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A direct,sensitive and high-throughput genus and species-specific molecular assay for large-scale malaria screening

Infectious Diseases of Poverty 2022年第2期11卷 11-22页

作者： Yaling Zhao Ye Zhao Yu Sun lihua fan Duoquan Wang Heng Wang Xiaodong Sun Zhi Zheng Institute of Basic Medical Sciences Chinese Academy of Medical SciencesSchool of Basic MedicinePeking Union Medical CollegeBeijingChina Yunnan Institute of Parasitic Diseases Yunnan Center for Malaria ResearchInstitute of Vector and Pathogen BiologyDali UniversityPuerYunnanChina National Institute of Parasitic Diseases Chinese Center for Disease Control and PreventionWHO Collaborating Center for MalariaSchistosomiasisand FilariasisKey Laboratory of Parasite and Vector BiologyMinistry of HealthShanghaiChina Blood Transfusion Department Peking Union Medical College HospitalChinese Academy of Medical SciencesBeijingChina

Background:Infectious disease diagnostics often requires sensitive molecular assays that identify at both genus and species levels.For large scale screening,such as malaria screening for elimination,diagnostic assay can be a challenge,as both the throughput and cost of the assay must be considered.The requirement of nucleic acid extraction hampers the throughput of most molecular assays.Co-amplification of multiple species or multiplex identification either can result in missed diagnosis or are too costly for large-scale screening.A genus-and species-specific diagnostic assay with simplified procedure,high sensitivity and throughput is still needed.This study aimed to develop a sensitive and high-throughput approach for large-scale infectious disease screening.Methods:We developed multi-section Capture and Ligation Probe PCR(mCLIP-PCR)for the direct detection of RNA without extraction and reverse transcription.Multiple tailed sandwich hybridization probes were used to bind at genus-and species-specific sections of the target RNA to cooperatively capture the target onto a 96-well plate.After enzymatic ligation of the bound probes,a single-stranded DNA formed at each section with distinct tail sequence at the ends.They were separately PCR-amplified with primers corresponding to tail sequences for genus or species identification.We applied the method to the active screening ofPlasmodium infections of 4,580 asymptomatic dried blood spot samples collected in malaria endemic areas and compared the results with standard qPCR using linear regression.Results:With multi-section cooperative capture but separate amplification strategy,we accurately identified genusPlasmodium and speciesP.falciparum andP.vivax without RNA extraction,with favorable sensitivities among the published reports.In the active screening,our method identified all 53 positive infections including two mixed infections,and twoP.vivax infections that were missed by standard qPCR.Conclusions:mCLIP-PCR provides a sensitive and high-throughput approach to la

关键词： Infectious disease Malaria Molecular screening CLIP-PCR Genus Species High-throughput

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Verification of Bacteroidales 16S rRNA markers as a complementary tool for detecting swine fecal pollution in the Yangtze Delta

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Journal of Environmental Sciences 2020年第4期32卷 59-66页

作者： lihua fan Xiaofeng Zhang Ruoxue Zeng Suhua Wang Chenchen Jin Yongqiang He Jiangbing Shuai Department of Food Science and Nutrition Zhejiang Key Laboratory for Agro-Food ProcessingZhejiang UniversityHangzhou 310016China Zhejiang Academy of Science and Technology for Inspection and Quarantine Hangzhou 310058China

To correctly assess and properly manage the public health risks associated with exposure to contaminated water,it is necessary to identify the source of fecal pollution in a watershed.In this study,we evaluated the efficacy of our two previously developed real time-quantitative PCR(qPCR)assays for the detection of swine-associated Bacteroidales genetic markers(gene 1-38,gene 3-53)in the Yangtze Delta watershed of southeastern China.The results indicated that the gene 1-38 and 3-53 markers exhibited high accuracy(92.5%,91.7%conditional probability,respectively)in detecting Bacteroidales spp.in water samples.According to binary logistic regression(BLR),these two swine-associated markers were well correlated(P<0.05)with fecal indicators(Escherichia coli and Enterococci spp.)and zoonotic pathogens(E.coli O157:H7,Salmonella spp.and Campylobacter spp.)in water samples.In contrast,concentrations of conventional fecal indicator bacteria(FIB)were not correlated with zoonotic pathogens,suggesting that they are noneffective at detecting fecal pollution events.Collectively,the results obtained in this study demonstrated that a swinetargeted qPCR assay based on two Bacteroidales genes markers(gene 1-38,gene 3-53)could be a useful tool in determining the swine-associated impacts of fecal contamination in a watershed.

