免疫磁珠法分离胆囊癌CD133阳性细胞及其生物学特性鉴定

目的 建立胆囊癌CD133+细胞分离纯化的方法,观察胆囊癌CD133+细胞和CD133-细胞生物学特性的差异.方法 流式细胞仪检测胆囊癌GBC-SD、SGC996细胞中CD133所占百分比,免疫磁珠法分选CD133+细胞亚群,免疫荧光法定位检测CD133蛋白表达,逆转录-聚合酶链反应(RT-PCR)检测CD133 mRNA表达,细胞计数试剂盒(CCK-8)法检测CD133+组和CD133-组细胞增殖能力及对5-氟尿嘧啶(5-Fu)耐药特性的差异,单克隆形成实验观察单个CD133+细胞生长特性.结果 胆囊癌GBC-SD、SGC996细胞中CD133+亚群所占相对百分比为(68.5±2.9)％、(0.3±0.2)％.GBC-SD组CD133 mRNA相对灰度值(0.6466±0.0259)显著高于SGC996组(0.2181±0.0108,P＜0.01).CD133蛋白定位于细胞膜表面,并在GBC-SD细胞中呈强表达.人胆囊癌GBC-SD细胞分选后,CD133+组CD133蛋白表达明显强于CD133-组.CD133+组中CD133+亚群所占比例为(90.4±0.9)％.CD133+组CD133mRNA相对灰度值(0.7734±0.0217)显著高于CD133-组(0.2146 ±0.0174,P＜0.01).CD133+组细胞增殖能力明显强于CD133-组(P＜0.01).经5-Fu(0.1μg/ml)处理后,CD133+组细胞生存率明显高于CD133-组(P＜0.05).单克隆形成实验结果示单个CD133+细胞可形成新的细胞克隆,且CD133+组[(36.25±2.99)％]细胞克隆球形成率高于CD133-组[(4.50±1.29)％,P＜0.01].体内成瘤实验结果示CD133+组与未分选组移植成瘤率分别为100％和60％,而CD133-组不成瘤.结论 免疫磁珠分选系统可成功分离高纯度的胆囊癌CD133+细胞,而且人胆囊癌CD133+细胞具有一定的增殖、耐药、自我更新及致瘤潜能.     

胃癌CD133阳性细胞亚群肿瘤起始细胞样特性的检测

目的 探讨人胃癌细胞CD133＋亚群的分选及其特性的鉴定.方法 对50例胃癌原发灶和癌旁胃黏膜组织标本行免疫组织化学染色及Western blot检测CD133蛋白的表达.采用流式细胞仪检测不同分化胃癌细胞系CD133所占百分比,免疫磁珠法分选CD133＋细胞亚群,悬浮培养并观察其生长特性,并检测CD133阳性细胞裸鼠皮下接种致瘤能力.单细胞克隆观察单个CD133＋细胞生长特性,半定量反转录聚合酶链反应法鉴定相关干细胞标记的表达.结果 CD133蛋白多定位于胃癌原发灶黏膜及黏膜下层的肿瘤细胞膜表面,其相对表达量高于癌旁胃黏膜组织（P＜0.05）.不同分化程度的人胃癌细胞系KATO-Ⅲ、SGC-7901、AGS及MKN-45中CD 133＋亚群所占相对百分比为（28±2）％、（17±2）％、（6±2）％及（4±2）％.人胃癌细胞系KATO-Ⅲ分选后阳性组中CD133＋亚群比例分别为（91±3）％;培养1周后,达到（95±2）％,并不断增殖形成细胞球.而且其增殖能力强于阴性细胞[群体倍增时间分别为（21±3）h和（40±8）h,P＜0.05].CD133阳性组和未分选组细胞裸鼠皮下接种时成瘤率分别为100％和80％;而CD133阴性组不成瘤,CD133阳性组移植瘤平均体积及重量均大于未分选组（P＜0.05,P＜0.05）.单克隆形成实验示单个CD133＋细胞可形成新的细胞克隆.半定量RT-PCR检测示其表达干细胞标记物Oct-4、Nanog、Sox-2、Musashi-1及EGFR.结论 体外可成功分离、纯化和扩增人胃癌细胞CD133＋亚群,其具有自我更新、增殖能力及较强的致瘤能力,并表达部分干细胞相关基因.     

