A prime-boost concept using a T-cell epitope-driven DNA vaccine followed by a whole virus vaccine effectively protected pigs in the pandemic H1N1 pig challenge model

Influenza A virus (IAV) vaccines in pigs generally provide homosubtypic protection but fail to prevent heterologous infections. In this pilot study, the efficacy of an intradermal pDNA vaccine composed of conserved SLA class I and class II T cell epitopes (EPITOPE) against a homosubtypic challenge was compared to an intramuscular commercial inactivated whole virus vaccine (INACT) and a heterologous prime boost approach using both vaccines. Thirty-nine IAV-free, 3-week-old pigs were randomly assigned to one of five groups including NEG-CONTROL (unvaccinated, sham-challenged), INACT-INACT-IAV (vaccinated with FluSure XP® at 4 and 7 weeks, pH1N1 challenged), EPITOPE-INACT-IAV (vaccinated with PigMatrix EDV at 4 and FluSure XP® at 7 weeks, pH1N1 challenged), EPITOPE-EPITOPE-IAV (vaccinated with PigMatrix EDV at 4 and 7 weeks, pH1N1 challenged), and a POS-CONTROL group (unvaccinated, pH1N1 challenged). The challenge was done at 9 weeks of age and pigs were necropsied at day post challenge (dpc) 5. At the time of challenge, all INACT-INACT-IAV pigs, and by dpc 5 all EPITOPE-INACT-IAV pigs were IAV seropositive. IFNγ secreting cells, recognizing vaccine epitope-specific peptides and pH1N1 challenge virus were highest in the EPITOPE-INACT-IAV pigs at challenge. Macroscopic lung lesion scores were reduced in all EPITOPE-INACT-IAV pigs while INACT-INACT-IAV pigs exhibited a bimodal distribution of low and high scores akin to naïve challenged animals. No IAV antigen in lung tissues was detected at necropsy in the EPITOPE-INACT-IAV group, which was similar to naïve unchallenged pigs and different from all other challenged groups. Results suggest that the heterologous prime boost approach using an epitope-driven DNA vaccine followed by an inactivated vaccine was effective against a homosubtypic challenge, and further exploration of this vaccine approach as a practical control measure against heterosubtypic IAV infections is warranted.     

Mapping Levels of Palliative Care Development in 198 Countries: The Situation in 2017

ContextPalliative care is gaining ground globally and is endorsed in high-level policy commitments, but service provision, supporting policies, education, and funding are incommensurate with rapidly growing needs.ObjectivesThe objective of this study was to describe current levels of global palliative care development and report on changes since 2006.MethodsAn online survey of experts in 198 countries generated 2017 data on 10 indicators of palliative care provision, fitted to six categories of development. Factor analysis and discriminant analysis showed the validity of the categorization. Spearman correlation analyses assessed the relationship with World Bank Income Level (WBIL), Human Development Index (HDI), and Universal Health Coverage (UHC).ResultsNumbers (percentages) of countries in each development category were as follows: 1) no known palliative care activity, 47 (24%); 2) capacity-building, 13 (7%); 3a) isolated provision, 65 (33%); 3b) generalized provision, 22 (11%); 4a) preliminary integration into mainstream provision, 21 (11%); 4b) advanced integration, 30 (15%). Development levels were significantly associated with WBIL (r_S = 0.4785), UHC (r_S = 0.5558), and HDI (r_S = 0.5426) with P < 0.001. Net improvement between 2006 and 2017 saw 32 fewer countries in Categories 1/2, 16 more countries in 3a/3b, and 17 more countries in 4a/4b.ConclusionPalliative care at the highest level of provision is available for only 14% of the global population and is concentrated in European countries. An 87% global increase in serious health-related suffering amenable to palliative care interventions is predicted by 2060. With an increasing need, palliative care is not reaching the levels required by at least half of the global population.     

思维导图在本科护生精神科实习带教中的应用

目的　探讨思维导图在本科护生精神科实习带教中的应用效果。方法　选取2018年6月至12月31名本科护生为试验组，采用思维导图进行带教；选取2019年4月至7月32名本科护生作为对照组，采用传统的带教方法进行带教。比较两组前后评判性思维能力、对带教方法的评价，以及两组护生的理论、操作考试成绩。采用SPSS 20.0进行t检验、卡方检验和秩和检验。结果　试验组在评判性思维能力量表各项得分明显提高，由带教前的（248.09±24.45）分提高至（331.03±14.31）分，且高于对照组（P<0.05）。试验组在带教方法肯定性评价各项得分均高于对照组，差异均有统计学意义（P<0.05）。试验组带教后理论成绩为（88.35±7.71）分、操作考试成绩为（86.45±5.03）分，对照组带教后理论成绩为（74.41±10.27）分、操作考试成绩为（81.02±6.64）分，两组比较差异有统计学意义（P<0.05）。结论　运用思维导图对精神科护生进行临床带教，提高了护生学习积极性，学习由被动变主动，同时促进了护生评判性思维能力培养，值得临床推广。     

