Development of a real-time PCR assay for the specific detection and identification of Streptococcus pseudopneumoniae using the recA gene

We sequenced the evolutionarily conserved genes 16S rRNA, atpD, tuf, and recA from Streptococcus pseudopneumoniae, Streptococcus pneumoniae, Streptococcus mitis, and Streptococcus oralis. Phylogenetic analysis revealed that recA provided good resolution between these species, including discrimination of the novel species S. pseudopneumoniae. By contrast, the more conserved 16S rRNA, tuf and atpD are not sufficiently discriminatory. Therefore, recA sequences were used to develop a real-time PCR assay with a locked nucleic acid-mediated TaqMan probe for the specific detection and identification of S. pseudopneumoniae. The PCR assay showed excellent specificity and a detection limit of <10 genome copies for the detection and identification of S. pseudopneumoniae strains, which makes it a promising tool for molecular identification and epidemiological studies. In conclusion, this article describes for the first time a PCR assay for the specific identification of S. pseudopneumoniae.     

Nanoprobes for in vitro diagnostics of cancer and infectious diseases

The successful and explosive development of nanotechnology is significantly impacting the fields of biology and medicine. Among the spectacular developments of nanobiotechnology, interest has grown in the use of nanomaterials as nanoprobes for bioanalysis and diagnosis. Herein, we review state-of-the-art nanomaterial-based probes and discuss their applications in in vitro diagnostics (IVD) and challenges in bringing these fields together. Major classes of nanoprobes include quantum dots (QDs), plasmonic nanoparticles, magnetic nanoparticles, nanotubes, nanowires, and multifunctional nanomaterials. With the advantages of high volume/surface ratio, surface tailorability, multifunctionality, and intrinsic properties, nanoprobes have tremendous applications in the areas of biomarker discovery, diagnostics of infectious diseases, and cancer detection. The distinguishing features of nanoprobes for in vitro use, such as harmlessness, ultrasensitivity, multiplicity, and point-of-care use, will bring a bright future of nanodiagnosis.     

人脂肪干细胞的体外培养鉴定及分化特性

目的：在体外从脂肪块中分离出脂肪干细胞（adipose derived stem/stromal cells,ASCs）,对其进行形态观察、干细胞鉴定、增殖和分化能力检测。方法：将腹部取皮术的皮下脂肪利用胶原酶消化法,进行体外分离培养,取第3代的细胞进行细胞爬片HE染色、流式鉴定、MTT、细胞周期检测等,利用成脂和成骨培养液诱导,油红O和茜素红染色鉴定诱导结果。结果：原代培养第1次换液时细胞多呈多角形和短梭形,第3代ASCs细胞爬片HE染色显示形态多为长梭形,呈漩涡状生长;流式鉴定显示：CD29＋,CD31-,CD34-,CD44＋,CD45-,CD49＋,CD106-,CD133-;MTT显示ASCs生长增殖活性强;细胞周期检测结果显示：G1=86.8%,G2=8.77%;成脂诱导后油红O染色阳性,对照组为阴性;成骨诱导后茜素红染色阳性,对照组为阴性。结论：人ASCs具有贴壁生长、多向分化以及干细胞表型等特征,且生长增殖活性强,是一种很有应用前景的间充质干细胞。     

HPLC-DAD-MS/MS对栀子中色素成分的研究

目的鉴定栀子中藏红花素类色素成分。方法色谱条件:Dikma Diamonsil C18色谱柱(4.6 mm×250 mm,5μm);0.15%甲酸水-甲醇梯度洗脱,流速0.6 mL/min,柱温30℃,检测波长440 nm。质谱条件:电喷雾离子源(ESI);负离子模式检测;扫描范围m/z 100～1 400。结合化学成分的紫外-可见吸收光谱特征,与多级质谱(MS/MS)裂解特征分析,对其结构进行确证。结果采用高效液相色谱-二级管阵列检测器(HPLC-DAD)鉴别了栀子中的31个藏红花素类色素成分,并根据MS/MS裂解规律推断了其中的23个化学成分。结论 HPLC-DAD-MS/MS可用于快速筛选和鉴定栀子中的藏红花素类色素成分。     

中医辨体施膳对糖尿病前期糖脂代谢的影响

目的:对糖尿病前期患者进行体质辨识的基础上研究中药膳食对体质的干预,并观察对糖脂代谢的影响。方法:将观察对象随机分为辨体施膳痰湿质观察组、湿热质观察组、阿卡波糖对照组,每组40例。每组采用相应的干预措施,干预6个月。结果:辨体施膳各组和对照组治疗前后空腹血糖及餐后血糖、血脂水平均有不同程度的降低,其中辨体施膳组血脂降低幅度大于对照组。结论:中医辨体施膳有可能经过多途径、多靶位来纠正病人偏颇的体质状态,达到调节和改善无症可辨的糖尿病前期患者的血糖、血脂指标。     

