树突状细胞在多发伤中的变化及临床意义

目的探讨多发伤树突状细胞(DC)变化及临床意义。方法从68例多发伤患者和22例健康人外周血中分别提取DC;通过流式细胞仪分别检测各组的DC数量,DC表面HLA-DR、CD80、CD86表达水平以及DC诱导的T细胞反应性增殖情况。同时检测各组中IL-6I、L-10的浓度。结果多发伤组DC细胞数〔(8.3±2.8)×106/L〕明显低于对照组〔(15.2±4.1)×106/L,P<0.01〕。多发伤组DC表面HLA-DR及CD80、CD86的表达水平与对照组相比明显降低(P<0.01)。DC诱导的T细胞增殖能力对照组明显强于创伤组(P<0.01)。多发伤组血清IL-6、IL-10的浓度显著升高,与对照组比较差异有非常显著性意义(P<0.01)。结论多发伤患者DC数量明显减少且功能下降,这可能与创伤后的免疫功能低下密切相关。     

rhG-CSF体内诱导造血干细胞移植供者外周血T细胞免疫耐受的初步研究

本研究探讨rhG CSF体内应用诱导健康供者外周血T淋巴细胞免疫耐受的机制。对 15例病人进行了外周血干细胞移植 ,借助三色和四色荧光标记技术 ,对供者rhG CSF动员前后外周血T细胞上共刺激分子CD2 8的表达、树突状细胞 (DC)亚群以及CD8 CD2 8- 抑制性T细胞的变化进行了流式细胞术测定。结果显示 ,rhG CSF动员后外周血采集物中CD3 CD2 8 细胞的相对数显著升高 (P <0 .0 1) ,CD2 8表达的平均荧光强度明显降低 (P<0 .0 5 ) ;CD8 CD2 8 细胞的相对数也显著升高 (P <0 .0 1)。但在T细胞上CD2 8总体表达的相对荧光强度无变化 (P >0 .0 5 )。动员前外周血中DC2的含量明显低于正常骨髓 (P <0 .0 1) ,动员后采集物中DC2的数量较动员前和正常骨髓均有显著增加 (P <0 .0 1) ,DC的数量也显著增加 (P <0 .0 1) ,DC1 DC2比值倒置 (P <0 0 1) ,而DC1在动员前后无变化 (P >0 .0 5 )。CD8 CD2 8- 细胞占有核细胞的百分比较动员前明显增加 (P <0 0 5 )。结论 :rhG CSF体内应用后 ,采集物中DC2和CD8 CD2 8- 抑制性T细胞数量的增加可能是外周血T细胞免疫耐受产生的重要机制。     

脐血树突状细胞的体外培养及免疫学特征

目的　探讨脐血树突状细胞 (DC)的体外诱导及其免疫学特征。方法　无菌条件下常规采集脐血 ,采用淋巴细胞分离液分离脐血单个核细胞 ,去除悬浮细胞后获得单核细胞。将获得的单核细胞分为两组 ,组 1:在含白介素 4(IL 4 )和粒细胞巨细胞集落刺激因子 (GM CSF)的DMEM液中培养。组 2 :在上述培养液中孵育 5天后 ,加入肿瘤坏死因子 (TNF α)。收集培养 7天的贴壁细胞 ,分别用流式细胞仪检测CD83、CD86 、CD54、CD1α和CD1 4等免疫因子。结果　①组 1培养 12～ 14天树突状细胞数含量约 2 0 %。组 2培养 7～ 10天即可见到明显树突状细胞达 30 %。②TNF α可明显增加干细胞早期增殖能力 ,两组相比P <0 0 5。③IL 4、GM CSF和TNF α可诱导脐血来源的单核细胞 (即DC前体细胞 )分化为成熟的DC ,两组均高表达DC特异性抗原CD83、CD86 、CD54、CD1α。结论　脐血中的DC前体细胞可在体外诱导成为功能性 (即成熟 )DC。GM CSF联合TNF α不仅可以促进细胞形态的生长 ,而且明显增加DC生成数量。故脐血有可能成为DC免疫治疗丰富的来源。     