化疗药物对胃癌CD133+细胞亚群的作用及其对相关凋亡基因表达的影响

目的 研究常规化疗药物对纯化的胃癌CD133+细胞亚群的作用及对相关凋亡基因Bcl-2和BAX的mRNA表达的影响,进而探讨胃癌耐药的作用机制.方法 采用免疫磁珠分选术分离纯化KATO-Ⅲ细胞株中CD133+和CD133细胞亚群.采用CCK-8法分析两种亚群细胞对不同化疗药物5-氟尿嘧啶(5-FU)、顺铂及依托泊苷(VP-16)敏感性的差别.行半定量聚合酶链反应分析化疗药物诱导后相关凋亡基因表达产物的表达差异.结果 KATO-Ⅲ细胞磁珠分选纯化后,经流式细胞仪分析结果显示:分选后CDI33+组所得细胞CD133+表达率为90％.5-FU、顺铂及VP-16分别作用12h后,CD133+组及CD133-组均开始出现凋亡的形态学改变.CCK-8检测发现CD133+组细胞生长抑制比率较CD133-组有一定程度的下降[5-FU组为:(30.56±1.99)％ ～ (88.60±1.95)％ vs(32.81±2.67)％ ～ (35.55±3.23)％,P =0.045;顺铂组为:(45.89±3.64)％ ～ (81.20±1.18)％ vs(50.21 ±3.22)％ ～(90.46±1.89)％,P=0.043; VP-16组为:(37.21±3.80)％ ～ (78.49±3.22)％ vs (35.55±3.23)％ ～ (89.32±3.54)％,P=0.048].3种化疗药物干预后,两组亚群细胞中均呈现凋亡抑制因子Bcl-2 mRNA表达降低,凋亡因子BAXmRNA表达升高.这些变化在CD133+组中较CD133组中更为明显.结论 胃癌CD133+细胞亚群对5-FU、顺铂及VP-16具有一定耐药潜能.可能是由于BAX基因上调及Bcl-2基因下调进而诱导胃癌CD133+细胞亚群凋亡,并由此而获得对肿瘤药物的抵抗性.     

CD44+-ESA+在结肠癌干细胞分选中的应用

目的 分选结肠癌细胞株SW480细胞中的CD133+-CD44+-ESA+亚群细胞,并观察其致瘤性.方法 用流式细胞仪分选SW480细胞中CD133+-CD44+-ESA+、CD133--CD44+-ESA+及CD133--CD44--ESA-亚群细胞.将这3组细胞分别接种于NOD/SCID小鼠,每组5只,观察肿瘤生长...     

胆管癌干细胞的分离、培养及鉴定

目的:从人胆管癌细胞系QBC-939中分离具有干细胞特征的肿瘤细胞,为后续胆管癌干细胞的研究提供材料。方法:用无血清培养法及流式细胞分选法从QBC-939细胞中分离得到具有干细胞特征的CD133+Ep CAM~(high)干细胞样细胞,继续在无血清培养基中培养,观察其成球能力,并比较CD133+Ep CAM~(high)干细胞样细胞与QBC-939细胞单克隆形成率、耐药性,增殖能力,干细胞相关核转录因子OCT-4、Bmi-1、E-cadherin蛋白表达情况,以及BALB/c小鼠皮下移植后的成瘤能力。结果:在无血清培养基中,CD133+Ep CAM~(high)干细胞样细胞具有较强的成球能力。与QBC-939细胞比较,CD133+Ep CAM~(high)干细胞样细胞克隆形成率、洛铂耐药性、增殖明显增加;干细胞相关核转录因子OCT-4、Bmi-1表达明显增加,而E-cadherin表达明显降低;皮下移植瘤形成率明显增加。以上差异均有统计学意义(均P0.05)。结论:从人胆管癌QBC-939细胞中分离的干细胞样细胞具有肿瘤干细胞特性,可用于胆管癌干细胞的研究。     