关键词： Bacteroidales genes markers Water pollution Real-time quantitative PCR Fecal indicators Zoonotic pathogens

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Traumatic extracranial internal carotid-jugular fistula leading to serious injury: a case report in forensic assessment

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Forensic Sciences Research 2020年第2期5卷 170-173页

作者： Xia Liu lihua fan Chongliang Ying Yibin Cheng Maowen Wang Department of Clinical Forensic Medicine Shanghai Key Laboratory of Forensic MedicineShanghai Forensic PlatformAcademy of Forensic ScienceShanghaiChina

Extracranial carotid artery injuries may produce severe haemorrhage,cerebral damage or arteriovenous fistula.Examples of traumatic extracranial carotid-jugular fistula are not frequently reported,especially in forensic medicine.We report a controversial case of an extracranial internal carotid-jugular fistula resulting from a stab wound to the neck.The degree of the injury was classified under“The Standard of Human Body Injury Assessment(2014)”(SIA)in China by forensic examiners.We believe this case report will provide information for the forensic assessment of similar cases.

关键词： Forensic sciences carotid artery trauma extracranial internal carotid-jugular fistula injury degree assessment

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Cr/Mo/W元素对镍基高温合金γ/γ′共格组织热稳定性的影响

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金属学报 2024年第4期60卷 453-463页

作者：凡莉花李金临孙九栋吕梦甜王清董闯大连理工大学材料科学与工程学院三束材料改性教育部重点实验室大连116024

近年来,随着航空航天发动机的持续发展,对其关键热端部件的承温能力要求越来越高,而高温合金共格组织热稳定性是影响其力学性能的关键因素,本工作在团簇成分式[Al-Ni_(12)](Al_(1)(Ti,Nb,Ta)_(0.5)(Cr,Mo,W)_(1.5))基础上,保持其他元素... 详细信息

近年来,随着航空航天发动机的持续发展,对其关键热端部件的承温能力要求越来越高,而高温合金共格组织热稳定性是影响其力学性能的关键因素,本工作在团簇成分式[Al-Ni_(12)](Al_(1)(Ti,Nb,Ta)_(0.5)(Cr,Mo,W)_(1.5))基础上,保持其他元素含量不变,改变Cr、Mo、W的含量设计了系列镍基高温合金(含Cr_(1.0)Mo_(0.5)的S1-CM、Cr_(1.0)W_(0.5)的S2-CW和含Cr_(0.7)Mo_(0.4)W_(0.4)的S3-CMW),研究Cr、Mo、W含量变化对合金γ/γ′共格组织热稳定性的影响。采用非自耗真空电弧炉制备合金铸锭,并对其在1300℃固溶15 h后在900℃下进行长期时效处理,并对时效态样品进行微观组织表征和力学性能测试。结果表明,这3个合金的微观组织均表现为高体积分数(f>70%)的γ′粒子均匀地分布在fcc-γ基体中。其中,S1-CM和S2-CW合金在900℃、50 h时效后γ′粒子形貌均呈椭球形,而S3-CMW合金则表现为方形,这是由于后者具有更负的γ/γ′点阵错配度(后者δ=-0.47%,前者δ=-0.25%~-0.33%)。500 h长期时效后,各合金中的γ′粒子形貌没有发生变化,且都具有较慢的粗化速率(K=10~18 nm^(3)/s);尤其S3-CMW合金表现出最高的γ/γ′共格组织稳定性(γ′粒子的粗化速率K=10.02 nm^(3)/s),且晶界处的第二相粒子析出明显低于其他合金。系列合金的硬度基本不随时效时间发生变化,也进一步表明了共格组织的稳定性;其中S3-CMW合金的显微硬度为397~418 HV,时效200 h后的室温压缩屈服强度为818 MPa。

关键词：镍基高温合金共格组织 γ′粒子粗化第二相析出

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无线射频压裂滑套系统模拟试验研究

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石油机械 2024年第4期52卷 27-32页

作者：张利巍孙露凯范立华东北石油大学物理与电子工程学院大庆油田井下作业分公司

将无线射频识别(RFID)技术应用到油水井压裂中,能够很好地解决现有压裂滑套存在的压裂级数受限、操作非智能等诸多问题。因井下工况复杂,理论分析与实际应用差别较大,为进一步系统地探究井下无线射频系统的开发应用,使用三维软件设计无... 详细信息

将无线射频识别(RFID)技术应用到油水井压裂中,能够很好地解决现有压裂滑套存在的压裂级数受限、操作非智能等诸多问题。因井下工况复杂,理论分析与实际应用差别较大,为进一步系统地探究井下无线射频系统的开发应用,使用三维软件设计无线射频滑套模拟试验台,对标签的运动状态进行理论分析,并建立室内模拟试验平台,系统地进行井下相关试验。试验结果表明:标签选择有铁氧体磁芯,且天线绕制形式选用单层螺线管线圈,可以提高整个系统的性能;在系统电感值相同的情况下,单层螺线管天线轴向长度对阅读器模块能识别的标签最大通过速度有积极的影响;标签天线线圈与阅读器天线线圈平面之间的角度也会影响标签的识别率。研究结果对井下无线射频系统的开发应用具有一定的指导作用。