人外周血树突状细胞的体外诱导培养

目的研究rhGM-CSF和rhIL-4培养体系对树突状细胞(DendriticCell,DC)的诱导培养。方法分离正常人或多发性骨髓瘤(MM)患者外周血中单个核细胞,体外培养树突状细胞。观察培养的DC的形态变化,并应用流式细胞仪分析DC表面的免疫分子的表达。结果100ng/mlrhGM-CSF和500U/mlrhIL-4能使正常人或多发性骨髓瘤患者外周血中单个核细胞分化发育为高表达CD80、CD86、HLAⅠ类和HLAⅡ类抗原的DC。结论rhGM-CSF和rhIL-4能使树突状细胞定向分化成为抗原提呈细胞,为DC疫苗治疗肿瘤感染性疾病奠定了基础。     

从K562细胞中提取热休克蛋白GP96及其对人树突状细胞分化和功能的影响

本研究旨在建立从K5 6 2细胞系中提取热休克蛋白GP96的方法 ,进而研究GP96对人树突状细胞分化和功能的影响。采用蛋白分离提纯技术 ,将K5 6 2细胞裂解 ,经饱和硫酸铵沉淀、亲和层析、离子交换层析后提纯出热休克蛋白GP96 ,用SDS PAGE凝胶电泳和Westernblot方法对其进行鉴定。将提纯的GP96加入从正常人外周血单个核细胞诱导培养的树突状细胞 (dendriticcell,DC)中 ,通过流式细胞术检测DC细胞表型变化 ,用MTT法测定混合淋巴细胞反应 (mixedlymphocytereaction ,MLR)。结果表明 :从 1× 10 1 0 K5 6 2细胞中能提纯出热休克蛋白GP96 6 0 - 80 μg ;加入GP96制备的HSP DC ,其细胞表面标志CD83、CD86和HLA DR的表达率比常规培养DC明显升高 ,CD1a表达率降低 (P <0 .0 5 ) ;HSP DC比常规培养DC具有更强的刺激混合淋巴细胞反应的能力 (P <0 .0 5 )。结论 :从K5 6 2细胞中可以提取出热休克蛋白GP96 ;热休克蛋白GP96可以促进树突状细胞分化成熟 ,使其具有更强的刺激同种异体人T淋巴细胞增殖的能力     

慢性乙肝患者CD4~+ CD25~+ T细胞对外周血树突状细胞成熟的作用

目的探讨慢性乙型肝炎(CHB)患者CD4+CD25+Treg细胞对外周血树突状细胞成熟的影响。方法对2013年6月至2013年12月就诊的15例CHB患者和18例健康对照各抽取外周血20 ml,分离外周血单个核细胞(PBMC),于体外培养获得树突状细胞(DC)及免疫磁珠分选获得CD4+CD25+Treg细胞;分别将CD4+CD25+Treg细胞和CD4+CD25-T细胞与DC共培养后采用流式细胞仪检测DC表面标志CD83、CD80、CD86、HLA-DR的表达。结果健康对照组,加入Treg后,相比于DC对照组及DC与CD4+CD25-T细胞共培养组,DC的CD83、CD80、CD86表达下降,差异有统计学意义。CHB患者组,加入Treg后,相比于DC对照组,DC的CD83、CD80、CD86表达下降,差异均有统计学意义;与DC和CD4+CD25-T细胞共培养组相比,Treg对DC的CD83、CD80表达抑制率较高,差异有统计学意义。CHB患者组的Treg对DC的CD83、CD80、CD86表达抑制率均高于健康对照组,差异均有统计学意义。结论 CHB患者外周血Treg细胞能够抑制DC的成熟,抑制效应强于健康对照。     

人外周血树突状细胞的体外诱导培养

目的研究rhGM-CSF和rhIL-4培养体系对树突状细胞(Dendritie Cell,DC)的诱导培养.方法分离正常人或多发性骨髓瘤(MM)患者外周血中单个核细胞,体外培养树突状细胞.观察培养的DC的形态变化,并应用流式细胞仪分析DC表面的免疫分子的表达.结果100ng/ml rhGM-CSF和500U/ml rhIL-4能使正常人或多发性骨髓瘤患者外周血中单个核细胞分化发育为高表达CD80、CD86、HLAⅠ类和HLAⅡ类抗原的DC.结论rhGM-CSF和rhIL-4能使树突状细胞定向分化成为抗原提呈细胞,为DC疫苗治疗肿瘤感染性疾病奠定了基础.     

