盐穗木金属硫蛋白HcMT在大肠杆菌中高效表达及金属离子结合能力的检测

旨在研究盐穗木金属硫蛋白Hc MT基因原核表达情况及结合金属离子的能力。从盐穗木中克隆获得的金属硫蛋白(Hc MT)基因亚克隆至p GEX-6p-2原核表达载体上,在大肠杆菌BL21中表达,分离纯化融白蛋白GST-Hc MT,通过原子吸收分光光度法测定融合蛋白GST-Hc MT结合金属离子的量以判断其与金属离子的结合能力。结果显示,诱导表达的融合蛋白GST-Hc MT分子量在34 k D左右,与预期一致,且通过Western blot进一步得到验证;原子吸收结果表明其具有结合Cu~(2+)、Zn~(2+)、Pb~(2+)、Cd~(2+)的能力,融合蛋白结合Cu~(2+)、Zn~(2+)、Pb~(2+)、Cd~(2+)的量分别是对照的25倍、10倍、4倍和2倍,其结合能力大小为Cu~(2+)Cd~(2+)Zn~(2+)Pb~(2+)。     

河南华溪蟹金属硫蛋白的GST融合表达及工程菌吸附重金属的研究

目的:构建河南华溪蟹金属硫蛋白(MT)的GST融合表达系统,诱导其可溶性表达,并检测转MT工程菌吸附重金属的能力。方法:采用基因工程技术,将河南华溪蟹MT的cDNA克隆至原核表达载体pGEX-6p-1,转化大肠杆菌BL21(DE3),SDS-PAGE鉴定融合蛋白的表达。在此基础上采用火焰原子吸收分光光度计测定工程菌对重金属离子Cu~(2+)、Cd~(2+)、Zn~(2+)的吸附能力。结果:pGEX-6p-1-MT重组质粒构建成功,SDS-PAGE分析表明GST-MT为可溶性表达,表达菌体对3种重金属的生物吸附能力均显著高于空菌体(P0.01)。结论:实现了华溪蟹MT的GST融合表达,转MT工程菌对Cu~(2+)、Cd~(2+)、Zn~(2+)具有显著吸附作用,且对Cd~(2+)的吸附作用最强。     

重金属铜、锌、镉复合胁迫对麻疯树幼苗生理生化的影响

该研究以Cu~(2+)、Zn~(2+)、Cd~(2+)单一胁迫为对照,探讨不同浓度的Cu~(2+)、Zn~(2+)、Cd~(2+)复合胁迫对麻疯树幼苗生理生化指标的影响。结果表明:随着Cu~(2+)、Zn~(2+)、Cd~(2+)浓度的增加,麻疯树幼苗叶片中的蛋白质(Pro)、丙二醛(MDA)含量均逐渐增加,其叶片叶绿素含量随着Zn~(2+)胁迫浓度的增加呈现出先降后升的趋势,在中等浓度(100 mg·L-1)的Zn~(2+)胁迫时含量最低、随着Cu~(2+)胁迫浓度的增加叶绿素含量先升高后降低,在Cu~(2+)浓度为200 mg·L-1时含量最高,达到1 200 mg·g-1FW; Cd~(2+)胁迫对叶绿素含量和根系活力无明显影响。根系活力在Zn~(2+)浓度为100 mg·L~(-1)时最强,随着Cu~(2+)浓度的增加而减弱。低浓度的Cu~(2+)、Zn~(2+)、Cd~(2+)对过氧化物酶活性和可溶性糖含量都具有促进作用。Cu~(2+)、Zn~(2+)、Cd~(2+)复合胁迫时对可溶性蛋白、叶绿素和丙二醛含量均无明显影响,随着复合胁迫时浓度的增加,可溶性糖含量和根系活力先增后减。这表明麻疯树对三种重金属的胁迫具有一定的抗性,过高浓度的胁迫会影响麻疯树幼苗生理生化的一些指标,但是麻疯树可以通过自身的防御系统使伤害降到最小。此外,重金属复合胁迫可以在一定程度上减轻单一胁迫对麻疯树幼苗造成的毒害作用。     

