精致教引导诚潜学——基于方剂学精品线下开放课程教改体会

积极响应校、省、国家教改要求,利用共享课程资源、开放式课程教育平台及智慧教学工具,以金课标准结合课程思政要求建设方剂学精品线下开放课程。教师从教改理念宣传、教学资源优化、教学过程设计、反馈交流指导方面全力打造精致课程,创建具有先进性、互动性、创新性的以学生为主的线上、线下混合式翻转课堂教学,引导学生端正态度潜心学习,实现知识、能力、情感素养的全面提升。     

Development and content validity of the Experienced Patient‐Centeredness Questionnaire (EPAT)—A best practice example for generating patient‐reported measures from qualitative data

IntroductionTo effectively foster patient‐centeredness (PC), it is crucial to measure its implementation. So far, there is no German measure to assess PC comprehensively. The aim of this study is to develop and select items for the Experienced Patient‐Centeredness (EPAT) Questionnaire, a patient‐reported experience measure (PREM). The EPAT intends to assess PC from the perspective of adult patients treated for different chronic diseases in inpatient and outpatient settings in Germany. Furthermore, we aim at providing a best‐practice example for developing PREMs from qualitative data.MethodsThe development process comprised a three‐phase mixed‐method design: (1) preparation, (2) item generation and (3) item selection and testing of content validity. We generated items using qualitative content analysis based on information from focus groups, key informant interviews and literature search. We selected items using relevance rating and cognitive interviews. Participants were patients from four chronic disease groups (cancer, cardiovascular disease, mental disorder, musculoskeletal disorder) and healthcare experts (e.g., clinicians, researchers, patient representatives).ResultsWe conducted six focus groups with a total of 40 patients, key informant interviews with 10 experts and identified 48 PREMs from international literature. After team discussion, we reached a preliminary pool of 152 items. We conducted a relevance rating with 32 experts and 34 cognitive interviews with 21 patients. We selected 125 items assessing 16 dimensions of PC and showed high relevance and comprehensibility.ConclusionsThe EPAT questionnaire is currently undergoing psychometric testing. The transparent step‐by‐step report provides a best practice example that other researchers may consider for developing PREMs. Integrating literature and experts with a strong focus on patient feedback ensured good content validity. The EPAT questionnaire will be helpful in assessing PC in routine clinical practice in inpatient and outpatient settings for research and quality improvement.Patient or Public ContributionPatients were not involved as active members of the research team. While developing the funding proposal, we informally reached out to several patient organizations who all gave us positive feedback on the study aims, thereby confirming their relevance. Those patient organizations endorsed the funding proposal with formal letters of support and supported recruitment by disseminating advertisements for study participation.     

MLL5基因敲除对小鼠结肠癌CT26细胞移植瘤生长的影响及其机制

目的：探讨混合谱系白血病5(MLL5)基因在小鼠结肠癌CT26细胞移植瘤生长中的作用及其分子机制。方法：利用CRISPR/Cas9技术构建MLL5基因缺失、MLL5和DDX58双基因缺失的结肠癌CT26细胞模型,用Sanger测序和WB法验证敲除效果。将基因敲除的CT26细胞接种到野生型BALB/c小鼠和免疫缺陷型NSG小鼠皮下,构建基因缺失结肠癌CT26细胞移植瘤小鼠模型,并观察移植瘤的生长及荷瘤小鼠的总生存期（OS）。结果：在野生型小鼠中,MLL5基因缺失的CT26细胞移植瘤生长速度显著性低于野生型癌细胞移植瘤,并延长荷瘤小鼠的OS(P<0.01);在NSG小鼠中,MLL5基因缺失对CT26细胞移植瘤的生长速度以及荷瘤小鼠的OS没有明显改变。MLL5基因缺失提高了癌细胞中视黄酸诱导基因1(RIG-1)蛋白水平,DDX58基因缺失可逆转MLL5基因缺失在CT26细胞移植瘤中的作用。结论：MLL5基因缺失可提高结肠癌CT26细胞中RIG-1蛋白水平、促进肿瘤免疫,从而抑制荷瘤小鼠肿瘤生长,提示MLL5可能成为结肠癌治疗的新靶点。     

补骨脂素混合结晶在S_(180)小鼠腹水和癌细胞内的浓度测定及药代动力学研究

补骨脂素混合结晶中补骨脂素在S_(180)小鼠腹水内属单室模型中呈双峰吸收。其动力学参数如下■l/2(h)为22.19;K_a(h~(-1))为12.3580;K(h-1)为0.0312;tlag(h)为0.0718;C_(max)(μg/ml)为6.3967;t_(max)(h)为0.5570;AUC(μg·h/ml)为175.0300。2亿S_(180)癌细胞内在0.25h时,其补骨脂素含量高达10.97μg/ml,腹水内补骨脂素含量既高且久,有利于肿瘤的冶疗。     

