Test Research on the Knock of a Common-Rail Diesel Engine Fueled with Diesel-Methanol Dual-Fuel

Experiments were conducted on a diesel-methanol dual-fuel(DMDF)engine modified by a six-cylinder,turbocharged,inter-cooled diesel engine.According to the number of diesel injection,the experiments are divided to two parts:the single injectionmode and double injectionmode.The results show that,at the double injectionmode,themaximumof pressure rise rate is small and the engine runs smoothly,however,knock still occurswhen the cocombustion ratio(CCR)is big enough.Under knock status,the power density of the block vibration concentrating at some special frequencies rises dramatically,and the special frequency of single injection mode(about 4.1 kHz)is lower than that of double injection mode(7–9 kHz).The cylinder pressure oscillations of knock status are very different fromthe non-knock status.Under knock status,cylinder pressure oscillations become more concentrated and fiercer at some special frequencies,and the same as the block vibration.The special frequency of single injection mode(3–6 kHz)is lower than that of double injection mode(above 9 kHz).

论海上拖航合同中的Knock for Knock条款

海上拖航合同中的Knock For Knock条款的效力在实践中一直存在争议。本文通过分析《海商法》第162条,结合Knock For Knock条款的性质进行讨论,得出结论《海商法》第162条可以作为订立Knock For Knock条款的依据,但是不能作为该条款在任何情形下均有效的依据。并且通过对Knock For Knock条款效力的进一步探讨,可知海上拖航中的承拖方不得以该条款对抗被拖方提出的由于承拖方的故意或重大过失行为造成其损失的赔偿请求。除此之外,本文将借鉴国外判例,结合我国国情,提出减少Knock For Knock条款实践纠纷的建议。

knock-in法建立Cre重组酶表达鼠系

目的 建立神经特异性表达的Cre重组酶鼠系 ,以便在特定的组织中 ,于某一特定的时间段 ,用条件性基因剔除技术在活体状态下研究基因功能。方法 用knock -in技术 ,以微小管相关蛋白 1a(Mapla)启动子控制Cre重组酶表达。结果 PCR检测结果表明 ,Cre重组酶基因已成功植入该鼠系中。结论 本研究为配合条件性基因剔除技术 。

慢病毒介导的CYP3A11基因knock-down小鼠的建立

目的构建和筛选小鼠P4503A11基因miR RNAi慢病毒载体,建立CYP3A11基因knock-down小鼠。方法利用Ravi Sachidanandam's Lab的在线软件设计沉默小鼠P4503A11基因的miRNA所对应的shRNA序列,PCR扩增后克隆到PCI-GFP-MiR质粒中,其再与慢病毒载体FUW进行重组。将psPAX2和pMD2.G两个载体和FUW-GFP-MiR-shRNA重组慢病毒载体共转染293FT细胞,包装成病毒。用包装好的病毒液感染293FT细胞后,并利用GFP蛋白表达水平进行滴度测定。将上述重组慢病毒载体分别转染FVB/N小鼠肝细胞,转染48h后,检测P4503A11基因mRNA表达水平的变化。选取干扰效果最好的FUW-GFP-MiR-shRNA1重组慢病毒载体进行制备基因knock-down小鼠模型。用显微注射法将浓缩的shRNA1病毒液注射至FVB/N小鼠12细胞期胚胎透明带下。将注射后发育至22细胞期的胚胎移植至假孕受体母鼠,得F0代小鼠。在紫外光照射下利用滤光片观察GFP在小鼠活体内的表达水平。结果DNA测序结果显示3个重组慢病毒载体均与所设计的shRNA序列一致。检测的3个浓缩前慢病毒悬液的滴度均≥106TU/ml,经过高速离心对病毒进行浓缩和纯化,其滴度达到109TU/ml以上。转染FUW-GFP-MiR-shRNA1、2的2组肝细胞的P4503A11基因mRNA表达水平相对于空白对照组均显著下降(P<0.05),而转染阴性对照FUW-GFP-MiR-shRNA-NC的肝细胞P4503A11基因mRNA表达水平相对于空白对照组无明显改变。选用FUW-GFP-MiR-shRNA1慢病毒载体制备的F0代3A11基因knock-down小鼠,在紫外光照射下,阳性小鼠可见较强的荧光。结论构建并筛选出小鼠P4503A11基因有效的靶向miR RNAi慢病毒载体,并成功建立CYP3A11基因knock-down小鼠。

