一种生物分子马达的冲激力模型

基于动力冲程模型并结合布朗棘轮模型的扩散机制,提出了分子马达的冲激力模型。该模型基于一系列具有时间或空间周期性的啄函数来模拟分子马达的做功冲击,得到了关于几率流（速度）的解析结果。计算结果与实验数据相符合。为了产生非零的几率流,新模型并不要求分子马达与轨道间的势能必须是不对称的,因而相对于布朗棘轮模型来说更加稳健。     

农用内燃机的工作原理及安装使用

内燃发动机是燃料在气缸内部进行燃烧产生的热能转化为机械能的一种机器。农用内燃发动机种类较多,按使用燃料分为柴油机、汽油机;按点火方式分为点燃式、压燃式;按冲程次数分为二冲程、四冲程;按气缸数目分为单缸、双缸、多缸;按气缸排列形式分为直列式、卧式、V型式;按冷却方式分为水冷式、风冷式。掌握农用内燃机的工作原理以及安装使用,对于延长农机使用寿命和农机安全生产作业意义重大。     

LKBⅢ型超薄切片机故障检修二例

自动切片时故障的排除瑞典LKBⅢ型超薄切片机,其样品臂的动作是上下垂直运动的形式,在切削过程中,样品臂因受热而成线性膨胀。当样品臂因重力作用下降进行切片后,在返程上升时,退刀磁铁通电,使刀台产生一向下弯曲动作,于是刀向后退回30微米,这样样品臂才能无阻地升到切削冲程最高位置。当退刀磁铁断电,刀即恢复原位继续进行切片。因     

光动力疗法在腹部恶性肿瘤的应用

近年来光动力治疗已经应用在腹部的恶性肿瘤的治疗上,并且取得了满意的疗效.对于部分腹部恶性肿瘤,相应的研究在实验室里也有报道.通过对光动力治疗的原理、机制、光敏剂,以及腹部恶性肿瘤光动力治疗相关的文献进行了综述,总结了光动力治疗应用在腹部恶性肿瘤的优势和相关问题.     

光动力疗法中氧的弥散模型仿真及其意义

组织中的氧是由血管中扩散而来,存在一定的差异。光动力疗法使用的卟啉类光敏剂主要是通过将能量转移到氧分子产生单线态氧来产生毒性物质,因此在光动力治疗中有氧的消耗,使组织中氧分布对光动力作用具有特殊意义。本文对氧在靶组织和正常组织中的分布、光动力效应对组织中氧含量的影响、以及氧含量对光动力效应的反作用等方面进行了综述。     

光动力疗法联合声动力疗法在抗肿瘤方面的研究进展

光动力疗法是一种使用光敏药物和激光活化治疗肿瘤疾病的方法。用特定波长的光辐照肿瘤部位,能使选择性聚集在肿瘤组织的光敏药物活化,引发光化学反应破坏肿瘤。然而,光动力疗法在临床上的应用却一直存在治疗深度受限的问题。本文分析了光动力疗法在临床应用中的局限性,并指出光动力疗法联合声动力疗法是一种可以克服光动力疗法治疗深度局限性的新型非侵入性治疗方法。     

湖泊水动力变化对沉水植物的影响研究综述

随着全球气候变化加剧及水利工程的快速发展,湖泊水动力状况发生了显著变化。通过影响湖泊水体和沉积物理化性质,水动力变化可以作用于沉水植物生存、生长与分布等方面。在长期适应进化过程中,沉水植物演化出了一系列有效的适应策略,能一定程度上克服水动力变化的负面影响。但当前湖泊水动力变化程度远超沉水植物适应上限,湖泊沉水植物消退已成为全球普遍现象。了解沉水植物适应水动力条件的过程有助于揭示湖泊沉水植被退化机制,为未来沉水植物的保护和恢复提供借鉴。因此,本文系统综述当前湖泊水动力变化成因,水动力变化对沉水植物的不利影响及沉水植物适应策略,包括：繁殖对策、形态学对策、生理对策等。同时,综述当前研究进展,今后还需大力加强沉水植物解剖学及物种忍耐力差异方面的研究。     

