MYH9在肾脏纤维化中表达及作用研究

目的:探讨MYH9(非肌性肌球蛋白重链9)在肾脏纤维化过程中的表达变化以及这种异常表达在疾病发展过程所发挥的作用。方法:通过免疫组化染色,Western blot以及实时PCR检测了MYH9在肾脏纤维化组织中表达水平的变化。同时在慢病毒介导下构建稳定过表达MYH9的肾脏纤维化HK-2细胞,并通过MTT实验、流式细胞仪检测MYH9对HK-2细胞增殖和细胞周期的影响。结果:在小鼠肾脏纤维化组织中MYH9表达水平明显高于正常小鼠肾脏组织(P0.05)。上调MYH9对肾脏纤维化细胞HK-2的增殖水平和细胞周期都呈现出明显的促进(P0.05)。结论:MYH9在肾脏纤维化过程中发挥了重要的促进作用,针对MYH9为靶点的研究为肾脏纤维化的治疗提供新的思路。     

名刊封面

《血液》2007.7.15巨大血小板的发生机制这张可以与古代宫廷华丽壁画相媲美的图片是一张相差显微镜照片,令人吃惊的是照片内容竟然是巨大血小板,它来自Myh9基因缺陷的小鼠胚胎干细胞。以往研究发现:Myh9的突变会引起一种先天性白细胞异常综合征和相关的巨大血小板综合征,而Myh9异常为什么会导     

MYH9与树突状细胞中TRIF-GEFH1-RhoB信号途径的相关性研究

为探讨Rho B(ras homolog family member B)靶蛋白MYH9(nonmuscle myosin heavy chain IIa)与TRIF-GEFH1-Rho B信号途径的关系,通过实时定量PCR技术、si RNA干扰技术、激光共聚焦显微镜、流式细胞术以及基因敲除小鼠等,分析了MYH9与TRIF(TIR domain-containing adapter inducing IFNβ)途径、GEFH1(guanine nucleotide-exchange factors H1)以及MHCII(major histocompatibility complex II)的关系。结果显示,在LPS(lipopolysaccharide)刺激后,MYH9的m RNA表达在野生型小鼠的树突状细胞(dendritic cells,DCs)中增加,在TRIF基因敲除小鼠的DCs中则未被上调。在野生型小鼠中MYH9的m RNA上调可被GEFH1的si RNA明显抑制(P0.01)。同时,在LPS刺激后,MYH9与MHCII在细胞内共定位。MYH9的si RNA还抑制了DCs中LPS介导的MHCII在细胞表面的表达(P0.05)。这些结果表明,MYH9与TRIF-GEFH1-Rho B信号途径存在相关性。     

Identification of a novel MYH9 mutation (p. V782Y) in a Chinese patient with thrombocytopenia: a case report

MYH9-related diseases (MYH9-RD) are a group of autosomal dominant diseases caused by mutations in the MYH9 gene, which are featured by thrombocytopenia, giant platelets and granulocyte cytoplasmic inclusion bodies. MYH9-RD patients generally suffer from bleeding syndromes, progressive kidney disease, deafness, or cataracts. Here, we reported on a case of MYH9-RD. A novel heterozygous mutation of MYH9 (c.2344-2345delGTinsTA, p.T782Y) was discovered by targeted sequencing technology. Immunofluorescence analysis of neutrophils confirmed abnormal aggregation of MYH9 protein. The results of this study should expand the MYH9 gene mutation spectrum and provide reference for subsequent researchers and genetic counseling.     

肌球蛋白重链基因在人类遗传性疾病中的研究进展

肌球蛋白超家族通过水解ATP,将化学能转化为机械能,在细胞迁移、肌肉收缩等多种生理活动中发挥重要的作用。其中,肌球蛋白Ⅱ类分子是肌细胞和非肌细胞中肌丝的重要组成成分。一个完整的肌球蛋白Ⅱ类分子是由2条肌球蛋白重链(myosin heavy chain, MyHC)和2对不同的轻链组成的六聚体。在人体中,存在多种MyHC亚型,分别由不同的MYH基因家族成员编码。迄今为止,人们已经发现MYH基因家族中多个成员的不同突变与人类遗传性疾病相关。其中,MYH2突变可以导致一类以眼肌麻痹为主要特征的骨骼肌疾病;MYH3和MYH8突变可以引起远端关节挛缩综合征;MYH7突变即可以引起骨骼肌疾病包括肌球蛋白沉积性肌病和Laing远端肌病,也与肥厚性心肌病的发生密切相关;MYH9突变可以导致一类以巨大血小板、血小板减少和中性粒细胞包涵体为特征的MYH9相关性疾病。本文简要介绍MYH基因的表达特点,着重阐述MYH基因与人类遗传性疾病之间的相关性及研究进展。     

