基于微流控核酸等温扩增的 登革病毒现场快速检测技术研究

[摘要]?目的?结合环介导等温扩增技术（loop-mediated isothermal amplification, LAMP）和微流控芯片技术，建立一种适合现场快速检测登革病毒的方法。方法?利用RT-LAMP，针对登革病毒基因组中3’非编码区中特异性序列进行扩增，建立基于微流控芯片技术的LAMP检测方法，优化检测体系，并对该方法的灵敏性和特异性进行评估。结果?基于LAMP 和微流控芯片技术的登革病毒检测方法，通过对病毒模板进行扩增，发现与其他病毒无交叉反应，特异性良好；同时，结果显示该方法对登革病毒检测灵敏性可达61.2 pg/μl，与实时荧光定量PCR仪所达到的检测灵敏性一致。结论?基于LAMP和微流控芯片技术的登革病毒检测方法具有操作简单、快速、对设备要求低等优势,并且灵敏性、特异性均较好，是一种便于开展现场快速检测的方法。     

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。     

A Pre-implementation Study of Volumetric Modulated Arc Therapy for Same-day Planning and Treatment of Vertebral Bone Metastases Within a Rapid-access Palliative Radiotherapy Programme

AimsWe aimed to develop a process for same-day contouring, planning, quality assurance and delivery of volumetric modulated arc therapy (VMAT) for vertebral bone metastases within our institution's rapid-access palliative radiotherapy programme.Materials and methodsTwo thoracic (T6–7, T3–7) and two lumbar (L2–3, L1–5) targets were contoured on computed tomography images acquired from an anthropomorphic phantom and five patient scans. Inverse planning aimed to provide coverage of a prescribed dose of 8 Gy with a combined lung V2Gy < 25% and a combined kidney mean dose <2 Gy. Serial plans were created to identify an efficient combination of six main planning variables specific to our treatment planning system: (i) voxel size (3 mm versus 5 mm), (ii) Monte Carlo statistical uncertainty (1% per calculation versus 3% per control point), (iii) fluence smoothing (medium versus high), (iv) number of iterations of segment shape changes during optimisation (1 versus 5), (v) dose calculation algorithm (Monte Carlo versus pencil beam) and (vi) number of arcs (single versus multiple). Contouring, planning, quality assurance and treatment delivery were timed.ResultsThe combination of planning variables deemed efficient and appropriate was: a 3 mm voxel size, statistical uncertainty of 1% per calculation, medium fluence smoothing, five iterations of segment shape changes, Monte Carlo dose calculation and single full arc delivery. Patient scan contouring times ranged from 7 to 9 min (T6–7), 11–13 min (T3–7), 5–7 min (L2–3) and 8–10 min (L1–5) and planning times ranged from 9 to 15 min (T6–7), 13–25 min (T3–7), 18–25 min (L2–3) and 21–31 min (L1–5). Physics quality assurance times ranged from 15 to 21 min and beam-on times ranged from 3 to 6 min.ConclusionsThe combined elements of VMAT for thoracic and lumbar vertebral bone metastases were completed in under 2 h. This new process makes same-day contouring, planning, quality assurance and treatment delivery of VMAT feasible within our rapid-access palliative radiotherapy programme.     

Redox active or thiol reactive? Optimization of rapid screens to identify less evident nuisance compounds

Compounds that exhibit assay interference or undesirable mechanisms of bioactivity are routinely encountered in assays at various stages of drug discovery. We observed that assays for the investigation of thiol-reactive and redox-active compounds have not been collected in a comprehensive review. Here, we review these assays and subject them to experimental optimization to improve their reliability. We demonstrate the usefulness of our assay cascade by assaying a library of bioactive compounds, chemical probes, and a set of approved drugs. These high-throughput assays should complement the array of wet-lab and in silico assays during the initial stages of hit discovery campaigns to pursue only hit compounds with tractable mechanisms of action.     