Cajal间质细胞在急进高原胃肠动力紊乱中的作用的展望

急进高原胃肠动力紊乱是高原胃肠应激反应的主要表现之一,腹胀、恶心、呕吐、腹泻、食欲减退等是其最突出的临床症状,目前有关其的研究多集中于临床及部分基础研究上,但在探讨有关高原胃肠动力紊乱形成机制的细胞分子生物学领域的研究则少见报道。而大量研究指出,慢波起源细胞Cajal间质细胞在胃肠动力调控中具有重要作用,并成为的研究的热点,那么Cajal间质细胞是否同样在急进高原胃肠动力紊乱中发挥同样重要的作用,这不但对从细胞分子生物学角度来解释急进高原胃肠动力紊乱的机制有着重要的意义,而且还可以对未来的临床干预提供新的思路。因此,本文拟对Cajal间质细胞在急进高原胃肠动力紊乱中的潜在作用作一综述。     

锁定加压钢板和动力加压钢板治疗肱骨中下段骨折的临床比较研究

目的:对比锁定加压钢板与动力加压钢板治疗肱骨中下段骨折患者的临床疗效。方法:抽取我院2010年8月~2013年12月收治的肱骨中下段骨折患者76例,按照随机数字表法分为锁定组(锁定加压钢板治疗)与动力组(动力加压钢板治疗),每组38例,随访1年。对比两组患者临床指标,治疗效果及并发症发生率。结果:锁定组患者手术时间、骨折愈合时间及住院时间均小于动力组,差异均有统计学意义(P0.05);锁定组患者优良率为89.47%,显著高于动力组的71.05%,差异有统计学意义(P0.05);锁定组的并发症发生率为10.53%,显著低于动力组的31.58%,差异有统计学意义(P0.05)。结论:锁定加压钢板应用于治疗肱骨中下段骨折患者疗效优于动力加压钢板治疗,它具有创伤小、恢复快的优点,且并发症发生率较低,值得推广。     

钙信号在光动力疗法中的产生及作用

光动力疗法是基于光敏剂选择性地积聚在肿瘤组织中,肿瘤接受光照后凋亡或坏死的一种细胞毒性治疗方法.光敏剂的亚细胞定位决定了细胞光敏损伤的初始位置,线粒体、内质网、细胞膜、溶酶体,细胞骨架等均可成为光敏损伤的靶点.细胞内Ca2 作为一个广泛意义上的信号分子,参与了多种信号转导途径,在光动力疗法诱导肿瘤细胞凋亡过程中起了重要作用.从光动力疗法造成的亚细胞损伤出发,探讨了光动力疗法中钙信号的产生机制,并简要介绍了钙信号在光动力疗法诱导肿瘤细胞凋亡中的作用机制.     

Millisecond-scale biochemical response to change in strain

Muscle fiber contraction involves the cyclical interaction of myosin cross-bridges with actin filaments, linked to hydrolysis of ATP that provides the required energy. We show here the relationship between cross-bridge states, force generation, and Pi release during ramp stretches of active mammalian skeletal muscle fibers at 20°C. The results show that force and Pi release respond quickly to the application of stretch: force rises rapidly, whereas the rate of Pi release decreases abruptly and remains low for the duration of the stretch. These measurements show that biochemical change on the millisecond timescale accompanies the mechanical and structural responses in active muscle fibers. A cross-bridge model is used to simulate the effect of stretch on the distribution of actomyosin cross-bridges, force, and Pi release, with explicit inclusion of ATP, ADP, and Pi in the biochemical states and length-dependence of transitions. In the simulation, stretch causes rapid detachment and reattachment of cross-bridges without release of Pi or ATP hydrolysis.     