Cu2+、Zn2+、Cd2+对黑水虻幼虫生长的影响 及在虫体和虫粪的积累

为研究不同金属离子对黑水虻幼虫生长的影响及在虫体和虫粪中的积累分布规律,通过在黑水虻幼虫饲料中加入不同浓度的Cu~(2+)、Zn~(2+)、Cd~(2+),分析观测了幼虫增重、虫体和虫粪中金属含量等情况。结果表明,适量的Cu~(2+)、Zn~(2+)、Cd~(2+)可以促进黑水虻幼虫的生长,Zn~(2+)浓度在400 mg/kg时黑水虻幼虫的体重最高。Cu~(2+)、Zn~(2+)、Cd~(2+)在黑水虻幼虫体内和虫粪中的残留随着饲料中Cu~(2+)、Zn~(2+)、Cd~(2+)浓度的升高而增加。Cu~(2+)、Zn~(2+)在虫体中的残留比例都小于50%,大部分残留在虫粪中,Cd~(2+)在10、20、40 mg/kg处理中虫体的残留比例大于50%,最高达到了91.3%,而在80、160 mg/kg处理中虫体残留量比例小于50%。各重金属在虫粪中的残留主要以残渣态存在,且各形态的含量顺序在高浓度处理中都为残渣态酸溶态水溶态碱溶态。说明饲料中适量金属离子的添加有利于黑水虻幼虫生长,但过量的添加会抑制黑水虻幼虫生长,并造成金属离子的积累。     

重金属累积对泽兰实蝇幼虫生长发育和保护酶的影响

为了弄清重金属通过寄主植物紫茎泽兰累积后对专性寄生天敌泽兰实蝇的生理影响,采用模拟Cd~(2+)、Pb~(2+)或Zn~(2+)污染的土壤培养法胁迫紫茎泽兰,让泽兰实蝇低龄幼虫寄生取食,利用原子吸收分光光度法测定泽兰实蝇老熟幼虫中Cd~(2+)、Pb~(2+)或Zn~(2+)的含量,同时测定分析泽兰实蝇老熟幼虫的体长、鲜重、能量物质和保护酶的变化。结果表明,实验处理的Cd~(2+)、Pb~(2+)或Zn~(2+)均可通过紫茎泽兰传递并累积到泽兰实蝇老熟幼虫体内,引起老熟幼虫一系列生理变化:长度和鲜重降低;总糖、总蛋白和总脂的含量降低,相应的热量值也降低;超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)等保护酶的活性降低。本研究为进一步阐明重金属污染对食物链各环节的影响提供了重要的理论依据。     

重金属镉(Cd)在植物体内的转运途径及其调控机制

重金属镉(Cd)的毒害效应与其由土壤向植物地上部分运输有关,揭示Cd~(2+)转运途径及其调控机制可为提高植物抗镉性以及镉污染的植物修复提供依据。对Cd~(2+)在植物体内的转运途径,特别是限制Cd~(2+)移动的细胞结构和分子调控机制研究进展进行了回顾。Cd~(2+)通过共质体和质外体途径穿过根部皮层进入木质部的过程中,大部分在皮层细胞间沉积,少部分抵达中柱后转移到地上部分。为了免受Cd~(2+)的危害,植物体产生了多种限制Cd~(2+)吸收和转移的生理生化机制:1)环绕在内皮层径向壁和横向壁上的凯氏带阻止Cd~(2+)以质外体途径进入木质部;2)螯合剂与进入根的Cd~(2+)螯合形成稳定化合物并区隔在液泡中;3)通过H+/Cd~(2+)离子通道等将Cd~(2+)逆向转运出根部。植物共质体和质外体途径转运重金属镉的能力以及两条途径的串扰尚待进一步明晰和阐明。     

耐镉真菌Paecilomyces lilacinus 6-20p对Cd~(2+)的吸附特性研究

开展了一株高耐镉真菌Paecilomyces lilacinus 6-20p对Cd~(2+)的吸附效果及影响因素的研究。结果表明获取Paecilomyces lilacinus 6-20p最大生长量的培养时间为108 h。Paecilomyces lilacinus 6-20p对培养液中Cd~(2+)的去除率随pH、培养温度、培养时间呈现先增高后降低的趋势,对Cd~(2+)吸附的最佳条件为pH 3.5,30℃培养108 h。培养液中Cd~(2+)初始浓度小于35.2 mg/L时,Paecilomyces lilacinus 6-20p的生长未受到抑制。当Cd~(2+)浓度大于35.2 mg/L时,其对培养液中Cd~(2+)的去除率随Cd~(2+)初始浓度增加而降低。pH、培养温度、培养时间以及Cd~(2+)初始浓度等因素均能显著影响Paecilomyces lilacinus 6-20p对Cd~(2+)的吸附效果,在菌株应用过程中应充分考虑这些因素。     

