金黄色葡萄球菌毒性休克综合征毒素1核酸适配体的筛选和鉴定

目的:筛选能特异性识别毒性休克综合征毒素1(TSST-1)的DNA适配体,为金黄色葡萄球菌感染的治疗奠定实验基础。方法:体外合成含有25个随机序列全长为63个碱基的单链DNA(ss DNA)文库,以TSST-1为靶标,利用指数富集的配体系统进化技术(SELEX),从ss DNA文库中筛选TSST-1适配体,利用生物信息学方法对适配体进行序列分析和结构预测,荧光定量法分析适配体的亲和力及特异性。结果:经过8轮筛选,文库的亲和力不断升高,获得了能识别TSST-1的DNA适配体T-7,它可选择性地与TSST-1结合,并测定其Kd值为103.8 nmol/L。结论:新型适配体T-7能选择性识别TSST-1,在金黄色葡萄球菌感染的治疗和诊断方面具有应用前景。     

运用分子生物学技术监测水环境中产毒蓝藻

有毒蓝藻在形成水华破坏水体环境的同时,也对人畜产生危害。运用分子生物学技术对水环境中产毒蓝藻进行监测,由于其简单、快速和经济等优点,逐渐成为各国学者关注的热点。本文从以下3个方面对其进行了综述:1)早期分子生物学技术在产毒蓝藻诊断中的运用;2)以产毒基因为靶标进行的产毒蓝藻诊断;3)运用基因芯片技术对产毒蓝藻进行检测。与传统的分子生物学技术相比,生物芯片技术具有体积小、重量轻、便于携带、分析自动快速、高通量等许多传统方法所不能比拟的优势。因此,该技术在蓝藻毒素检测中的运用必将给目前的产毒蓝藻的检测带来一场新的革命。     

中国沿海新报道的三种产毒拟菱形藻

为了明确我国海域拟菱形藻属(Pseudo-nitzschia)物种的产毒特征, 从中国沿海建立了15个拟菱形藻单克隆培养株系, 利用高效液相色谱-质谱联用法对其多莫酸特征进行检测, 在10个株系中检测到多莫酸。结合光学显微镜下的群体特征和透射电镜下的超微形态学特征, 以及基于核糖体转录间隔区的分子系统学数据, 确认上述10个产毒株系分别隶属于3个物种:尖细拟菱形藻P. cuspidata、伪柔弱拟菱形藻P. pseudodelicatissima、伪善拟菱形藻P. fraudulenta, 其中伪善拟菱形藻是我国的新记录种。建立尖细拟菱形藻的11个尖细拟菱形藻株系, 其中3个株系未检出多莫酸, 其余8个株系有检出, 单细胞产毒水平为0.4—5.5 fg。建立伪柔弱拟菱形藻株系2个, 1个未检出多莫酸, 另1个株系的单细胞产毒量为1 fg。建立伪善拟菱形藻株系2个, 纯种培养株系均未检出多莫酸。利用卤虫(Artemia salina)对部分藻株进行混培诱导, 其中尖细拟菱形藻(MC4049)和伪柔弱拟菱形藻(MC3015)的产毒水平略有下降, 单细胞产毒水平分别由2、1 fg降至0.2、0.4 fg, 而伪善拟菱形藻(MC4074)的产毒能力则有显著改变, 单细胞产毒水平由未检出上升至17.5 fg。研究丰富了我国产毒拟菱形藻的物种多样性, 明确了其物种信息和产毒水平, 可为后续深入研究提供基础数据。     

荧光原位杂交技术和流式细胞仪用于环境样品中产毒及非产毒微囊藻的定量监测

全球范围内,高频次、大范围暴发的蓝藻水华对淡水水体环境造成严重影响.微囊藻因其在生长特别是衰亡过程中向水体释放微囊藻毒素而威胁人类健康.因此,分析其产毒株及非产毒株在环境样品中的组成,建立产毒蓝藻的预报及评价体系显得极为重要.本文采用荧光原位杂交技术结合流式细胞技术实现对环境样品中产毒藻株的鉴别与定量.针对目标基因mcyA设计的、以地高辛标记的双链DNA探针可有效应用于产毒微囊藻FACHB905和PCC7806的鉴别.分别对来自滇池、太湖和关桥的11个样品进行分析显示,该方法与传统的形态学鉴定及PCR方法有较好的匹配.荧光原位杂交技术与流式细胞相结合可有效鉴别产毒与非产毒微囊藻,尤其可以对野外样品中产毒与非产毒藻株进行简便、可视化地鉴别,从而达到对产毒微囊藻水华早期预警的目的.     