人结直肠癌干细胞休眠与增殖阶段细胞形态学的研究

目的初步研究人结直肠癌干细胞休眠与增殖阶段细胞形态的变化。方法流式细胞仪从新鲜人结直肠癌组织中分选EpCAMhigh／CD44＋／CD133＋细胞亚群,并通过裸鼠（NOD／SCID）成瘤实验鉴定其干细胞特性：采用三维培养,WST．1绘制人结直肠癌干细胞的生长曲线;流式细胞仪测定P27及Ki一67的表达水平,区分人结直肠癌干细胞休眠和增殖时相;细胞免疫荧光显示休眠期与增殖期人结直肠癌干细胞形态的变化。结果EpCAMhigh／CD44＋／CDl33‘细胞亚群在人结直肠癌组织中占1．6％．经NOD／SCID成瘤实验证实为人结直肠癌干细胞。人结直肠癌干细胞生长曲线呈“S”型;前3d生长缓慢,细胞为静止阶段（休眠期）,P27表达水平逐渐增加,Ki．67表达偏低;从第4d开始,细胞进入增殖期,Ki-67表达逐渐增加,P27表达降低。细胞免疫荧光染色显示：休眠期人结直肠癌干细胞圆而大,伪足少;而增殖期细胞伪足增多,呈现增殖和侵袭的状态。结论肿瘤的复发、转移可能与肿瘤干细胞的生长状态发生改变有一定关系;人结直肠癌干细胞增殖期与休眠期相比。呈现出明显增殖和侵袭能力：这为治疗人结直肠癌及其他肿瘤的复发和转移提供了新的切人点。     

Notch信号通路在肺癌干细胞中的表达及其对增殖的影响

目的 观察Notch信号通路在肺癌干细胞中的表达及阻断Notch信号通路对肺癌干细胞增殖的影响.方法 以CD133作为肿瘤干细胞表面标志,采用流式细胞仪高速分选技术从人肺腺癌细胞株A549中分离肺癌干细胞及普通肿瘤细胞,实时荧光定量聚合酶链反应(FQ-PCR)及Western blot检测肺癌干细胞和普通肿瘤细胞内的Notch信号通路的表达水平;细胞计数试剂盒(CCK-8)检测肺癌干细胞和普通肿瘤细胞的体外增殖能力,并绘制生长曲线;使用γ-分泌酶抑制剂(DAPT)阻断Notch信号通路传导,观察肺癌干细胞和普通肿瘤细胞的体外生长差异.结果 流式分选前检测CD133阳性细胞(肺癌干细胞)占所有A549细胞的百分比为(0.40±0.11)％,而流式分选后所得细胞中,CD133阳性细胞占百分比为(95.00±0.63)％,两者差异有统计学意义(P＜0.01).Notch通路在肺癌干细胞及普通肿瘤细胞中均表达,Notch1及Notch2在肺癌干细胞中的表达显著低于在普通肿瘤细胞内的表达,差异有统计学意义(P ＜0.05);Hes1仅在普通肿瘤细胞中表达,在肺癌干细胞中未检测到表达.肺癌干细胞与普通肿瘤细胞的增殖能力差异无统计学意义(P＞0.05),而在DAPT干预48 h后,肺癌干细胞和普通肿瘤细胞的体外增殖均受到了抑制,肺癌干细胞的生长抑制率[(33.7±1.9)％]显著高于普通肿瘤细胞的生长抑制率[(21.5±3.4)％],差异有统计学意义(P＜0.05).结论 较之普通肿瘤细胞,阻断Notch通路传导对肺癌干细胞的生长抑制效果更强,这可能与DAPT抑制了Notch的过高表达对肺癌细胞的负反馈作用有关.     

树突状细胞瘤苗联合吉西他滨对肝癌干细胞的杀伤效应

目的:探讨负载CD133+肝癌细胞抗原的树突状细胞(DC)联合吉西他滨(GEM)在体外对肝癌干细胞的杀伤效应。方法:取对数生长期的人肝癌细胞系Huh-7以CD133作为分子标志进行流式分选,得到肝癌干细胞。将人外周血单个核细胞(PBMC)在体外诱导分化为树突状细胞(DC)。DC负载Huh-7细胞和CD133+细胞抗原后与T细胞共育得到特异性细胞毒性T细胞,将CD133+细胞作为靶细胞进行细胞毒性试验。实验组按处理因素分为:GEM组,CD133+-CTL组,GEM+CD133+-CTL组,Huh7-CTL组和GEM+Huh7-CTL组。CCK-8法检测杀伤率,然后比较各组间差异。结果:对CD133+细胞的杀伤效应以GEM+CD133+-CTL组最强,与其他各组比较差异均有统计学意义(P0.05)。单独就DC瘤苗杀伤率,CD133+-CTL组高于Huh7-CTL组(P0.05)。结论:CD133+肝癌干细胞裂解产物致敏的DC瘤苗联合化疗药物可以有效杀伤肝癌干细胞,进而可能降低肝癌术后和肝癌肝移植后的转移和复发率。     