MYH9 基因多态性与肾脏疾病关系研究进展

非肌性肌球蛋白重链9基因,编码非肌性肌球蛋白重链ⅡA,既往研究其与May-Hegglin异常(May-Hegglin anomaly,MHA)、Fechtner综合征(Fechtner syndrome,FTNS)、Sebastian综合征(Sebastian syndrome,SBS)、Epstein综合征(Epstein syndrome,EPS)和Alport样综合征相关。2008年首次用混合连锁不平衡绘图(MALD)方法证实其与非糖尿病终末期肾病及局灶节段性肾小球硬化症相关,后陆续有该基因与高血压肾病、C1q肾病等的相关报道。本文综合国外该基因与肾脏病的相关研究,对其与肾脏病的关系研究进展做一概括,对今后的研究起一定的帮助作用。     

RhoB分子靶蛋白MHY9的鉴定

鉴定能与活性RhoB分子结合的靶蛋白。制备GST融合的活性RhoB蛋白(GST-RhoB),与LPS刺激的DC2.4细胞裂解物混合后实施pull-down实验,沉降复合物通过SDS-PAGE分析、金染色后,对与GST-RhoB结合的蛋白条带进行基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)分析。然后制备小鼠的树突状细胞,LPS刺激12 h后,进行荧光标记的抗体染色。激光共聚焦显微镜下观察RhoB与MYH9的细胞内定位。通过MALDI-TOF质谱分析,鉴定到一个新的可以与RhoB的活性形式结合的马达分子MYH9。激光共聚焦显微镜的结果表明,RhoB在LPS刺激前后均与MYH9在树突状细胞内共定位。该研究首次发现MYH9可与活性RhoB结合,可能是RhoB下游的一个靶蛋白。     

兰州百合小孢子母细胞减数分裂异常现象的观察

对兰州百合(Lilium davidii var.unicolor)小孢子母细胞减数分裂异常进行了研究,发现存在不等二价体、同源染色体早分离、染色体桥、不均等分离、滞后染色体、核外染色体、微核等。分析了这些异常形成的可能机制及对正常小孢子形成的影响。人工花粉萌发实验表明：小孢子母细胞减数分裂异常是导致花粉败育的主要原因。认为兰州百合长期行无性繁殖引起染色体结构变异,导致减数分裂异常。     

百合花粉母细胞减数分裂异常现象观测分析

对“Siberia”百合小孢子母细胞减数分裂异常进行了系统的细胞学观察,发现“Siberia”百合小孢母细胞减数分裂中存在不等二价体、倒位环、染色体桥、滞后染色体、微核、不均等分离、染色体显著减少的核、游离核、异常四分体、无核花粉等异常现象。研究表明：“Siberia”百合长期进行无性繁殖引起染色体结构变异,导致减数分裂异常,小孢子母细胞减数分裂异常是导致花粉败育的主要原因。     

9;21相互易位携带者生育多例9p部分三体患者细胞遗传学分析

在河南南阳收集到一个家系4代23人, 其中6人患先天性智力障碍, 具有轻度的面部和小母指畸形等特点, 先证者伴随有癫痫的发生。采用常规的外周血培养染色体G带分析, 发现先证者的核型为：46, XY, der(21) t(9; 21) (9p22.2; 21q22.3)pat, 是部分9p三体。对该家系其他成员的染色体进行分析, 发现所有患者均为部分9p三体, 异常染色体均来自9号与21号染色体平衡易位携带者染色体相互易位的异常分离, 因此这是一个部分9p三体综合征家系。而重复区段发生在9号染色体短臂远端一半区域(9pter→9p21)内, 该区是关键区, 导致智力障碍和面容轻微畸形。     

The design of a biochip: a self-assembling molecular-scale memory device

A design for a biochip memory device based on known materials and existing principles is presented. The fabrication of this memory system relies on the self-assembly of the nucleic acid junction system, which acts as the scaffolding for a molecular wire consisting of polyacetylene-like units. A molecular switch to control current is described which is based on the formation of a charge-transfer complex. A molecular-scale bit is presented which is based on oxidation-reduction potentials of metal atoms or clusters. The readable 'bit' which can be made of these components has a volume of 3 x 10(7) A3, and should operate at electronic speeds over short distances.     

