Semi-quantification

半量化
  • 文章类型: Journal Article
    背景:间质性肺异常(ILA)与疾病的进一步进展有关,死亡风险增加,老年人的肺功能下降,这值得足够的关注。
    目的:本研究的目的是使用低剂量CT(LDCT)量化中国非吸烟无症状城市队列中间质性肺异常(ILA)的程度,并分析与年龄相关的病理变化。
    方法:我们回顾性分析了733名受试者的临床数据和胸部LDCT图像,这些受试者根据年龄分为3组:18-39岁、40-59岁和≥60岁。此外,我们选择了40例肺大泡切除术后存档的蜡包埋肺组织块,并对相同年龄组进行了分类。ILA的四个代表性CT征象,包括小叶间隔增厚(ILST),小叶内间质增厚(ILIT),毛玻璃不透明度(GGO),和网状阴影(RS),根据受影响面积的百分比进行半量化。比较不同性别和年龄组ILA4种CT征象的评分和分布。分析与年龄相关的病理改变。
    结果:ILA发现主要在下叶和胸膜下区域。所有40岁以下受试者的四种CT征象的半定量评分均为0.然而,在40岁以上的受试者中,分数随着年龄的增长而逐渐增加,尽管他们中的大多数仍然很低。肺泡的大小增加,肺泡数量减少,肺泡隔膜变薄,ATII细胞的数量随着年龄的增长而增加。不同年龄组间差异有统计学意义(χ2=50.624,P=0.033;χ2=80.000,P=0.043;χ2=33.833,P=0.000;χ2=13.525,P=0.031)。肺泡隔巨噬细胞数量和胶原纤维百分比增加,而弹性纤维的百分比随着年龄的增长而下降。不同年龄组间差异无统计学意义(χ2=19.817,P=0.506;χ2=52.419,P=0。682;χ2=54.868,P=0.518)。
    结论:当上述四个CT征象位于中央上部区域时,分数有中等或高分,确定潜在的病理原因至关重要。ILA可能是慢性肺损伤的结果。
    BACKGROUND: Interstitial lung abnormalities (ILA) are associated with further disease progression, increased mortality risk, and decline in lung function in the elderly, which deserves enough attention.
    OBJECTIVE: The objective of this study was to quantify the extent of interstitial lung abnormalities (ILA) in a non-smoking asymptomatic urban cohort in China using low-dose CT (LDCT) and to analyze the age-related pathological changes.
    METHODS: We retrospectively analyzed clinical data and chest LDCT images from a cohort of 733 subjects who were categorized into 3 groups: 18-39, 40-59, and ≥60 years old according to age. Furthermore, we selected 40 cases of wax-embedded lung tissue blocks archived after pulmonary bullectomy and the same age groups were categorized. Four representative CT signs of ILA, including interlobular septal thickening (ILST), intralobular interstitial thickening (ILIT), ground-glass opacity (GGO), and reticular shadow (RS), were semi-quantified based on the percentage of the affected area. The scores and distribution of four CT signs of ILA were compared between different sex and age groups. The age-related pathological changes were analyzed.
    RESULTS: The ILA findings were found predominantly in the lower lobes and the subpleural region. The semi-quantitative scores of four CT signs in all subjects under 40 were 0. However, in subjects over 40 years old, the scores gradually increased with age, although most of them remained low. The size of the alveoli increased, the number of alveoli decreased, the alveolar septum became thinner, and the number of ATII cells increased with age. A statistically significant difference was observed among the different age groups (χ2=50.624, P=0.033; χ2=80.000, P=0.043; χ2=33.833, P=0.000; χ2=13.525, P=0.031). The macrophage population and the percentage of collagen fibers in the alveolar septum increased, while the percentage of elastic fibers decreased with age. There was no significant difference among the different age groups (χ2=19.817, P=0.506; χ2=52.419, P=0. 682; χ2=54.868, P=0.518).
    CONCLUSIONS: When the four CT signs mentioned above are in the upper central area, and the score has a medium or high score, it is crucial to determine the underlying pathological causes. ILA may be the result of chronic lung injury.
