Evaluation of Commercial Diagnostic Assays for the Specific Detection of Avian Influenza A (H7N9) Virus RNA Using a Quality-Control Panel and Clinical Specimens in China

A novel avian influenza A H7N9-subtype virus emerged in China in 2013 and threatened global public health. Commercial kits that specifically detect avian influenza A (H7N9) virus RNA are urgently required to prepare for the emergence and potential pandemic of this novel influenza virus. The safety and effectiveness of three commercial molecular diagnostic assays were evaluated using a quality-control panel and clinical specimens collected from over 90 patients with confirmed avian influenza A (H7N9) virus infections. The analytical performance evaluation showed that diverse influenza H7N9 viruses can be detected with high within- and between-lot reproducibility and without cross-reactivity to other influenza viruses (H1N1 pdm09, seasonal H1N1, H3N2, H5N1 and influenza B). The detection limit of all the commercial assays was 2.83 Log₁₀ copies/μl [0.7 Log₁₀TCID₅₀/mL of avian influenza A (H7N9) virus strain A/Zhejiang/DTID-ZJU01/2013], which is comparable to the method recommended by the World Health Organization (WHO). In addition, using a WHO-Chinese National Influenza Center (CNIC) method as a reference for clinical evaluation, positive agreement of more than 98% was determined for all of the commercial kits, while negative agreement of more than 99% was observed. In conclusion, our findings provide comprehensive evidence for the high performance of three commercial diagnostic assays and suggest the application of these assays as rapid and effective diagnostic tools for avian influenza A (H7N9) virus in the routine clinical practice of medical laboratories.     

Oxidative stress mediates chemical hypoxia-induced injury and inflammation by activating NF-κb-COX-2 pathway in HaCaT cells

Hypoxia of skin is an important physiopathological process in many diseases, such as pressure ulcer, diabetic ulcer, and varicose ulcer. Although cellular injury and inflammation have been involved in hypoxia-induced dermatic injury, the underlying mechanisms remain largely unknown. This study was conducted to investigate the effects of cobalt chloride (CoCl₂), a hypoxia-mimicking agent, on human skin keratinocytes (HaCaT cells) and to explore the possible molecular mechanisms. Exposure of HaCaT cells to CoCl₂ reduced cell viability and caused overproduction of reactive oxygen species (ROS) and oversecretion of interleukin-6 (IL-6) and interleukin-8 (IL-8). Importantly, CoCl₂ exposure elicited overexpression of cyclooxygenase-2 (COX-2) and phosphorylation of nuclear factor-kappa B (NF-κB) p65 subunit. Inhibition of COX-2 by NS-398, a selective inhibitor of COX-2, significantly repressed the cytotoxicity, as well as secretion of IL-6 and IL-8 induced by CoCl₂. Inhibition of NF-κB by PDTC (a selective inhibitor of NF-κB) or genetic silencing of p65 by RNAi (Si-p65), attenuated not only the cytotoxicity and secretion of IL-6 and IL-8, but also overexpression of COX-2 in CoCl₂-treated HaCaT cells. Neutralizing anti-IL-6 or anti-IL-8 antibody statistically alleviated CoCl₂-induced cytotoxicity in HaCaT cells. N-acetyl-L-cysteine (NAC), a well characterized ROS scavenger, obviously suppressed CoCl₂-induced cytotoxicity in HaCaT cells, as well as secretion of IL-6 and IL-8. Additionally, NAC also repressed overexpression of COX-2 and phosphorylation of NF- B κ p65 subunit induced by CoCl₂ in HaCaT cells. In conclusion, our results demonstrated that oxidative stress mediates chemical hypoxia-induced injury and inflammatory response through activation of NF-κB-COX-2 pathway in HaCaT cells.     

Cumulative association between age-related macular degeneration and less studied genetic variants in PLEKHA1/ARMS2/HTRA1: a meta and gene-cluster analysis

The objective of this study is to examine the cumulative effect of the less studied genetic variants in PLEKHA1/ARMS2/HTRA1 on age-related macular degeneration (AMD). We performed an extensive literature search for studies on the association between AMD and the less studied genetic variants in PLEKHA1/ARMS2/HTRA1. Multiple meta-analyses were performed to evaluate the association between individual genetic variants and AMD. A gene-cluster analysis was used to investigate the cumulative effect of these less studied genetic variants on AMD. A total of 23 studies from 20 published papers met the eligibility criteria and were included in our analyses. Several genetic variants in the gene cluster are significantly associated with AMD in our meta-analyses or in individual studies. Gene-cluster analysis reveals a strong cumulative association between these genetic variants in this gene cluster and AMD (p < 10^?5). However, two previously suspected SNPs in ARMS2, including rs2736911, the SNP having the largest number of studies in our meta-analyses; and rs3793917, the SNP with the largest sample size, were not significantly associated with AMD (both p’s > 0.12). Sensitivity analyses reveal significant association of AMD with rs2736911 in Chinese but not in Caucasian, with c.372_815del443ins54 in Caucasian but not in Chinese, and with rs1049331 in both ethnic groups. These less studied genetic variants have a significant cumulative effect on wet AMD. Our study provides evidence of the joint contribution of genetic variants in PLEKHA1/ARMS2/HTRA1 to AMD risk, in addition to the two widely studied genetic variants whose association with AMD was well established.     

