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靶向CD99的CAR-T细胞扩增优化研究
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作者 王伊玄 于淼 +6 位作者 赵家旋 赵芬芳 曾毅 王友湧 祝海川 张同存 史江舟 《中国癌症杂志》 CAS CSCD 北大核心 2024年第7期639-649,共11页
背景与目的:嵌合抗原受体T(chimeric antigen receptor T,CAR-T)细胞疗法对血液和淋巴系统肿瘤的治疗效果显著,但对实体瘤效果较差,这与靶点选择的因素有关。针对尤因肉瘤(Ewing sarcoma,ES),CD99可作为CAR-T细胞潜在的靶点。由于T细胞... 背景与目的:嵌合抗原受体T(chimeric antigen receptor T,CAR-T)细胞疗法对血液和淋巴系统肿瘤的治疗效果显著,但对实体瘤效果较差,这与靶点选择的因素有关。针对尤因肉瘤(Ewing sarcoma,ES),CD99可作为CAR-T细胞潜在的靶点。由于T细胞自身表达CD99蛋白,靶向CD99的CAR-T细胞存在体外扩增能力有限的问题。本研究旨在通过增加CD99敲低的短发夹RNA(short hairpin RNA,shRNA)、优化慢病毒转导的感染复数(multiplicity of infection,MOI)、筛选培养CAR-T细胞的培养基及培养容器,以获得CD99 CAR-T细胞制备的最优条件。方法:筛选shRNA序列,提高CD99 CAR-T细胞的扩增能力。采用不同的MOI、培养基和培养容器,分别检测在不同条件下CAR-T细胞的转导效率、细胞存活率、增殖能力、特异性杀伤能力及干扰素-γ(interferon-γ,IFN-γ)释放水平等,筛选出最优的细胞制备条件。结果:通过shRNA敲低得到的KO-CD99 CAR-T细胞扩增水平显著高于CD99 CAR-T细胞[(16.40±0.40)vs(6.33±1.53),P<0.01]。转导MOI为0.25~1.0、培养基为OptiVitro时细胞的扩增效果最优。在透气瓶中培养的CAR-T细胞扩增倍数显著高于在培养袋中培养的细胞[MOI=0.25:(50.23±3.32)vs(13.02±4.82);MOI=0.50:(49.96±0.83)vs(18.25±2.88);MOI=1.00:(48.27±5.08)vs(13.16±6.26);P<0.01],且细胞分型更优、特异性杀伤更高。结论:通过shRNA技术得到的KO-CD99 CAR-T细胞可实现稳定扩增。从扩增条件优化结果看,MOI为0.25~1.00,培养基为OptiVitro,培养容器为透气瓶的条件下,KO-CD99 CAR-T细胞可获得更优的扩增能力、更多比例的记忆T细胞,为后续开展CD99 CAR-T细胞治疗ES的临床试验奠定了坚实基础。 展开更多
关键词 CD99 嵌合抗原受体T细胞 扩增优化 KO-CD99
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靶向CD30的CAR-T细胞慢病毒转导条件优化研究
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作者 田高辉 张琴星 +5 位作者 史江舟 赵芬芳 王宁 赵家旋 卢玉琳 徐瑶 《中国癌症杂志》 CAS CSCD 北大核心 2023年第7期646-654,共9页
背景与目的:嵌合抗原受体T(chimeric antigen receptor T,CAR-T)细胞技术在血液肿瘤治疗领域已被广泛应用,慢病毒转导是CAR-T细胞制备的关键环节,与CAR-T细胞的质量密切相关,因此慢病毒转导过程涉及的各项参数仍需进一步优化。本研究旨... 背景与目的:嵌合抗原受体T(chimeric antigen receptor T,CAR-T)细胞技术在血液肿瘤治疗领域已被广泛应用,慢病毒转导是CAR-T细胞制备的关键环节,与CAR-T细胞的质量密切相关,因此慢病毒转导过程涉及的各项参数仍需进一步优化。本研究旨在探讨携带CD30抗体序列的慢病毒转导T细胞的感染复数(multiplicity of infection,MOI)、温育密度、转导前活化时间和转导体系高度对抗CD30 CAR-T细胞的影响,优化CAR-T细胞的转导条件,提高转导效率和CAR-T细胞功能。方法:采用不同的MOI、温育密度、转导前活化时间和转导体系高度等对人外周血来源的T细胞进行转导优化,转导后分别检测抗CD30 CAR-T细胞的增殖能力、转导效率、细胞存活率和体外杀伤效率等,以确定最优的T细胞转导条件。结果:MOI为1.00、1.50和3.00时转导效率和有效细胞数显著高于0.00、0.25和0.50组。在温育密度大于1.0×10^(7)个/mL时,温育密度对T细胞转导效率无影响。活化时间为72 h组细胞存活率低于80%,显著低于其他组;24、48 h的转导效率显著高于0、8、16 h组;48 h组CAR-T细胞的增殖速率显著高于24 h组。转导体系高度为0.16 mm时转导效率和增殖倍数都显著高于0.53 mm组,但对CAR-T细胞的体外杀伤效率无影响。结论:通过对CAR-T细胞功能的综合评估,确定慢病毒的最佳转导条件为MOI=1、温育密度为1.0×10^(7)个/mL、转导前活化48h、转导体系高度为0.16mm。 展开更多
关键词 慢病毒载体 抗CD30嵌合抗原受体T细胞 转导条件
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水下圆形重力式网箱缆绳张力对波、流的响应特性 被引量:7
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作者 刘圣聪 黄六一 +5 位作者 倪益 程晖 赵芬芳 王欣欣 梁振林 KINOSHITA Takeshi 《水产学报》 CAS CSCD 北大核心 2019年第6期1518-1526,共9页
通过模型试验研究了周长40 m、网高10 m的HDPE圆形升降式网箱沉降在水下时的缆绳张力。试验采用狄克逊准则,模型大比例尺λ=10和小比例尺λa=λd=2;网箱采用网格形式锚泊系统。试验时前主缆绳水平方向与波流夹角为0°,试验流速为0、... 通过模型试验研究了周长40 m、网高10 m的HDPE圆形升降式网箱沉降在水下时的缆绳张力。试验采用狄克逊准则,模型大比例尺λ=10和小比例尺λa=λd=2;网箱采用网格形式锚泊系统。试验时前主缆绳水平方向与波流夹角为0°,试验流速为0、5、10和15 cm/s,波浪周期范围1.3~2.2 s,波高范围10~30 cm。在各试验条件下对主缆绳和系框绳以及浮框绳张力进行了测试,以受力最大的缆绳为研究对象,分析了缆绳最大张力与波流因子之间的关系。结果显示:①在纯流作用下,前主缆绳受力最大;在纯波作用下,后侧主缆绳受力最大,最大张力随波高增大呈线性增长趋势,随周期变化的影响较小。②在波流作用下,高流速、低波高、高波周期条件时前主缆绳受力最大;在低流速、高波高、低波周期时后侧主缆绳受力最大;前主缆绳受水流作用影响较大,后侧主缆绳受波浪作用影响较大。③在波流作用下,各缆绳最大张力随波高、流速增大而增大,随周期增大而减小;后侧主缆绳最大张力无量纲量随波陡增大且基本呈线性增长趋势。 展开更多
关键词 升降式重力式网箱 缆绳张力 波浪 水流
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基于模型试验准则的重力式圆形网箱水动力比较研究 被引量:5
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作者 黄六一 倪益 +3 位作者 程晖 王欣欣 赵芬芳 梁振林 《渔业现代化》 2018年第2期6-12,共7页
模型试验准则是水槽模型试验的核心问题,关系到最终推算实物系统结果的准确性。以中国常用的高密度聚乙烯(HDPE)圆形重力式网箱为研究对象,采用田内准则和狄克逊准则为模型试验准则,以大尺度比10∶1、小尺度比2∶1设计制作了网箱模型,... 模型试验准则是水槽模型试验的核心问题,关系到最终推算实物系统结果的准确性。以中国常用的高密度聚乙烯(HDPE)圆形重力式网箱为研究对象,采用田内准则和狄克逊准则为模型试验准则,以大尺度比10∶1、小尺度比2∶1设计制作了网箱模型,在循环动力水槽中分别进行网箱水动力特性模型试验,比较在水流作用下两种准则的试验结果及其差异性。结果显示:网箱整体和网箱箱体的阻力随着流速增加而增加,二者呈幂函数关系;网箱整体阻力系数(C)随着雷诺数(Re)的增加而减小,二者的关系为C=17.587Re^(-0.354)(R^2=0.941);C随着Re的增加而减小,二者的关系为C=30.526 Re^(-0.424)(R^2=0.902);在实际流速30~130 cm/s范围内,田内准则的试验数据比狄克逊准则的试验数据稍小,二者的阻力差约为-5%。由试验结果可知,田内准则和狄克逊准则均可用于网箱模型水流试验。研究结果可为网箱模型实验准则的选择和圆形网箱水动力学计算提供理论依据和基本参数。 展开更多
