Very high density of CHO cells in perfusion by ATF or TFF in WAVE bioreactor™. Part I. Effect of the cell density on the process

High cell density perfusion process of antibody producing CHO cells was developed in disposable WAVE Bioreactor? using external hollow fiber filter as cell separation device. Both “classical” tangential flow filtration (TFF) and alternating tangential flow system (ATF) equipment were used and compared. Consistency of both TFF‐ and ATF‐based cultures was shown at 20–35 × 10⁶ cells/mL density stabilized by cell bleeds. To minimize the nutrients deprivation and by‐product accumulation, a perfusion rate correlated to the cell density was applied. The cells were maintained by cell bleeds at density 0.9–1.3 × 10⁸ cells/mL in growing state and at high viability for more than 2 weeks. Finally, with the present settings, maximal cell densities of 2.14 × 10⁸ cells/mL, achieved for the first time in a wave‐induced bioreactor, and 1.32 × 10⁸ cells/mL were reached using TFF and ATF systems, respectively. Using TFF, the cell density was limited by the membrane capacity for the encountered high viscosity and by the pCO₂ level. Using ATF, the cell density was limited by the vacuum capacity failing to pull the highly viscous fluid. Thus, the TFF system allowed reaching higher cell densities. The TFF inlet pressure was highly correlated to the viscosity leading to the development of a model of this pressure, which is a useful tool for hollow fiber design of TFF and ATF. At very high cell density, the viscosity introduced physical limitations. This led us to recommend cell densities under 1.46 × 10⁸ cell/mL based on the analysis of the theoretical distance between the cells for the present cell line. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:754–767, 2013     

In Vivo Quantitative Assessment of Myocardial Structure,Function, Perfusion and Viability Using Cardiac Micro-computed Tomography

The use of Micro-Computed Tomography (MicroCT) for in vivo studies of small animals as models of human disease has risen tremendously due to the fact that MicroCT provides quantitative high-resolution three-dimensional (3D) anatomical data non-destructively and longitudinally. Most importantly, with the development of a novel preclinical iodinated contrast agent called eXIA160, functional and metabolic assessment of the heart became possible. However, prior to the advent of commercial MicroCT scanners equipped with X-ray flat-panel detector technology and easy-to-use cardio-respiratory gating, preclinical studies of cardiovascular disease (CVD) in small animals required a MicroCT technologist with advanced skills, and thus were impractical for widespread implementation. The goal of this work is to provide a practical guide to the use of the high-speed Quantum FX MicroCT system for comprehensive determination of myocardial global and regional function along with assessment of myocardial perfusion, metabolism and viability in healthy mice and in a cardiac ischemia mouse model induced by permanent occlusion of the left anterior descending coronary artery (LAD).     

一组有或无CT 脑灌注" 点征"的自发性脑内出血患者临床效果观察

目的:观察有或无CT脑灌注"点征"自发性脑内出血(sICH)患者的临床疗效。方法:以本院2013年5月~2015年5月就诊的100例s ICH,均接受基线CT脑灌注检查,依据是否有"点征"将患者分为观察组(有"点征")与对照组(无"点征"),24内行CT平扫复查,比较两组影像学结果及临床结果。并通过Logistic多因素分析影响s ICH患者预后不良(死亡)危险因素。结果:观察组24h内CT平扫复查血肿增长6 mL以上、血肿增长相对值33%及以上比率均明显高于对照组(P0.05);观察组血肿进展发生率、3个月内死亡率分别为63.04%、36.96%,显著高于对照组的14.81%、7.41%(P0.05)。Logistic回归分析发现sICH患者预后不良独立危险因素包括基线血肿体积、"点征"。结论:有或无CT脑灌注"点征"自发性脑内出血患者血肿进展、预后不同,基线血肿体积、"点征"为sICH患者预后不良的独立危险因素。     

