细胞因子诱导的杀伤细胞抗肿瘤机制及应用

摘要： 细胞因子诱导的杀伤细胞(CIK)是一群体外诱导的以CD3⁺CD56⁺ T细胞为主的异质细胞群,具有效应CD8⁺ T细胞的TCR特异性和非MHC限制性抗肿瘤活性的特点。采用荷瘤小鼠模型进行CIK临床前研究,结果表明CIK对实体肿瘤和血液系统肿瘤均有明显的抗肿瘤效应。临床研究证实CIK可用于治疗多种恶性肿瘤,显著延长患者的生存期且无毒副作用。用免疫学与基因工程方法增强CIK疗法的特异性和细胞毒活性,寻求其疗效判断的生物学标志及CIK治疗的纳入与排除标准是目前研究的热点。     

细胞因子诱导杀伤细胞抗淋巴瘤细胞作用的研究

目的:探讨体外细胞因子诱导杀伤(CIK)细胞对淋巴瘤细胞的杀伤作用.方法:采集健康人外周血单个核细胞(PBMC),经IL-2、CD3Mc-Ab、IL-1、IFN-γ诱导,制备CIK细胞,同时以淋巴因子激活的杀伤细胞(LAK细胞)作为对照,流式细胞仪检测CIK细胞的免疫表型,LDH释放法检测其对淋巴瘤细胞株的杀伤活性.结果:CIK细胞与LAK细胞增殖在前期无明显差异,CIK细胞在培养14d后大量增殖,21d后扩增能力显著高于LAK细胞(P＜0.05);表型分析表明CIK细胞主要由CD3+和CD56+双阳性细胞构成;同一效靶比时,CIK细胞对淋巴瘤细胞株的杀伤活性明显高于LAK细胞(P＜0.05).结论:CIK细胞对淋巴瘤细胞株的杀伤作用强于LAK细胞,具有较强的抗淋巴瘤细胞活性,是肿瘤过继免疫治疗中更为有效的杀瘤效应细胞.     

健康人CIK细胞输注对恶性肿瘤患者细胞免疫功能的影响

目的:研究用健康人细胞因子诱导的杀伤细胞(CIK细胞)输注前后恶性肿瘤患者外周血细胞免疫功能的变化.方法:分离健康人外周血单个核细胞(PBMC),予rhIFN-γ、rhIL-2、rhIL-1β、CD3单克隆抗体(CD3mAb)诱导CIK细胞,回输至病人体内;比较肿瘤患者用CIK细胞治疗前后PBMC对K562的杀伤活性和外周血T淋巴细胞亚群绝对计数的变化情况.结果:肿瘤患者治疗前PBMC杀伤活性和淋巴细胞亚群绝对计数均低于健康人,治疗后除NK细胞绝对计数未见明显改变外均有所提高,但仍比健康人低.结论:健康人CIK细胞治疗能提高肿瘤患者自身抗肿瘤能力.     

细胞因子诱导杀伤细胞过继免疫治疗血液系统恶性肿瘤的研究进展

细胞因子诱导杀伤(CIK)细胞具有T淋巴细胞与自然杀伤(NK)细胞的表型及功能特征.其体外培养细胞扩殖率高、不受主要组织相容性复合物(MHC)限制,在体外对血液系统恶性肿瘤细胞有显著的细胞毒性.这些特性均使得CIK细胞过继免疫治疗抗血液系统恶性肿瘤活性逐步成为相关研究的热点.多项临床试验结果表明,CIK细胞过继免疫治疗在血液系统恶性肿瘤患者中具有可行性及有效性;通过CIK细胞过继免疫治疗结合标准治疗方案已被证实在抗血液系统恶性肿瘤中发挥协同作用.CIK细胞过继免疫治疗在血液系统恶性肿瘤的治疗中能够提高患者完全缓解(CR)率、延长生存时间并提高生活质量.笔者拟就CIK细胞过继免疫治疗血液系统恶性肿瘤的相关研究进展进行综述.     