关键词 HDPE圆形网箱 水动力 田内准则 狄克逊准则
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重症患者尿管相关性尿路感染的病原菌分布及耐药性分析 被引量:11
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作者 赵芬芬 李萍 王花 《山西医药杂志》 CAS 2018年第1期21-23,共3页
目的探讨重症患者尿管相关性尿路感染(CAUTI)的病原菌分布和耐药性,指导临床合理用药。方法收集重症医学科1 214例留置尿管患者的尿液标本,采用法国生物梅里埃公司VITEK-2Compact全自动微生物分析仪进行病原菌鉴定和药敏试验。结果 1 21... 目的探讨重症患者尿管相关性尿路感染(CAUTI)的病原菌分布和耐药性,指导临床合理用药。方法收集重症医学科1 214例留置尿管患者的尿液标本,采用法国生物梅里埃公司VITEK-2Compact全自动微生物分析仪进行病原菌鉴定和药敏试验。结果 1 214例留置尿管住院患者中,合并尿路感染者有315例,感染率为25.95%;共分离出病原菌338株,主要以革兰阴性菌(84.0%)为主,但非发酵菌感染和革兰阳性菌感染率较高。当前主要尿路感染病原菌为大肠埃希菌(46.4%)、铜绿假单胞菌(13.0%)、鲍曼不动杆菌(11.5%)、屎肠球菌(6.8%)和粪肠球菌(4.7%),均存在多重耐药情况。结论 CAUTI病原菌复杂,耐药严重;结合CAUTI的病原菌种类及耐药谱针对性用药,才能取得较好疗效。 展开更多
关键词 重症监护 尿管相关性尿路感染 病原菌 耐药性
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Studies on Factors Influencing Hydrodynamic Characteristics of Plates Used in Artificial Reefs 被引量:7
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作者 TANG Yanli YANG Wenzhao +3 位作者 SUN Liyuan zhao fenfang LONG Xiangyu WANG Gang 《Journal of Ocean University of China》 SCIE CAS CSCD 2019年第1期193-202,共10页
As a simplified model of artificial reefs, a series of plate models punched with square or circular openings are designed to investigate the effects of openings on the hydrodynamic characteristics of artificial reefs.... As a simplified model of artificial reefs, a series of plate models punched with square or circular openings are designed to investigate the effects of openings on the hydrodynamic characteristics of artificial reefs. The models are grouped by various opening numbers and opening-area ratios. They are physically tested in a water flume or used in the numerical simulation to obtain the drag force in the uniform flow with different speeds. The simulation results are found in good agreement with the experimental measurements. By the non-dimensional analysis, the drag coefficient specified to each model is achieved and the effects of openings are examined. It is found that the key factor affecting the drag coefficient is the open-area ratio. Generally, the drag coefficient is a linear function of the open area ratio with a minus slope. The empirical formulae for the square and circular openings respectively are deduced by means of the multiple regression analysis based on the measured and numerical data. They will be good references for the design of new artificial reefs. As a result of numerical simulation, the vorticity contours and pressure distribution are also presented in this work to better understand the hydrodynamic characteristics of different models. 展开更多
关键词 artificial REEFS drag COEFFICIENT HYDRODYNAMIC characteristics model EXPERIMENT
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A Comparative Study on Hydrodynamic Performance of Double Deflector Rectangular Cambered Otter Board 被引量:6
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作者 XU Qingchang FENG Chunlei +7 位作者 HUANG Liuyi XU Jiqiang WANG Lei ZHANG Xun LIANG Zhenlin TANG Yanli zhao fenfang WANG Xinxin 《Journal of Ocean University of China》 SCIE CAS CSCD 2018年第5期1218-1224,共7页
