主动脉夹层血管壁中氧化应激蛋白NOX1 表达及意义

目的:探讨主动脉夹层(aortic dissection,AD)血管组织中氧化应激蛋白NOX1的表达及意义。方法:收集2014年~2015年在武汉大学人民医院AD患者升主动脉血管组织标本(AD组)及多器官捐献患者升主动脉血管组织标本(对照组)各12例。采用维多利亚蓝染色观察主动脉中膜弹性纤维形态结构;应用SP免疫组织化学法及Western blot法对标本组织中的NOX1进行检测分析。结果:AD组主动脉中膜弹性纤维形态和排列不规则、破碎、丢失,结构紊乱。免疫组织化学显示NOX1阳性表达见于主动脉壁平滑肌细胞胞质中,AD组与对照组比较,NOX1表达明显升高。Western blot蛋白印迹示AD组NOX1表达明显增高,差异有统计学意义(P0.05)。结论:NOX1在AD主动脉中膜中表达上调,可能在AD的发生中发挥作用。     

Stanford A型主动脉夹层血管壁及外周血中热休克蛋白90的表达及意义

目的:探索热休克蛋白90(Heat shock protein 90, HSP90)在Stanford A型主动脉夹层(Aortic dissection, AD)血管壁组织中的表达及其与平滑肌细胞(Smooth muscle cells, SMCs)表型标志物的相关性。方法:收集本院急性Stanford A型主动脉夹层患者病变主动脉壁组织和外周血标本(AD组,20例),心脏移植供体等来源的正常主动脉壁组织和外周血标本(正常组,10例);ELISA法检测血浆HSP90含量;免疫组化染色检测HSP90的表达差异及定位;Western blot检测组织标本中蛋白的表达差异;应用Spearman分析HSP90和SMCs表型标志物表达的相关性。结果:AD患者血浆中HSP90的含量显著高于正常组(142.38±40.16ng/mL vs. 54.99±24.46 ng/mL,P<0.01)。相对于正常主动脉壁组织,主动脉夹层血管壁组织中HSP90表达明显升高,且主要分布在血管壁中层细胞的胞浆中,分泌型SMCs标志物OPN的表达明显增多,而收缩型标志物α-SMA表达则显著减少。HSP90和α-SMA的蛋白表达呈负相关(R2=0.677,P<0.01),和OPN呈正相关(R²=0.572,P<0.01)。结论:主动脉夹层患者的血管壁组织及外周血中的HSP90含量明显升高,参与了主动脉夹层的发生发展。     

α-平滑肌肌动蛋白在人胸主动脉夹层中的表达

目的:探讨胸主动脉壁中α-平滑肌肌动蛋白(α-SMA)的表达与胸主动脉夹层(TAD)的关系。方法:采集人TAD的动脉壁组织和正常人胸主动脉壁组织,采用Western Blotting和免疫组织化学方法检测α-SMA在组织中的表达程度。结果:在DA组中,α-SMA的表达明显减少,血管平滑肌细胞(VSMC)以增殖表型为主。结论α-SMA的减少主要发生在血管中膜层的VSMC中,细胞发生表型变化,导致中膜弹性变差,发生夹层病变。因此,α-SMA可能在TAD的发病中具有重要作用,值得进一步深入探讨。     

华法令诱导大鼠体内主动脉的钙化作用

目的:研究华法令对大鼠体内主动脉的钙化作用。方法:采用华法令(3 mg/g饲料)的饲料喂养诱导大鼠体内动脉钙化。实验分3组(n=6),对照组、6 W钙化组、12 W钙化组。Von Kossa染色观察钙结节形成,邻甲酚酞络合酮比色法定量测定钙沉积含量。采用TUNEL染色法检测大鼠主动脉组织内血管平滑肌细胞(Vascular Smooth Muscle Cell,VSMC)凋亡。Western blot法检测大鼠主动脉组织中生长抑制特异蛋白6(Growth Arrest Specific Gene 6 Protein,Gas 6)蛋白表达水平。结果:钙化组钙结节、钙沉积含量及VSMC凋亡均高于对照组(P0.01)有显著差异。钙化形成与凋亡表达呈显著正相关(R2=0.8853,P0.0001);钙化组Gas 6蛋白表达量较对照组下调(P0.01)有显著差异。结论:华法令通过下调Gas 6蛋白诱导大鼠体内主动脉钙化形成。     

