双重实时荧光PCR快速鉴别人参和西洋参＊

目的 建立一种能够快速鉴定人参和西洋参的双重实时荧光PCR方法。方法 通过对人参属及其近似种基因序列的分析比对，设计特异性的引物探针，建立双重实时荧光PCR检测方法。PCR反应体系为Premix Ex Taq （Probe qPCR） 10 μL，人参上下游引物各0.5 μL，西洋参上下游引物各0.3 μL，人参与西洋参特异性探针各0.4 μL，DNA模板2 μL，灭菌去离子水补至20 μL。反应条件为95℃预变性30 s；然后以95℃变性5 s，60℃退火延伸30 s进行40个循环。应用该方法测试了27份DNA样品，包括7份人参、6份西洋参、4份人参与西洋参混合样、10份其他人参属样品及其他常见中药材样品的DNA，以确定该方法的特异性。将人参与西洋参样品DNA混合后10倍稀释成不同浓度后进行检测，用以确定该方法的灵敏度。结果 人参及西洋参样品均在特定的荧光通道下出现典型的阳性扩增曲线，人参与西洋参混合样品均同时出现两条阳性扩增曲线，其它对照样品及空白对照均没有出现阳性扩增曲线。灵敏度检测结果显示该方法检测人参及西洋参的最低检测限均为2 pg DNA/反应。结论 本实验建立的双重实时荧光PCR方法能够同时快速、准确、灵敏地鉴别出人参和西洋参。     

2013-2017年我国四种恶性肿瘤住院费用水平及结构变动度分析

背景　恶性肿瘤给家庭、社会带来了沉重的医疗、经济负担，极易导致部分家庭“因病致贫”或放弃治疗，目前相关研究多集中于单一病种、分散地域的研究，仍缺乏对于全国范围与多病种恶性肿瘤住院费用变化及结构构成的考量。目的　分析2013-2017年我国4种恶性肿瘤住院费用水平以及影响住院费用的主要项目和结构变动情况，为控制医疗费用上涨、深化新医改提供参考依据。方法　本研究数据来源于《2014中国卫生和计划生育统计年鉴》《2015中国卫生和计划生育统计年鉴》《2016中国卫生和计划生育统计年鉴》《2017中国卫生和计划生育统计年鉴》以及《2018中国卫生健康统计年鉴》，样本跨度为2013-2017年。统计“30种疾病人均住院费用”中的胃恶性肿瘤、肺恶性肿瘤、食管恶性肿瘤以及膀胱恶性肿瘤的数据，4种恶性肿瘤的人均住院费用包括药费、检查费、治疗费、手术费和手术材料费。2019年4-8月，采用结构变动度法分析我国2013-2017年4种恶性肿瘤的住院费用的结构变动情况〔结构变动值（VSV）、结构变动度（DSV）、结构变动贡献率〕。结果　2013-2017年，4种恶性肿瘤的人均住院费用逐年上升，其中胃恶性肿瘤的人均住院费用始终最高，且肺恶性肿瘤的人均住院费用上升幅度最大。2013-2017年，在4种恶性肿瘤住院各项费用的占比中，药费占比最高且总体逐年下降。从4种恶性肿瘤住院各项费用的实际变化来看，药费在2013-2014年有所上升，2014-2017年逐年下降；检查费在2013-2014年下降，2014-2017年缓慢上升；手术费与手术材料费在2013-2017年逐年上升。2013-2017年，在4种恶性肿瘤住院各项费用中均是药费的VSV最大；4种恶性肿瘤药费、检查费的VSV均呈负向变化，手术费和手术材料费的VSV均呈正向变化，治疗费的VSV增减均不明显。2013-2017年，4种恶性肿瘤住院费用的DSV从大到小依次为肺恶性肿瘤、胃恶性肿瘤、食管恶性肿瘤、膀胱恶性肿瘤。2013-2017年，4种恶性肿瘤住院各项费用中均是药费的结构变动贡献率最大，治疗费的结构变动贡献率最小；除药费外，胃恶性肿瘤、肺恶性肿瘤住院各项费用中均是手术材料费和手术费的结构变动贡献率次之，食管恶性肿瘤住院各项费用中手术费、检查费的结构变动贡献率次之，膀胱恶性肿瘤住院各项费用中检查费、手术材料费的结构变动贡献率次之。结论　2013-2017年我国4种恶性肿瘤手术费的结构变动贡献率虽然较为理想，但药费、治疗费仍是住院费用结构的重点调整对象；同时为有效降低恶性肿瘤的人均住院费用，应当加强控制手术材料费与检查费；而胃恶性肿瘤与肺恶性肿瘤患者的疾病经济负担严重，若要缓解应加强疾病的早期预防与住院费用管控。     

