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knock-in法建立Cre重组酶表达鼠系
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作者 董铭 于澎 +1 位作者 饶明俐 于涌 《哈尔滨医科大学学报》 CAS 2002年第1期35-37,共3页
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... 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 。 展开更多
关键词 CRE重组酶 微小管相关蛋白1a knock-in法
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Estimation of Knock Wheel Noise of Rail Transport with an Acoustic Intensity Method
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作者 Kolykhalin Vitaly 《Journal of Mechanics Engineering and Automation》 2012年第8期465-469,共5页
The application of railway wheel noise compensation method is considered. The effect can be obtained by shift a pair of usual parallel rail joints on a pair of offset rail joints, for example, the right rail joint is ... The application of railway wheel noise compensation method is considered. The effect can be obtained by shift a pair of usual parallel rail joints on a pair of offset rail joints, for example, the right rail joint is shifted concerning the left rail joint on the certain distance. The distance is depend on the speed of train and promotes the correlation interconnection between excited acoustical pulses. The intensity wheel knock spectrograms corresponding of different distance between rail joints registered by the microphone are estimated. The spectrograms considerably differed from each other in low frequency band are analyzed. The result of practical using the acoustic noise compensation method is proved. 展开更多
关键词 Control noise ACOUSTIC RAILWAY MICROPHONE wheel knock.
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慢病毒介导的CYP3A11基因knock-down小鼠的建立 被引量:1
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作者 庞浩 刘昌峨 +4 位作者 刘勤 王露露 郭科男 王勇 魏泓 《第三军医大学学报》 CAS CSCD 北大核心 2010年第5期422-426,共5页
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... 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。 展开更多
关键词 细胞色素P-450 CYP3A knock-DOWN 微RNAS 慢病毒 小鼠
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Effects of SIRT1 Gene Knock-Out via Activation of SREBP2 Protein-Mediated PI3K/AKT Signaling on Osteoarthritis in Mice 被引量:7
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作者 于斐 曾晖 +4 位作者 雷鸣 肖德明 李伟 袁昊 林健静 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2016年第5期683-690,共8页
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 norm... 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. 展开更多
关键词 cartilage knock articular degeneration cruciate medial collagen ligament staining aggravate
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Theoretical Analysis of Noise of Piston Knocking Cylinder Wall in Automotive Engines 被引量:1
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作者 肖云魁 李世义 +2 位作者 曹亚娟 杨万成 冯汉生 《Journal of Beijing Institute of Technology》 EI CAS 2005年第3期284-288,共5页
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... 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. 展开更多