膀胱移行细胞癌中树突状细胞浸润及其表面分子表达变化的临床意义

【目的】探讨膀胱移行细胞癌 (BTCC)中树突状细胞 (DC)浸润及其表面分子表达变化的临床意义。【方法】采用流式细胞术检测 12例BTCC病例 (浅表性癌Ta 1期 5例 ,浸润性癌T2～ 4期 7例 ;高中分化癌G1～ 2 4例 ,低分化癌G3 8例 )和 10例正常膀胱组织中树突状细胞CD1a、CD80及CD86的表达 ,并分析CD1a、CD80及CD86的表达与肿瘤病理参数之间的关系。【结果】①在BTCC和正常膀胱组织中CD1a阳性树突状细胞的数量没有显著差别 ,但在BTCC组织中DC的共刺激分子CD80、CD86的表达下调 ;②在BTCC中CD1a阳性DC的数量与临床分期、组织学分级无关 (P >0 .0 5 ) ,但DC的共刺激分子CD80、CD86的表达与BTCC临床分期、组织学分级呈负相关 (P <0 .0 5 )。【结论】BTCC组织中DC的共刺激分子CD80、CD86表达下调可能是BTCC发生免疫逃逸的原因之一     

血管内皮生长因子对CD34+干/祖细胞来源的树突细胞分化及功能的影响

目的　探讨血管内皮生长因子 (VEGF)对人脐血CD34 干 /祖细胞来源的树突细胞(dendriticcell,DC)分化和功能的影响。方法　利用免疫磁珠分离法 (MACS)分离纯化脐血CD34 造血干 /祖细胞 ,并在体外将其诱导扩增为DC ,观察VEGF在培养早期和晚期对DC分化和功能的影响。观察培养过程中细胞增殖方式 ,用流式细胞术检测DC表面分化相关抗原CD1α、CD83、CD80、CD5 4、HLA DR等的表达 ,混合淋巴细胞反应法测定DC体外刺激同种异体T细胞增殖的能力 ,ELISA法检测DC培养上清中IL 12的含量。结果　在细胞增殖方面 ,培养第 1天加入VEGF(2 5ng/ml)可显著促进细胞增殖 ,第 14天收获的总细胞数量较对照组增高 (1.5 1± 0 .2 3)倍 (P =0 .0 0 1) ,而培养第 9天加入VEGF则未出现明显的促细胞增殖效应 (P >0 .0 5 ) ;在细胞分化和功能方面 ,培养第 1天加入VEGF明显抑制DC的分化和功能 ,第 1天加VEGF组和对照组DC分化抗原的表达CD1a分别为(33.0 0± 2 .12 ) %和 (81.2 0± 6 .93) % ,CD83分别为 (42 .2 3± 1.15 ) %和 (87.98± 9.79) % ,CD80分别为 (42 .93± 1.32 ) %和 (94 .5 3± 0 .87) % ,HLA DR分别为 (37.93± 5 .30 ) %和 (74 .15± 3.74 ) % (P值均 <0 .0 0 1) ,同时CD14的表达较对照组明显升高 ;刺激同种异体T淋巴     

外周血树突状细胞的制备和表型分析

目的：探讨体外分离人外周血单个核细胞（PBMC）和诱导生成树突状细胞（dendriticcell,DC）。方法：从外周血分离PBMC,加入细胞因子诱导DC,流式细胞仪测定CD86和HLA-DR,分析其成熟度和激活度。结果：HLA—DR和CD86在PBMC中几乎无表达,经GM-CSF和IL-4诱导后低表达;加入TNF-α后高表达。结论：GM-CSF和IL-4诱导培养PBMC,可获得大量不成熟DC,加入TNF-α后,DC成熟度高,激活性好。     

Determination of creatine kinase isoenzyme MB activity in serum using immunological inhibition of creatine kinase M subunit activity. Activity kinetics and diagnostic significance in myocardial infarction.