Surgeons’ Preoperative Work Burden Has Increased Before Total Joint Arthroplasty: A Survey of AAHKS Members

BackgroundImplementation of rapid recovery protocols and value-based programs in total joint arthroplasty (TJA) has required changes in preoperative management, such as optimization, education, and coordination. This study aimed to quantify the work burden associated with preoperative TJA care.MethodsTwo web-based surveys were distributed to surgeon members of the American Association of Hip and Knee Surgeons. The first questionnaire (265 respondents) consisted of questions related to preoperative patient care in TJA and the associated work burden by orthopedic surgeons and their financially dependent health care providers. The second survey (561 respondents) consisted of questions related to relative change in preoperative patient care work burden since 2013.ResultsGreater than 98% of survey respondents reported providing some level of preoperative medical optimization to their patients. The mean amount of reported time spent by the surgeon and/or a qualified health care provider in preoperative activities not included in work captured in current procedural terminology or hospital billing codes was 153 minutes. The mean amount of reported time spent by ancillary clinical staff in preoperative activities was 177 minutes. Most surgeons reported an increase in work burden for total knee (86%) and total hip (87%) arthroplasty since 2013, with a large portion reporting a 20% or greater increase in work (knee 66%, hip 64%).ConclusionTo provide quality arthroplasty care with marked reductions in complication rates, lengths of stay, and readmissions, members of the American Association of Hip and Knee Surgeons report a substantial preoperative work burden that is not included in current coding metrics. Policy makers should account for this time in coding models to continue to promote pathway improvements.     

快速康复护理在腹腔镜下全子宫切除中的应用

目的探讨在腹腔镜下全子宫切除术中应用快速康复护理措施对患者围手术期指标和患者并发症的影响。方法将在本院行腹腔镜下全子宫切除术的70例患者随机分为两组,各35例,对照组35例给予常规护理,观察组35例给予快速康复护理,对比两组围手术期相关指标、手术前后VAS评分、并发症发生情况以及患者护理满意度。结果观察组胃肠蠕动时间[(9.32±1.67)h]、肛门排气时间[(4.67±1.37)h]、下床活动时间[(6.69±1.24)h]、术后进食时间[(6.21±0.55)h]和住院时间[(4.25±1.20)d]均显著短于对照组的[(12.87±2.08)h]、[(7.28±1.13)h]、[(9.75±3.29)h][(9.33±0.61)h]、[(7.25±1.38)d],P<0.05;观察组术后1 d和术后3 d的VAS评分分别为(4.01±1.28)分、(1.23±0.35)分,显著低于对照组的(6.80±1.32)分、(2.62±0.41)分,P<0.05;观察组和对照组术后并发症发生率(2.86%vs 17.14%)和患者护理满意度(97.14%vs.80.00%)比较,观察组均显著优于对照组P<0.05。结论对于行腹腔镜下全子宫切除术的患者,给予快速康复护理模式可促进患者早期康复,减少术后疼痛程度,降低术后并发症发生率。     

The Association between Reasons for a Rapid Response Team Alert and Immediate Patient Management in Total Hip Arthroplasty Patients

BackgroundThe purpose of this study is to evaluate the value and efficacy of rapid response teams (RRTs) for different triggering events in total hip arthroplasty (THA) patients.MethodsA retrospective review of all RRT events at a single, tertiary referral center from 2014 to 2016 was performed. Inclusion criteria were defined as patients >18 years old that underwent primary or revision THA. Information queried included demographics, primary reason for RRT, Charlson Comorbidity Index (CCI), underlying etiology, whether any changes in management occurred, and whether the patient was uptriaged.ResultsIn total, 168 RRTs were called on 153 hip arthroplasty patients (mean age 65.2 ± 14.1 years; mean body mass index 32.3 ± 4.8, 66% female). Length of stay in RRT for primary and revision THA was 3.4 and 6.2 days, respectively. This was significantly longer than the length of stay for primary THA patients (2.4 days, P < .001) and revision THA patients (4.6 days, P = .005) that did not require an RRT. There were no mortalities. RRTs for hypotension/presyncope (11%) and for syncope (11%) resulted in significantly fewer changes in management (P < .01) than tachycardia (77%), hypoxia (57%), AMS (79%), and other (47%). RRTs for hypotension/presyncope (28%), syncope (15%), and hypoxia (30%) resulted in significantly fewer patients being uptriaged (P < .001) than tachycardia (81%). Hypotension/presyncope was found to be significantly more commonly due to volume depletion (67%) (P < .001) than other etiologies. Hypoxia was significantly more commonly due to atelectasis (57%) and opioids/oversedation (30.4%) (P = .037). AMS/delirium was also significantly more commonly caused by opioids/over-sedation (71%) (P < .001).ConclusionIn patients undergoing THA, RRTs for hypotension/presyncopal symptoms and syncope were significantly less likely to result in changes in management or uptriaging compared to tachycardia. The most common etiologies were potentially preventable, including volume depletion and opioid use.