黑色菜豆(Phaseolus sp.)凝集素的纯化和性质

黑色菜豆(phaseolussp.)种子中含有对人A型血专一凝集的凝集素。用猪胃粘蛋白-Sepharose 4B作亲和吸附剂和Sephadex G-200凝胶过滤,可以纯化这种凝集素。纯化的凝集素在pH8.9,Tris-EDTANa_2-borate缓冲液的PAGE中,呈现单一蛋白带;酚-硫酸法测得总糖含量为3.22％。在SDS-PAGE中发现其分子由两种亚基所组成,亚基分子量分别为38,000和35,000。当凝集素浓度分别为0.98μg/ml和1.95μg/ml时能强烈地凝集人A型和AB型血细胞。在凝集素浓度高达500μg/ml时,B型血细胞能发生弱凝集反应,但对O型血和兔红细胞则完全不发生凝集反应。其凝集活性可被GalNAC、L-Fuc、猪甲状腺球蛋白和卵粘蛋白所抑制。该凝集素对人外周血中淋巴细胞的转化率达80％,细胞分裂比率高达37.1％;氨基组成分析表明,凝集素分子中Asp和Glu含量较高,而cys和Met含量很低。     

苦参凝集素的分离纯化及部分性质研究

从苦参 (SophoraflavescensAit.)根浸出液经硫酸铵分级 ,得苦参凝集素 (SFL)粗品 ,再经DEAE_Sepharose、SephadexG_1 5 0和HPLC层析 ,获得具有强凝集活性的SFL样品 ,用PAGE和SDS_PAGE检测均为单一蛋白染色带。SDS_PAGE显示SFL分子仅有一条肽链 ,SephadexG_1 0 0和SDS_PAGE测得其分子量均为 32kD。当SFL浓度为 0 .97μg/mL时能凝集兔红细胞 ,无血型专一性 ,其凝血活性可被甘露糖和果糖抑制 ,麦芽糖和葡萄糖有弱的抑制作用 ,凝集素分子含有 2 .89%的中性糖 ;当SFL量为 6 2 μg时 ,对棉花枯萎病菌 (FusariumvasinfectumAtk .)、小麦赤霉病菌(Gibberllasaubinetii (Mont.)Sacc .)和水稻稻瘟病菌 (PiriculariaoryzaeCav)菌丝体的生长发育有明显地抑制作用。用Edman法在蛋白测序仪上测出SFL的N端肽链 30个氨基酸的排列顺序为 :T/A/VDXLXFTFSDFDP NGEDLLFQGDAHVTSNN。     

香螺血清、肌肉和唾液腺凝集素凝集性能的初步研究

研究香螺(Naptunea cumingi crosse)肌肉、血清和唾液腺凝集素对7 种脊椎动物红细胞、人的4 种红细胞、4 种单细胞藻类和11 种微生物细胞的凝集性能,同时进行pH敏感性、热稳定性、糖抑制性、EDTA、金属离子以及盐浓度影响试验.结果表明,3 种凝集素对各种细胞的凝集效应存在差异,以家兔红细胞的凝集效果最佳.肌肉和血清凝集素在pH<7.0时均失活,唾液腺凝集素的热稳定性最强,温度为90 ℃仍具有活性.此外,不同的糖溶液对3 种凝集素凝集效果的影响不同,在EDTA浓度为2.00～0.25 mmol/L范围内,3 种凝集素的凝集效价均受到不同程度的抑制.在金属离子影响试验中发现,Zn2+能明显提高肌肉和血清凝集素的凝集活力,而对唾液腺凝集素有抑制的作用.当盐浓度为12～18 g/L时,可增加香螺肌肉凝集素的凝集效价,而当浓度为24～60 g/L时却会抑制肌肉凝集素的凝集活性.     