Simulation Research on the Effect of Cooled EGR, Supercharging and Compression Ratio on Downsized SI Engine Knock

Knock in spark-ignition（SI） engines severely limits engine performance and thermal efficiency. The researches on knock of downsized SI engine have mainly focused on structural design, performance optimization and advanced combustion modes, however there is little for simulation study on the effect of cooled exhaust gas recirculation（EGR） combined with downsizing technologies on SI engine performance. On the basis of mean pressure and oscillating pressure during combustion process, the effect of different levels of cooled EGR ratio, supercharging and compression ratio on engine dynamic and knock characteristic is researched with three- dimensional KIVA-3V program coupled with pressure wave equation. The cylinder pressure, combustion temperature, ignition delay timing, combustion duration, maximum mean pressure, and maximum oscillating pressure at different initial conditions are discussed and analyzed to investigate potential approaches to inhibiting engine knock while improving power output. The calculation results of the effect of just cooled EGR on knock characteristic show that appropriate levels of cooled EGR ratio can effectively suppress cylinder high-frequency pressure oscillations without obvious decrease in mean pressure. Analysis of the synergistic effect of cooled EGR, supercharging and compression ratio on knock characteristic indicates that under the condition of high supercharging and compression ratio, several times more cooled EGR ratio than that under the original condition is necessarily utilized to suppress knock occurrence effectively. The proposed method of synergistic effect of cooled EGR and downsizing technologies on knock characteristic, analyzed from the aspects of mean pressure and oscillating pressure, is an effective way to study downsized SI engine knock and provides knock inhibition approaches in practical engineering.

Effects of SIRT1 Gene Knock-Out via Activation of SREBP2 Protein-Mediated PI3K/AKT Signaling on Osteoarthritis in Mice

This study investigated the effects of SIRT1 gene knock-out on osteoarthritis in mice, and the possible roles of SREBP2 protein and the PI3K/AKT signaling pathway in the effects. Mice were randomly divided into a normal group and a SIRT1 gene knock-out group（6 mice in each group）. In these groups, one side of the knee anterior cruciate ligament was traversed, and the ipsilateral medial meniscus was cut to establish an osteoarthritis model of knee joint. The countralateral synovial bursa was cut out, serving as controls. The knee joint specimens were then divided into four groups： SIRT1^（＋/＋） control group（group A, n=6）; SIRT1^（＋/＋） osteoarthritis group（group B, n=6）; SIRT1^（–/–） control group（group C, n=6）; SIRT1^（–/–） osteoarthritis group（group D, n=6）. HE staining, Masson staining, Safranin O-Fast Green staining and Van Gieson staining were used to observe the morphological changes in the articular cartilage of the knee. Immunohistochemical staining was employed to detect the expression of SIRT1, SREBP2, VEGF, AKT, HMGCR and type Ⅱ collagen proteins. SA-β-gal staining was utilized to evaluate chondrocyte aging. The results showed clear knee joint cartilage destruction and degeneration in the SIRT1^（–/–） osteoarthritis group. The tidal line was twisted and displaced anteriorly. Type Ⅱ collagen was destroyed and distributed unevenly. Compared with the SIRT1^（＋/＋） osteoarthritis group and SIRT1^（–/–） control group, SIRT1 protein expression was not obviously changed in the SIRT1^（–/–） osteoarthritis group（P〉0.05）, while the expression levels of the SREBP2, VEGF and HMGCR proteins were significantly increased（P〈0.05） and the levels of AKT and type Ⅱ collagen proteins were significantly decreased（P〈0.05）. SIRT1 gene knock-out may aggravate cartilage degeneration in osteoarthritis by activating the SREBP2 protein-mediated PI3K/AKT signalling pathway, suggesting that SIRT1 gene may play a protective role against osteoarthritis.