关键词 piston knocking cylinder wall second order motion fault diagnosis
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Simulation Research on the Effect of Cooled EGR,Supercharging and Compression Ratio on Downsized SI Engine Knock 被引量:1
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作者 SHU Gequn PAN Jiaying +1 位作者 WEI Haiqiao SHI Ning 《Chinese Journal of Mechanical Engineering》 SCIE EI CAS CSCD 2013年第2期341-350,共10页
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 advance... 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. 展开更多
关键词 exhaust gas recirculation compression ratio knock pressure wave equation pressure oscillation
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Comparing successful gene knock-in efficiencies of CRISPR/Cas9 with ZFNs and TALENs gene editing systems in bovine and dairy goat fetal fibroblasts 被引量:12
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作者 LIU Hui LIU Chang +5 位作者 ZHAO Yu-hang HAN Xue-jie ZHOU Zheng-wei WANG Chen LI Rong-feng LI Xue-ling 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2018年第2期406-414,共9页
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 effe... 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. 展开更多
关键词 myostatin(MSTN) β-casein(CSN2) bovine fetal fibroblasts CRISPR/Cas9 dairy goat fetal fibroblasts eGFP hFat-1 knock-in mutation efficiency TALENs ZFNs
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CRISPR/Cas9 knock-in介导的输卵管特异性表达转基因鸡研究进展
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作者 刘灵康 郑喜邦 《中国家禽》 北大核心 2020年第3期75-81,共7页
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。... 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。 展开更多
关键词 转基因鸡 原始生殖细胞 CRISPR/Cas9 基因敲入 输卵管特异性表达
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运用Mask Pro和Knock Out进行指纹等痕迹物证图像处理的尝试
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作者 王德江 李福臻 《吉林公安高等专科学校学报》 2002年第4期37-40,共4页
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。
关键词 knock 0ut软什 MASK Pro软什 图像处理 计算机应用 指纹 痕迹学
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Knock prediction for dual fuel engines by using a simplified combustion model
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作者 费少梅 刘震涛 严兆大 《Journal of Zhejiang University Science》 EI CSCD 2003年第5期591-594,共4页
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 fue... 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. 展开更多
关键词 Dual fuel engines Combustion model knock prediction
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Test Research on the Knock of a Common-Rail Diesel Engine Fueled with Diesel-Methanol Dual-Fuel
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作者 Chao Zhu Zhuopei Liu +1 位作者 Hao Chen Yangyang Li 《Energy Engineering》 EI 2023年第5期1081-1105,共25页
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 p... 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). 展开更多
关键词 Common rail diesel engine diesel-methanol dual-fuel knock block vibration pressure oscillation