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  • 文章类型: Journal Article
    这项研究建立了一种用于快速鉴定的高通量多重遗传检测测定法(HMGA),直接从临床非侵入性口腔样本中对幽门螺杆菌进行半定量和毒力分析。
    收集了2021年至2022年上海242名患者的胃粘膜和口腔样本。所有样品均通过幽门螺杆菌的常规临床试验检测,Sanger测序结果不一致。设计并优化了一种可在4小时内提供结果的新的多重PCR检测方法,该方法涉及靶向16SrRNA基因的荧光染料标记的特异性引物,幽门螺杆菌的半定量基因ureC和10个毒力基因。通过模拟阳性口腔样品的连续10倍稀释度进行半定量,并进一步比较不同临床样本中的幽门螺杆菌载量。使用毒力基因vacAs1,vacAm1和vacAm2的混合质粒来评估不同基因型的性能。10个毒力基因在胃黏膜中的一致性,唾液,比较幽门螺杆菌阳性患者的漱口水和牙菌斑.
    非侵入性HMGA对于检测幽门螺杆菌和人类内部参考基因β-珠蛋白的所有12个靶标具有高度特异性,对所有靶基因的敏感性可达10拷贝/μL。与常规临床试验和测序相比,非侵入性HMGA具有较高的灵敏度(>0.98),特异性,准确度,PPV,直接检测口腔样本中H.pylori的NPV和kappa系数。此外,通过检测ureC的峰面积水平,证实胃黏膜中的H.pylori负荷显著高于三种口腔样本(p<0.05)。我们还发现,在不同的口腔样本中,有45.0%(91/202)的患者具有不同的H.pylori毒力基因。唾液和胃黏膜各毒力基因阳性检出率均达到78%以上(p<0.05)。
    非侵入性HMGA被证明是快速鉴定幽门螺杆菌的可靠方法,口腔样本中10个毒力基因的半定量和直接检测,为幽门螺杆菌的无创检测提供了新的思路。
    This study established a high-throughput multiplex genetic detection assay (HMGA) for rapid identification, semi-quantification and virulence analysis of Helicobacter pylori directly from the clinical non-invasive oral samples.
    The gastric mucosa and oral samples were collected from 242 patients in Shanghai from 2021 to 2022. All the samples were detected by routine clinical tests for H. pylori and Sanger sequenced for inconsistent results. A new multiplex PCR assay providing results within 4 hours was designed and optimized involving fluorescent dye-labeled specific primers targeted 16S rRNA gene, semi-quantitative gene ureC and 10 virulence genes of H. pylori. Semi-quantification was carried out by simulating the serial 10-fold dilutions of positive oral samples, and the H. pylori loads in different clinical samples were further compared. The mixed plasmids of virulence genes vacA s1, vacA m1 and vacA m2 were used to evaluate the performance on different genotypes. The consistency of 10 virulence genes in gastric mucosa, saliva, mouthwash and dental plaque of H. pylori-positive patients was compared.
    The non-invasive HMGA was highly specific for detection of all 12 targets of H. pylori and human internal reference gene β-globin, and the sensitivity to all target genes could reach 10 copies/μL. Compared with routine clinical tests and sequencing, non-invasive HMGA has a high level (>0.98) of sensitivity, specificity, accuracy, PPV, NPV and kappa coefficient for direct detection of H. pylori in oral samples. Moreover, by detecting peak area levels of ureC, it was confirmed that the H. pylori loads in gastric mucosa were significantly higher than those of the three kinds of oral samples (p<0.05). We also found that 45.0% (91/202) of patients had different H. pylori virulence genes in different oral samples. The concordance of positive detection rates of each virulence gene between saliva and gastric mucosa was more than 78% (p<0.05).
    The non-invasive HMGA proved to be a reliable method for the rapid H. pylori identification, semi-quantification and detection of 10 virulence genes directly in oral samples, providing a new idea for non-invasive detection of H. pylori.
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  • 文章类型: Journal Article
    我们最近开发了冠状动脉树描述和病变评估(CatLet或Hexu,由He和Xu发明)血管造影评分系统,which,考虑到冠状动脉解剖结构的多样性,冠状动脉的狭窄程度,心肌区域由患病的冠状动脉包围,可用于预测急性心肌梗死患者的临床结果(可在www.catletscore.com).其在临床实践和冠状动脉疾病研究中的价值正在建立。在过去的两年里,这种新型血管造影评分系统的基本原则没有实质性改变,尽管确实发生了轻微的调整.鉴于这些调整和在日常使用中获得的评分经验,我们认为有必要对这些观点进行详细阐述,以便有兴趣的读者能够在临床实践和科学研究中更好地使用这种CatLet或Hexu血管造影评分系统.