Inheritance and mapping of the ore gene controlling the quantity of β-carotene in cucumber (Cucumis sativus L.) endocarp

The metabolic precursor of vitamin A, ??-carotene, is essential for human health. The gene(s) controlling ??-carotene quantity (Q??C) has been introgressed from Xishuangbanna gourd (XIS, possessing ??-carotene; Cucumis sativus L. var. xishuangbannanesis Qi et Yuan; 2n?=?2x?=?14) into cultivated cucumber (no ??-carotene; Cucumis sativus L.). To determine the inheritance of Q??C in cucumber fruit endocarp, F1 progeny and a set of 124 F7 recombinant inbred lines (RILs) derived from the cultivated cucumber line CC3 and XIS line SWCC8 were evaluated for Q??C during 2009 and 2010 in Nanjing, China. Segregation analysis revealed that endocarp Q??C of greenhouse-grown fruit was controlled by a single recessive gene. Further, marker analysis indicated the gene controlling Q??C was linked to seven SSR markers on linkage group 3, where their order was SSR20710?CSSR19511?CSSR15419?CSSR07706?Core?CSSR23231?CSSR11633?CSSR20270. These markers and the putative candidate gene were mapped to cucumber chromosome 3DS. An evaluation of 30 genetically diverse cucumber lines indicated that marker SSR07706 has utility in further genetic analyses of the Q??C orange endocarp gene, designated ore. Moreover, the markers defined herein may have utility for marker-assisted selection directed towards the development of cucumber germplasm with high fruit ??-carotene content.     

Two new flavonol glycosides from Gymnema sylvestre and Euphorbia ebracteolata

Two new flavonol glycosides, namely kaempferol 3-O-beta-D-glucopyranosyl-(1-->4)-alpha-L-rhamnopyranosyl-(1-->6)-beta-D-galactopyranoside (1) and quercetin 3-O-6"-(3-hydroxyl-3-methylglutaryl)-beta-D-glucopyranoside (2), have been isolated from the aerial parts of Gymnema sylvestre and Euphorbia ebracteolata, respectively. Their structures were determined on the basis of chemical and spectroscopic methods.     

适合于咸宁基地生产的螺旋藻品系的选育研究

1　引　　言　　为综合利用螺旋藻资源,我国第一座利用高原碱水湖(ｐＨ8.6～9.3)、光、温、地热等自然资源[2]的中试基地在云南程海湖畔建成,并通过改造温棚和利用当地的地热在武汉咸宁建成第一个利用地热控温、半封闭、全循环、高洁净的螺旋藻生产基地.本项研究旨在选育一株适合于该基地特有自然条件生长的优良品系.2　自然概况与研究方法2.1　基地概况　　咸宁基地是由4个玻璃温室改建成的螺旋藻生产工厂,面积为1×104ｍ2.该基地结合当地自然条件和自然资源有如下特征:1)温度控制系统:热水资源(来自地热水)、冷水资源(来自流过工厂的小河)…     

不同海拔地区高原鼠兔探究性和静止代谢率的差异

环境的差异影响动物种群生活史对策的进化。对地理分布范围广的物种而言,栖息地环境差异大,不同种群的生活史对策亦存在显著差异。个性特征作为生活史的一部分,反映了动物对环境的适应模式。前期研究表明,不同海拔区域的高原鼠兔的生活史对策存在显著差异,然而,其个性特征及与能量代谢的关联性是否也存在差异,目前尚不清楚。2018年9-10月,分别在海拔3268 m的贵南县和3980 m的玛沁县测定当地高原鼠兔的探究性与静止代谢率。结果发现,高海拔雄性高原鼠兔的探究性显著低于低海拔雄性个体;高海拔雌性高原鼠兔体重矫正静止代谢率显著高于低海拔雌性个体。高海拔地区,雌性高原鼠兔的探究性和体重矫正静止代谢率呈显著正相关,雄性高原鼠兔的探究性和体重呈显著负相关;低海拔地区,高原鼠兔的探究性和体重矫正静止代谢率或体重之间均无显著相关性。该结果 表明,调整个性特征和能量代谢及其两者的关联性也是高原鼠兔适应不同海拔栖息地环境差异的生活史对策之一。     

One-Step Reverse-Transcription FRET-PCR for Differential Detection of Five Ebolavirus Species

Ebola is an emerging infectious disease caused by a deadly virus belonging to the family Filoviridae, genus Ebolavirus. Based on their geographical distribution, Ebolavirus has been classified into total five species so far, mainly Zaire, Sudan, Taï Forest, Bundibugyo and Reston. It is important to be able to differentiate the Ebolavirus species as they significantly differ in pathogenicity and more than one species can be present in an area. We have developed a one-step step-down RT-PCR detecting all five Ebolavirus species with high sensitivity (1 copy of Ebolavirus DNA, 10 copies of RNA and 320 copies of RNA spiked in 1 ml whole blood). The primers and FRET-probes we designed enabled us to differentiate five Ebolavirus species by distinct T_m (Zaire: flat peaks between 53.0°C and 56.9°C; Sudan: 51.6°C; Reston: flat peaks between 47.5°C and 54.9°C; Tai Forest: 52.8°C; Bundibugyo: dual peaks at 48.9°C and 53.5°C), and by different amplicon sizes (Zaire 255bp, Sudan 211bp, Reston 192bp, Taï Forest 166bp, Bundibugyo 146bp). This one-size-fit-all assay enables the rapid detection and discrimination of the five Ebolavirus species in a single reaction.