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论海上拖航合同中的Knock for Knock条款
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作者 刘姝彤 唐青玉 《珠江水运》 2023年第12期33-35,共3页
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... 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。 展开更多
关键词 海上拖航合同 knock For knock条款 适拖义务
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雾化吸入后高频率叩背联合诱发性肺量计训练对老年肺癌术后排痰和肺康复的影响 被引量:1
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作者 吴杏尧 刘明慧 +1 位作者 李心慧 曹慧娇 《现代医学》 2025年第1期141-146,共6页
目的:探讨雾化吸入后高频率叩背联合诱发性肺量计训练对老年肺癌术后排痰和肺康复的影响。方法:选取2021年3月至2024年3月在本院行手术治疗的老年肺癌患者112例,采用随机数字表法将患者分为对照组与试验组。对照组(56例)采用传统低频率... 目的:探讨雾化吸入后高频率叩背联合诱发性肺量计训练对老年肺癌术后排痰和肺康复的影响。方法:选取2021年3月至2024年3月在本院行手术治疗的老年肺癌患者112例,采用随机数字表法将患者分为对照组与试验组。对照组(56例)采用传统低频率叩背联合呼吸训练干预;试验组(56例)采用高频率叩背联合诱发性肺量计训练干预,比较两组术后排痰情况、肺康复效果及生活质量。结果:试验组干预后日均排痰量为(24.23±4.67)mL,多于对照组的(17.90±4.12)mL(P<0.05);痰液黏稠度低于对照组(P<0.05)。呼气流量峰值(PEF)为(461.28±19.37)L·min^(-1)、用力肺活量(FVC)为(3.09±0.61)L、第1秒用力呼气容积(FEV1)为(77.41±7.35)%,均大于对照组的(420.09±16.35)L·min^(-1)、(2.79±0.78)L、(70.88±9.44)%(P<0.05)。试验组Borg呼吸困难评分为(2.07±0.49)分,低于对照组的(2.89±0.51)分(P<0.05)。肺部并发症总发生率为3.57%,低于对照组(14.29%)(P<0.05)。生活质量评分高于对照组(P<0.05)。结论:雾化吸入后高频率叩背联合诱发性肺量计训练在改善老年肺癌术后患者排痰效果,减轻呼吸困难程度,促进肺功能康复,降低肺部并发症发生风险,提高患者生活质量方面具有显著优势,值得临床推广。 展开更多
关键词 肺癌 综合护理 叩背 诱发性肺量计训练 排痰 肺功能
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Improved Learning and Memory of Pavlovian Context Fear Conditioning in Histidine Decarboxylase Knock- Out Mice
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作者 Liu Luying Shen Dongxia Jin Chunlei Zhuge Zhengbing Gong Yingxia Zhu Yuanyuan Chen Zhong 《中国药理通讯》 2005年第4期30-31,共2页
关键词 巴甫洛夫学说 学习功能 记忆能力 组氨酸脱羧酶 小鼠 动物实验
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柴油机空转加速敲缸异响故障诊断与改进实践 被引量:1
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作者 李飞 《机械管理开发》 2025年第9期155-157,共3页
针对柴油机空转加速时易出现敲缸异响故障,通过对敲缸异响故障原理与诊断的初步分析,进而分别从预喷油量、喷油提前角及轨压上升速率三个方面出发,提出柴油机空转加速敲缸异响故障的改进方案。由研究可知,预喷油量对敲缸异响的影响相对... 针对柴油机空转加速时易出现敲缸异响故障,通过对敲缸异响故障原理与诊断的初步分析,进而分别从预喷油量、喷油提前角及轨压上升速率三个方面出发,提出柴油机空转加速敲缸异响故障的改进方案。由研究可知,预喷油量对敲缸异响的影响相对较小,而喷油提前角与轨压上升速率对敲缸异响的影响相对较大,其中,当预喷油量为1.0 mg、喷油提前角(CA)处于5°~11°区间内、轨压上升速率减小到100 MPa/s时,柴油机性能最佳,空转加速时产生的敲缸异响越小,有效改善柴油机运行状态,使柴油机在机械设备工作中发挥出更大的作用。 展开更多
关键词 柴油机 敲缸异响 故障诊断 改进方案
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自有外汇“对敲”行为的刑法认定
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作者 汪明亮 《上海政法学院学报(法治论丛)》 2025年第4期100-111,共12页
自有外汇“对敲”行为是否构成非法经营罪存在截然相反的观点。界定自有外汇“对敲”行为性质应坚持形式判断与实质判断相结合理念、刑法最后手段性理念、社会效果与政治效果理念、治罪与治理并重理念。自有外汇“对敲”行为的刑法认定... 自有外汇“对敲”行为是否构成非法经营罪存在截然相反的观点。界定自有外汇“对敲”行为性质应坚持形式判断与实质判断相结合理念、刑法最后手段性理念、社会效果与政治效果理念、治罪与治理并重理念。自有外汇“对敲”行为的刑法认定必须考察“对敲型”非法经营罪的四个核心要件,即具有刑法意义上的非法经营行为、变相买卖外汇是变相倒买倒卖外汇、具有营利目的、属于地下钱庄或换汇中介。自有外汇“对敲”行为不具有“对敲型”非法经营罪的核心要件,不构成非法经营罪。可以通过反向行刑衔接,妥善解决自有外汇“对敲”行为的法律责任。 展开更多
关键词 自有外汇“对敲”行为 非法经营罪 司法理念 构成要件 反向行刑衔接
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宿主蛋白HERC4影响口蹄疫病毒复制的作用研究 被引量:1
