基于COI条形码的市售蝉蜕及其混淆品DNA分子鉴定研究

目的 利用COI序列对中药材蝉蜕Cicadae Periostracum及其混淆品进行DNA条形码鉴定研究,为蝉蜕药材的快速准确鉴定提供新的技术手段。方法 收集全国市售蝉蜕药材、基地自采蝉蜕及其混淆品总45份样本,同时采集江苏徐州黑蚱养殖基地不同树种中蝉卵样品5份作为对照。提取DNA,进行PCR扩增及双向测序,测序结果采用DNAMAN和SnapGene进行拼接校对,运用MEGA11.0进行比对分析,计算种内及种间Kimura 2-Parameter(K2P)遗传距离并构建邻接法（neighbor-joining,NJ）系统发育树。结果 黑蚱种内遗传距离远小于种间最小遗传距离,NJ树结果显示,黑蚱蝉蜕样品全部聚集为独立的一支,支持率为99%。混淆品样品分别聚集为单独的一支,支持率均大于90%,表明基于COI序列的DNA条形码技术可以有效鉴别蝉蜕药材真伪。结论 应用COI序列作为DNA条形码能够准确有效地鉴别蝉蜕及其混淆品。     

Headache Professional Societies: Ethical Challenges and Suggested Solutions

Codes of professional conduct continue to be an essential component of maintaining the integrity of individuals, academic institutions, and medical societies. We review ethical issues of professional conduct focusing on conflicts of interest (COI). We explain how to manage or mitigate COI in the context of professionals involved in headache medicine and its medical specialty societies. We identify the roles of institutional, medical society, and governmental regulation in the protection of patients and maintaining the integrity of physicians and others involved in patient care.     

虫卵基因序列分析鉴定斯氏并殖吸虫病

目的探索运用分子生物学技术分析虫卵基因序列鉴定并殖吸虫病。方法从云南省的并殖吸虫病流行区采集溪蟹,常规分离囊蚴,经形态学鉴定后感染实验终宿主家猫,从猫粪便中分离虫卵,对虫卵进行详细的形态学观察和鉴定。用匀浆器研磨虫卵提取基因组DNA,PCR扩增出虫卵中完整的核糖体基因第二间隔区（ITS2）和部分线粒体细胞色素C氧化酶亚单位I基因（COI）,测序获得该基因的核苷酸序列。将核苷酸序列输入GenBank中进行Blast比较,通过ITS2和COI的同源性来判断所感染的并殖吸虫种类。结果取自猫粪便中的虫卵形态符合并殖吸虫卵的形态特征。该虫卵的ITS2基因序列与GenBank中的斯氏并殖吸虫的基因序列同源性为99%,COI基因序列的同源性也高达98%。结果鉴定为斯氏并殖吸虫虫卵,证明家猫感染的是斯氏并殖吸虫病。结论通过并殖吸虫卵的基因序列分析,不仅可以快速诊断并殖吸虫病,而且还能准确地鉴定感染者所感染的并殖吸虫虫种。     

DNA barcodes confirm the presence of a single member of the Anopheles maculipennis group in Morocco and Algeria: An. sicaulti is conspecific with An. labranchiae

Anopheles labranchiae Falleroni is the only member of the Maculipennis Group known to occur in northern Africa; however, confusion exists as to the taxonomic status of its junior synonym, An. sicaulti Roubaud (type locality: near Rabat, Morocco). Based on morphological and behavioural distinctions, it has been suggested that Moroccan populations have been isolated from other North African populations by the Atlas Mountains, and that Moroccan populations may represent An. sicaulti, originally described as a variety of An. maculipennis Meigen. DNA barcodes (658 bp of the mitochondrial COI gene) obtained from 89 An. maculipennis s.l. collected in Morocco (n = 79) and Algeria (n = 10) in 2007 and 2008 were used to determine if Moroccan populations are genetically isolated from those east of the Atlas Mountains (Algeria), and whether there is molecular evidence to support the presence of more than one member of the Maculipennis Group in the region. No evidence for speciation was found between Moroccan and Algerian populations, or within populations in northern Morocco. Moreover shared COI haplotypes between Algeria and Morocco indicate ongoing gene flow between populations in these countries, suggesting that the Atlas Mountains are not a boundary to gene flow in An. labranchiae. The synonymy of An. sicaulti with An. labranchiae is confirmed. That An. labranchiae comprises the same species in these North African countries is important for malaria control.     