Galectin-3在CD133+肺腺癌细胞中表达增高诱导CD8+T细胞凋亡

目的 检测肺腺癌CD133+细胞中Galectin-3的表达.方法 磁珠分选出10例患者肺腺癌中CD133+细胞.流式细胞术检测分选效率,荧光实时定量聚合酶链反应( fqRT-PCR)和Western blot检测CD133+细胞中Galectin-3的表达,并在体外检测了CD133+细胞上清诱导CD8+T细胞凋亡的能力.结果 流式细胞术结果表明磁珠分选出的细胞中CD133+细胞数为90％.fqRT-PCR 和Western blot结果发现Galectin-3在CD133+细胞中的表达量分别为CD133-细胞中的1.24倍和1.5倍,差异有统计学意义(P＜0.05).凋亡检测结果显示CD133+细胞的上清诱导CD8+T细胞凋亡率为(27.1±2.6)％,并且这种诱导凋亡的能力可被乳糖以及抗Galectin-3的抗体中和.结论 Galectin-3在肺腺癌CD133+细胞中高表达,并具有很强的生物学活性,在体外具有明显的诱导CD8+T细胞凋亡的功能.     

人胰腺癌干细胞差异性基因的表达

目的 分选、鉴定人胰腺癌干细胞,运用基因芯片技术分析其差异性基因的表达.方法 运用流式分选技术分选胰腺癌干细胞(CD24+CD44+ESA+),NOD/SCID鼠移植瘤试验进行肿瘤干细胞特性鉴定.采用Affymetrix U133 plus2.0人类全基因组表达谱芯片对胰腺癌干细胞和非干细胞进行差异基因筛选.结果 分选得到人胰腺癌CD24+CD44+ESA+亚群细胞,占所有细胞的0.8%;5×103个CD24+CD44+ESA+细胞就能成瘤(2/4),而阴性细胞1×105才能成瘤(1/4);CD24+CD44+ESA+具有一定的自我更新和分化能力.基因芯片杂交获得6553(11.99%)条差异基因,胰腺癌干细胞中5255(9.61%)条上调表达,1298(2.37%)条下调表达.其中差异基因涉及细胞凋亡、细胞周期、代谢、细胞线粒体结构和耐药等多个方面.结论 胰腺癌于细胞具有自身特征性基因表达谱,为进一步从干细胞层面研究胰腺癌发病机制及靶向治疗奠定基础.     

Is the recovery profile of mivacurium independent of the rate of decay of its plasma concentration in patients with normal plasma cholinesterase activity?

Background: The duration of action of muscle relaxants is poorly correlated to the rate of decay of their plasma concentration. The plasma concentration of mivacurium may rapidly decrease below its active concentration because of the extensive hydrolysis of mivacurium. By inflating a tourniquet on one upper limb for 3 min after the administration of atracurium, mivacurium or vecuronium, we studied the influence of the initial decline of their plasma concentration on their effect. Methods: In 50 patients anaesthetised with thiopental, isoflurane and fentanyl, the effect of bolus doses of 0.15 or 0.25 mg . kg^?1 mivacurium (MIV 15, MIV 25), 0.3 or 0.5 mg . kg^?1 atracurium (ATR 30, ATR 50) and 0.06 or 0.1 mg . kg^?1 vecuronium (VEC 06, VEC 10) were measured on both arms (evoked response of the adductor pollicis to train-of-four stimulation every 12 s), a tourniquet being applied on one arm just before and during 3 min after the muscle relaxant bolus. Results: Tourniquet inflation of 3 min almost abolished the neuromuscular effect of mivacurium. In the vecuronium groups and in the ATR 50 group, tourniquet inflation did not modify the maximum degree of depression of the twitch response. Also, the duration of action of vecuronium was unaffected by the tourniquet. In the ATR 30 group, times to return of the twitch response to 25% (duration 25%) and 75% (duration 75%) of control response were significantly shorter in the cuffed arm, 23 min vs 27 min, and 41 min vs 45 min, respectively. In the ATR 50 group, only duration 25% was significantly shorter in the cuffed arm (41 min vs 45 min). Conclusion: The results suggest that the rate of decline of the plasma concentration of mivacurium is so rapid, that a very low and almost clinically ineffective concentration is present as soon as 3 min after its administration. The results also indicate that the recovery from a mivacurium-induced neuromuscular blockade is not influenced by the rate of decay of its plasma concentration in patients with genotypically normal plasma cholinesterase.     