Simulations of the myosin II motor reveal a nucleotide-state sensing element that controls the recovery stroke

During the recovery stroke, the myosin motor is primed for the next power stroke by a 60° rotation of its lever arm. This reversible motion is coupled to the activation of the ATPase function of myosin through conformational changes along the relay helix, which runs from the Switch-2 loop near the ATP to the converter domain carrying the lever arm. Via a hydrogen bond between the side-chain of Asn475 on the relay helix and the Gly457/Ser456 peptide group on the Switch-2, the rotation of the converter domain is coupled to the formation of a hydrogen bond between Gly457 and γ-phosphate that is essential for ATP hydrolysis. Here, molecular dynamics simulations of Dictyostelium discoideum myosin II in the two end conformations of the recovery stroke with different nucleotide states (ATP, ADP·Pi, ADP) reveal that the side-chain of Asn475 breaks away from Switch-2 upon ATP hydrolysis to make a hydrogen bond with Tyr573. This sensing of the nucleotide state is achieved by a small displacement of the cleaved γ-phosphate towards Gly457 which in turn pushes Asn475 away. The sensing plays a dual role by (i) preventing the wasteful reversal of the recovery stroke while the nucleotide is in the ADP·Pi state, and (ii) decoupling the relay helix from Switch-2, thus allowing the power stroke to start upon initial binding to actin while Gly457 of Switch-2 keeps interacting with the Pi (known to be released only later after tight actin binding). A catalytically important salt bridge between Arg238 (on Switch-1) and Glu459 (on Switch-2), which covers the hydrolysis site, is seen to form rapidly when ATP is added to the pre-recovery stroke conformer and remains stable after the recovery stroke, indicating that it has a role in shaping the ATP binding site by induced fit.     

Mechanokinetics of rapid tension recovery in muscle: the Myosin working stroke is followed by a slower release of phosphate

Crystallographic and biochemical evidence suggests that the myosin working stroke that generates force in muscle is accompanied by the release of inorganic phosphate (Pi), but the order and relative speed of these transitions is not firmly established. To address this problem, the theory of A. F. Huxley and R. M. Simmons for the length-step response is averaged over elastic strains imposed by filament structure and extended to include a Pi-release transition. Models of this kind are applied to existing tension-recovery data from length steps at different phosphate concentrations, and from phosphate jumps upon release of caged phosphate. This body of data is simulated by the model in which the force-generating event is followed by Pi release. A version in which the Pi-release transition is slow provides a better fit than a version with rapid Pi release and a slow transition preceding force generation. If Pi is released before force generation, the predicted rate of slow recovery increases with the size of the step, which is not observed. Some implications for theories of muscle contraction are discussed.     

A kinetic model that explains the effect of inorganic phosphate on the mechanics and energetics of isometric contraction of fast skeletal muscle

A conventional five-step chemo-mechanical cycle of the myosin–actin ATPase reaction, which implies myosin detachment from actin upon release of hydrolysis products (ADP and phosphate, Pi) and binding of a new ATP molecule, is able to fit the [Pi] dependence of the force and number of myosin motors during isometric contraction of skeletal muscle. However, this scheme is not able to explain why the isometric ATPase rate of fast skeletal muscle is decreased by an increase in [Pi] much less than the number of motors. The question can be solved assuming the presence of a branch in the cycle: in isometric contraction, when the force generation process by the myosin motor is biased at the start of the working stroke, the motor can detach at an early stage of the ATPase cycle, with Pi still bound to its catalytic site, and then rapidly release the hydrolysis products and bind another ATP. In this way, the model predicts that in fast skeletal muscle the energetic cost of isometric contraction increases with [Pi]. The large dissociation constant of the product release in the branched pathway allows the isometric myosin–actin reaction to fit the equilibrium constant of the ATPase.     