The use of pH indicators to identify suitable environments for freezing samples in aqueous and mixed aqueous/nonaqueous solutions

The colours of frozen solutions containing pH indicators are shown to provide a test for changes in pH in the solvent environment which occur on freezing. Yeast alcohol dehydrogenase loses activity on freezing in phosphate buffer (a buffer in which pH indicator colour changes shows a marked decrease in pH on freezing) but when frozen in bis-tris, Hepes, or N-glycylglycine buffers (all of which show little change in the colour of universal pH indicator and hence of pH on freezing) is stable on freezing. The effects of freezing in different buffer systems on the rate of decomposition of NADPH, and on the rate hydrolysis of 4-nitrophenyl acetate, are rationalised in terms of the pH shifts in these buffers which were determined using universal pH indicator. It is proposed that a major reason for the instability of samples on freezing is the pH changes which occur when some systems are frozen. From the results a general scheme for selecting the best environment for safely freezing samples is proposed which is based on the use of pH indicators.     

Structure and function of an acetylcholine receptor.

Structural analysis of an acetylcholine receptor from Torpedo californica leads to a three-dimensional model in which a "monomeric" receptor is shown to contain subunits arranged around a central ionophoretic channel, which in turn traverses the entire 110 A length of the molecule. The receptor extends approximately 15 A on the cytoplasmic side, 55 A on the synaptic side of the membrane. The alpha-bungarotoxin/agonist binding site is found to be approximately 55 A from the entrance to the central gated ion channel. A hypothesis for the mechanism of AcChR is presented which takes into account the structural and kinetic data, which is testable, and which serves as a focus for future studies on the agonist-induced structure change in AcChR.     

Defence glands in the giant spiny phasmid Extatosoma tiaratum

Adults of Extatosoma tiaratum liberate a strong smell when handled. The smell is associated with a colourless, volatile liquid which is apparently harmless to man. It is ejected on to the thorax of the insect, and on to the hands of the investigator, from a pair of tubercles located on the anterior border of the prothorax. The tubercles have a slit-like aperture through which the secretion is sprayed on attack. The secretion is produced by two glands in the thorax, each comprising a pair of tubes which unite just beneath the tubercle. The tubes consist of a thick, glandular epithelium lined with a thin layer of cuticle; most of the gland is enveloped in a layer of muscle. The defence reaction is discussed in relation to that of other phasmids.     

Age structure effects in predator-prey interactions

A mathematical model of predator-prey interactions is proposed which incorporates both age structure in the predators and density dependence in the prey. The properties of the model are investigated by a linearized analysis, which enables the conditions for stability to be formulated. The analysis indicates that for a substantial domain of parameter space, a stable equilibrium is possible with the prey well below its carrying capacity. The effect of violating the stability conditions on the behaviour of the model was investigated by computer simulation. Two further types of behaviour are noted in which coexistence is possible. The first is a two point limit cycle in which young and old predators occur in alternate time periods. The second involves a limit cycle in which the component population trajectories lie on closed curves in phase space.     