This is a new method for the determination of creatine kinase isoenzyme MB activity in serum. The method uses direct activity measurement of creatine kinase B subunit activity after blocking of CK-M subunit activity by inhibiting antibodies. The test takes no longer than 15 min. The method yields an intra-serial C.V. of 2.0-12.9%, and a C.V. from day to day of 5.5%. The detection limit is 3.4 U/l creatine kinase MB. In the 95 cases with proven myocardial infarction several types of creatine kinase MB activity kinetics could be determined. The percentage of creatine kinase MB of peak CK-total is 6-25%, with a mean of 11.1%. The amount of creatine kinase MB with respect to total CK activity after reinfarction is higher than the amount after initial infarction.     

俯卧位通气对高海拔地区肺复张治疗无效急性呼吸窘迫综合征患者氧合的影响

目的 探讨俯卧位通气对高海拔地区肺复张术(RM)治疗无效急性呼吸窘迫综合征(ARDS)患者的治疗作用.方法 从海拔2260m的地区医院筛选RM治疗无效的41例ARDS患者[平均氧合指数( PaO2/FiO2)较RM前升高＜20％视为RM无效],依不同病因分为肺内源性ARDS组(ARDSp组)和肺外源性ARDS组(ARDSexp组),每组再按信封法随机分为俯卧位组和仰卧位组,即ARDSp俯卧位组(11例)、ARDSp仰卧位组(9例)、ARDSexp俯卧位组(10例)、ARDSexp仰卧位组(11例).在通气前及通气1、2、3、4h监测动脉血氧分压( PaO2)、PaO2/FiO2、静态顺应性(Cst)、气道阻力(Raw)的变化.结果 通气lh时,ARDSexp俯卧位组PaO2/FiO2( mm Hg,l mm Hg=0.133 kPa)即较通气前显著升高(157.4±40.6比129.3±48.7,P＜0.05),并随通气时间延长呈持续增高趋势,4h达峰值(219.1 ±41.1);且ARDSexp俯卧位组通气3h内PaO2/FiO2较其他3组显著增高,另3组间则差异无统计学意义.ARDSp俯卧位组、ARDSexp俯卧位组通气4h时PaO2/FiO2均较相应仰卧位组显著增高(208.8±39.7比127.4±47.1,219.1±41.1比124.9±50.8,均P＜0.05).4组通气前后Cst无显著改变,各组间差异也无统计学意义.ARDSp俯卧位组通气4h时Raw(cmH2O·L-1·s-1)较通气前显著降低(6.8±1.7比10.7±1.8,P＜0.05),且明显低于其他3组;其他3组各时间点Raw组内及组间比较差异均无统计学意义.结论 俯卧位通气作为ARDS机械通气重要策略之一,可以改善RM无效高原ARDS患者的氧合,为抢救患者赢得宝贵的时间.     