双向排斥反应联体动物模型的建立

目的 探讨在异基因成年小鼠之间建立联体模型用以研究双向排斥反应。方法 54只雄性C57BL/6(H-2b,C57)小鼠和54只雌性Balb/c(H-2d)小鼠配对分为3组:1组(注射生理盐水)、2组(注射脾细胞+环磷酰胺)、3组(注射脾细胞+环磷酰胺+环孢素A)。于术中经尾静脉注入对方的脾细胞;术前3d和术后1周每日分别腹腔注射环孢素A(30mg·kg·d^-1);术后2d腹腔注射环磷酰胺(150mg/kg)1次。对经以上处理的部分联体小鼠在术后1周后分开,相互植皮。观察联体维持时间,并分别检测其混合淋巴细胞反应和诱导迟发型超敏反应(DTH)。结果 第3组联体时间明显延长;第3组联体小鼠DTH,2、3组显示明显抑制,但3组小鼠抑制程度明显比第2组小鼠深。结论 应用免疫抑制剂,可成功建立双向排斥反应联体动物模型。     

重组E.coli. LLO/OVA明显增强小鼠CD11c细胞活性及抗肿瘤免疫

目的 探讨重组E.coli. LLO/OVA 诱导C57BL/6小鼠机体免疫的途径,观察其免疫后小鼠机体抑制B16-OVA黑色素瘤的效果.方法 磁珠分离E.coli. LLO/OVA及E.coli. OVA免疫后小鼠脾脏CD11c、CD4 和CD8 T细胞并检测CD11c细胞诱导同源CD4 和CD8 T增殖水平和细胞因子分泌程度;流式细胞检测小鼠脾脏内肿瘤抗原OVA257-264 SIINFEKL特异的细胞毒T细胞含量.比较两种E.coli. 免疫后,恶性黑色素瘤B16-OVA在小鼠肺内形成瘤结节的数量.结果 与E.coli. OVA相比, E.coli. LLO/OVA免疫后小鼠脾脏CD11c细胞诱导同源CD4 T细胞增殖作用增强、IL-2分泌增高;同时诱导CD8 T细胞增殖和IFN-γ分泌的作用也明显增强;OVA257-264 SIINFEKL特异的CD8 T细胞含量也明显增高;小鼠肺内形成B16-OVA瘤结节平均数明显减少. 结论 E.coli. LLO/OVA有效地诱导小鼠CD11c细胞活化,增强其对CD4 T细胞增殖和IL-2分泌以及对CD8 T细胞增殖、IFN-γ分泌的作用,诱导了更多的OVA特异的CD8 T细胞,使机体产生了更强的抗肿瘤免疫.     

Shrinkage estimation and the use of additional information when calibrating in the presence of random effects

Let x denote a precise measurement of a quantity and Y an inexact measurement, which is, however, less expensive or more easily obtained than x. We have available a calibration set comprising clustered sets of (x,Y ) observations, obtained from different sampling units. At the prediction step, we will only observe Y for a new unit, and we wish to estimate the corresponding unknown x, which we denote by ξ. This problem has been treated under the assumption that x and Y are linearly related. Here, we expand on those results in three directions: First, we show that if we center ξ about a known value c, for example, the mean x‐value of the calibration set, then the proposed estimator now shrinks to c. Second, we examine in detail the performance of the estimator, which was proposed when one or more (x,Y ) observations can be obtained for the new subject. Third, we compare the Fieller‐like confidence intervals, previously proposed, with t‐like intervals based on asymptotic moments of the point estimate. We illustrate and evaluate our procedures in the context of a data set of true bladder‐volumes (x) and ultrasound measurements (Y). Copyright © 2013 John Wiley & Sons, Ltd.     

Latent class models for medical diagnostic tests in multicenter trials

Medical diagnostic tests must enjoy appropriate validity and high reliability in order to qualify as adequate assessment tools. Without a gold standard test, available medical diagnostic tests are not perfect; hence, the reliability of such tests must be evaluated precisely. Kappa coefficient statistics are often utilized to assess reliability of tests when there are two or more medical diagnostic tests. However, the statistics are imprecise for a typical case when the prevalence rate of a target disease is unknown. Although latent class models could be used to assess reliability, the models cannot estimate reliability in the case of two tests, due to unidentifiability or the lack of degrees of freedom. An alternative approach to assess reliability for the case of two tests is stratifying a two‐by‐two contingency table under the assumption that sensitivities and specificities between the two tests be equal over all strata and that prevalence rates in the strata be different from each other. Because stratification is basically a multi‐sample analysis, it should not be applied to the situation where subsamples (i.e., centers) are randomly selected from a larger population. In this article, a type of mixed‐effect model is proposed to evaluate the reliability of two tests for trials in randomly selected multiple centers. Several types of distributions for prevalence rates over subpopulations are considered. Simulation studies show that our proposed method performs nicely. Analysis of real data is also reported. Copyright © 2013 John Wiley & Sons, Ltd.     

PCR for detection of respiratory viruses: seasonal variations of virus infections

Real-time PCR and related methods have revolutionized the laboratory diagnosis of viral respiratory infections because of their high detection sensitivity, rapidness and potential for simultaneous detection of 15 or more respiratory agents. Results from studies with this diagnostic modality have significantly expanded our knowledge about the seasonality of viral respiratory diseases, pinpointed the difficulties to make a reliable etiologic diagnosis without the aid of an unbiased multiplex molecular assay for respiratory viruses, and revealed previously unknown details as to possible infections with multiple agents as aggravating factors. The scope of this article is to review and discuss this new knowledge and its implications for diagnostic strategies and other measures essential for the clinical management of respiratory viral infections and for epidemiological surveillance of seasonal respiratory infections.