几种因子对离体豌豆叶绿体蛋白质合成的影响

利用制备的豌豆完整叶绿体研究了离体条件下蛋白质合成的条件。结果表明：叶绿体蛋白质合成的饱和光强为４５０μｍｏｌ ｍ＾－２ｓ＾－１,合成的速率在最初５ｍｉｎ内最大,此后随时间延长而合成速率下降;Ｋ＾＋对蛋白质合成有促进作用,其最适浓度为３０￣４０ｍｍｏｌ／Ｌ,进一步增加浓度其促进使用反而降低;Ｍｇ＾２＋在１ｍｍｏｌ／Ｌ以下对蛋白质合成有轻微的促进作用,当浓度超过１．５ｍｍｏｌ／Ｌ则开始产生明显的抑制     

一氧化氮抑制AngⅡ介导的心肌肥大反应的信号机制

本文主要利用培养的新生大鼠心肌细胞,从细胞学及分子生物学角度研究一氧化氮（NO）信号系统在AngⅡ介导的心肌肥大反应中的作用及机制。实验以心肌细胞蛋白合成速率、心房钠尿肽（ANP）的表达作为心肌肥大反应的指标,以硝酸盐及亚硝酸盐含量反映心肌细胞NO水平,以免疫印迹法测定MKP－1蛋白表达,以RT－PCR测定eNOS mRNA水平。结果发现：（1）L－精氨酸（L－Arg)10,100μmol/L分别增加心肌细胞NO水平16％及31％,L－Arg(100μmol/L）还可增加心肌细胞eNOS mRNA表达,其作用可被NOS抑制剂L－NAME所抑制;（2）L－Arg(100μmol/L）可降低AngⅡ(0.1μmol/L）诱导的心肌细胞ANP mRNA表达水平和蛋白合成速率,而在L－Arg处理之前用针对MKP－1的反义寡核苷酸转染心肌细胞,蛋白合成速率明显增加,可取消L－Arg的抑制作用,甚至超过AngⅡ组;（3）L－Arg(100μmol/L）明显增加MKP－1蛋白表达,比对照组增加225％,NOS抑制剂L－NAME及蛋白激酶G（PKG）抑制剂KT－5823皆可抑制L－Arg诱导的MKP－1蛋白表达,分别抑制88％、83％,而AngⅡ能增加L－Arg诱导的MKP－1的表达,较对照组增加365％,增强了L－Arg的作用。以上结果表明,NO抑制AngⅡ介导心肌肥大反应的机制可能是通过激活PKG,促进MKP－1的表达,从而增加MAPK去磷酸化实现的。     

在非光合条件下外源蔗糖能促进蚕豆的气孔开放

蚕豆植株经暗饥饿处理40h后,取其下表皮,再用超声波“原位分离”下表皮上的保卫细胞对,然后在无菌、非光合条件下,用外源蔗糖处理蚕豆下表皮上的保卫细胞对,考察其对气孔开放的效应。结果发现,在1d的无菌培养过程中,蔗糖显著促进了气孔的开放。100mmol/L的蔗糖在10mmol/L的MES-NaOH/KOH(pH6.1)缓冲液中,开度分别增加2.0/2.6μm;在1μmol/L的气孔开放促进剂fusicoccin(FC)的存在下,100mmol/L的蔗糖在MES-NaOH/KOH(pH6.1)缓冲液中分别增加开度5.0/5.5μm。不同浓度（5～200mmol/L）的蔗糖处理结果表明气孔开度的增加与蔗糖浓度呈一定的正相关,浓度为100mmol/L的蔗糖处理表现出最大促进作用。同时还初步观察到,蔗糖可维持保卫细胞存活率和叶绿体的完整性。     

NaCl胁迫对海蓬子(Salicornia bigelovii Torr.)离子区室化、光合作用和生长的影响

用含有0、100、300、600mmol/L的NaCl的Hoagland培养液处理海蓬子幼苗,处理一定时间后测定其鲜重、干重、离子含量、Na /K 比值、Na 在细胞中的分布、光合速率、叶绿体超微结构等的变化.结果表明一定浓度NaCl处理促进了海蓬子的生长,300mmol/L左右的NaCl是海蓬子生长的最适盐度.盐处理条件下海蓬子主要将Na 、Cl-积累在地上部,且主要储存在液泡中.随盐处理浓度的增加海蓬子地上部的Na /K 比增大;光合速率在低盐度下随盐浓度的升高而增大,高盐度下受到抑制;叶绿体超微结构在高盐度下受到部分破坏.     