关键词：油水井压裂滑套无线射频识别标签试验台天线

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Study on the Pharmacological Identification of Polygonum capitatum

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Agricultural Biotechnology 2022年第1期11卷 58-59,72页

作者： Wei WEI Huagang LIU Huarong HE lihua fan Qiuyan LU Dan LAN Haicheng WEN Songji WEI Guangxi University of Chinese Medicine Nanning 530001China Guangxi Xingui Environmental Protection Technology Group Co. Ltd.Nanning 530001China

[Objectives]This study was conducted to establish a microscopic identification method of Polygonum capitatum Buch.Ham.ex D.Don.[Methods]The cross sections were identified by microscopic identification.[Results]The stem cross section of P.capitatum is round-like,and shows pericyclic fibers forming a ring,strongly lignified,many vascular bundles,and a hollow pith part.There are many starch granules in the powder,and single granules are more common;and fibers are mostly bundled or scattered,lignified or non-lignified,and calcium oxalate cluster crystals are common.There are many pollen grains,obtuse triangular or round-like,and some of them have three germination apertures.They have fine thorn-like protrusions on the outer wall,the surface of which has reticulate carvings.[Conclusions]The results of microscopic identification are reliable and can be used as the basis for identification of P.capitatum.

关键词： Polygonum capitatum Microscopic identification Pharmacology

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Mirror-like tongue is an important predictor of acute heart failure:a cohort study of acute heart failure in Chinese patients

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Journal of Traditional Chinese Medicine 2023年第6期43卷 1243-1251页

作者： CHEN Yunhu YIN Moqing fan lihua JIANG Xuechun ZHANG Tao ZHU Xingyu XU Hongfeng Department of Cardiology Taicang TCM Hospital Affiliated to Nanjing University of Chinese MedicineSuzhou 215400China Department of Cardiology Changzhou Hospital affiliated with Nanjing University of Chinese MedicineChangzhou 213003China Department of Clinical Pharmacy Taicang TCM Hospital Affiliated to Nanjing University of Chinese MedicineSuzhou 215400China

OBJECTIVE:To reveal that mirror-like tongue,observed via a noninvasive inspection,is a powerful indicator of the severity and prognosis of patients with acute heart failure(AHF).METHODS:This was an observational,prospective study.A total of 408 patients who met the inclusion criteria and were diagnosed with AHF for the first time at Taicang Traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine from August 2019 to January 2021 were selected as the research subjects.According to their tongue fur,the patients were divided into group A(mirror-like tongue group)and group B(non-mirror-like tongue group).The clinical characteristics and incidence of major adverse cardiovascular events(MACEs)within 1 year of follow-up were systematically compared between the two groups.RESULTS:Sixty-five patients were included in group A,and 343 patients were included in group B.There were 32 males and 33 females in group A and 168 males and 175 females in group B.The average age of the overall population was 64 years old,and the average age of group A was significantly higher than that of group B(74 vs 62 years,P<0.001).Compared with that in group B,the left ventricular ejection fraction(LVEF)in group A was significantly lower(35%vs 42%,P<0.001),and the log N-terminal pro B-type natriuretic peptide(NT-proBNP)level was significantly higher(4.0 vs 3.4,P<0.001).The proportion of the combined application of vasoactive drugs in group A was significantly higher than that in group B(64%vs 38%,P<0.001).Group B had a higher proportion of coronary angiography(29.5%vs 16.9%,P=0.038).Group A was more inclined to require mechanical ventilation than group B(33.9%vs 22.5%,P=0.049).The length of hospital stay in group A was significantly longer than that in group B(13.1 vs 7.6,P<0.001).The incidence of MACEs,such as recurrence of AHF,new myocardial infarction and stroke,in group A within one year was higher than that in group B(P=0.007,0.009,<0.001).The incidence of cumulative MACEs in group A was significantly high

关键词： heart failure risk factors fur Yin deficiency cohort studies

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微生物源示踪技术在食品链中进行污染源追溯的研究进展

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食品科学 2024年第7期45卷 332-339页

作者：樊丽华王江雪欧宗沅张艳茹张璐瑶刘诗艺周自梅李国梁陕西科技大学食品科学与工程学院陕西西安710021

在食品链中,粪便微生物污染是诱发食源性疾病的重要途径之一,也是食品安全领域备受关注的问题。利用微生物源示踪(microbial source tracking,MST)技术对粪便微生物来源和污染情况进行追踪是解决该问题的有效手段之一。MST方法一般分为... 详细信息