In the present work,the hydrodynamic performance of the double deflector rectangular cambered otter board was studied using wind tunnel experiment,flume tank experiment and numerical simulation.Results showed that the... In the present work,the hydrodynamic performance of the double deflector rectangular cambered otter board was studied using wind tunnel experiment,flume tank experiment and numerical simulation.Results showed that the otter board had a good hydrodynamic performance with the maximum lift-to-drag ratio(K_(MAX) = 3.70).The flow separation occurred when the angle of attack(AOA) was at 45?,which revealed that the double deflector structure of the otter board can delay the flow separation.Numerical simulation results showed a good agreement with experiment ones,and could predict the critical AOA,which showed that it can be used to study the hydrodynamic performance of the otter board with the advantage of flow visualization.However,the drag coefficient in flume tank was much higher than that in wind tunnel,which resulted in a lower lift-to-drag ratio.These may be due to different fluid media between flume tank and wind tunnel,which result in the big difference of the vortexes around the otter board.Given the otter boards are operated in water,it was suggested to apply both flume tank experiment and numerical simulation to study the hydrodynamic performance of otter board. 展开更多
关键词 wind tunnel experiment flume tank experiment numerical simulation double deflector rectangular cambered otter board hydrodynamic performance
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Experimental Investigation of Local Scour Around Artificial Reefs in Steady Currents
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作者 TANG Yanli WEI Shuangwu +2 位作者 YANG Mingda WANG Xinxin zhao fenfang 《Journal of Ocean University of China》 SCIE CAS CSCD 2022年第2期445-456,共12页
Artificial reef is a man-made object that is deployed purposefully on the seafloor to restore the offshore fishery resources and the ecological environment.To secure its ecological effects,it is important to study the... Artificial reef is a man-made object that is deployed purposefully on the seafloor to restore the offshore fishery resources and the ecological environment.To secure its ecological effects,it is important to study the possible instability of artificial reefs,like drifting and reversing caused by burial and scour in different seafloor conditions.In the present study,experiments of local scour around an artificial reef are carried out in steady currents.The effect of the open-area ratios and the open-hole heights of the cubic reefs,and the bottom angles of the triangular reefs on the time-scale of the scour process and the equilibrium scour depth are investigated.The results indicate that for the cubic artificial reef,the scour depth decreases with the increasing open-area ratios,and increases with the increasing open-hole heights.In the present study,the optimal prototype of the cubic reef with an open-area ratio of 0.49 and open-height of 0.7 m produces the minimum scour depth.For the triangular reef,the scour depth increases when the bottom angle increases.Moreover,based on the experimental results,empirical equations of the effects of the cut-opening and the bottom angle on the maximum equilibrium scour depth are proposed.The formulas will provide theoretical support and practical guidance for the optimized design and construction of artificial reefs. 展开更多
关键词 local scour artificial reefs open-area ratios open-hole heights bottom angles
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Effects of Trawl Selectivity and Genetic Parameters on Fish Body Length Under Long-Term Trawling
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作者 YU Yang SUN Peng +4 位作者 CUI He SHENG Huaxiang zhao fenfang TANG Yanli CHEN Zelin 《Journal of Ocean University of China》 SCIE CAS 2015年第5期835-840,共6页