How myosin motors power cellular functions: an exciting journey from structure to function: based on a lecture delivered at the 34th FEBS Congress in Prague, Czech Republic, July 2009

Molecular motors such as myosins are allosteric enzymes that power essential motility functions in the cell. Structural biology is an important tool for deciphering how these motors work. Myosins produce force upon the actin-driven conformational changes controlling the sequential release of the hydrolysis products of ATP (Pi followed by ADP). These conformational changes are amplified by a 'lever arm', which includes the region of the motor known as the converter and the adjacent elongated light chain binding region. Analysis of four structural states of the motor provides a detailed understanding of the rearrangements and pathways of communication in the motor that are necessary for detachment from the actin track and repriming of the motor. However, the important part of the cycle in which force is produced remains enigmatic and awaits new high-resolution structures. The value of a structural approach is particularly evident from clues provided by the structural states of the reverse myosin VI motor. Crystallographic structures have revealed that rearrangements within the converter subdomain occur, which explains why this myosin can produce a large stroke in the opposite direction to all other myosins, despite a very short lever arm. By providing a detailed understanding of the motor rearrangements, structural biology will continue to reveal essential information and help solve current enigma, such as how actin promotes force production, how motors are tuned for specific cellular roles or how motor/cargo interactions regulate the function of myosin in the cell.     

Strain-dependent modulation of phosphate transients in rabbit skeletal muscle fibers.

When inorganic phosphate (Pi) is photogenerated from caged Pi during isometric contractions of glycerinated rabbit psoas muscle fibers, the released Pi binds to cross-bridges and reverses the working stroke of cross-bridges. The consequent force decline, the Pi-transient, is exponential and probes the kinetics of the power-stroke and Pi release. During muscle shortening, the fraction of attached cross-bridges and the average strain on them decreases (Ford, L. E., A.F. Huxley, and R.M. Simmons, 1977. Tension responses to sudden length change in stimulated frog muscle fibers near slack length. J. Physiol. (Lond.). 269:441-515; Ford, L. E., A. F. Huxley, and R.M. Simmons, 1985. Tension transients during steady state shortening of frog muscle fibers. J. Physiol. (Lond.). 361:131-150. To learn to what extent the Pi transient is strain dependent, muscle fibers were activated and shortened or lengthened at a fixed velocity during the photogeneration of Pi. The Pi transients observed during changes in muscle length showed three primary characteristics: 1) during shortening the Pi transient rate, Kpi, increased and its amplitude decreased with shortening velocity; Kpi increased linearly with velocity to > 110 s-1 at 0.3 muscle lengths per second (ML/s). 2) At a specific shortening velocity, increases in [Pi] produce increases in Kpi that are nonlinear with [Pi] and approach an asymptote. 3) During forced lengthening Kpi and the amplitude of the Pi transient are little different from the isometric contractions. These data can be approximated by a strain-dependent three-state cross-bridge model. The results show that the power stroke's rate is strain-dependent, and are consistent with biochemical studies indicating that the rate-limiting step at low strains is a transition from a weakly to a strongly bound cross-bridge state.     

Rate of phosphate release after photoliberation of adenosine 5'-triphosphate in slow and fast skeletal muscle fibers.

Inorganic phosphate (Pi) release was determined by means of a fluorescent Pi-probe in single permeabilized rabbit soleus and psoas muscle fibers. Measurements of Pi release followed photoliberation of approximately 1.5 mM ATP by flash photolysis of NPE-caged ATP in the absence and presence of Ca2+ at 15 degrees C. In the absence of Ca2+, Pi release occurred with a slow rate of 11 +/- 3 microM . s-1 (n = 3) in soleus fibers and 23 +/- 1 microM . s-1 (n = 10) in psoas fibers. At saturating Ca2+ concentrations (pCa 4.5), photoliberation of ATP was followed by rapid force development. The initial rate of Pi release was 0.57 +/- 0.05 mM . s-1 in soleus (n = 13) and 4.7 +/- 0.2 mM . s-1 in psoas (n = 23), corresponding to a rate of Pi release per myosin head of 3.8 s-1 in soleus and 31.5 s-1 in psoas. Pi release declined at a rate of 0.48 s-1 in soleus and of 5.2 s-1 in psoas. Pi release in soleus was slightly faster in the presence of an ATP regenerating system but slower when 0.5 mM ADP was added. The reduction in the rate of Pi release results from an initial redistribution of cross-bridges over different states and a subsequent ADP-sensitive slowing of cross-bridge detachment.     