铜锌超氧化物歧化酶的重组研究——Ⅰ.铜锌的去除和重组

在制备去Cu、Zn的酶蛋白方法上作了改进并提出新的制备途径,在此基础上对牛红细胞铜锌超氧化物歧化酶进行金属辅基去除和重组研究。通过对酶活性、吸收光谱、圆二色谱和电子自旋共振波谱变化的观察,比较系统地探究了天然酶(Holo-SOD),去Cu、Zn后的酶蛋白(Apo-SOD)和加入Cu~(2+)、Zn~(2+)重组后的酶(Reco-SOD)的性质,以及Cu和Zn对酶活性及功能方面的作用,其主要结果有以下几点:1.用改进法制备去除Cu、Zn的酶蛋白样品可以避免EDTA-Cu~(2+)络合物的污染,残存酶活力仅为天然酶的0.4%,在pH3.8～4.2条件下,加入Cu~(2+)、Zn~(2+)重组后,不仅酶活性可以恢复到98%,而且其UV、CD和ESR图谱均可以恢复到天然酶的状态。2.Cu为酶催化活性所必需。当过量的Zn~(2+)先加入酶蛋白中,然后再补加Cu~(2+),重组酶的活力恢复有所下降。而Zn~(2+)和Cu~(2+)单独加到酶蛋白中去时,Zn~(2+)对酶蛋白的UV图谱没有影响,Cu~(2+)则引起UV吸收大幅度上升。3.同时去除Cu、Zn后的酶蛋白(E_2E_2-SOD)比单独只去Cu后的酶蛋白(E_2Zn_2-SOD)的重组恢复更加困难。     

狭叶香蒲分解体系对镉污染水体修复的影响

选用狭叶香蒲(Typha angustifolia)凋落叶及其微生物分解者为修复材料,探索水生植物与微生物对重金属镉离子的联合修复作用。以水生浮游动物隆线溞(Daphnia carinata)为受试对象,研究了不同浓度Cd~(2+)(0.1、1.0、10μg·L-1)对其生存、生长和繁殖的影响。选择其中0.1μg·L-1作为Cd~(2+)污染浓度,分别将自然驯化以及Cd~(2+)驯化15天和21天的蒲叶放于含Cd~(2+)(0.1μg·L-1)的培养基中,测定隆线溞体长、第一次怀卵日、后代个数、繁殖率以及驯化后微生物分解者的群落组成。结果表明:Cd~(2+)添加显著降低了隆线溞的存活率,其中10μg·L-1Cd~(2+)的毒害作用最强,隆线溞的存活率仅有40%;自然驯化和加镉驯化的蒲叶能够显著改善Cd~(2+)干扰下隆线溞的个体生长与群体繁殖状况,且修复效果会随着驯化时间的延长而提高。个体的第一次生产时间提前1~2天,总产幼数提高22%~81%,群体繁殖率提高3.07~4.7倍,从而使食物链得到快速修复。真菌多样性分析表明,Dactylaria和Massarina在真菌分解者中占据优势地位,推测该属真菌具有修复Cd~(2+)污染的生态功能。     

香根草对镉毒害的生理响应——采用傅里叶变换红外光谱法(FTIR)

为阐明香根草(Vetiveria zizanioides)对镉(Cd~(2+))胁迫的耐性机理及其对重金属Cd~(2+)的积累特征,通过水培试验,采用傅里叶变换红外光谱法(FTIR)研究在不同Cd~(2+)浓度处理时香根草根、茎、叶化学组分的变化,同时测定香根草叶Cd~(2+)的亚细胞分布和其重金属Cd~(2+)含量。结果表明,其根组织在2927 cm~(-1)处峰高先下降后上升,表明在低Cd~(2+)(Cd~(2+)3 mg/L)处理条件下香根草分泌的有机酸不断螯合Cd~(2+),造成羧酸O—H的减少,但随着Cd~(2+)含量的升高,其羧酸螯合力变弱,有机酸含量渐渐升高;茎组织在1631 cm~(-1)峰高处先上升后下降,表明在低Cd~(2+)(Cd~(2+)3 mg/L)处理条件下香根草产生氨基酸、多肽和蛋白质等物质,通过渗透调节来增强抗逆性,但随着Cd~(2+)含量的升高,蛋白质二级结构中肽键间氢键的结合力受影响较大。当Cd~(2+)处理浓度为7 mg/L时,香根草生长10 d后,地上部分及根组织富集Cd~(2+)量分别高达212.49 mg/kg和290.59 mg/kg。香根草地上部分的Cd~(2+)含量随Cd~(2+)处理浓度的增加而增加。香根草叶片富集的Cd~(2+)主要分布于细胞质中,其次为细胞壁,而在线粒体和叶绿体中含量最低。     

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.