甘薯凝集素的提取及性质研究

抗蔓割病甘薯的叶片组织浸取液,经硫酸铵分级沉淀,甲壳素柱层析及葡聚糖凝胶过滤得到一种在PAGE或SDS－PAGE上均呈现单一蛋白带的甘薯凝集素。该凝集素没有血型专一性及被测动物红细胞专一性,其凝集活性可被N－乙酰葡萄糖胺或岩藻糖所抑制。甘薯凝集素在75℃加热10min,即丧失全部凝集活性,其凝集活性依赖于Ca^2 和Mg^2 ,Mn^2 则无作用。经Sephadex G-100和SDS－PAGE测定,凝集素相对分子质量为63000,中性糖含量为6.21%,该凝集素对蔓割病菌有抑制作用,是一种酸性糖蛋白。     

棉花凝集素的纯化及性质研究

抗枯萎、抗黄萎病的棉种的浸取液,经硫酸铵分级,纤维素柱、亲和层析后,再经分子筛过滤,获得在ＰＡＧＥ、ＳＤＳ－ＰＡＧＥ或ＨＰＬＣ柱上均呈现单一蛋白带的棉花凝集素。该凝集素只凝集兔红细胞,对人Ａ、Ｂ或Ｏ型血细胞均不凝集,其凝集活性可被半乳糖或猪甲状腺球蛋白等所抑制。棉花凝集素在６５℃加热５ ｍ ｉｎ,即丧失全部凝集活性;它的凝集活性强烈地依赖于Ｃａ２＋ ;Ｍｎ２＋ 对活性也有促进作用,Ｍｇ２＋ 则无作用。经凝胶过滤或ＳＤＳ－ＰＡＧＥ测定,凝集素的分子量为６３０００,Ｎ－末端氨基酸为Ｖａｌ。凝集素含有１．５％ 的中性糖,是一种促有丝分裂原,细胞转化率为５０．３％ 。     

用兔红细胞膜制备亲和树脂纯化红芸豆红细胞凝集素的方法

建立运用兔红细胞膜制备亲和树脂来纯化红芸豆中红细胞凝集素的方法。红芸豆经过浸提,(NH4)2SO4沉淀,红细胞膜亲和树脂吸附、洗脱得到红细胞凝集素(PHA-E)试样。采用电泳法测定其纯度、相对分子质量和等电点。用体积分数2%的兔红细胞悬液测定试样凝血活力及影响凝血因素。经PAGE分析PHA-E试样为单带,SDS-PAGE分析显示亚基相对分子质量为3.2×104,等电点为6.5。研究发现,促使50%兔红细胞产生凝集的试样蛋白质最低质量浓度为4μg/mL,单糖不影响PHA-E凝血活力,EDTA抑制其凝血活力,Zn2+促进其凝血。     

南鹤虱凝集素的研究

本文对伞形科16种植物(药用部分)是否存在凝集素进行了筛选。发现野胡萝卜果实与芹菜果实中有凝集素。野胡萝卜果实(中药名称南鹤虱)抽提液的凝血活性较芹菜强。 采用CM-纤维素及DEAE-纤维素对南鹤虱凝集素进行分离纯化。用聚丙烯酰胺凝胶电泳鉴定其纯度。用SDS-聚丙烯酰胺凝胶电泳测定亚基分子量为32600。 此凝集素对热及酸较稳定,对碱略差。对人的A、B、O血型无专一性。能凝集兎、小鼠、牛及鸡的血,但不凝集蟾蜍的血。 此凝集素的凝血活性能被D-木糖及N-乙酰葡萄糖胺轻微地抑制。 经shiff试剂检测表明南鹤虱凝集素为一糖蛋白。糖含量为3.4％。此凝集素分子中的谷氨酸门冬氨酸、精氨酸及絲氨酸的含量较高。     