Logical Probability and Risk Assessment

A risk assessment is intended to provide a statement of current knowledge which is intended to inform a decision-maker of the current state of knowledge in response to a particular concern. Because answering the concerns of decision-makers often requires inferences to be drawn, doubt often arises over how the inference is to be drawn. In quantitative risk assessment, where a mathematical equation or model is used to draw the inference, this uncertainty is referred to as model uncertainty. A two-step process, which is referred to as logical probability, is proposed as a technique for representing model uncertainty in a risk assessment. The first step involves assigning model weights in which the degree of evidential support for each of the alternative models is considered. The second step involves assigning a unique interval in the range of 0 to 1 for each model which reflects the models' weight, to form a probability distribution. While the second step is straightforward, the first step is not. Assigning model weights requires consideration of any line of evidence that may reasonably impact the validity of the assertion of a model. While the development of a procedure for doing so may be expected to be a process which reflects the subjective preferences of whomever is involved in creating it, there are some historical precedents on which to build. Foremost among these are (1) the use of a correlation coefficient or other goodness-of-fit criteria to measure the degree of correspondence between a given model and a set of observations which are used as evidence to support it, and (2) preference given to models which are simpler, which may be ascertained as the number of adjustable parameters the model contains. Additional principles, which have little or no tradition to stand on, must be used to reflect the impact of other empirically supported beliefs on model preference. The procedure proposed is comparable to the procedure known as decision analysis, in which probabilities are assigned to alternative models based on expert or subjective input. The principal difference in the present case is that it is suggested that principles which transcend the decision at hand should be sought and articulated in order to generate a consistent measure of uncertainty arising from interpretation.     

Measurement of gas free body volume by a diving device.

A device is presented for determination of gas free body volume, by which total body volume as well as lung volume together with intestinal gas can be measured. The measurement is performed under water. The subject to be measured keeps his head in an airfilled cylinder which allows free breathing during the measurement procedure under water. The subject sits on a seat which is connected with a vertical "probe" that is submerged in a water basin. This "body density probe" carries several measuring rods of different diameter on its upper end, which lead to an increase of buoyancy when sinking deeper into the water after additional weights have been put on the device. These rods lead to new equilibrium states which are also dependent on the compressable gas volume attached to the probe. Three equilibrium states with different added weights give a physical definition of gas free body volume, which can be calculated from a number of accurately measurable single items.     

Parallel recognition of idealised line characters

Summary The aim is to design a machine which is able to learn a number of idealised characters and to recognise them, irrespective of their size, position and context on an infinite retina. If the number of characters which such a machine can possibly learn to recognise is astronomical, it is not practical to use separate templates for every possible character. It is more economical to use, instead, templates for various parts, called features, of characters. In recognising a number of characters simultaneously, without scanning, the question arises of how to tell which feature belongs to which character of figure on the retina. In particular, if a given character is not present but all its features are included in nonsense figures simultaneously present on the retina then the machine must not indicate the presence of the given character. The technique which overcomes this difficulty employs overlapping features which must be mutually consistent for recognition. This consistency technique is assessed by comparison with a more conventional technique, and the work is restricted to closed line characters which are not subject to deformations or mutilations.     

The human embryo: a scientist's point of view

...Thus, my judgement is that a human embryo is not a human person, and so we may do experiments on it which involve killing it. But my judgement is also that a human embryo has the potential to become a human being. The consequence of this attribute is that it imposes limits on the kinds of experiments which may be performed on human embryos. It is this which sets the boundaries. Experiments which may harm the embryo while still allowing it subsequently to realise its potential, and become a person, should not be permitted. It is the potentiality of the human embryo which governs our behaviour towards it. Its potential makes it special, and radically different from any other human tissue. This potential which the early embryo has means that great respect must always be accorded it, and great thought and care must surround any dealings with it....     

Ecology and behaviour of domestic sheep (Ovis aries): a review

Behaviour of sheep is complex and poorly understood. It is only within the past 20 years that a concerted effort has been made to elucidate those aspects of behaviour which have most relevance to sheep raising under open-range conditions. The present report attempts to summarize and review published work on the behaviour and ecology of sheep. The whole question of the significance of behaviour and the extent to which it is possible to shape it is a field which is open for study. This paper is an attempt to summarize the work on sheep behaviour and on those aspects of physiology which impinge. Consideration is given to the evolution of Bovids, scope of domestication and world distribution of sheep since these aspects have a major influence on the sheep with which we deal today. Animal requirements for food, water, shelter, protection from predators, and control of disease and parasites have been largely met by man and the forces for selection which operated among the progenitors of domestic sheep have been largely negated. Despite this, genetic selection still goes on. Behavioural adaptation to a wide variety of environmental and nutritional regimes is a common feature among free-ranging sheep. Reproductive success, a matter of considerable commercial concern is often poor. Lamb survival is of critical importance. Factors involved in neonatal mortality are reviewed in detail. Finally, the now extensive literature on sheep behaviour, physiology and management is collated in a bibliography with over 150 literature citations.