Application of recombinase polymerase amplification method for rapid detection of infectious laryngotracheitis virus

Infectious laryngotracheitis is a significant respiratory disease of chickens that causes huge economic losses due to high morbidity and mortality and reduced egg production. A real-time recombinase polymerase amplification (RPA) assay was developed to accurately detect ILTV. The specific probe and primer sets were carefully designed and screened. The real-time RPA assay was carried out at 39 °C for 30 min, and results were obtained within 15 min. The results of the specificity assay showed no fluorescence signals with other avian-related viruses. The sensitivity of the assay was 1 × 10² copies/μL. The low CV value showed that the assay was reproducible. A total of 115 clinical samples were tested using the real-time RPA assay and the real-time PCR assay in parallel; the coincidence rates of the two detection methods were 100%. The results indicated that the real-time RPA assay is a specific, sensitive, rapid, and useful tool for epidemiological studies and clinical diagnosis, especially in the field and in resource-poor areas.     

四妙勇安汤方源探析

四妙勇安汤由金银花、玄参、当归、甘草4味药物组成,为《古代经典名方目录(第一批)》中100个经典名方之一。经溯源发现,四妙勇安汤源于《石室秘录》,后由《古今图书集成·医部全录》《验方新编》等书引用。从古代文献记载来看均有方无名,"四妙勇安汤"之名,最早见于1956年《中医治疗动脉栓塞性坏疽症的成效》,是由当时记者吕民报道河北省释迦宝山用"四妙勇安汤"治疗当地的动脉栓塞性坏疽时冠名。四妙勇安汤从方药组成与剂量上看,从《石室秘录》开始即是"金银花三两,当归二两,生甘草一两,玄参三两",历代版本《方剂学》确定四妙勇安汤金银花、玄参、当归、甘草的比例就是3∶3∶2∶1。而查阅文献,释迦宝山临证所用的四妙勇安汤由"玄参132 g,当归99 g,银花66 g,甘草33 g"组成,金银花、玄参、当归、甘草的比例变成2∶4∶3∶1。从治疗时间上看,原方记载的7日愈或是10日愈,而释迦宝山将其用到了三四个月,甚至五六个月。研究认为,古籍中的四妙勇安汤,应该是用于疾病的初期,尽早发现和治疗;而释迦宝山修改过的剂量,是广泛用于脱骨疽的中后期,甚至出现坏疽的严重病情所使用的,因此服药时间长,剂量大。且四妙勇安汤临证不仅限于治疗脱骨疽,也用于大头疮等,现代该方的使用已经大为拓展。相关研究已证实四妙勇安汤具有抗炎、稳定斑块、降脂、保护血管、改善血液流变学、抗凝、抑制血栓形成和促纤溶等作用,后续应开展君臣佐使辨析,对其临床应用范围重新进行界定。     

用高分辨G带和人工细菌染色体荧光原位杂交技术分析中国儿童孤独症患者的染色体改变

目的 检测中国儿童孤独症患者的特征性染色体改变。方法 应用高分辨G带和人工细菌染色体(bacterial artificial chromosome，BAC)荧光原位杂交(flourescence in situ hybridization，FISH)分析68例中国儿童孤独症患者的染色体改变。结果 用G带分析观察到有染色体改变的4例患者，分别为1例t(4；6)(q23-24；p21)、1例21p 和2例9号染色体臂间倒位。BAC FISH进一步证实易位病例，而且更精确[t(4；6)(q25-26；p21-1)]；涉及7号、15号、2号染色体的BAC FISH均未观察到文献中报道的染色体改变；而9号染色体的臂间倒位和21p 因无BAC克隆而无法证实。结论 用G带和BAC FISH发现少数中国孤独症患者有染色体改变，但远没有文献中报道的10％～48％那么高。BAC FISH有助于精确地确定染色体易位断裂点。     

Linkage Disequilibrium and Haplotype Architecture for two ABC Transporter Genes (ABCC1 and ABCG2) in Chinese Population: Implications for Pharmacogenomic Association Studies

Information about linkage disequilibrium (LD) patterns and haplotype structures for candidate genes is instructive for the design and analysis of genetic association studies for complex diseases and drug response. ABCC1 and ABCG2 are genes coding for two multidrug resistance (MDR) associated transporters; they are also related to some pathophysiological traits. To pinpoint the LD profiles of these MDR genes in Chinese, we systemically screened 27 unrelated individuals for single nucleotide polymorphisms (SNPs) in the coding and regulatory regions of these genes, and thereby characterized their haplotype structures. Despite marked variations in haplotype diversity, LD pattern and intragenic recombination intensity between the two genes, both loci could be partitioned into several LD blocks, in which a modest number of haplotypes accounted for a high fraction of the sampled chromosomes. We concluded that each locus has its own genomic LD profile, but that they still share a common segmental LD architecture with low haplotype diversity. Our data will benefit genetic association studies of complex traits and drug response possibly related to these genes.     