益气健脾汤联合腹腔热灌注化疗用于结直肠癌术后的临床疗效及对患者血清TK1、COX-2、sICAM-1水平的影响

目的:探讨益气健脾汤联合腹腔热灌注化疗用于结直肠癌术后的临床疗效及对患者血清胸苷激酶1 (TK1)、环氧化酶-2(COX-2)与可溶性细胞间黏附分子-1(sICAM-1)水平的影响。方法:选择本院2014年2月到2016年2月收治的结直肠癌患者62例进行回顾性分析,采用系统随机的方法降所有患者分为对照组和治疗组,每组各31例。对照组采用腹腔热灌注化疗进行治疗,治疗组采用益气健脾汤联合腹腔热灌注化疗进行治疗。观察和比较两组患者治疗后的临床疗效、生活质量改善情况、24个月内生存率、治疗前后的血清TK1、COX-2、s ICAM-1水平的变化及不良反应的发生情况。结果:治疗后,观察组患者总有效率为93.55%,明显高于对照组(77.42%,P0.05);两组患者的血清TK1、COX-2、s ICAM-1水平较治疗前显著降低(P0.05);且观察组血清TK1、COX-2、s ICAM-1水平明显低于对照组(P0.05);观察组患者生活质量改善总有效率为95.55%,明显高于对照组(77.78%,P0.05);治疗组患者24个月内生存率为67.74%,明显高于对照组患者(32.26%,P0.05)。两组不良反应的发生情况比较差异无明显统计学意义(P0.05)。结论:益气健脾汤联合腹腔热灌注化疗用于结直肠癌术后的临床疗效显著,可以有效改善患者的生存质量和预后,且安全性高,可能与其显著降低血清TK1、COX-2、s ICAM-1水平有关。     

胃癌根治术后腹腔热灌注化疗对患者血清CEA、CA19-9水平及免疫功能的影响

目的:探讨胃癌根治术后腹腔灌注化疗对患者血清CEA(血清癌胚抗原,carcinoembryonic antigen)、CA19-9(糖链抗原19-9,carbohydrate antigen19-9)水平及免疫功能的影响。方法:回顾性2015年2月至2017年4月我院收治的胃癌患者临床资料,依据接受治疗方案不同分为全身静脉化疗组(对照组)和全身静脉化疗联合腹腔热灌注化疗组(观察组),每组各41例。检测和比较两组患者化疗前(治疗前)与化疗1个月后(治疗后)血清肿瘤标志物CEA、CA19-9与免疫功能指标水平的变化,治疗后毒副作用发生情况及治疗前后生活质量的改善情况。结果:治疗前,两组间血清CEA、CA19-9、CD3~+、CD4~+、CD8~+、CD4+/CD8~+水平比较差异均无统计学意义(P0.05);观察组治疗后血清CEA、CA19-9及CD8~+水平显著低于对照组,CD3~+、CD4~+、CD4~+/CD8~+水平显著高于对照组,差异有统计学意义(P0.05);两组骨髓抑制、恶心呕吐、腹痛腹泻及肠梗发生率比较差异均不显著无统计学意义(P=0.478,0.668,0.315,0.552);观察组生活质量改善总有效率为85.37%,显著高于对照组(70.73%,P=0.017)。结论:与单纯全身化疗相比,胃癌根治术后腹腔灌注化疗可更有效降低患者血清CEA、CA19-9水平,改善患者免疫功能,提高其生活质量,且安全性较高。     

急性心肌梗死高龄患者心肌灌注水平影响左室重构

为了探讨高龄急性心肌梗死(acute myocardial infarction, AMI)患者心脏超声特点,分析左室重构(left ventricle remodel, LVR)与心肌灌注水平的相关性,本研究选取2016年2月至2017年10月在广西医科大学第一附属医院治疗的高龄AMI患者104例,根据患者年龄分为A组49例(60~79岁)和B组55例(≥80岁),比较两组心脏超声指标,采用声学造影积分指数(contrast score index, CSI)评估两组术后心肌灌注水平。结果表明,B组后下壁心肌梗死比例为27.27%,明显高于A组(p<0.05);B组和A组前壁、下壁、前壁+下壁心肌梗死比例差异无统计学意义(p>0.05);B组左心室射血分数(left ventricular ejection fraction, LVEF)为(45.29±12.14)%,明显低于A组(p<0.05),左心房内径和左心室内径分别为(46.10径和左心室) mm和(57.29径和左心室内) mm,明显高于A组(p<0.05);B组经皮冠状动脉介入治疗(percutaneous coronary intervention,PCI)术后6个月CSI为(0.68±0.20),明显低于A组(p<0.05);B组术后左心房内径和左心室内径分别为(50.01±8.10) mm和(64.10±7.02) mm,明显高于A组(p<0.05);左心室内径与CSI呈负相关(r=-0.312, p<0.05)。综上表明,≥80岁患者与60~79岁患者心脏超声特点有所差异,年龄超过80岁的患者心功能以及PCI术后心肌灌注水平较差;心肌灌注水平与左室重构有一定相关性。