蛋白核小球藻凝集素的分离纯化及部分性质研究

蛋白核小球藻藻粉的PBS抽提液经硫酸铵二步分级沉淀 ,再经DEAE Sepharose和SephadexG 10 0层析 ,从中分离纯化得到蛋白核小球藻凝集素 (CPL)。经测定 ,该凝集素为单个亚基的蛋白质 ,相对亚基分子量为 1 4× 10 4 — 1 5× 10 4 ,分子中不含糖。在氨基酸组成中 ,苯丙氨酸 (Phe)的含量最高 ,其次是天冬氨酸 (Asp)和谷氨酸 (Glu) ,不含组氨酸 (His)。CPL能够凝集兔、绵羊及鸽子红细胞 ,其中对兔红细胞的凝集活性最大 ,最低浓度为 6 88μg/mL ,对鸡、鸭及人红细胞 (A型、O型及B型 )无凝集活性。卵黏蛋白和 7种单糖对CPL的凝血活性具有抑制作用。CPL具有很好的热稳定性 ,在 90℃处理 10min不失活。     

染病刺参凝集素的分离纯化与性质比较

目的：研究刺参免疫机制,探寻其发病机理。方法：刺参（Apost／chopusjapon／cus）经磷酸盐缓冲溶液抽提、20—75％硫酸铵分级沉淀、DEAE—SepharoseF．F．和SephadexG一100柱层析,得到刺参凝集素（A几）。结果：经SDS—PAGE显示单一条带,亚基相对其分子量为31000。分别提取感染Tenacibaculum属和Vibrionaks属两种刺参烂皮病病原菌的刺参凝集素,感染Temcibaculum属病菌刺参的凝集素产量较大,但凝集兔红细胞的作用均不被所测试的单糖和寡糖所抑制,而仅被牛甲状腺球蛋白所抑制。凝集兔红细胞的作用均不被二价金属离子Ca2＋、Mg2＋、Mn2＋、Zn2＋及EDTA所抑制。在pH4．0。10．14系列缓冲液中均有活性,其中在pH4．0—7．5时活性最大。该凝集活性在90℃加热30min后凝集活性仅损失50％。结论：推断菌株Tenac／bacu／um可诱导刺参体内产生大量的凝集素。     

从绿藻礁膜（Monostroma nitidum Wittr）分离半乳糖专一的凝集素

礁膜（Monostroma nitidum Wittr）经 25%～80%硫酸铵分级、DEAE-纤维素52离子交换层析和Sephadex G-200凝胶过滤层析,得到纯化礁膜凝集素（Monostroma nitidum lectin,MNL）,在SDS-PAGE上显示单一蛋白染色带. 用Sephadex G-200层析测得其分子质量为66.6 kD, 用SDS-PAGE测得其分子质量为66.2 kD．该凝集素可以凝集人A、B、AB、O型红细胞,且凝集活性相同. 在对人（A、B、AB、O）、兔、鲤、鲫、鼠、羊、鸡、狗的红细胞凝集作用中,兔凝集作用最强．该凝集素在pH 4.00～10.53范围内均有活性,但在pH 5.20～9.40范围内活性最大．经100 ℃热处理30 min后,该凝集素对兔红细胞血凝活性保留25%,活性最大的温度范围为25～55 ℃．MNL被EDTA抑制,最小抑制浓度为3.13 mmol/L,但对 Ca^２＋和Mg^２＋不敏感．该凝集素凝集兔红细胞的作用不被D -果糖、D-甘露糖、D-葡萄糖、蔗糖、麦芽糖、γ-球蛋白、牛甲状腺球蛋白所抑制,但被D- 半乳糖和乳糖抑制,最小抑制浓度分别为5 mmol/L和2.5 mmol/L．     

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.     

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     

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.     

鸡传染性法氏囊病病毒研究进展

传染性法氏囊病(Infection bursal disease, IBD)是由鸡传染性法氏囊病毒(Infectious bursal disease virus, IBDV)引起的鸡和火鸡的一种高度接触性传染病,给世界各国的禽养殖业带来了巨大损失.自IBDV发现至今新的变异株不断出现,分子结构的改变导致病毒致病力的改变及宿主对疫苗应答的改变,使得传统的疫苗已不能控制其流行,因此各国学者对其基因组结构和功能进行了广泛深入的研究,并积极研制新型有效的疫苗以达到防治的目的.     

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.     

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).     

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.