中医健康状态辨识模式的研究现状与展望

健康状态的辨识是把握健康的前提。当前学界已存在专家辨识模式、标准辨识模式、数字辨识模式、智能辨识模式以及微观辨识模式等。联合多种辨识方法,构建健康状态辨识体系,形成常态与动态结合、主观与客观结合、人机互参的中医健康状态个体化辨识方法是研究的趋势所在。文章对未来的研究方向进行展望,探讨了多元辨识模式、远程辨识模式、终身辨识模式、自动辨识模式的思路方法,以期促进全民健康事业,助力"健康中国"战略。     

支持向量机法及其在中药研究中的应用

在中医药现代化进程中，有效地结合现代科学技术手段对传统中医药进行更为合理的分析是一个重要研究方向。计算机科学技术与人工智能算法在中医药研究领域已取得长足发展，尤其是分类算法所具有的快速鉴别和分析能力，使其在复杂的中医药研究体系中发挥举足轻重的作用。对支持向量机分类算法及其结合其他算法在解决中医药相关领域问题方面及成果进行综述，以相关智能算法为基础探讨中医药研究过程中的诸多难题，为推动中医药现代化进程提供支撑。     

后外侧结构重建在新鲜股骨颈骨折人工股骨头置换术中的应用

目的探讨后外侧结构重建对后外侧入路人工股骨头置换术术后早期关节脱位的影响。方法选取2016年9月至2017年8月于我院行后外侧入路初次人工股骨头置换术的股骨颈骨折患者60例,根据术中是否修补关节囊及外旋肌群分为重建组(33例:舌形切开关节囊,术中将关节囊及外旋肌群原位缝合在大转子后方及臀中肌肌腱附着处)和对照组(27例:切除关节囊后,术中未进行外旋肌群修复重建)。比较两组的手术情况及术后近期关节功能情况。结果重建组的手术时间为(45.0±15.3) min,长于对照组的(35.0±12.4) min (P <0.05)。重建组术腔引流量为(200.0±80.0) m L,少于对照组的(420.0±120.6) m L (P <0.05)。重建组的早期脱位率为0.000%(0例),与对照组的7.407%(2例)比较无统计学差异(P>0.05)。重建组术后Harris评分为(92.0±3.4)分,高于对照组的(88.2±5.0)分(P <0.05)。结论在后外侧入路人工股骨头置换过程中行后外侧结构重建能够有效减少术腔引流量,提高髋关节Harris评分,对维持髋关节软组织平衡具有重要意义。     

Automated classification of depression from structural brain measures across two independent community‐based cohorts

Major depressive disorder (MDD) has been the subject of many neuroimaging case–control classification studies. Although some studies report accuracies ≥80%, most have investigated relatively small samples of clinically‐ascertained, currently symptomatic cases, and did not attempt replication in larger samples. We here first aimed to replicate previously reported classification accuracies in a small, well‐phenotyped community‐based group of current MDD cases with clinical interview‐based diagnoses (from STratifying Resilience and Depression Longitudinally cohort, ‘STRADL’). We performed a set of exploratory predictive classification analyses with measures related to brain morphometry and white matter integrity. We applied three classifier types—SVM, penalised logistic regression or decision tree—either with or without optimisation, and with or without feature selection. We then determined whether similar accuracies could be replicated in a larger independent population‐based sample with self‐reported current depression (UK Biobank cohort). Additional analyses extended to lifetime MDD diagnoses—remitted MDD in STRADL, and lifetime‐experienced MDD in UK Biobank. The highest cross‐validation accuracy (75%) was achieved in the initial current MDD sample with a decision tree classifier and cortical surface area features. The most frequently selected decision tree split variables included surface areas of bilateral caudal anterior cingulate, left lingual gyrus, left superior frontal, right precentral and paracentral regions. High accuracy was not achieved in the larger samples with self‐reported current depression (53.73%), with remitted MDD (57.48%), or with lifetime‐experienced MDD (52.68–60.29%). Our results indicate that high predictive classification accuracies may not immediately translate to larger samples with broader criteria for depression, and may not be robust across different classification approaches.     