在食品链中,粪便微生物污染是诱发食源性疾病的重要途径之一,也是食品安全领域备受关注的问题。利用微生物源示踪(microbial source tracking,MST)技术对粪便微生物来源和污染情况进行追踪是解决该问题的有效手段之一。MST方法一般分为依赖数据库型方法(library-dependent methods,LDMs)和非依赖数据库型方法(library-independent methods,LIMs)2种方法,其中LIMs分为依赖培养型和非依赖培养型。在非依赖培养型方法中,以16S rRNA和宿主-微生物互作基因为特异性分子标记物建立的MST方法在食品链的污染调查中得到了广泛应用。本文对MST的基本原理及特点进行简要介绍,并着重总结近几年基于宿主特异性分子标记物的MST方法在食品链中进行污染溯源的研究进展,同时结合目前MST技术所具有的独特优势及现存不足对MST技术进行展望。

关键词：微生物源示踪方法食源性疾病粪便污染依赖数据库非依赖数据库宿主特异性分子标记

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早发型帕金森病家系相关基因的筛查和危险度分析

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海南医学院学报 2024年第12期30卷 888-896页

作者：许茜范丽华林昆明吴方真周美倪慧心刘海鑫范自立高正涛林瑶福建中医药大学中西医结合学院福建福州350122 三明市中西医结合医院神经内科福建三明365000 三明市第一医院中医科福建三明365000 福建省第二人民医院风湿内分泌科福建福州350001

目的:通过早发型帕金森病(early‑onset Parkinson’s disease,EOPD)遗传家系成员的高通量基因筛查,发现疾病相关突变位点并检测其在人群中对疾病的影响。方法:本研究对早发型PD遗传家系中19名成员进行单核苷酸多态性芯片检测和全外显子... 详细信息

目的:通过早发型帕金森病(early‑onset Parkinson’s disease,EOPD)遗传家系成员的高通量基因筛查,发现疾病相关突变位点并检测其在人群中对疾病的影响。方法:本研究对早发型PD遗传家系中19名成员进行单核苷酸多态性芯片检测和全外显子基因分型,并通过对家系外散发人群58人进行测序验证,筛选具有较高风险的突变位点。结果:芯片连锁分析发现3个染色体区段、全外显子测序和共表达分析筛选出24个可能与疾病关联的基因。其中5个位点基因型与患病结局之间的Spearman相关系数r>0.6且P<0.05,表示具有关联。通过家系外散发患者测序验证,Ddx56基因(OR=10.923)和Aspn基因(OR=8.198)的突变对患病结局产生显著影响(P<0.05)。结论:本研究通过临床患者进行多层次突变位点筛查分析,发现外显子区域的5个突变基因Pdxdc1、Ddx56、Aspn、Rbm28和Shisa9可能与帕金森病密切相关。

关键词：早发型帕金森病全外显子测序连锁分析

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基于直接TaqMan-PCR的无创检测脂质代谢相关SNP基因分型技术

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基础医学与临床 2024年第6期44卷 858-865页

作者：孙元宏谢吕范利华郑直中国医学科学院基础医学研究所、北京协和医学院基础学院、生物化学与分子生物学系北京100005

目的建立一种基于直接TaqMan-PCR的无创多重单核苷酸多态性分型技术用于高通量检测与脂质代谢相关的rs688和rs964184位点,以满足风险人群快速筛查的需求。方法采用提取的DNA对设计的TaqMan探针进行PCR退火延伸温度和扩增程序的优化;将... 详细信息

目的建立一种基于直接TaqMan-PCR的无创多重单核苷酸多态性分型技术用于高通量检测与脂质代谢相关的rs688和rs964184位点,以满足风险人群快速筛查的需求。方法采用提取的DNA对设计的TaqMan探针进行PCR退火延伸温度和扩增程序的优化;将口腔拭子放入样品处理液后进行短暂震荡和离心,取少量上清进行直接qPCR扩增分析;探索探针冻融稳定性和样品保存时间对分型结果的影响。结果本方法仅需对口腔拭子样本简单处理后即可进行qPCR分析,并且可在室温情况下保存4 d。优化后的方法能够较好地区分rs688和rs964184位点的野生纯合子、突变纯合子及杂合子。结论建立了一种快速便捷、高通量、低成本的SNP分型新技术,为大规模筛查携带易感基因的风险人群提供了高效且准确的新方法。

关键词：直接PCR 核酸提取 SNP基因分型高通量

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