    这种新型血管造影评分系统的基本原理包括17心肌节段模型,竞争性血液供应法,和流量守恒定律。
    对这种新颖的血管造影评分系统进行的调整包括:(I)使用基础水平的左心室短轴来表征六种类型的右冠状动脉大小;(II)标记为X和S的部分在左前降支的表征中具有统一的预设差异;(III)在某些情况下添加了标记为“”的部分,以解释罕见的后分支血管中的边缘差异。CatLet或Hexu血管造影评分系统在权重分配中严格遵循流量守恒定律,并进一步强调和详细的病变评分校正。
    在CatLet或Hexu血管造影评分系统上对这些调整和评分经验的阐述将有助于提高其在心血管领域的应用。这种新颖的血管造影评分系统的实用性已得到初步验证,其未来值得期待。
    UNASSIGNED: We have recently developed the Coronary Artery Tree description and Lesion EvaluaTion (CatLet or Hexu, invented by He and Xu) angiographic scoring system, which, considering the coronary anatomy in its diversity, the stenosis degree of a coronary artery, and the myocardial territory subtended by the diseased coronary artery, can be utilized to predict clinical outcomes for patients with acute myocardial infarction (available at www.catletscore.com). Its values for clinical practice and coronary artery disease research are building upon. Over the past two years, the principles underlying this novel angiographic scoring system do not materially change although slight adjustments have really happened. Given these adjustments and the scoring experience gained in daily use, we think that it is necessary to elaborate on these points so that readers with interest are capable to better use this CatLet or Hexu angiographic scoring system both in clinical practice and in scientific research.
    UNASSIGNED: The principles underlying this novel angiographic scoring system include the 17-myocardial segmental model, law of competitive blood supply, and law of flow conservation.
    UNASSIGNED: The adjustments made to this novel angiographic scoring system include: (I) the short axis of the left ventricle at the basal level is used to characterize the six types of right coronary artery size; (II) segments marked with \'X and \'S have a unified preset difference of one segment as adopted in the characterization of left anterior descending artery; (III) segments marked with \'+ have been added to explain the rare variability in the obtuse marginal branches or in the posterolateral vessels in some cases. The CatLet or Hexu angiographic scoring system strictly follows the law of flow conservation in weighting assignment, and the lesion scoring correction has been additionally emphasized and detailed.
    UNASSIGNED: The elaboration on these adjustments and scoring experience gained on the CatLet or Hexu angiographic scoring system will help to boost its use in cardiovascular field. The utilities of this novel angiographic scoring system have been preliminarily validated and its future is deserving of being anticipated.
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  • 文章类型: Journal Article
    背景:农药抗性是无脊椎害虫化学控制中长期存在且日益严重的问题。分子诊断方法可以通过准确有效地检测抗性突变及其频率来促进农药抗性管理。在这项研究中,竞争性等位基因特异性PCR(KASP)方法,一种高通量SNP基因分型技术,首次被验证为表征农药抗性基因座基因型的有用方法。我们重点研究了halmithrips的nAChRα6亚基基因中G275E的多角菌抗性突变。
    结果:在接受测试的341名T.palmi个体中,98.24%基因分型成功,与Sanger测序结果100%一致。然后,我们定量混合已知基因型的基因组DNA,以建立21种DNA混合物,其抗性等位基因频率为0%至100%,步长为5%。线性判别分析(LDA)显示,原始分组病例的75.8%被正确分类;六组成员没有重叠(抗性等位基因频率:0%,5%,10%-75%,80-85%,90-95%,和100%)。当我们为LDA选择11个10%步长的合并样本时,84.4%的原始分组病例被正确分类;七组成员没有重叠(0%,10%,20%-30%,40%-70%,80%,90%,100%)。结果表明,KASP应用于合并样品可能会提供耐药性的半定量估计。
    结论:我们的研究指出KASP对于影响农药抗性的基因型的高通量基因分型和群体抗性等位基因频率的半定量评估的适用性。
    BACKGROUND: Pesticide resistance is a long-standing and growing problem in the chemical control of invertebrate pests. Molecular diagnostic methods can facilitate pesticide resistance management by accurately and efficiently detecting resistant mutations and their frequency. In this study, the kompetitive allele specific PCR (KASP) approach, a technology for high-throughput single nucleotide polymorphism (SNP) genotyping, is validated as a useful method for characterizing genotypes at a pesticide-resistance locus for the first time. We focus on the spinetoram resistance mutation of G275E in the nicotinic acetylcholine receptor alpha 6 (nAChR α6) subunit gene of Thrips palmi.