脱落酸对发育中花生胚萌发和贮藏蛋白质合成的影响

发育中的花生胚在无激素固体培养基上高体培养时提前萌发,其发芽力随胚的成熟增加而提高。果针入土后40d胚的发芽率达100％。禹体培养过程中,外源ABA能够阻止花生胚提前萌发和促进胚的发育。胚成熟前期,较低浓度的ABA(10~(-5)mol/L)便抑制胚的萌发;而在成熟中期以后,则要求较高浓度的ABA(10~(-4)mol/L)才能抑制胚的萌发。ABA对成熟前期胚的贮藏蛋白质合成无影响,而对成熟中期至后期胚的贮藏蛋白质合成起促进作用。ABA维持花生胚贮藏蛋白质合成和积累的作用表现在转录水平上。     

外源钙对镉胁迫下南美蟛蜞菊毛状根生长、抗氧化酶活性和镉吸收的缓解效应

为了探讨外源钙对重金属镉(Cd)缓解南美蟛蜞菊Wedelia trilobata毛状根毒害的生理机理,采用溶液培养法研究了重金属Cd单独及其与Ca组合对南美蟛蜞菊毛状根生长、抗氧化酶超氧化物岐化酶(SOD)和过氧化物酶(POD)活性及对Cd2+吸收的影响。结果表明,Cd≤50μmol/L时促进毛状根生长;高于100μmol/LCd则抑制其生长,使其侧根短小,根尖变褐或变黑。与对照相比,不同浓度Cd培养的毛状根POD活性、SOD活性和MDA含量都比对照明显提高,但高于100μmol/L Cd培养的毛状根可溶性蛋白含量均比对照降低。与仅添加200μmol/L或300μmol/L Cd的毛状根相比,Cd和10~30 mmol/L Ca组合培养可促进毛状根生长,使其主、侧根变粗;提高其可溶性蛋白含量;降低其MDA含量、POD活性及SOD活性。原子吸收分光光度法测定结果表明,南美蟛蜞菊毛状根能吸收和吸附重金属Cd2+,当Cd2+浓度为100μmol/L时毛状根对Cd2+的吸收量最大,而Cd2+浓度为300μmol/L时毛状根对Cd2+的吸附量最大。外源加入10~30 mmol/L Ca2+可显著减少毛状根对Cd2+的吸收和吸附,并可调节其抗氧化酶活性,降低其膜脂过氧化水平而解除重金属镉对毛状根生长的抑制或毒害。     

腺苷减少豚鼠心室肌细胞内游离钙浓度

目的：研究腺苷对豚鼠心室肌细胞内游离钙浓度（[Ca^2＋]i）的影响并探讨其可能机制。方法：用激光共聚焦显微镜探测细胞内游离钙浓度,结果用相对荧光强度（（FI-FI0）／FI0,％;FI0：对照;FI：给药）表示。结果：①在正常台氏液和无钙台氏液中,腺苷（10,50,100μmol／L）浓度依赖性地降低[Ca^2＋];。②含30mmol／L KCl的台氏液（高钾台氏液）能够增加[Ca^2＋]i。腺苷（10,50,100μmol／L）能够显著抑制KCl引起的[Ca^2＋]i的增加。③预先应用选择性腺苷AI受体拮抗剂DPCPX（1μmol／L）,可大部分取消腺苷（100μmol／L）在高钾台氏液中的作用。腺苷（100μmol／L）在高钾台氏液的作用也可被预先应用一氧化氮（No）合酶抑制剂L-NAME（1mmol／L）所部分减弱。④腺苷（100μmol／L）能明显抑制无钙台氏液中由低浓度ryanodine引起的[Ca^2＋];增加。⑤当细胞外液钙浓度由1mmol／L增加到10mmol／L而诱发心室肌细胞钙超载时,部分心室肌细胞产生可传播的钙波,腺苷（100μmol／L）可降低钙波发生的频率和持续时间,最终阻断钙波并降低[Ca^2＋];。结论：腺苷可通过抑制外钙内流和减少肌浆网内钙释放从而降低[Ca^2＋],其减少外钙内流可能是由于腺苷A1受体介导的电压依赖性Ca^2＋通道的抑制,NO可能参与这一过程。     