两种培养基对细胞因子诱导杀伤细胞体外扩增的影响

目的:比较AIM V、OpTmizer两种培养基对培养细胞因子诱导的杀伤细胞(cytokine-induced killercells,CIK细胞)的增殖、表型及细胞毒功能的影响。方法:分离7位志愿者外周血单核细胞(peripheral bloodmononuclear cells,PBMC),分别应用AIM V、OpTmizer两种培养基体外常规诱导CIK细胞,于培养第3、7、10、12、14、17、21天时,计算CIK细胞扩增倍数;通过流式细胞仪分析CD3、CD8、CD56分子的表达;通过LDH释放实验比较两种培养的CIK细胞对K562的细胞毒作用。结果:培养的10、12、14、17、21 d,OpTmizer扩增倍数明显高于AIM V(P<0.05);平均CD56~+及CD3~+CD56~+细胞频数,两组均高于基线(P<0.05);平均CD3-CD56~+细胞频数,OpTmizer组高于基线(ρ<0.05);对K562细胞的细胞毒作用,两组比较差异没有显著性。结论:OpTmizer扩增CIK细胞效率明显高于AIMV,而扩增的细胞亚群及对K562细胞的杀伤作用两组基本相同,OpTmizer比AIM V更适合于体外诱导培养CIK细胞。     

细胞因子诱导的杀伤细胞与肿瘤浸润淋巴细胞体外细胞毒活性比较

目的比较用多种细胞因子诱导的杀伤细胞(CIK)与癌性胸水中肿瘤浸润淋巴细胞(TIL)的细胞毒活性,为临床应用提供依据。方法用不同的细胞因子分别诱导培养CIK和癌性胸水TIL,按常规法计数细胞增殖量,采用四甲基偶氮唑盐(MTT)比色法检测其细胞毒活性。结果诱导培养14d后,CIK细胞毒活性为(76.65±16.78)%,增殖倍数为520±102;TIL细胞毒活性为(55.31±11.02)%,增殖倍数为182±78。结论CIK细胞对K562细胞的细胞毒活性在14d时达峰值,而TIL细胞在21d时才达峰值,且CIK细胞毒活性明显高于TIL细胞,增殖细胞数量也比TIL细胞高。CIK可替代TIL细胞进行胸腔注射治疗癌性胸水。     

细胞因子诱导的杀伤细胞与肿瘤浸润淋巴细胞体外细胞毒活性比较

目的比较用多种细胞因子诱导的杀伤细胞(CIK)与癌性胸水中肿瘤浸润淋巴细胞(TIL)的细胞毒活性,为临床应用提供依据.方法用不同的细胞因子分别诱导培养CIK和癌性胸水TIL,按常规法计数细胞增殖量,采用四甲基偶氮唑盐(MTT)比色法检测其细胞毒活性.结果诱导培养14 d后,CIK细胞毒活性为(76.65±16.78)%,增殖倍数为520±102;TIL细胞毒活性为(55.31±11.02)%,增殖倍数为182±78.结论CIK细胞对K562细胞的细胞毒活性在14 d时达峰值,而TIL细胞在2l d时才达峰值,且CIK细胞毒活性明显高于TIL细胞,增殖细胞数量也比TIL细胞高.CIK可替代TIL细胞进行胸腔注射治疗癌性胸水.     

肿瘤自体CIK细胞免疫治疗前后T细胞亚群的变化

细胞因子诱导的杀伤细胞（CIK细胞）具有增殖快、杀瘤活性强和杀瘤谱广的特点。目前，用于临床治疗肿瘤的报告较少。本文用CIK细胞过继性免疫治疗肿瘤患者，探讨其对细胞免疫的影响。方法：用CS-3000得到肿瘤患者未梢血单个核细胞（PBMC），用含有IFN-r、IL-2和CD3单抗的培养液进行培养扩增，于培养的第11天和第12天回输给病人，并检测CIK治疗前后患者的外周血T细胞亚群的变化。结果：38例肿瘤患砉输注CIK细胞一疗程后，患者外周血CD3^＋、CD4^＋、CD8^＋T淋巴细胞都较治疗前明显提高（P〈0．01）。结论：用自身CIK细胞过继性回输治疗能明显提高肿瘤患者细胞免疫功能。     