    RESULTS: Of the 341 individuals of Thrips palmi tested, 98.24% were successfully genotyped, with 100% concordance with Sanger sequencing results. We then quantitatively mixed genomic DNA of known genotypes to establish 21 DNA mixtures with a resistant allele frequency ranging from 0 to 100% at steps of 5%. The linear discriminant analysis (LDA) showed that 75.8% of original grouped cases were correctly classified; six groups had no overlap in membership (resistant allele frequency: 0%, 5%, 10-75%, 80-85%, 90-95%, and 100%). When we chose 11 pooled samples with 10% steps for LDA, 84.4% of original grouped cases were correctly classified; seven groups had no overlap in membership (0%, 10%, 20-30%, 40-70%, 80%, 90%, 100%). The results indicated that KASP applied to pooled samples may provide a semi-quantitative estimate of resistance.
    CONCLUSIONS: Our study points to the suitability of KASP for high-throughput genotyping of genotypes affecting pesticide resistance and semi-quantitative assessments of resistance allele frequencies in populations. © 2023 Society of Chemical Industry.
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  • 文章类型: Journal Article
    Zearalenone (ZEN) is a prevalent mycotoxin that needs intensive monitoring. A semi-quantitative and quantitative immunochromatographic assay (ICA) was assembled for investigating ZEN contamination in 187 samples of cereal and their products from China in 2019. The semi-quantitative detection model had a limit of detection (LOD) of 0.50 ng/mL with visual judgment and could be completely inhibited within 5 min at 3.0 ng/mL ZEN. The quantitative detection model had a lower LOD of 0.25 ng/mL, and ZEN could be accurately and digitally detected from 0.25-4.0 ng/mL. The ICA method had a high sensitivity, specificity, and accuracy for on-site ZEN detection. For investigation of the authentic samples, the ZEN-positive rate was 62.6%, and the ZEN-positive levels ranged from 2.7 to 867.0 ng/g, with an average ZEN-positive level being 85.0 ng/g. Of the ZEN-positive samples, 6.0% exceeded the values of the limit levels. The ZEN-positive samples were confirmed to be highly correlated using LC-MS/MS (R2 = 0.9794). This study could provide an efficiency and accuracy approach for ZEN in order to achieve visual and digitized on-site investigation. This significant information about the ZEN contamination levels might contribute to monitoring mycotoxin occurrence and for ensuring food safety.
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  • 文章类型: Journal Article
    Although passive sampling is widely accepted as an excellent tool for environmental monitoring, their integration with suspect or non-targeted screening by high-resolution mass spectrometry has been limited. This study describes the application of the organic-diffusive gradients in thin-films (o-DGT) passive sampler as a tool for accurate measurement of both targeted and suspect polar organic contaminants (primarily pharmaceuticals) in wastewater. First, performance of o-DGT was assessed alongside the polar organic chemical integrative sampler (POCIS) and active sampling at two wastewater treatment facilities using targeted analyses. Overall, water concentrations measured by o-DGT, POCIS, and 24-hr integrative active samples were in good agreement with each other. There were exceptions, including a systematic difference between o-DGT and POCIS at certain sites that we propose was a result of site-specific conditions and a difference in sampling rates between the two techniques. The second component of this work involved suspect screening of the o-DGT extracts using high-resolution, high mass accuracy quadrupole time-of-flight mass spectrometry (QTOF). Lamotrigine, venlafaxine, and des-methylvenlafaxine were three suspect compounds identified and selected as proof-of-concept case studies to determine the feasibility and accuracy of o-DGT for estimating water concentrations based upon predicted sampling rates using a previously validated o-DGT diffusion model. Semi-quantification of the suspect compounds was conducting using an average surrogate response factor based on the suite of compounds measured by the targeted analyses. This, combined with the modelled sampling rates provided time-weighted average wastewater concentrations of the identified suspects within a factor of 2 of the true value, confirmed by isotope dilution with mass labelled internal surrogates. To the knowledge of the authors, this work is the first to demonstrate the utility of the o-DGT passive sampler as a potential environmental screening tool that can be integrated into the rapidly advancing field of non-targeted high resolution mass spectrometry.