基于COI与18S rRNA基因序列的4种肉食螨DNA条形码研究

目的　基于线粒体细胞色素C氧化酶亚基I（COI）与核糖体18S小亚基（18S rRNA）基因序列,分析确定4种肉食螨合适的DNA条形码,从而进一步完善肉食螨DNA条形码数据库。方法　2018年5月—2019年7月于安徽省阜阳、芜湖、铜陵等3市小型面粉作坊采集分离肉食螨样本,经形态学鉴定后,提取单个螨基因组DNA,经PCR扩增、克隆和测序获得COI与18S rRNA基因序列,所获序列进行BLAST比对。结合已报道肉食螨序列,用ClustalX 1.83软件进行多序列比对,基于MEGA X软件进行序列分析并计算遗传距离,采用最大似然法构建系统发育树。结果　马六甲肉食螨、转开肉食螨、鳞翅触足螨分子鉴定结果与形态学一致,而GenBank中缺少网真扇毛螨相关序列。4种肉食螨COI与18S rRNA 基因序列（A + T）分别为69.6%和55.1%,碱基替换数分别为137对和46对。基于COI与18S rRNA基因序列变异位点分析,发现4种肉食螨种间变异位点分别为154 ~ 321个和58 ~ 99个。4种肉食螨COI与18S rRNA基因序列种内遗传距离均≤ 0.020,种间遗传距离分别为0.235 ~ 0.583和0.078 ~ 0.114。基于COI与18S rRNA基因序列的系统发育树显示,4种肉食螨均能各自聚为一支,支持率均达100%,与形态学鉴定结果一致。结论　线粒体COI基因序列作为4种肉食螨DNA条形码优于18S rRNA基因序列,更适用于探索肉食螨的属、种低阶元亲缘关系。     

Undisclosed Financial Conflicts of Interest of Authors of Clinical Drug Trials Published in Influential Medical Journals: A Cohort Study

The International Committee of Medical Journal Editors requires authors to disclose all financial conflicts of interest (COI) that can be perceived as influencing the related trials. Undisclosed financial COI may influence the perception of the authors' scientific impartiality and erode the public trust in the reported results. Data regarding completeness of COI disclosure in high-impact–factor general medicine journals are limited. We compared payments disclosed by US-based physicians who were first or last authors of clinical drug trials published between August 2016 and August 2018 in the New England Journal of Medicine, JAMA, and Lancet, to payments reported by industry to the Centers for Medicare & Medicaid Services Open Payments Database. Of 247 included authors, 198 (80%) have not disclosed some or all received payments. The median undisclosed sum was $8409 (US Dollars) (interquartile range [IQR] $123 to $44,890). Most authors (n=170, 69%) have received more than $10,000 per year (median $120,403, IQR $58,905 to $242,014). The median undisclosed sum for these authors was $26,530 (IQR $7462 to $71,562). Median undisclosed sums for authors of papers from studies performed with and without industry funding were $20,899 (IQR $4191 to $59,883) and $149 (IQR $0 to $3276), respectively. In 10 (8%) of 125 industry-funded trials, the first or last author had not disclosed personal payments from the study sponsor (median $9741, IQR $4508 to $101,484). These findings could raise concerns about the authors' equipoise toward the trial results and influence the public perception of the credibility of reported data. Health care professionals, reviewers, and journal editors should demand more transparent reporting of financial COI.     