Theoretical Analysis of Noise of Piston Knocking Cylinder Wall in Automotive Engines

Based on the loading conditions of engine, applying difference method to solve the hydrodynamic lubrication equation of piston skirt movement, the force acting on piston skirt and the moment on wrist pin were obtained. A computer program for simulating the piston second order motion was conducted to calculate the lateral motion of the upper part and the bottom part of piston skirts of the engine of automotive model CA1091. From the simulated result, the maximal impacting phase and the maximal impacting region of the piston were obtained. The result can be used for designing engine, diagnosing the noise of piston knocking cylinder wall and explaining many practical fault phenomena in theory.

Comparing successful gene knock-in efficiencies of CRISPR/Cas9 with ZFNs and TALENs gene editing systems in bovine and dairy goat fetal fibroblasts

This study aimed to compare the efficiencies of clustered regulatory interspaced short palindromic repeat（CRISPR）/Cas9-mediated gene knock-ins with zinc finger nucleases（ZFNs） and transcription activator-like effector nucleases（TALENs） in bovine and dairy goat fetal fibroblasts. To test the knock-in efficiency, a set of ZFNs and CRISPR/Cas9 plasmids were designed to edit the bovine myostatin（MSTN） gene at exon 2, while a set of TALENs and CRISPR/Cas9 plasmids were designed for editing the dairy goat β-casein gene at exon 2. Donor plasmids utilizing the ZFNs, TALENs, and CRISPR/Cas9 cutting sites were constructed in theGFP-PGK-Neo R plasmid background, including a 5′ and 3′ homologous arm flanking the genes humanized Fat-1（h Fat-1） or enhanced green fluorescent protein（eGFP）. Subsequently, the ZFNs, TALENs, or CRISPR/Cas9 and thehFat-1 or eGFP plasmids were co-transfected by electroporation into bovine and dairy goat fetal fibroblasts. After G418（Geneticin） selection, single cells were obtained by mouth pipetting, flow cytometry or a cell shove. The gene knock-in events were screened by PCR across the homologous arms. The results showed that in bovine fetal fibrobalsts, the efficiencies of ZFNs-mediated eGFP andhFat-1 gene knock-ins were 13.68 and 0%, respectively. The efficiencies of CRISPR/Cas9-mediated eGFP andhFat-1 gene knock-ins were 77.02 and 79.01%, respectively. The eGFP gene knock-in efficiency using CRISPR/Cas9 was about 5.6 times higher than when using the ZFNs gene editing system. Additionally, thehFat-1 gene knock-in was only obtained when using the CRISPR/Cas9 system. The difference of knockin efficiencies between the ZFNs and CRISPR/Cas9 systems were extremely significant（P〈0.01）. In the dairy goat fetal fibroblasts, the efficiencies of TALENs-mediated eGFP andhFat-1 gene knock-ins were 32.35 and 26.47%, respectively. Theefficiencies of eGFP and hFat-1 gene knock-ins using CRISPR/Cas9 were 70.37 and 74.29%, respectively. The knock-in efficiencies difference between the TALENs and CRISPR/Cas9 systems were extremely significant（P〈0.01）. This study demonstrated that CRISPR/Cas9 was more efficient at gene knock-ins in domesticated animal cells than ZFNs and TALENs. The CRISPR/Cas9 technology offers a new era of precise gene editing in domesticated animal cell lines.

运用Mask Pro和Knock Out进行指纹等痕迹物证图像处理的尝试

如何滤除复杂背景,以获取清晰的痕迹物证图像,除应用计算机进行图像处理外,还应借用Photo shop作为平台,尝试运用Mask Pro和Knock Out两款特效插件对指纹图像进行处理,效果会更加理想。