PBL与思维导图相结合教学模式在临床教学中的应用

目的探讨PBL与思维导图相结合教学模式在临床教学中的应用。方法本文研究对象为在我院见习的临床本科生,为大理大学2015级一个班级,共55名学生,数据收集时间为2019年1月;按照授课方式的差异将学生分为实验班与对照班,实验班27名学生,对照班28名学生;对照班实施常规教学法,实验班实施PBL与思维导图相结合教学法。结果实验班教学结束测试得分为(85.60±3.46)分,对照班教学结束测试得分为(77.48±3.12)分,差异具有统计学意义(P<0.05);实验班教学方式满意度评分为(90.35±4.12)分,对照班教学方式满意度评分为(80.25±4.05)分,差异具有统计学意义(P<0.05)。结论在临床教学中采用PBL与思维导图相结合教学模式,能够提高学生的专业知识掌握水平与对教学模式的认可度。     

Effect of interictal epileptiform discharges on EEG-based functional connectivity networks

ObjectiveFunctional connectivity networks (FCNs) based on interictal electroencephalography (EEG) can identify pathological brain networks associated with epilepsy. FCNs are altered by interictal epileptiform discharges (IEDs), but it is unknown whether this is due to the morphology of the IED or the underlying pathological activity. Therefore, we characterized the impact of IEDs on the FCN through simulations and EEG analysis.MethodsWe introduced simulated IEDs to sleep EEG recordings of eight healthy controls and analyzed the effect of IED amplitude and rate on the FCN. We then generated FCNs based on epochs with and without IEDs and compared them to the analogous FCNs from eight subjects with infantile spasms (IS), based on 1340 visually marked IEDs. Differences in network structure and strength were assessed.ResultsIEDs in IS subjects caused increased connectivity strength but no change in network structure. In controls, simulated IEDs with physiological amplitudes and rates did not alter network strength or structure.ConclusionsIncreases in connectivity strength in IS subjects are not artifacts caused by the interictal spike waveform and may be related to the underlying pathophysiology of IS.SignificanceDynamic changes in EEG-based FCNs during IEDs may be valuable for identification of pathological networks associated with epilepsy.     

Reproducibility of Computerized Measurements of QT Interval from Multiple Leads at Rest and During Exercise

Background: Accurate measurement of the QT interval is important for diagnosing long QT syndrome (LQTS), and in research on determinants of ventricular repolarization time. We tested automatic analysis of QT intervals from multiple ECG leads on chest. Methods: Eleven healthy volunteers and 10 genotyped LQTS patients were tested at rest and during exercise with a bicycle ergometer twice 1–31 months apart. Electrocardiograms were recorded with the body surface potential mapping system, and 12 precordial channels were selected for analysis. Averaged QT peak and QT end intervals were determined with an automated algorithm, and the difference QT end minus QT peak (Tp‐e) was calculated. Repeatability was assessed by coefficient of variation (CV) between measurements. Results: Within one test at rest the QT end intervals were highly repeatable with CV 0.6%. In repeated tests CV was 4.4% for QT end interval and 3.5% when the QT interval was corrected for heart rate. In exercise test at specified heart rates, mean CV was 3.0% for QT end and 2.9% for QT peak interval. The CV of Tp‐e interval was 10.2% at rest, and 9.3% in exercise test. Reproducibility was comparable between healthy subjects and LQTS patients. Conclusions: The BSPM system with automated analysis produced accurate and highly repeatable QT interval measurements. Reproducibility was adequate also over prolonged time periods both at rest and in exercise stress test. The method can be applied in studying duration of ventricular repolarization time in different physiologic and pharmacologic interventions.     

T辅助细胞在疫苗研制中的作用

发展感染性疾病疫苗之关键挑战在于利用确定的抗原以刺激产生能引起保护作用的合适的免疫反应。肽类疫苗的运用得到了极大的关注，其意义在于，已知不同的多表位构成单一结构以诱导出所希望的免疫反应所表现出的灵活性。这一般比利用减毒的活疫苗要安全并且相对而言比制造亚单位疫苗要容易。然而，多肽疫苗的发展面临巨大挑战。这一方法在诱导遗传背景复杂的人群免疫反应方面受到限制，这与主要组织相溶性复合物(MHC)多态性有关。因同样的理由，肽类免疫应答常因缺乏适当的辅助T淋巴细胞(HIL)而引导出不充分的细胞毒素T淋巴细胞(CTL)和抗体反应。另一个运用线性肽链结构的可能缺点是：为了引导出合适抗体反应，表面免疫球蛋白受体簇对于激活静息的B细胞就成为必须因素。由WHC多肽性引起的问题可由运用不加区别的T细胞表位来解决。从麻疹病毒F蛋白(氨基酸288到302)中得到的不加区别的T细胞表位和鼠的确定结合在多种MHC分子上的辅助T细胞表位(v1EB，aa191-209)已被定性并且被用于能极大激发免疫应答的结构中，以克服单一限制型免疫应答的缺陷。合成的，非自然Pan DR表位(PADRE)具有退化的结合几种通常HLA—DR的能力，能以绝对效价和抗体反应质量两种形式来增强激发短肽链的免疫应答。另外，一些所谓的从流感病毒血凝素(HA)来的“不加区别的”T细胞表位，恶性疟疟原虫红细胞前期抗原和分枝杆菌蛋白被报道能激发广泛的免疫应答。为了不加区别地结合于几种同型和同种异型的MHCⅡ类分子，这些肽类应显示出部重叠MHC结合形式或应利用保存于配体中的固定位点和应缺失等位基因特异性固定残基，以防止结合于其它Ⅱ类分子。了解MHCⅡ类分子对肽链的不加区别及特异性识别的生物物理学基础将为在疫苗设计中突破遗传限制的策略提供分子水平的依据。