表达Irkut病毒糖蛋白的重组复制缺陷型人5型腺病毒的构建与小鼠免疫试验

本研究利用复制缺陷型人5型腺病毒载体,表达了我国蝙蝠狂犬病毒分离株IRKV-THChina12的G基因,并将重组病毒命名为rAd5-IRKV-G。Western blot和间接免疫荧光试验证明,IRKV-THChina12的G基因在重组腺病毒感染的293AD细胞中得到表达。将rAd5-IRKV-G对小鼠分别进行腹腔和肌肉免疫,同时设立wt-rAd5对照,结果显示两种免疫方式均能诱导小鼠产生抗IRKV中和抗体,且rAd5-IRKV-G组与wt-rAd5组差异显著(P≤0.05),腹腔免疫组与肌肉免疫组无显著差异(P0.05)。结果表明,Irkut病毒糖蛋白可以作为疫苗抗原,而且构建的重组腺病毒具有较好的免疫原性。     

FHL1在胸主动脉瘤发病机制中的初步研究

目的:研究FHL1蛋白在胸主动脉瘤发病机制中的作用。方法:利用Western Blotting分析胸主动脉瘤患者与正常人主动脉组织中FHL1蛋白表达的情况,利用免疫组织化学检测FHL1蛋白在主动脉组织中的定位,并进一步分析该蛋白在两组中的表达情况,结合文献报道分析FHL1蛋白在胸主动脉瘤发病机制中的作用。结果:Western Blotting、免疫组织化学分析均表明FHL1蛋白在胸主动脉瘤患者主动脉组织表达水平较正常人明显降低,FHL1蛋白主要定位于主动脉血管平滑肌细胞的细胞质中。结论:FHL1蛋白在胸主动脉瘤患者主动脉组织中明显降低,这可能导致主动脉血管平滑肌细胞增殖能力下降,从而在胸主动脉瘤的发病中发挥重要的作用。     

细胞内铜/锌超氧化物歧化酶在人胸主动脉夹层的表达研究

目的:探讨细胞内铜/锌超氧化物岐化酶(copper zinc superoxide dismutase,Cu/Zn-SOD,SOD-1)在人胸主动脉夹层(humanthoracic aortic dissection,hTAD)中的表达情况及其在hTAD中的可能作用。方法:蛋白质印迹法(Western blot,WB)检测SOD-1在TAD和正常人胸主动脉(NA)中膜组织中的表达情况,免疫组织化学染色(immunohistochemistry,IHC)验证SOD-1在动脉壁中的表达和定位。结果:蛋白质印迹和免疫组化染色均显示SOD-1在TAD组表达量较NA组减低(P<0.05);免疫组化染色进一步显示,SOD-1主要位于主动脉壁中膜平滑肌细胞的胞质内,其在夹层主动脉壁中膜撕开处表达缺失。结论:SOD-1在TAD中表达量减少,可能由于参与氧化应激引起的脂质过氧化和炎症反应,以及细胞外基质(extracellular matrix,ECM)的降解等机制所致。     

神经干细胞移植对AD模型大鼠SNAP-25表达的影响

目的：观察神经干细胞对AD大鼠海马周围微环境中SNAP-25 表达及其认知功能的影响。方法：取成年雄性Wistar大鼠30 只,随机分为对照组、AD模型组、细胞移植组,每组10 只。采用凝聚态Abeta1-42 注射到大鼠海马组织内建立阿尔茨海默病(AD)大 鼠动物模型,通过Y 迷宫测试大鼠学习记忆能力和Western blot技术检测大鼠海马组织内SNAP-25 的表达。结果：Y 迷宫测试结 果显示术后4 周时AD模型组和细胞移植组大鼠学习记忆均低于对照组,与AD模型组比较,细胞移植组大鼠学习记忆能力明显 高于AD模型组,差异有统计学意义（P＜ 0.05）;Western blot 检测结果显示术后4 周时AD模型组和细胞移植组大鼠海马组织内 SNAP-25 蛋白表达量均低于对照组,与AD 模型组比较,细胞移植组大鼠海马组织SNAP-25 蛋白表达量高于AD 模型组差异有 统计学意义（P＜0.05）。结论：移植的NSCs 可改善AD 大鼠的学习和记忆能力,其机制可能是通过改变海马区周围的微环境并上 调了海马组织内SNAP-25 表达。     