CRISPR/Cas9 knock-in介导的输卵管特异性表达转基因鸡研究进展

利用动物生物反应器生产重组蛋白是一种具有应用前景的生物技术。鸡输卵管生物反应器是理想的动物生物反应器之一,其优点在于表达的外源蛋白能够分泌到蛋清中,可避免蛋白提取过程中对鸡本身造成伤害,同时蛋清成分简单,便于后期的纯化。目前利用慢病毒结合原始生殖细胞(PGCs)制备转基因鸡被认为是最可行的方法,但因外源基因随机整合且生殖系传递效率较低,使转基因鸡研究受到技术上的限制。而2013年问世的CRISPR/Cas9基因敲入(CRISPR/Cas9 knock-in)技术能够使外源基因精准定向插入基因组特异性位点,这对生产输卵管特异性转基因鸡具有重大意义。文章综述了鸡输卵管反应器的研究进展、CRISPR/Cas9 knock-in技术在输卵管特异性表达转基因鸡研究和鸡育种领域的应用现状,并指出了目前存在的问题和相应的解决办法。

Knock prediction for dual fuel engines by using a simplified combustion model

The present work used a methane-air mixture chemical kinetics scheme consisting of 119 elementary reaction steps and 41 chemical species to develop a simplified combustion model for prediction of the knock in dual fuel engines. Calculated values by the model for natural gas operation showed good agreement with corresponding experimental values over a broad range of operating conditions.

多导睡眠监测设备和肠鸣音设备之间数据同步采集方法研究

目的提出一种基于敲击脉冲信号的两个独立设备数据同步采集的方法,以实现数据的同步采集。方法利用多导睡眠监测设备的鼾声传感器与肠鸣音设备相互敲击产生瞬态脉冲信号作为同步标记信号,基于同步标记信号的三阶累积量实现同步标记信号的识别和对齐,并采用多组同步标记信号评价数据同步的精度。结果多导睡眠监测设备的鼾声传感器和肠鸣音设备相互敲击,产生了清晰明确的同步标记脉冲信号,结合时域特征和三阶累积量实现了同步标记信号的精确识别和对齐。经过多组同步标记信号识别验证表明,此种同步方法可保证同步精度范围在5 ms内。结论该数据同步采集方法简单易行,可保证多导睡眠监测设备和肠鸣音设备的高精度同步采集,同时也为其他完全独立的多个设备数据同步采集提供参考。

Injuries Associated with Auto-Tricycle Crashes in an African City: Incidence and Pattern

Purpose: The aim of this study was to determine the incidence and pattern of injuries resulting from auto-tricycle crashes among patients in a tertiary referral centre in Ghana. Methods: Data were retrospectively extracted from hospital records of patients who got involved in auto-tricycle crashes and presented to the Accident and Emergency Centre of the Komfo Anokye Teaching Hospital (KATH), over a one-year period using a structured questionnaire. The gathered data were then entered into an electronic database and then analysed with SPSS version 20.0. Results: The incidence of injury following auto-tricycle crashes over the one-year period was 5.9% (95% CI: 4.9% - 7.0%) with a case fatality rate (FR) of 3.8% (95% CI: 1.3% - 8.7%). All the mortalities resulted from head and neck injuries and none of the patients involved wore a crash helmet. Only 5% of those studied wore crash helmets and were all drivers. Closed fractures accounted for 58% of the injuries, followed by open fractures, 28%. The most commonly fractured bones were the tibia/fibula, followed by the femur and then radius/ulna. The most common mechanism of injury was auto-tricycle toppling over (29%). Passengers were the most injured (48%), followed by drivers (37%) and pedestrians (15%). Most (72%) injuries among participants involved a single body part. On the injury severity scale, most (61%) of patients had minor trauma and 38% had major trauma. Conclusion: Auto-tricycle crashes account for 5.9% of injuries at the study site with a case fatality rate of 3.8%. Passengers had a higher injury rate (48%) than drivers (37%). Fractures of the tibia/fibula were most commonly associated with auto-tricycle crashes. Injuries to the head and neck were responsible for the deaths in the study participants and non-use of a crash helmet was associated with mortalities.