红菇属两种凝集素的分离纯化与比较研究

以红菇属大白菇Russula delica和美丽红菇Russula lepida子实体为材料,利用离子交换柱层析、凝胶过滤层析的手段,分离获得新的凝集素RDL和RLL。结合凝胶过滤层析和SDS-PAGE的手段,确定RDL和RLL分别是分子量为60kDa和32kDa的双亚基蛋白,其N-末端部分氨基酸序列分别为GLKLAKQFAL和VWYIVAIKTDVPRTT。性质研究表明,RDL在20－70℃、低于25mmol/L HCl或12.5mmol/L NaOH下稳定,其凝集活性可以被邻硝基苯酚-β-D呋喃型半乳糖苷（25mmol/L）和菊糖（50mmol/L）所抑制;RDL具有抑制人肝癌Hep G2和人乳腺癌MCF7细胞增殖以及HIV-1反转录酶（RT）的活性,其半抑制浓度IC50分别为0.88μmol/L、0.52μmol/L和0.26μmol/L。RLL在20－70℃、低于12.5mmol/L HCl或NaOH下稳定,其凝集活性可以被菊糖（25mmol/L）和邻硝基苯酚-β-D呋喃型半乳糖苷（100mmol/L）所抑制;RDL具有抑制人肝癌Hep G2和人乳腺癌MCF7细胞增殖的活性,其半抑制浓度IC50分别为1.60μmol/L和0.90μmol/L,但不具有抑制HIV-1 RT的活性。     

Calpain对细胞骨架蛋白tau降解作用的研究(英)

Calpain是钙依赖性中性蛋白酶,根据其对钙敏感性的不同,可分为m-和μ-calpain两型.分别用不同浓度CaCl₂溶液孵育Wistar大鼠脑皮质匀浆液,并用蛋白质印迹和定量图像分析技术检测不同亚型calpain对tau蛋白的降解作用.研究发现：在37℃用1 mmol/L Ca²⁺孵育底物15 min,可见tau蛋白明显降解,并在分子质量为29 ku处出现tau蛋白降解片段;当Ca²⁺浓度为5 mmol/L时,tau蛋白几乎全部被降解;这种tau蛋白降解可被calpain特异性抑制剂完全逆转.进一步的研究发现,分别用μ-calpain抑制剂（0.05 μmol/L calpastatin）,m-calpain抑制剂（100 μmol/L calpain inhibitor Ⅳ）或总calpain抑制剂（552 μmol/L calpeptin）与1 mmol/L Ca²⁺共同孵育Wistar大鼠脑皮质匀浆液,Ca²⁺激活的tau蛋白降解分别被抑制8.6%,92.5%和97.8%.结果表明一定浓度的Ca²⁺可同时激活μ-calpain和m-calpain,这两种亚型calpain均参与降解tau蛋白,但m-calpain的作用比μ-calpain更强.     

白藜芦醇降低大鼠心室肌细胞内游离钙浓度

实验旨在研究白藜芦醇(resveratrol)对大鼠心室肌细胞内钙浓度(intracellular calcium concentratoin,[Ca2+]i)的影响.应用激光共聚焦显微镜技术记录心室肌细胞内的钙荧光强度.结果表明在正常台氏液和无钙台氏液中,白藜芦醇(15～60μmol/L)呈浓度依赖性地降低[Ca2+]i.蛋白酪氨酸磷酸酶抑制剂正钒酸钠(sodium orthovanadate,1.0 mmol/L)和L型Ca2+通道激动剂Bay K8644(10 μmol/L)可部分抑制正常台氏液中白藜芦醇的效应.但NO合酶阻断剂L-NAME(1.0 mmol/L)对白藜芦醇的作用无影响.白藜芦醇也能明显抑制无钙台氏液中由低浓度ryanodine(1.0 nmol/L)引起的[Ca2+]i增加.当细胞外液钙浓度由1 mmol/L增加到10 mmol/L而诱发心室肌细胞钙超载时,部分心室肌细胞产生可传播的钙波,白藜芦醇(60 μmol/L)可降低钙波的传播速度和持续时间,最终阻断钙波.结果提示,白藜芦醇能够降低心室肌细胞内游离钙浓度,此作用可能与其抑制电压依赖性Ca2+通道、酩氨酸激酶和肌浆网内钙释放有关.     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

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.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.