Membranes in Artificial Organs

Abstract: Membrane processes play a pivotal and enabling role in modern replacement therapy for acute and chronic organ failure and in the management of immunologic diseases. In fact, virtually all contemporary extracorporeal blood purification methods employ membrane devices, and the next generation of artificial organs and tissue engineering therapies are almost certain to be similarly grounded in membrane technology. In this short essay, we comment on the similarities and differences among synthetic membranes and their natural counterparts and also provide a critical overview of the demographics and technology of hemodialysis, hemofiltration, apheresis, oxygenation, and emerging membrane technologies and applications.     

Dilemma of Membrane Biocompatibility and Reuse

Abstract: Numerous articles have been published on the multiple use of dialyzers and on the effect of different reprocessing chemicals and techniques on the dialyzer biocompatibility and performance. The results often appear contradictory, especially those comparing standard biocompatibility parameters. Despite this confusion, a discerning review of the published works allows certain limited conclusions to be drawn. Reprocessing of used hemodialyzers changes the biocompatibility profile of a dialyzer as defined by the parameters complement activation. leukopenia, and cytokine release. The effect of reprocessing depends on the chemicals and reprocessing technique applied and also on the type of membrane polymer being subjected to the reprocessing procedure. Reports of pyrogenic reactions indicate that the flux of the membrane also influences how suitable it is for safe reuse. An increased risk of allergic and pyrogenic reactions appears to be associated with dialyzer reuse. Furthermore, there has been a lack of investigations into the immunologic effect of the layer of adsorbed and chemically altered proteins that remains on the inner surface of reprocessed dialyzers. We conclude that the clinical benefit of dialyzer reuse cannot be generally accepted from a biocompatibility point of view.     

The analgesic effect of racemic ketamine in patients with chronic ischemic pain due to lower extremity arteriosclerosis obliterans

Background : Ketamine in sub-dissociative doses has been shown to have analgesic and phantom-Limb pain, where conventional treatment has often failed. Chronic ischemic pain due to lower extremity arteriosclerosis obliterans often responds poorly to analgesics, and the pain-generating mechanisms are not well understood.
Methods : Eight patients with rest pain in the lower extremity due to arteriosclerosis obliterans were given sub-dissociative doses of 0.15, 0.30, or 0.45 mg/kg racemic ketamine and morphine 10 mg as a 5-min infusion on four separate days in a cross-over, double-blind, randomised protocol. Plasma levels of (S)- and (R)-ketamine and their nor-metabolites were analysed with an enantioselective high-performance liquid chromatography (HPLC) method. Pain levels were evaluated with a visual analogue scale (VAS).
Results : Individual pain levels were highly variable during and after all the infusions but the pooled pain levels showed a dose-dependent analgesic effect of ketamine with a transient but complete pain relief in all patients at the highest dose (0.45 mg/ kg). Side-effects, mainly disturbed cognition and perception, were pronounced and dose-dependent. Morphine 10 mg had an analgesic peak at 20 min and 5/8 patients had complete pain relief. The remaining 3 patients also had high baseline pain scores, indicating a higher analgesic potency for the 0.30 and 0.45 mg/ kg ketamine doses than for morphine 10 mg.
Conclusion : We have demonstrated a potent dose-dependent analgesic effect of racemic ketamine in clinical ischemic pain. Due to a narrow therapeutic window, this analgesic effect is probably best utilised in combination with other analgesics.     