浙江省杭州市入侵生物福寿螺种群遗传学研究

目的　对浙江省杭州市西湖区入侵生物福寿螺的分布现状及遗传标记进行分析，了解该地区福寿螺的扩散趋势。方法　2017年在杭州市西湖区各种水体（如湖泊、河流、湿地）及沿岸陆地选择5个采样点，现场采集福寿螺标本，对采集的标本进行形态学鉴定。同时取成螺足部肌肉组织提取总DNA进行COI基因扩增，并进行单倍型多样性、核酸多样性分析。此外，对GenBank中的福寿螺单倍型及本研究获得的单倍型进行系统进化树分析，以了解其近缘福寿螺。结果　在杭州市西湖区池塘、河流、湿地等生境均发现福寿螺。根据形态特征，采集的标本初步鉴定为福寿螺。从上述福寿螺DNA样本中共获得16条序列，属于3个单倍型，分别为Hap1、Hap2、Hap3。单倍型中Hap1、Hap3频率较高，Hap2频率相对较低。以上表明本次采集的标本包括小管福寿螺和斑点福寿螺两种。Hap1与来自阿根廷及中国广东省和香港特别行政区的小管福寿螺具有较近的亲缘关系，Hap2与来自阿根廷、日本、中国广东省广州市的小管福寿螺具有较近的亲缘关系，Hap3与来自阿根廷、巴西的斑点福寿螺具有较近的亲缘关系。结论　杭州市西湖区存在小管福寿螺和斑点福寿螺。斑点福寿螺扩散至西湖区可能是由多次引入导致或随水流扩散蔓延而致。     

基于可视化环介导等温扩增技术快速检测中药材蜈蚣真伪＊

目的 建立少棘巨蜈蚣的环介导等温扩增（loop-mediated isothermal amplification，LAMP）的方法，以实现对市场蜈蚣药材真伪的快速检测。方法 针对少棘巨蜈蚣COI序列设计专属性LAMP引物；考察恒温条件下最佳反应时间；通过对DNA模板的梯度稀释，确定LAMP反应对蜈蚣药材的最低检测浓度；对少棘巨蜈蚣及其伪品进行LAMP扩增反应，以考察LAMP引物对少棘巨蜈蚣的特异性。结果 筛选得到了一套特异性较好可用于检测少棘巨蜈蚣的LAMP引物，LAMP反应最佳反应时间为30 min，皮克级别的DNA就能被显著检出。结论 LAMP反应对于少棘巨蜈蚣的检测显示了良好的特异性，使用简便，有用于指导市场流通过程中蜈蚣药材的快速检测的潜力。     

Developing social presence in online learning among nurses: Exploration of the community of inquiry models domain of social using a qualitative descriptive design

There is marked increase in the demand for online learning in nursing curriculums. However, online learning has been criticised for being detached and impersonal resulting in low levels of engagement. In a world that is rapidly shifting to online learning as a result of the global COVID-19 pandemic creating quality online learning environments has become more pressing. The Community of Inquiry (COI) is a well-established model used to frame online environments in many disciplines including nursing. The COI model proposes that social, cognitive and teacher presences must exist in the online learning space for learning to occur. It is argued that social presence remains less understood despite being one of the essential elements. Therefore, the objectives of this study were to understand how facilitators and participants engage with and use a social presence in an online seminar. It is concluded that when facilitators include the use of a social presence in the design of the online learning space in-depth engagement with online content also increases. This can happen rapidly (less than an hour) and continue for several weeks. It is deduced that a social presence enhances a sense of realness among participants of online learning and through an increased sense realness engagement increases.     

基于DNA条形码技术鉴定青海柴达木盆地小型兽类

目的 为弥补传统形态分类方法不足,提高小型兽类现场鉴定成功率和准确性。方法 将采自青海省柴达木盆地总计3目5科12属145只小型兽类进行COI基因序列检测。结果 分析所测全部COI基因序列,种内遗传距离0.01%~2.2%,种间遗传距离10.9%~15.6%,种间遗传距离显著大于种内遗传距离。属间遗传距离16.2%~21.3%,科间遗传距离16.2%~27.0%, NJ树显示同种个体聚为高支持度的单一分支,在14个形态种中共发现17个分子种。讨论 研究结果表明DNA条形码技术能纠正传统形态学鉴定中的错误, 提高动物分类鉴定的准确性。