液质联用技术鉴定清血八味片化学成分研究

目的:采用高效液相色谱-三重四级杆质谱(HPLC-MS/MS)测定清血八味片中的化学成分。方法:在负离子条件下采用Halo C_(18)色谱柱(2.1 mm×100 mm,2.7μm),以0.1%甲酸-10 mmol/L甲酸铵水和乙腈为流动相进行梯度洗脱,质谱采用Scan模式和MRM模式,检测清血八味片中的化学成分。结果:共鉴定出紫草中成分7种、土木香中成分3种、人工牛黄中成分4种、栀子中成分7种、瞿麦中成分7种、甘草中成分12种。结论:该方法快速可靠,操作简便,可用于清血八味片的质量控制研究,为临床药物使用提供一定的依据。     

Developmental validation of the ANDE™ rapid DNA system with FlexPlex™ assay for arrestee and reference buccal swab processing and database searching

A developmental validation was performed to demonstrate reliability, reproducibility and robustness of the ANDE System with the FlexPlex assay, including an integrated Expert System, across a number of laboratories and buccal sample variations. Previously, the related DNAscan™/ANDE 4C Rapid DNA System using the PowerPlex^®16 assay and integrated Expert System Software received NDIS approval in March 2016. The enhanced ANDE instrument, referred to as ANDE 6C, and the accompanying 6-dye, 27-locus STR assay, referred to as FlexPlex, have been developed to be compatible with all widely used global loci, including the expanded set of the CODIS core 20 loci.Six forensic and research laboratories participated in the FlexPlex Rapid DNA developmental validation experiments, testing a total of 2045 swabs, including those obtained from 1387 unique individuals. The goal of this extensive and comprehensive validation was to thoroughly evaluate and document the ANDE System and its internal Expert System to reliably genotype reference buccal swab samples in a manner compliant with the FBI’s Quality Assurance Standards and the NDIS Operational Procedures.The ANDE System, including automated Expert System analysis, generated reproducible and concordant results for buccal swabs when testing various instruments at different laboratories by a number of different operators. When testing a number of non-human DNAs, including oral bacteria, the ANDE System and FlexPlex assay demonstrated limited cross-reactivity. Potential PCR inhibitors were evaluated as part of the validation and no inhibition was detected. Reproducible and concordant profiles were generated from buccal swab samples collected with a limit of detection appropriate for buccal swab collections from arrestees. The precision and resolution of the System met industry standards for detection of microvariants and single base resolution.The integrated Expert System appropriately demonstrated the ability to correctly pass or fail profiles for CODIS upload without human review. During this comprehensive developmental validation, the ANDE System successfully interpreted over 2000 samples tested with over 99.99% concordant alleles. The data package described herein led to the ANDE System with the FlexPlex assay receiving NDIS approval in June 2018.     

Current status of MALDI-TOF mass spectrometry in clinical microbiology

Mass spectrometry (MS) is a type of analysis used to determine what molecules make up a sample, based on the mass spectrum that are created by the ions. Mass spectrometers are able to perform traditional target analyte identification and quantitation; however, they may also be used within a clinical setting for the rapid identification of bacteria. The causative agent in sepsis is changed over time, and clinical decisions affecting the management of infections are often based on the outcomes of bacterial identification. Therefore, it is essential that such identifications are performed quickly and interpreted correctly. Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometer is one of the most popular MS instruments used in biology, due to its rapid and precise identification of genus and species of an extensive range of Gram-negative and -positive bacteria. Microorganism identification by Mass spectrometry is based on identifying a characteristic spectrum of each species and then matched with a large database within the instrument. The present review gives a contemporary perspective on the challenges and opportunities for bacterial identification as well as a written report of how technological innovation has advanced MS. Future clinical applications will also be addressed, particularly the use of MALDI-TOF MS in the field of microbiology for the identification and the analysis of antibiotic resistance.     

基于18SrRNA序列的云南大叶千斤拔与细叶千斤拔根内丛枝菌根真菌鉴定

目的:调查云南省大叶千斤拔(Flemingia macrophylla)与细叶千斤拔(Flemingia lineata)根内丛枝菌根真菌(Arbuscular Mycorrhizal Fungi,AMF)群落结构多样性。方法:使用巢式-PCR、克隆、限制性片段长度多态性(Restriction Fragment Length Polymorphism,RFLP)分析及测序技术。结果:共获得558个含有丛枝菌根真菌18S rRNA片段的克隆子,经RFLP分析后得到83个RFLP类型,DNA序列分析可将其划分为23个可操作分类单元(Operational Taxonomic Units,OTUs),分属于5个科,Glomeraceae为优势类群。在MaarjAM数据库中进行比对后,23个OTUs可鉴定为18个虚拟分类分子种,分布于13种不同的生境。经统计分析大叶千斤拔与细叶千斤拔根内丛枝菌根真菌群落组成比较,差异有统计学意义(P<0.001)。结论:比较大叶千斤拔与细叶千斤拔根内丛枝菌根真菌群落差异,结合丛枝菌根真菌在生态系统中的分布特点,为种植千斤拔属植物的选址提供有力地环境指标数据,并为筛选促生菌株提供依据。