Long-term fishing pressure affects the biological characteristics of exploited fish stocks. The biological characteristics of hairtail(Trichiurus lepturus) in the East China Sea are unable to recover because of long-t... Long-term fishing pressure affects the biological characteristics of exploited fish stocks. The biological characteristics of hairtail(Trichiurus lepturus) in the East China Sea are unable to recover because of long-term trawling. Fishing induces evolutionary effects on the fish's biological characteristics. Evidence of these changes includes small size at age, a shift to earlier age structure, and early maturation. Natural and artificial selection usually affect the fish's life history. Selection can induce different chances of reproduction, and individual fish can give a different genetic contribution to the next generation. In this study, analysis of time-dependent probability of significance and test of sensitivity were used to explore the effects of fish exploitation rate, mesh size, and heritability with long-term trawling. Results showed that fishing parameters were important drivers to exploited fish population. However, genetic traits altered by fishing were slow, and the changes in biological characteristics were weaker than those caused by fishing selection. Exploitation rate and mesh size exhibited similar evolutionary trend tendency under long-term fishing. The time-dependent probability of significance trend showed a gradual growth and tended to be stable. Therefore, the direction of fishing-induced evolution and successful management of fish species require considerable attention to contribute to sustainable fisheries in China. 展开更多
关键词 trawling pressure fish biological characteristics fishery management
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Two-Dimensional Numerical Simulation of Flow Around Three-Stranded Rope
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作者 WANG Xinxin WAN Rong +2 位作者 HUANG Liuyi zhao fenfang SUN Peng 《Journal of Ocean University of China》 SCIE CAS 2016年第4期627-636,共10页
Three-stranded rope is widely used in fishing gear and mooring system. Results of numerical simulation are presented for flow around a three-stranded rope in uniform flow. The simulation was carried out to study the h... Three-stranded rope is widely used in fishing gear and mooring system. Results of numerical simulation are presented for flow around a three-stranded rope in uniform flow. The simulation was carried out to study the hydrodynamic characteristics of pressure and velocity fields of steady incompressible laminar and turbulent wakes behind a three-stranded rope. A three-cylinder configuration and single circular cylinder configuration are used to model the three-stranded rope in the two-dimensional simulation. The governing equations, Navier-Stokes equations, are solved by using two-dimensional finite volume method. The turbulence flow is simulated using Standard κ-ε model and Shear-Stress Transport κ-ω(SST) model. The drag of the three-cylinder model and single cylinder model is calculated for different Reynolds numbers by using control volume analysis method. The pressure coefficient is also calculated for the turbulent model and laminar model based on the control surface method. From the comparison of the drag coefficient and the pressure of the single cylinder and three-cylinder models, it is found that the drag coefficients of the three-cylinder model are generally 1.3–1.5 times those of the single circular cylinder for different Reynolds numbers. Comparing the numerical results with water tank test data, the results of the three-cylinder model are closer to the experiment results than the single cylinder model results. 展开更多
关键词 cylinder turbulent Reynolds turbulence laminar circular hydrodynamic governing incompressible Navier
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