Digital imaging among medical facilities

The Department of Veterans Affairs (VA) in the USA operates a network of 172 medical centres which all utilize a hospital information system (HIS) which has been developed and is currently maintained by the VA. During the past several years, an image management and communication module has been developed, installed and clinically utilized at the Washington DC and Maryland VA Medical Centres. This image management and communication system, referred to as the decentralized hospital computer program (DHCP) imaging system, is fully integrated with a commercial picture archiving and communication system (PACS). The system is utilized to capture, archive, and display all images generated within the hospital including radiology, nuclear medicine, pathology, endoscopy, bronchoscopy, and dermatology, intraoperative photographs, ECG data, and a limited number of paper documents. The ultimate goal of the project is to have all patient text and image data available at any clinical workstation to any authorized user anywhere within the network of medical centres. Clinical requirements for an imaging workstation include ease of use, rapid and reliable access to the complete set of patient information, and images which are of acceptable quality to meet the requirements of the user and the subspecialty. Patient confidentiality and data security must be safeguarded at all times. Integration of the images with the remainder of the patient's database was found to be critical to the success of the project. The experience at the Washington and Maryland facilities suggests that an imaging system that is successfully integrated with a hospital information system can provide substantial clinical and economic benefits both within and among medical centres. Clinical acceptance and utilization of the system has been excellent, particularly in diagnostic radiology where DHCP Imaging has been interfaced to a commercial PAC system. Based upon this initial experience, the VA has begun to deploy the system throughout its large network of medical centres.     

Myocardial elastography at both high temporal and spatial resolution for the detection of infarcts

Myocardial elastography is a novel method for noninvasively assessing regional myocardial function, with the advantages of high spatial and temporal resolution and high signal-to-noise ratio (SNR). In this paper, in-vivo experiments were performed in anesthetized normal and infarcted mice (one day after left anterior descending coronary artery [LAD] ligation) using a high-resolution (30 MHz) ultrasound system (Vevo 770, VisualSonics Inc., Toronto, ON, Canada). Radiofrequency (RF) signals of the left ventricle (LV) in longitudinal (long-axis) view and the associated electrocardiogram (ECG) were simultaneously acquired. Using a retrospective ECG gating technique, 2-D full field-of-view RF frames were acquired at an extremely high frame rate (8 kHz) that resulted in high-quality incremental displacement and strain estimation of the myocardium. The incremental results were further accumulated to obtain the cumulative displacements and strains. Two-dimensional and M-mode displacement images and strain images (elastograms), as well as displacement and strain profiles as a function of time, were compared between normal and infarcted mice. Incremental results clearly depicted cardiac events including LV contraction, LV relaxation and isovolumetric phases in both normal and infarcted mice, and also evidently indicated reduced motion and deformation in the infarcted myocardium. The elastograms indicated that the infarcted regions underwent thinning during systole rather than thickening, as in the normal case. The cumulative elastograms were found to have higher elastographic SNR (SNR(e)) than the incremental elastograms (e.g., 10.6 vs. 4.7 in a normal myocardium, and 6.0 vs. 2.4 in an infarcted myocardium). Finally, preliminary statistical results from nine normal (m = 9) and seven infarcted (n = 7) mice indicated the capability of the cumulative strain in differentiating infracted from normal myocardia. In conclusion, myocardial elastography could provide regional strain information at simultaneously high temporal (>/=0.125 ms) and spatial ( approximately 55 microm) resolution as well as high precision ( approximately 0.05 microm displacement). This technique was thus capable of accurately characterizing normal myocardial function throughout an entire cardiac cycle, at the same high resolution, and detecting and localizing myocardial infarction in vivo.     

Clinical Pharmacokinetics of Morphine

Morphine, the most widely used mu-opioid analgesic for acute and chronic pain, is the standard against which new analgesics are measured. A thorough understanding of the pharmacokinetics of morphine is required in order to safely and effectively use this analgesic in a wide variety of patients with different levels of organ function. A MEDLINE search was conducted to identify literature published between 1966 and January 2002 relevant to the pharmacokinetics of morphine. These publications were reviewed and the literature summarized regarding unique and clinically important elements of morphine disposition relative to its parenteral administration (including intravenous, intramuscular, subcutaneous, epidural and intrathecal administration), absorption profile (immediate release, controlled release, and sublingual/buccal, and rectal administration), distribution, and its metabolism/ excretion. Special populations, including infants, elderly, and those with renal/liver failure, have a unique morphine pharmacokinetic profile that must be taken into account in order to maximize analgesic efficacy and reduce the risk of adverse events.     

