研究利用Real-time PCR技术建立的定量猪源性成分的检测方法可以用来检测猪源性成分在混合样品中的百分含量。

猪NLRP3基因Real-time PCR检测方法的建立及应用

根据NCBI NLRP3 cds(NM-001256770)序列设计1对特异性引物,构建标准品质粒pMD18T-NLRP3-189。通过反应条件优化、稳定性、敏感性等试验,成功建立了NLRP3的SYBR GreenⅠreal-time PCR检测方法,用该方法定量检测猪肺炎支原体(Mycoplasma hyopneumoniae,Mhp)感染后的猪肺组织中NLRP3表达水平。最佳引物浓度为0.15μmol/L,建立的标准曲线方程为y=-3.5196x+36.968,E值为0.96,最低检测量为84个拷贝,熔解曲线有单一峰,批内变异系数CV为0.067%~0.184%,批间变异系数为0.221%~0.594%,重复性好。该方法检测仔猪在感染Mhp后肺组织中的NLRP3 mRNA表达水平,感染组与健康组相比差异显著(P<0.01),NLRP3表达水平明显增高,在感染后14 d可达到高峰,平均mRNA拷贝数达18000左右,可持续42 d。表明该检测方法稳定性良好,精确度高,特异性强,为进一步研究猪炎症小体NLRP3的激活及其相关炎症反应机制提供了技术支持。