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作者 郑东东 冯涛 +4 位作者 郝荣增 杨彪 晏子龙 齐晓兰 郑海学 《中国兽医科学》 北大核心 2025年第4期435-440,共6页
本研究旨在探究宿主蛋白HERC4对口蹄疫病毒(FMDV)复制的影响,首先利用CRISPR/Cas9技术在PK15细胞中构建了HERC4基因敲除细胞系,通过DNA测序和蛋白免疫印迹试验证明构建的基因敲除细胞系成功,然后在BHK21细胞中转染不同剂量的HERC4蛋白... 本研究旨在探究宿主蛋白HERC4对口蹄疫病毒(FMDV)复制的影响,首先利用CRISPR/Cas9技术在PK15细胞中构建了HERC4基因敲除细胞系,通过DNA测序和蛋白免疫印迹试验证明构建的基因敲除细胞系成功,然后在BHK21细胞中转染不同剂量的HERC4蛋白表达质粒,制备了HERC4基因过表达细胞,随后在HERC4基因敲除细胞和过表达细胞中,分别利用蛋白免疫印迹、实时荧光定量和病毒滴度测定方法检测了HERC4蛋白对FMDV复制的影响,结果表明,HERC4蛋白对FMDV复制有影响,敲除可以抑制病毒复制,过表达可以促进病毒复制。该研究结果为后续开展HERC4蛋白调控FMDV复制机制研究奠定了基础。 展开更多
关键词 口蹄疫病毒 宿主蛋白HERC4 基因敲除 基因过表达
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喷射模式对高压缩比甲醇发动机爆震特性影响的试验研究
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作者 于立信 尹晓军 +3 位作者 王锡斌 段浩 胡二江 曾科 《西安交通大学学报》 北大核心 2025年第7期108-117,共10页
为深入探究高压缩比甲醇发动机的爆震抑制方法,在一台压缩比为15∶1的单缸自然吸气点燃式甲醇发动机上,开展了化学计量比混合气条件下喷射模式对爆震特性影响的试验研究。首先,在进气道喷射模式下,研究了负荷、点火时刻和过量空气系数... 为深入探究高压缩比甲醇发动机的爆震抑制方法,在一台压缩比为15∶1的单缸自然吸气点燃式甲醇发动机上,开展了化学计量比混合气条件下喷射模式对爆震特性影响的试验研究。首先,在进气道喷射模式下,研究了负荷、点火时刻和过量空气系数对爆震特性的影响;其次,在缸内直喷模式下,研究了喷射时刻对爆震特性的影响;最后,在节气门全开状态下,研究了多次喷射策略对爆震的抑制效果。结果表明:采用进气道喷射模式时,甲醇发动机在中高负荷下均发生了爆震燃烧,在指示平均有效压力(p_(i))为0.7 MPa时,出现低频率、高强度的强烈爆震现象,最大爆震强度达8.42 MPa;采用缸内直喷模式时,发动机爆震现象被显著抑制,当p_(i)为0.91 MPa时,最大爆震强度下降到0.43 MPa;合理的两次喷射策略相比单次喷射策略可进一步降低爆震强度,同时保持较高的p_(i)。研究验证了缸内直喷结合两次喷射策略在抑制高压缩比甲醇发动机爆震现象方面的有效性,为甲醇发动机的发展和应用提供了理论依据和数据支撑。 展开更多
关键词 甲醇 高压缩比 爆震特性 爆震抑制 喷射策略
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烟草脂类合成关键基因NtLPAT的功能验证
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作者 吉白璐 孙艺文 +12 位作者 刘万峰 钱亚新 蒋彩虹 耿锐梅 刘旦 程立锐 杨爱国 黄立钰 李晓旭 蒲文宣 高军平 张强 文柳璎 《作物学报》 北大核心 2025年第9期2527-2537,共11页
溶血磷脂酰基转移酶(LPAT)是负责催化溶血磷脂酸(LPA)和脂肪酰基-辅酶A(Acyl-CoA)酯化生成磷脂酸(PA)合成途径的关键酶,尚不清楚烟草中LPAT的基因功能。本研究从K326中克隆得到NtLPAT基因,利用CRISPR/Cas9技术获得NtLPAT敲除植株ntlpat... 溶血磷脂酰基转移酶(LPAT)是负责催化溶血磷脂酸(LPA)和脂肪酰基-辅酶A(Acyl-CoA)酯化生成磷脂酸(PA)合成途径的关键酶,尚不清楚烟草中LPAT的基因功能。本研究从K326中克隆得到NtLPAT基因,利用CRISPR/Cas9技术获得NtLPAT敲除植株ntlpat,对其农艺性状和抗病性以及外观质量进行鉴定,结合脂质组、转录组分析NtLPAT的功能。结果表明,NtLPAT受青枯病菌和黑胫病病菌的诱导。ntlpat株高降低,对黄瓜花叶病毒病(CMV)与青枯病的抗性提高。脂质组分析表明,ntlpat中甘油酯代谢发生了重排,三酰甘油含量显著降低,组成类囊体膜的甘油糖脂含量(MGDG,DGDG)随之上升,并且鞘脂和磷脂酰肌醇含量也发生变化。转录组分析表明,ntlpat的光合作用、碳固定代谢、鞘脂合成和磷脂酰肌醇信号转导途径的相关基因发生重编程,因此推测NtLPAT具有脂酰基转移酶活性,调控三酰甘油的从头合成途径,并参与细胞信号传导,影响烟草的生长和对CMV、青枯病菌的抗性,研究结果为挖掘烟草株型及抗性改良提供基因资源和试验依据。 展开更多
关键词 溶血磷脂酰基转移酶 烟草 基因敲除 脂质组学 转录组学
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缸内直喷氢气发动机爆震抑制方法模拟研究
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作者 黄海碧 卢华林 +3 位作者 涂展飞 覃玉峰 林铁坚 黄豪中 《车用发动机》 北大核心 2025年第6期1-8,共8页
受爆震限制,氢气发动机的压缩比普遍低于柴油发动机,因此克服高压缩比下爆震燃烧限制对氢气发动机的应用具有重要意义。本研究通过数值模拟,研究了高压缩比条件下喷射策略结合点火时刻对缸内直喷氢气发动机爆震的抑制潜力。结果表明:压... 受爆震限制,氢气发动机的压缩比普遍低于柴油发动机,因此克服高压缩比下爆震燃烧限制对氢气发动机的应用具有重要意义。本研究通过数值模拟,研究了高压缩比条件下喷射策略结合点火时刻对缸内直喷氢气发动机爆震的抑制潜力。结果表明:压缩比为15时,随着喷射压力的增大,爆震强度(KI)先减小后增大;喷射压力为6 MPa时,KI最小,此时火花塞附近氢气分布更为富集,混合气较易点燃,火焰核心更为稳定;爆震强度随着喷射正时的变化呈非线性,当喷射正时为-100°时,缸内混合气分布更均匀,燃烧重心(CA50)更接近上止点,KI降到最低;仅通过调整喷射压力或喷射正时,最低KI值均未能降至1 MPa以下,需要延迟点火提前角以降低KI值。 展开更多
关键词 直喷氢气发动机 高压缩比 喷射策略 爆震强度
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