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  • 文章类型: Journal Article
    Peptides could have specific tastes or bioactivities depending on the length and sequence of amino acids. Till date it remains unknown what peptides are formed during the white tea manufacturing process and whether they contribute to the flavor or bio-activities of white tea. As a first step to address these questions, we applied ultra-high pressure liquid chromatography coupled with quadrupole-orbitrap ultra-high resolution mass spectrometry (UPLC-Quadrupole-Orbitrap-UHRMS) to monitor peptides dynamic changes during the withering process. A total of 196 abundant peptides were identified. Most of them were oligopeptides within a molecular weight of 1000 Da. Four of them were randomly selected, synthesized peptides were applied for further confirmation and quantification. Sequence analysis suggested that some of them were potential taste contributors. Proteinase cleave site analysis identified two separate periods of active proteins degradation at 0-12 h and 30-42 h of the withering processes. Further analysis of cleavage sites also suggested that protein degradation during withering steps were random rather than a stepwise reaction.
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  • 文章类型: Journal Article
    Qingre Xiaoyanning capsule is a famous traditional Chinese medicine prescription which consisted of Sarcandrae Herba (also named Caoshanhu in China) water extract for the frequent treatment of inflammation and immunity related diseases. Until now, the in vivo bioactive components of Qingre Xiaoyanning capsule have not yet been fully addressed. In this study, a total of 42 xenobiotics including 20 prototypes and 22 metabolites were identified in rats after oral administration of Qingre Xiaoyanning capsule using ultra-high performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry. Subsequently, isofraxidin and rosmarinic acid, two bioactive components with high exposure in rat plasma, were quantitatively analyzed, while another 20 major absorbed components were semi-quantitatively measured, to investigate together the pharmacokinetics behavior of Qingre Xiaoyanning capsule. Taken together, this study provided comprehensive knowledge of in vivo disposal of this prescription, which could help reveal the potential bioactive components, and would be conducive to further pharmacological mechanism research as well as quality control approach improvement of Qingre Xiaoyanning capsule and Sarcandrae Herba related prescriptions.
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  • 文章类型: Journal Article
    A quick, easy, effective method followed by ultra-high-pressure liquid chromatography coupled with linear ion trap-Orbitrap tandem mass spectrometry (UHPLC-LTQ-Orbitrap MS) was developed for the simultaneous identification and quantification of the metabolites produced by amentoflavone (AMF) in human intestinal bacteria from human feces. The method validated for quantification of AMF concerning precision, accuracy, recovery, matrix effect, stability and limits showed acceptable results. Compared with blank human intestinal bacteria chromatography, three metabolites were identified based on high-accuracy protonated precursors and multi-stage mass spectrometry (MSn ) using the proposed strategy. At the same time, a new method was developed for semi-quantification of three metabolites. We describe the trend over 24 h of concentration-time curves for AMF and its metabolites. Moreover, the main metabolic pathway of AMF was clarified in human intestinal bacteria. The method was validated and successfully applied to the detection and quantification of AMF and its metabolites.
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  • 文章类型: Journal Article
    Paper-based devices have been broadly used for the point-of-care detection of dengue viral nucleic acids due to their simplicity, cost-effectiveness, and readily observable colorimetric readout. However, their moderate sensitivity and functionality have limited their applications. Despite the above-mentioned advantages, paper substrates are lacking in their ability to control fluid flow, in contrast to the flow control enabled by polymer substrates (e.g., agarose) with readily tunable pore size and porosity. Herein, taking the benefits from both materials, the authors propose a strategy to create a hybrid substrate by incorporating agarose into the test strip to achieve flow control for optimal biomolecule interactions. As compared to the unmodified test strip, this strategy allows sensitive detection of targets with an approximately tenfold signal improvement. Additionally, the authors showcase the potential of functionality improvement by creating multiple test zones for semi-quantification of targets, suggesting that the number of visible test zones is directly proportional to the target concentration. The authors further demonstrate the potential of their proposed strategy for clinical assessment by applying it to their prototype sample-to-result test strip to sensitively and semi-quantitatively detect dengue viral RNA from the clinical blood samples. This proposed strategy holds significant promise for detecting various targets for diverse future applications.
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