手转胎头术失败的原因与分娩结局临床分析

目的 探讨手转胎头术失败的原因与分娩结局.方法 选择2008年1月至2010年12月于我院住院分娩的持续性枕横位、枕后位产妇198例,根据行手转胎头术后结果分为成功组126例、失败组72例.比较两组分娩结局,对比分析失败原因.结果 失败组胎儿体质量≥3500 g的发生率[76.4％(55/72)]明显高于成功组[31.7％(40/126)],差异有统计学意义(x2=30.177,P=0.001)、失败组宫缩乏力发生率[58.3％(42/72)]高于成功组[38.1％ (48/126)],差异有统计学意义(x2=7.569,P=0.006)、失败组骨盆临界或轻度狭窄发生率[38.9％ (28/72)]高于成功组[23.8％(30/126)],差异有统计学意义(x2 =5.030,P=0.002)、失败组手转胎头时机不当(宫口开大＜6 cm、胎头位于坐骨棘上及宫口开大8～10 cm、胎头位于坐骨棘下≥2 cm)发生率[61.1％(44/72)]高于成功组[38.9％(49/126)],差异有统计学意义(x2=9.084,P=0.003).失败组母儿并发症(产后出血、产褥病率、胎儿窘迫、新生儿窒息)发生率高于成功组(x2 =9.586,P=0.002、x2=9.334,P=0.002、x2=5.910,P=0.015、x2=5.240,P=0.022)、失败组剖宫产发生率[72.2％(52/72)]明显高于成功组[34.1 ％(43/126),x2=26.641,P=0.001)].结论 手转胎头术能使难产变顺产,降低剖宫产率,减少母儿并发症,但须积极预防、处理导致手转胎头术失败的原因,对矫正失败后继续矫正及试产应慎重.     

Evidence-Based Pain Management and Palliative Care in Issue Four for 2009 of The Cochrane Library

ABSTRACT

The Cochrane Library of Systematic Reviews is published quarterly. Issue 4 for 2009 contains 4027 complete reviews, 1906 protocols for reviews in production, and 11447 one-page summaries of systematic reviews published in the general medical literature. In addition, there are citations of 600,000 randomized controlled trials, and 12,200 cited papers in the Cochrane methodology register. The health technology assessment database contains over 7500 citations. This edition of the Library contains 90 new reviews, of which 19 have potential relevance for practitioners in pain and palliative medicine.     

Wie zuverlässig sind Angaben zu nichtentzündlichen ausgedehnten Schmerzen?

ZusammenfassungFragestellung Es wurde geprüft, wie sich der Differenziertheitsgrad zweier Schmerzmessmethoden auf Angaben zur Ausgedehntheit klinischer Schmerzen auswirkt. Zugleich wurde der Referenzzeitraum variiert, über den die Patienten berichten sollten.Methode Erfasst wurde der Einfluss zu Lasten der Befragungsdifferenziertheit durch den Vergleich zweier Körperschema-Bildvorlagen. Drei Referenzzeiträume (Schmerz aktuell, letzte Woche, letztes halbes Jahr) wurden vorgegeben.Ergebnisse Patienten mit ausgedehnten Schmerzen gaben bei differenzierter Befragung um so mehr Schmerzen an, je weiter die Schmerzen zurück lagen und je größer der Berichtszeitraum war. Patienten mit gelenknahen Schmerzen gaben bei hoch differenzierter Befragung weniger ausgedehnte Schmerzen in der Vergangenheit an als bei globaler Einschätzung. Patienten mit Rückenschmerzen berichteten bei differenzierter Befragung zum aktuellen Schmerz über weniger ausgedehnte Schmerzen als bei globaler Befragung.Schlussfolgerung Die Angaben zur Schmerzausdehnung variieren vor allem bei Patienten mit ausgedehnten Schmerzen in Abhängigkeit von der Differenziertheit der Befragung. In diesen Fällen ist die Wahrscheinlichkeit erhöht, dass sich die Beschwerdesymptomatik zumindest teilweise erst in der Reaktion auf die situativen Befragungsbedingungen konstituiert und daher nicht auf andere Befragungsbedingungen generalisiert werden kann.