1ω-3不饱和脂肪酸对视网膜新生血管的实验研究

目的:研究ω-3不饱和脂肪酸对鼠视网膜新生血管的作用。方法:32只7日龄Wistar白鼠分为4组。每组各8只,分别与母鼠共同饲养,制备早产儿视网膜病变(ROP)视网膜新生血管动物模型,其中A组为ROP模型对照组,B、C、D组为ω-3PUFAs喂养组,分别按母鼠公斤体重给与ω-3PUFAs2.5mg/kg/d、7.5mg/kg/d及15mg/kg/d喂养。采用免疫组化观察小鼠中突破视网膜内界膜CD31的特异性表达,Western印迹法检测视网膜标本中RGS5的表达。结果:免疫组化观察视网膜CD31的特异性表达,A、B、C、D组突破视网膜内界膜的新生血管内皮细胞核数分别为(17.25±1.04)个、(15.75±1.83)个、(9.75±1.49)个、(8.5±1.69)个;免疫印迹法检测各实验组视网膜RGS5结果:A组与C组、A组与D组比较差异均有显著意义((P<0.001),其中A、B组相比较;C、D组相比较差异无显著意义(P>0.05)。结论:一定剂量的摄入量后对于新生鼠视网膜新生血管病变将起到一定的防治作用。     

BMP7基因沉默抑制钙盐诱导猪主动脉瓣膜间质细胞成骨分化

目的:探究小干扰RNA(small interference RNA,siRNA)介导的骨形态发生蛋白7(bone morphogenetic protein7,BMP7)基因沉默对钙盐诱导猪主动脉瓣膜间质细胞成骨分化的影响及机制,为钙化性主动脉瓣膜病(calcific aortic valve disease,CAVD)的干预及治疗提供理论依据。方法:非CAVD瓣膜组织(non-CAVD组)取自手术治疗的主动脉夹层患者,CAVD瓣膜组织(CAVD组)取自因钙化性主动脉瓣狭窄而进行主动脉瓣膜置换术的患者,采用免疫组化和Western blot法检测non-CAVD组和CAVD组中BMP7、Runt相关转录因子2(Runx2)的蛋白质表达水平。选取健康家猪处死后即刻于无菌条件下取主动脉瓣叶,采用胶原酶连续消化法分离主动脉瓣膜间质细胞,观察其形态特征,并用免疫荧光染色行表型鉴定。采用脂质体转染法将BMP7-siRNA转染猪主动脉瓣膜间质细胞,采用qPCR和Western blot法验证BMP7表达的变化;利用钙盐培养基诱导细胞成骨分化,建立体外主动脉瓣膜间质细胞钙化模型后,采用ALP染色和茜素红S染色实验分别检测细胞早期及晚期成骨分化能力;采用qPCR和Western blot法分别检测细胞成骨相关基因及蛋白质Runx2、OCN和OPN的表达情况。并用Western blot法检测BMP7下游信号通路中Smad1/5/8的磷酸化水平。结果:BMP7和Runx2蛋白在CAVD组中表达明显高于non-CAVD组。成功分离出原代猪主动脉瓣膜间质细胞,α-平滑肌肌动蛋白(α-SMA)及波形蛋白(vimentin)染色阳性,血管性血友病因子(von willebrand factor,vWF)染色阴性。转染BMP7-siRNA后猪主动脉瓣膜间质细胞中BMP7的mRNA和蛋白质水平均明显下调,早期及晚期成骨分化能力均明显降低。沉默BMP7基因的表达,可下调Runx2、OCN和OPN的基因及蛋白质表达,且磷酸化的Smad1/5/8(p-Smad1/5/8)蛋白水平明显降低。结论:BMP7基因沉默抑制钙盐诱导的主动脉瓣膜间质细胞的成骨分化能力,BMP7/Smads信号通路可能在该过程中发挥重要作用。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Physiological characteristics of various species of strains of carboxydobacteria