Intravenous endotoxin does not increase tissue extravasation of albumin in rats

Background : It is unclear whether activation of the inducible nitric oxide synthase (iNOS) increases or decreases the extravasation of plasma.
Methods : Chloralose anaesthetised male Wistar rats received E. coli lipopolysacharide (LPS), 3 mg kg^-1 i.v., or the corresponding volume of saline, 3 or 5 h before the end of the experiment. Mean arterial pressure (MAP) and heart rate (HR) were recorded. Tissue clearance of radio-labelled albumin, during the last 2 h of each experiment, was determined by a double-isotope method. In separate animals, the serum concentration of nitrite and nitrate was determined, 5 h after LPS or the solvent.
Main Results : LPS initially decreased MAP and lastingly increased HR. In the 3-h LPS animals (n=8), tissue plasma clearance was lower in the heart and calf muscle and increased only in diaphragm, compared to corresponding control animals (n=8). In the 5-h LPS rats, clearance was lowered (n=8) in the entire gastrointestinal tract and in testes, compared to controls (n=8). The serum nitrite/nitrate concentration was higher in animals given LPS (n=6) than in controls (n=6).
Conclusion : After LPS, tissue clearance of albumin was not increased in any major tissue, in spite of increased serum levels of NO end products. Apparently, after activation of iNOS, the augmented release of NO is not necessarily associated with increased albumin extravasation.     

Adrenaline markedly improves thoracic epidural analgesia produced by a low-dose infusion of bupivacaine, fentanyl and adrenaline after major surgery: A randomised, double-blind, cross-over study with and without adrenaline

Background: Basic pharmacological research indicates that there are synergistic antinociceptive effects at the spinal cord level between adrenaline, fentanyl and bupivacaine. Our clinical experience with such a mixture in a thoracic epidural infusion after major surgery confirms this. The objectives of the present study were to evaluate the effects on postoperative pain intensity, pain relief and side effects when removing adrenaline from this triple epidural mixture. Methods: A prospective, randomised, double-blind, cross-over study was carried out in 24 patients after major thoracic or abdominal surgery. Patients with only mild pain when coughing during a titrated thoracic epidural infusion of about 10 ml · h^?1 of bupivacaine 1 mg · ml^?1, fentanyl 2 μg · ml^?1, and adrenaline 2 μg · ml^?1 were included. On the 1^st and 2^nd postoperative days each patient was given a double-blind epidural infusion, at the same rate, with or without adrenaline. The effect was observed for 4 h or until pain when coughing became unacceptable in spite of a rescue analgesic procedure. Rescue analgesia consisted of up to two epidural bolus injections per hour and i.v. morphine if necessary. All patients received rectal paracetamol 1 g, every 8 h. Fentanyl serum concentrations were measured with a radioimmunoassay technique at the start and end of each study period. Main outcome measures were extent of sensory blockade and pain intensity at rest and when coughing, evaluated by a visual analogue scale, a verbal categorical rating scale, the Prince Henry Hospital pain score, and an overall quality of pain relief score. Results: The number of hypaesthetic dermatomal segments decreased (P <0.001) and pain intensity at rest and when coughing increased (P <0.001) when adrenaline was omitted from the triple epidural mixture. This change started within the first hour after removing adrenaline. After 3 h pain intensity when coughing had increased to unacceptable levels in spite of rescue analgesia (epidural bolus injections and i.v. morphine). Within 15–20 min after restarting the triple epidural mixture with adrenaline, pain intensity was again reduced to mild pain when coughing. Serum concentration of fentanyl doubled from 0.22 to 0.45 ng · ml^?1 (P <0.01), and there was more sedation during the period without adrenaline. Conclusions: Adrenaline increases sensory block and improves the pain-relieving effect of a mixture of bupivacaine and fentanyl infused epidurally at a thoracic level after major thoracic or abdominal surgery. Serum fentanyl concentrations doubled and sedation increased when adrenaline was removed from the epidural infusion, indicating more rapid vascular absorption and systemic effects of fentanyl.     

Reduction of Plasma Fibrinogen, Immunoglobulin G, and Immunoglobulin M Concentrations by Immunoadsorption Therapy with Tryptophan and Phenylalanine Adsorbents

Abstract Immunoadsorption (1A) therapy with tryptophan (TR-350) or phenylalanine (PH-350) adsorbents has been used to reduce the concentration of serum antibodies in human lymphocyte antigen (HLA)-immunized patients. Other forms of plasma purification have been reported to reduce the level of fibrinogen, which affects the blood properties. In this study we investigated the effects of IA therapy using both adsorbents on plasma fibrinogen and immunoglobulins G and M in 13 patients (8 patients were treated with TR-350, and 5 patients were treated with PH-350). During each session 1 plasma volume (2.8 ± 0.4 L of plasma) was processed through the immunocolumn and then returned to the patient together with the blood cells. Compared with the pretreatment values, the plasma fibrinogen, IgG, and IgM concentrations were significantly reduced after IA therapy (p < 0.01 for TR-350; p < 0.04 for PH-350). There was a positive correlation between the degree of reduction of plasma proteins and the number of IA treatments given. A nonpara-metric test (Wilcoxon's signed-rank test or the Mann-Whitney test) was used for statistical analysis. We conclude from our study that IA therapy effectively lowers the plasma levels of fibrinogen, IgG, and IgM and thus can be considered a valuable alternative to other blood purification methods.     