肿瘤细胞淋巴细胞混合培养诱导产生胶质瘤特异性CTL的实验研究

目的 通过自身肿瘤激活诱导产生高活性的胶质瘤特异性细胞毒T淋巴细胞(G-S-CTL)。方法 应用淋巴细胞、肿瘤细胞混合培养(MLTC)方法，诱导产生G-S-CTL，采用3H-TdR释放法及免疫荧光法检测G-S-CTL的细胞毒活性及免疫表型。结果 G-S-CTL与自身LAK相比，抗肿瘤活性大为提高，CD3 、CD4 、CD8 和CD25 细胞明显增多。结论 G-S-CTL抗肿瘤活性及免疫表型类似于胶质瘤浸润的淋巴细胞(GIL)，对脑胶质瘤过继免疫治疗具有一定的实用价值。     

不同培养基中CIK的生物学特性及其对肿瘤细胞的杀伤效果

目的比较健康人和肿瘤患者细胞因子诱导的杀伤细胞（cytokine-induced killer,CIK）在淋巴细胞无血清培养基及普通无血清培养基中的细胞形态、体外扩增速度、细胞表型及其对肿瘤细胞株的杀伤活性。方法取健康人和肿瘤患者外周血各3例,经密度梯度离心获得外周血单个核细胞（PBMC）,分别在淋巴细胞无血清培养基及普通无血清培养基中,加入基因重组人干扰素γ（rhIFN-γ）、抗人CD3单克隆抗体（OKT3）和基因重组人白细胞介素2（rhIL-2）,体外诱导CIK细胞,比较细胞的形态、扩增速度、细胞表型及杀伤活性。结果在不同培养基中健康人外周血CIK细胞的扩增速度、流式细胞学检测的CD3＋CD8＋和CD3＋CD56＋细胞百分比以及对K562和HL-60肿瘤细胞株的杀伤活性均显著高于肿瘤患者,差异有统计学意义（P〈0.05）;两种来源的PBMC在不同的培养基中经过诱导获得的CIK细胞在增殖速度、流式细胞学分析结果及杀伤活性方面差异亦有统计学意义（P〈0.05）。结论在不同的培养基中肿瘤患者的PBMC经诱导培养获得的CIK细胞可有显著差别,为临床进一步提高CIK细胞的治疗效果提供了实验依据。     

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.     

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.     

Molecular characterization of virulence and antimicrobial resistance profile of Shigella species isolated from children with moderate to severe diarrhea in northeastern Brazil

Molecular characterization of virulence and antimicrobial resistance profiles were determined for Shigella species isolated from children with diarrhea in Fortaleza, Brazil. Fecal specimens were collected along with socioeconomic and clinical data from children with moderate to severe diarrhea requiring emergency care. Shigella spp. were isolated by standard microbiological techniques, and we developed 4 multiplex polymerase chain reaction assays to detect 16 virulence-related genes (VRGs). Antimicrobial susceptibility tests were performed using disk diffusion assays. S. flexneri and S. sonnei were the predominant serogroups. S. flexneri was associated with low monthly incomes; more severe disease; higher number of VRGs; and presence of pic, set, and sepA genes. The SepA gene was associated with more intense abdominal pain. S. flexneri was correlated with resistance to ampicillin and chloramphenicol, whereas S. sonnei was associated with resistance to azithromycin. Strains harboring higher numbers of VRGs were associated with resistance to more antimicrobials. We highlight the correlation between presence of S. flexneri and sepA, and increased virulence and suggest a link to socioeconomic change in northeastern Brazil. Additionally, antimicrobial resistance was associated with serogroup specificity in Shigella spp. and increased bacterial VRGs.     

护理干预对面部中重度寻常型痤疮的临床疗效影响分析

目的研究护理干预对面部中重度寻常型痤疮的临床疗效影响。方法选取本院在2014年4月~2016年7月诊治的136例面部中重度寻常型痤疮患者,随机分为研究组与对照组,每组68例;所有患者均依据其情况给予对应的治疗,其中对照组在治疗期间给予常规护理,研究组在对照组的基础上再给予综合性护理干预,比较两组的治疗效果及护理满意度情况等。结果患者在接受治疗和护理后,研究组中度与重度患者的治疗效果较对照组均明显提高(P0.05),研究组护理满意度较对照组明显增高(P0.05)。结论对面部中重度寻常型痤疮患者在其治疗期间给予综合性护理干预,具有良好的效果。