Large enthesophytes in teenage skulls: Mechanical,inflammatory and genetic considerations

BackgroundThe literature implies that large enthesophytes are exclusive to genetically predisposed individuals and to Spondyloarthropathies sufferers. Accordingly, the aim of this investigation and report was to assess the involvement of genetic predisposition, inflammatory and/or mechanical influences in the development of large enthesophytes in a sample population of teenagers presenting with large enthesophytes emanating from the external occipital protuberance.MethodsAnalysis was based on four teenage males (13–16 year-old) possessing 14.5–30.5 mm enthesophytes projecting from the external occipital protuberance. This study included assessment of radiographs, MRI scans, blood-work, history, the SF-36 health survey, and the comparison of these data with the relevant literature to describe the interrelationships between the presence of enlarged external occipital protuberance, forward head protraction, active inflammation and/or genetic factors.FindingsKnown genetic markers (e.g. HLA-B27) were not detected by allele-specific primers and both ESR and CRP tests were negative. Additionally, MRI analyses failed to detect active localised inflammation at the external occipital protuberance and surrounding structures. The health survey yielded normal parameters for all participants. All participants displayed significantly large Forward Head Protraction values (>40 mm), and interviews with participants and their parents indicated that concerns related to posture were prevalent since early childhood.InterpretationThis report suggests that mechanical load has an important role in enthesophyte development, irrespective the involvement of inflammatory or genetic factors.     

层析法提纯A、C、Y、W135群脑膜炎球菌荚膜多糖工艺的建立

目的 建立A、C、Y、W135群脑膜炎球菌荚膜多糖层析纯化工艺。方法 采用陶瓷羟基磷灰石和DEAE Sepharose FF层析柱，在PBS体系中纯化A群多糖。以0.5%脱氧胆酸钠预处理C群多糖，用陶瓷羟基磷灰石层析柱在PBS体系中纯化C群多糖。以含有脱氧胆酸钠的平衡缓冲液溶解Y和W135群多糖，再用Capto Adhere和Capto DEAE层析柱串联纯化。纯化的多糖经Sephadex G-25 Medium层析柱脱盐后冻干，按中国药典2015年版三部的要求进行检定。结果  经过层析纯化，A、C、Y、W135群多糖的蛋白质含量分别降至3.7、4.2、5.4和5.3 mg/g，核酸含量分别降至1.2、3.0、1.1和0.8 mg/g，均符合药典要求。此外，磷、唾液酸和O-乙酰基含量等指标亦均符合药典标准。结论 建立了A、C、Y、W135群脑膜炎球菌荚膜多糖的层析纯化工艺。     

CBL结合循证医学思维在临床免疫学检验实验课教学中的应用

目的 探讨案例教学法 (case-based learning，CBL) 结合循证医学思维在临床免疫学检验实验课教学中的应用价值。方法 选取首都医科大学 2016 级本科临床检验专业 A 班学生为对照组，B 班学生为实验组，实验组采用常规教学方法结合 CBL 结合循证医学思维的教学方法进行教学，对照组只采用常规教学方法教学，最后以笔试、问卷调查方式进行分析。结果 90% 以上的学生认为 CBL 结合循证医学思维的教学方法新颖、生动，有助于对理论知识的理解和记忆，理论联系实际。对照组考试平均分数为 (84.5±5.1) 分，实验组为 (88.1±4.3) 分；对照组病例分析题 ( 总分 40 分 ) 平均分为 (27.2±3.7) 分，实验组为 (36.3±4.1) 分。结论 CBL 结合循证医学思维的教学方法培养学生提出、分析和解决问题的能力，能帮助学生更好地掌握临床免疫学检查技术。