Utilisation des médicaments prokinétiques en réanimation : indications et limites ?

Enteral feeding is often limited by gastric and intestinal motility disturbances in critically ill patients, particularly in patients with shock. So, promotility agents are frequently used to improve tolerance to enteral nutrition. This review summaries the pathophysiology, presents the available pharmacological strategies, the clinical data, the counter-indications and the principal limits. The clinical data are poor. No study demonstrates a positive effect on clinical outcomes. Metoclopramide and erythromycin seems to be the more effective. Considering the risk of antibiotic resistance, the first line use of erythromycin should be avoided in favor of metoclopramide.     

Anesthésie générale chez l’enfant : quid des pratiques en 2010 ?

Introduction

The practice of pediatric anesthesia requires a regular update of scientific knowledge and technical skills. To provide the most adequate Continuing Medical Education programs, it is necessary to assess the practices of pediatric anesthesiologists. Thus, the objective of this survey was to draw a picture of the current clinical practices of general anesthesia in children, in France.

Material and methods

One thousand one hundred and fifty questionnaires were given to anesthesiologists involved in pediatric cases. These questionnaires collected information on various aspects of clinical practice relative to induction, maintenance, recovery from general anaesthesia and also classical debated points such as children with Upper Respiratory Infection (URI), emergence agitation, epileptoid signs or anaesthetic management of adenoidectomy. Differences in practices between CHG (general hospital), CHU (teaching hospital), LIBERAL (private) and PSPH (semi-private) hospitals were investigated.

Results

There were 1025 questionnaires completed. Fifty-five percent of responders worked in public hospitals (CHG and CHU); 77% had a practice that was 25% or less of pediatric cases. In children from 3 to 10 years: 72% of respondents used always premedication and two thirds performed inhalation induction in more than 50% of cases. For induction, 53% used sevoflurane (SEVO) at 7 or 8%. Respondents from LIBERAL used higher SEVO concentrations. Tracheal intubation was performed with SEVO alone (37%), SEVO and propofol (55%) and SEVO with myorelaxant (8%), 93% of respondents used a bolus of opioid. For maintenance, the majority of respondents used SEVO associated with sufentanil; desflurane and remifentanil were more frequently used in CHU. Two thirds of respondents used N₂O. Depth of anesthesia was commonly assessed by hemodynamic changes (52%), end tidal concentration of halogenated (38%) or automated devices based on EEG (7%). In children with URI, 98% of respondents used SEVO for anesthesia. To control the airway 42% used a tracheal tube, 30% a laryngeal mask and 20% a facial mask. Emergence agitation was an important concern for two thirds of respondents, while epileptoid signs were considered as important by only 20%. Eighty-nine percent of respondents practiced anesthesia for adenoidectomy. Anesthesia was induced by inhalation of SEVO 7–8% (41%), 6% (39%) or 4% (12%), 66% put an intravenous line (less frequently in LIBERAL). 67% of the responders managed adenoidectomy without any device to control the airway (more frequently in LIBERAL), 32% administrated a bolus of opioid (less frequently in LIBERAL).

Discussion

This survey demonstrated that the practices regarding general anesthesia in children are relatively homogenous. Most of the differences appeared between LIBERAL and the others structures; the anaesthetic management for adenoidectomy illustrates these findings.     

Intérêt d’une rééducation précoce pour les patients neurologiques

Rehabilitation improves the functional prognosis of patients after a neurologic lesion, and tendency is to begin rehabilitation as soon as possible. This review focuses on the interest and the feasibility of very early rehabilitation, initiated from critical care units. It is necessary to precisely assess patients’ impairments and disabilities in order to define rehabilitation objectives. Valid and simple tools must support this evaluation. Rehabilitation will be directed to preventing decubitus complications and active rehabilitation. The sooner rehabilitation is started; the better functional prognosis seems to be.