Seven strains of aerobic carbon monoxide-oxidizing bacteria (carboxydebacteria) when growing on CO as sole source of carbon and energy had doubling times which ranged from 12–42 h. The activity profiles obtained after discontinuous sucrose density gradient centrifugation indicated that the CO-oxidizing enzymes are soluble and the hydrogenases are membrane-bound in all strains examined. The CO-oxidizing enzymes of Pseudomonas carboxydohydrogena, Pseudomonas carboxydoflava, Comamonas compransoris, and the so far unidentified strains OM2, OM3, and OM4 had a molecular weight of 230,000; that of Achromobacter carboxydus amounted to 170,000. The molecular weights of the CO-oxidizing and H₂-oxidizing enzymes turned out to be identical. The cell sonicates were shown to catalyze the oxidation of both CO and H₂ with methylene blue, thionine, phenazine methosulfate, toluylene blue, dichlorophenolindophenol, cytochrome c or ferricyanide as electron acceptors. Methyl viologen, benzyl viologen, FAD⁺, FMN⁺, and NAD(P)⁺ were not reduced. The spectrum of electron acceptors was identical for all strains tested. Neither free formate, hydrogen nor oxygen gas were involved in the CO-oxidation reaction. Methylene blue was reduced by CO at a 1:1 molar ratio. The results indicate that CO-oxidation by carboxydobacteria is catalyzed by identical or similar enzymes and that the reaction obeys the equation CO+H₂OCO₂+2H⁺+2e^- as previously shown for Pseudomonas carboxydovorans.Dedicated to Otto Kandler remembering almost three decades of enjoyable cooperation     

Modulation of allostery of pyruvate kinase by shifting of an ensemble of microstates

Since the introduction of the concepts of allostery about four decades ago, much advancement has been made in elucidating the structure-function correlation in allostery. However, there are still a number of issues that remain unresolved. In this review we used mammalian pyruvate kinase (PK) as a model system to understand the role of protein dynamics in modulating cooperativity. PK has a triosephosphate isomerase (TIM)(α/β)₈ barrel structural motif. PK is an ideal system to address basic questions regarding regulatory mechanisms about this common (α/β)₈ structural motif. The simplest model accounting for all of the solution thermodynamic and kinetic data on ligand-enzyme interactions involves two conformational states, inactive E^T and active E^R. These conformational states are represented by domain movements. Further studies provide the first evidence for a differential effect of ligand binding on the dynamics of the structural elements, not major secondary structural changes. These data are consistent with our model that allosteric regulation of PK is the consequence of perturbation of the distribution of an ensemble of states in which the inactive E^T and active E^R represent the two extreme end states. Sequence differences and ligands can modulate the distribution of states leading to alterations of functions. The future work includes: defining the network of functionally connected residues; elucidating the chemical principles governing the sequence differences which affect functions; and probing the nature of mutations on the stability of the secondary structural elements, which in turn modulate allostery.     

（鱼句）亚科花（鱼骨）型鱼类骨骼系统的比较

对我国花型Hemibarbuspattern鱼类作了骨骼系统比较,结果表明,此类型鱼类脑颅较长,副蝶骨平直或稍弯曲,眶蝶骨腹纵嵴发达(铜鱼Coreius septentrionalis例外),下颞窝和咽突中等大,基枕骨后突发达;脑颅中的上筛骨的后突、侧筛骨的外筛突,蝶耳骨的外突、上耳骨的后突、围眶骨和后颞窝等均有明显的差异;咽颅中的舌颌骨、尾舌骨、鳃盖骨和下咽齿的列数等又有显著的区别;附肢骨骼中的腰带骨、脊椎骨中的复合神经骨和第4椎骨腹侧的悬器等也有不同之处。据此,这些差异和区别可作为属间或种间的分类依据。