携带TSST-1和PVL基因的金黄色葡萄球菌的耐药特点、分布特征及与致病性的关系

目的:探讨携带中毒休克综合征毒素-1(TSST-1)和杀白细胞毒素(PVL)基因的金黄色葡萄球菌的耐药特点、分布特征及其与致病性的关系。方法:收集金黄色葡萄球菌临床分离菌株93株,采用聚合酶链反应(PCR)检测TSST-1基因和PVL基因,采用琼脂扩散法检测金黄色葡萄球菌菌株对青霉素(PEN)、苯唑西林(OXA)、头孢噻吩(CEF)、氨苄西林(AMP)、头孢噻肟(CTX)、阿莫西林/克拉维酸(AMC)、亚胺培南(IPM)、克拉霉素(CLR)、万古霉素(VAN)、环丙沙星(CIP)、庆大霉素(GM)、左氧氟沙星(LVX)和利福平(RA)13种抗菌药物的耐药性。结果:耐甲氧西林金黄色葡萄球菌(MRSA)占总数的88.2%,甲氧西林敏感金黄色葡萄球菌(MSSA)占总数的11.8%。TSST-1+菌株在MRSA、MSSA中分别占12.2%、0,差异无统计学意义(P0.05);PVL+菌株在MRSA、MSSA中分别占40.2%、9.1%,差异有统计学意义(P0.05)。MRSA存在明显的耐药性,且表现出多药耐药性,而携带TSST-1与PVL基因的MRSA耐药性更严重。结论:MRSA在金黄色葡萄球菌中的分离率高,耐药性严重,携带TSST-1与PVL基因的MRSA耐药性与致病力增加。     

黄曲霉群菌种产生黄曲霉毒素B_1的调查研究

本文对来自我国20个省、市、自治区的不同基物上分离的和中国科学院微生物研究所菌种保藏室以及其他单位提供的黄曲霉群菌种,经随机选取82株进行了黄曲霉毒素B_1的测定,证明在测试的9个已知分类群中产生黄曲霉毒素B_1的菌种只限于寄生曲霉和黄曲霉,另外4株种名未定者也能产生此种毒素。在黄曲霉中产毒菌株约占30％(28.3％),其在GAN(葡萄糖硝酸铵)和大米培养基中的黄曲霉毒素B_1的最高产量分别为133,333.3和160,000.0ppb。总的来说,大体上可以反映在我国一般基物上黄曲霉产毒菌株存在的现状。在实验过程中,还对黄曲霉群菌种在GAN和大米培养基中黄曲霉毒素B_1的产量和产毒菌株数作了比较。发现在大米培养基中黄曲霉毒素B_1的产量高于GAN,而且测试的黄曲霉产毒菌株在这两种培养基中均各有不能产毒的菌株,因此,在测定产毒菌株时,若仅采用其中一种产毒培养基,往往会有漏掉产毒菌株的可能性。     

黄曲霉群菌种产生黄曲霉毒素B1的调查研究

本文对来自我国20个省、市、自治区的不同基物上分离的和中国科学院微生物研究所菌种保藏室以及其他单位提供的黄曲霉群菌种,经随机选取82株进行了黄曲霉毒素B_1的测定,证明在测试的9个已知分类群中产生黄曲霉毒素B_1的菌种只限于寄生曲霉和黄曲霉,另外4株种名未定者也能产生此种毒素。在黄曲霉中产毒菌株约占30％(28.3％),其在GAN(葡萄糖硝酸铵)和大米培养基中的黄曲霉毒素B_1的最高产量分别为133,333.3和160,000.0ppb。总的来说,大体上可以反映在我国一般基物上黄曲霉产毒菌株存在的现状。在实验过程中,还对黄曲霉群菌种在GAN和大米培养基中黄曲霉毒素B_1的产量和产毒菌株数作了比较。发现在大米培养基中黄曲霉毒素B_1的产量高于GAN,而且测试的黄曲霉产毒菌株在这两种培养基中均各有不能产毒的菌株,因此,在测定产毒菌株时,若仅采用其中一种产毒培养基,往往会有漏掉产毒菌株的可能性。     

黑曲霉对黄曲霉生长、产毒及黄曲霉毒素B1的影响

目的研究黑曲霉对黄曲霉生长、产毒的抑制作用及对AFB1的降解作用。方法将黑曲霉分别与黄曲霉、AFB1共同培养,定期测定培养液pH、菌丝体干重、黄曲霉孢子数、AFB1含量。结果黑曲霉与黄曲霉混合培养时,黄曲霉孢子数、AFB1含量均比单独培养的低,2组之间差异有统计学意义(P<0.05),抑制率达到68.06%~91.52%;加入黑曲霉后,AFB1含量降低,实验组与对照组之间差异有统计学意义(P<0.05),降解率为46.19%。结论黑曲霉既能抑制黄曲霉生长、产毒,又能降解AFB1。     

黄曲霉产毒菌株的快速筛选实验

黄曲霉产毒菌株的常规鉴定方法多采用待检菌株经产毒培养后用薄层层析法测定其产毒能力。这种方法需要较完善的实验条件,操作也较为繁琐。1975年以来,为鉴定从谷物、食品和饲料中分离出的大批黄曲霉菌株的产毒性能,我们曾参考了     

水华蓝藻产毒特性的PCR检测法

特异引物对（TOX 1P／1F;TOX 2P／2F）用于检测微囊灌毒素合成酶基因mcyB片段在38种水华蓝藻中的分布情况。结果显示,所有能产生微囊灌毒素的微囊藻都有特异扩增条带,非产毒株则没有,几种常规的毒性检测方法验证了PCR方法所获结果的准确性。本研究发展了以全细胞PCR法检测mcyB片断,说明全细胞PCR检测法适用于不同来源的蓝藻材料。结果证明以DNA为基因鉴别产毒和非产毒微囊藻及其他水华蓝藻的方法是可行的和实用的。     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.