利用ISSR分子法标记鉴别新会茶枝柑

目的研究利用ISSR分子标记方法在核酸分子水平上鉴别新会茶枝柑。方法从28条ISSR引物中筛选合适的引物,对从新会茶枝柑、砂糖桔、阳山桔、贡柑、芦柑共16个新鲜叶片样品中提取的总DNA进行PCR扩增及电泳分析,寻找特征位点。结果新会茶枝柑所有样品与其中2条ISSR引物扩增出较为明显的特征条带,可区别于其他品种。结论 ISSR作为一种简便有效的分子标记方法,可用于新会茶枝柑的鉴别。     

新疆克拉玛依社区居民酒滥用情况及自测健康状况调查

目的 调查新疆社区居民酒滥用情况的发生率和酒滥用人群的健康状况.方法 采用多阶段随机抽样法,使用自行设计的社会人口学特征调查表、酒精使用障碍筛查量表(AUDIT)和自测健康评定量表(SIIRMS)对1992例社区居民进行调查,采用单因素分析和秩和检验等方法,计算酒滥用的发生率、在不同人群中的分布情况和酒滥用者的自测健康状况.结果 筛查出酒滥用者298例,总体人群发生率为15.1％(男性26.4％,女性4.8％),其中以男性、中年、高文化程度、在职、行政管理类工作、高收入水平、家中常住人口数较多者高发,各组间差异具有统计学意义,推测职场人群为酒滥用的高危人群.酒滥用者的自测健康状况明显差于普通人群(P＜0.01),尤其表现在心理健康和社会健康方面.结论 新疆地区酒滥用发生率较高,需要得到重视,建议针对高危人群开展进一步的筛查和早期干预工作,达到预防和控制的目的.     

Evaluation of a PCR-RFLP-ITS2 assay for discrimination of Anopheles species in northern and western Colombia

Anopheles mosquitoes are routinely identified using morphological characters of the female that often lead to misidentification due to interspecies similarity and intraspecies variability. The aim of this work was to evaluate the applicability of a previously developed PCR–RFLP–ITS2 assay for accurate discrimination of anophelines in twelve localities spanning three Colombian malaria epidemiological regions: Atlantic Coast, Pacific Coast, and Uraba-Bajo Cauca-Alto Sinu region. The evaluation of the stability of the PCR–RFLP patterns is required since variability of the ITS2 has been documented and may produce discrepancies in the patterns previously reported. The assay was used to evaluate species assignation of 939 mosquitoes identified by morphology. Strong agreement between the morphological and molecular identification was found for species Anopheles albimanus, Anopheles aquasalis, Anopheles darlingi and Anopheles triannulatus s.l. (p ≥ 0.05, kappa = 1). However, disagreement was found for species Anopheles nuneztovari s.l., Anopheles neomaculipalpus, Anopheles apicimacula and Anopheles punctimacula (p ≤ 0.05; kappa ranging from 0.33 to 0.80). The ITS2–PCR–RFLP assay proved valuable for discriminating anopheline species of northern and western Colombia, especially those with overlapping morphology in the Oswaldoi Group.     

新型洛菲不动杆菌的分离与鉴定

目的对从呼吸道感染的分泌物中分离的1株革兰阴性球杆样细菌进行研究,探讨其种系发生及分类学特征。方法利用形态、生理、生化特性等表型性状进行初步鉴定,通过细菌16S rRNA基因测序及基因比对明确其种系来源。结果分离菌为革兰阴性球杆状菌,生化反应不活泼,对环丙沙星耐药;BLAST序列比对显示该分离菌与洛菲不动杆菌的相似度为99.2%,与洛菲不动杆菌标准株不同的是在底物利用试验上有8项结果存在差异。结论根据表型性状和种系发生结果证实该菌为洛菲不动杆菌,鉴于该菌株与模式株在生化特性方面存在一定差异,初步考虑该菌株为洛菲不动杆菌的新亚型。     

哈尔滨市污水处理厂原水中安氏隐孢子虫的分子鉴定

目的对哈尔滨市污水处理厂原水中隐孢子虫进行分子鉴定。方法提取污水处理厂原水中的隐孢子虫基因组DNA,对其18S rRNA基因进行巢式PCR扩增,通过序列分析确定隐孢子虫虫种,并与GenBank的参考序列进行同源性分析。结果从哈尔滨市污水处理厂原水中分离鉴定出安氏隐孢子虫,该虫株18S rRNA基因序列与文献报道的安氏隐孢子虫序列(EU926592)同源性为100%。结论哈尔滨市污水处理厂原水中存在安氏隐孢子虫,对当地畜牧业的发展构成严重威胁,但对当地人群危害较小。