The Effect of Ignition Parameters on the Combustion Characteristics of an Aviation Piston Engine

A cylinder combustion simulation model was established for a two-stroke aviation piston engine used in a small unmanned aerial vehicle. The influence of different ignition system parameters on the combustion process of aviation kerosene was studied using this model. The research results showed that under the working conditions of 5500 r/min and 50% throttle opening, as the ignition energy increased, the peak values of average cylinder pressure and average temperature increased, and the combustion duration shortened, The advance of the combustion center of gravity increases the tendency of the engine to knock. Under the same operating conditions, as the ignition timing advances, the peak values of average pressure and average temperature in the cylinder increase, gradually approaching the top dead center, and the tendency of engine detonation increases more significantly.

基于CRISPR/Cas9技术构建IDO1基因敲除的THP-1细胞株及其表型研究

目的 采用CRISPR/Cas9技术构建吲哚胺-2,3-双加氧酶1(IDO1)基因敲除的THP-1细胞株,为研究IDO1在巨噬细胞中的作用提供细胞模型。方法 设计靶向IDO1基因的3条向导RNA(guide RNA,gRNA),分别构建IDO1-gRNA重组质粒,酶切及测序鉴定后,包装成Lenti-IDO1-gRNA慢病毒。利用Cas9慢病毒感染THP-1细胞获得稳定表达Cas9蛋白的细胞株,再经Lenti-IDO1-gRNA慢病毒感染敲除IDO1基因,有限稀释法获得单克隆细胞株。T7E1酶切检测gRNA打靶效率,PCR产物测序和Western blot鉴定IDO1基因敲除效果。CCK8法检测IDO1基因敲除对THP-1细胞增殖活性的影响,流式细胞术检测对巨噬细胞标志物CD11b、CD68和CD14表达的影响,中性红法检测巨噬细胞吞噬功能。结果 成功构建了3种IDO1-gRNA重组质粒,3条gRNA均能有效编辑IDO1基因,以gRNA2编辑效率最高。经PCR产物测序和Western blot验证获得了3株THP-1 IDO1-KO单克隆细胞株,并发现IDO1基因敲除可抑制THP-1细胞增殖,下调THP-1巨噬细胞CD11b、CD68表达,上调CD14表达,增强THP-1巨噬细胞吞噬功能。结论 成功构建IDO1基因敲除的THP-1细胞株;IDO1对THP-1细胞增殖活性、分化调节以及THP-1巨噬细胞吞噬功能调节有重要作用,为后续进一步探讨IDO1基因在巨噬细胞中的功能及机制研究奠定基础。

CRISPR/Cas9系统敲除山羊CDK2基因载体构建及转染效率检测

细胞周期蛋白依赖性蛋白激酶-2(CDK2)是细胞通过和进入S期所必须的细胞周期调节激酶,与许多癌细胞的生长有关。然而,目前在山羊上关于CDK2基因敲除及相关功能的研究尚未见报道。研究采用CRISPR/cas9系统,首先设计CDK2基因的sgRNA片段,在合成CDK2 sgRNA核苷酸序列后,构建能够同时表达sgRNA和Cas9 D10A的质粒PYSY-CDK2 sgRNA,连接并转化至大肠杆菌DH5α感受态细胞,对转化子进行验证,最后转染HEK293T细胞,检测细胞荧光和细胞增殖。酶切和测序结果证明,CDK2基因敲除载体构建正确;荧光显微镜检测显示,细胞转染效率在75%以上;细胞增殖检测表明,山羊CDK2敲除质粒不会对人源细胞增殖产生影响。说明采用CRISPR/Cas9构建的载体,可为后续获得山羊CDK2基因缺失型细胞系,研究支原体肺炎感染引发的细胞凋亡分子机制提供理论依据。

高温高负荷下Miller发动机扭矩提升的控制策略

为了解决Miller循环混动发动机在高温高负荷下爆震倾向明显增加而导致扭矩损失严重的问题,该研究通过台架试验,对进排气相位、喷油比例和喷油相位等关键控制参数进行了优化,并根据高温高负荷下发动机的工作特性,在发动机控制逻辑中引入温度修正。结果表明:进排气相位优化等措施成功抑制了爆震;优化后的发动机在1600 r/min高温全负荷工况下扭矩提升了18.5%;并且实现了对进排气相位、喷油参数和点火角的高温补偿,以及不同温度下控制参数的解耦,显著改善了高温高负荷下的动力性能衰减。