Changes in the ATPase activity of insect fibrillar flight muscle during calcium and strain activation probed by phosphate-water oxygen exchange

During ATP hydrolysis by Ca2+-activated chemically skinned fibers from the flight muscle of the giant waterbug Lethocerus indicus, there is extensive phosphate-water oxygen exchange. For unstrained fibers the pattern of exchange shows that there is more than one pathway for hydrolysis, due to the ATPase activity of cross-bridges. Multiple pathways are an established property of both vertebrate actomyosin and fibers. The pattern of exchange can be fitted by two pathways: one with low exchange because the step(s) controlling Pi release are rapid, the other with high exchange and slow Pi release. The high-exchange pathway is responsible for most of the increase in ATPase activity on Ca2+ activation. On strain activation, only the high-exchange pathway is present, accounting for all the ATPase increase and responsible for force generation. In fully activated fibers, the cross-bridges which hydrolyze ATP and generate force behave uniformly with respect to oxygen exchange. The exchange pattern shows that the rate of Pi release changes dramatically over a very narrow strain increase. Step(s) controlling Pi release are at least partially rate-limiting for the overall ATPase reaction. The results are discussed in relation to models for strain activation and the identity of force-generating states.     

Characterization of phosphate oxygen exchange reactions catalyzed by myosin through measurement of the distribution of 18-O-labeled species

The change in the distribution of the phosphate species containing 0 to 4 18O oxygens per Pi was investigated during medium Pi equilibrium HOH exchange catalyzed by myosin subfragment 1. At 25 degrees C, a Pi molecule once bound loses an average of 3.9 of its original 4 oxygens prior to release which means that at least 100 reversals of the exchange reaction must have occurred. At 0 degrees C, only 3.4 of the 4 oxygens are lost prior to release indicating an average of 17 reversals. Distribution patterns are consistent with equivalent participation in the exchange reactions of all 4 oxygens of bound Pi. The intermediate exchange of Pi oxygens during hydrolysis of 18O-labeled ATP by myosin has also been investigated. The distribution of the product Pi species shows that there is an ATPase component in myosin preparations which hydrolyzes ATP without intermediate exchange. Presence of this component, which is likely a contaminating ATPase, provides a simple explanation of the apparent nonequivalence of phosphate oxygens which has been observed. When correction is made for this contaminant, characteristics of the myosin intermediate Pi equilibrium HOH exchange are similar to those of myosin subfragment 1 medium exchange, and intermediate exchange data are in much closer agreement with other kinetic measurements.     

Conformational flexibility of the Cys 697-Cys 707 segment of myosin subfragment-1. Distance distributions by frequency-domain fluorometry

The separation between Cys 697 (SH1) and Cys 707 (SH2) of the heavy chain of myosin subfragment-1 was previously measured by fluorescence resonance energy transfer with a donor linked to SH1 and an acceptor to SH2. In the present study the distribution of the distances between the two thiols was recovered from frequency-domain fluorometry. In the native state and in the presence of ligands such as MgADP, pyrophosphate, orthovanadate (Vi) and actin, we found wide distributions of the separations between SH1 and SH2 (11-16 A) comparable to that found in the random-coil state (20 A). These results suggest that the SH1-SH2 segment has a high degree of conformational flexibility even in native S1. The flexibility is not much affected by the physiological state of S1. However, the ligands MgADP, Vi and MgADP + Vi decrease significantly the mean SH1-SH2 distance from 27 to 17 A with the effect of MgADP+ Vi being the most pronounced. The anisotropy decay of donor-labeled S1 is biphasic with two rotational correlation times. The long component is decreased by these ligands from 289 to 93 ns, suggesting a more compact symmetric structure of S1 in the presence of the ligands. The complex S1(MgADP)Vi has been shown to be a stable analogue of S1(MgADP)Pi, an unstable intermediate that is generated in the actomyosin ATPase cycle during muscle contraction. Since the power stroke of muscle is accompanied by release of Pi from S1(MgADP)Pi, the present results are consistent with a model in which force generation can be accompanied by transition of S1 from a highly symmetric or compact structure to a more extended structure.