IL-8单克隆抗体靶向偶联SonoVue微气泡造影剂的方法学研究

目的 制备IL-8单克隆抗体与SonoVue声学微气泡偶联的声学造影剂,并探讨其偶联的方法学.方法 采用SPDP法将IL-8单克隆抗体与SonoVue声学微气泡进行共价偶联,玻片凝集法检验抗体与SonoVue微气泡造影剂是否偶联成功.结果 SPDP交联法制备的携IL-8单克隆抗体的免疫微气泡与二抗(羊抗鼠IgG血清)进行的玻片凝集反应,结果呈阳性,即偶联成功.结论 应用SPDP交联法能够有效地将IL-8单克隆抗体结合到声学微气泡造影剂表面,为进一步研究靶向性声学微气泡造影剂的制备与应用奠定了基础.     

抗体偶联白蛋白超声造影剂的制备及体外靶向性研究

目的制备单克隆抗体修饰的白蛋白微气泡，并探讨其体外靶向性作用。方法采用N琥珀酰亚胺基-3-（2吡啶二硫）-丙酸酯（SPDP）交联法将鼠抗人血管细胞粘附分子（VCAM-1）单克隆抗体与白蛋白微气泡进行共价偶联，采用玻片凝集法检验抗体是否与白蛋白微气泡交联；体外培养高表达VCAM-1抗原的损伤血管内皮细胞，通过抗体修饰微气泡与该细胞的特异性结合作用检验该微气泡的靶向性作用。结果SPDP交联法制备的免疫微气泡的玻片凝集反应呈阳性；损伤内皮细胞（细胞膜表面高表达VCAM-1抗原）周围可见大量免疫微气泡聚集，而正常内皮细胞（细胞膜表面无VCAM-1抗原表达）表面未见明显免疫微气泡附着。结论应用SPDP交联法能够将抗体有效地偶联到白蛋白微气泡表面，体外实验证实了其与靶细胞的特异性结合作用。     

靶向性声学造影剂与血管内皮细胞相互作用的实验研究

目的 制备携抗人VCAM-1单克隆抗体的白蛋白声学造影剂，观察其与损伤血管内皮细胞的相互作用，探讨评价血管内皮功能的新方法。 方法 采用交联法将抗人VCAM-1单克隆抗体共价偶联到自制氟碳气体为核心的白蛋白微气泡表面，制备靶向性声学微气泡；倒置显微镜下分别观察普通白蛋白微气泡、靶向性微气泡与正常内皮细胞、损伤内皮细胞的结合作用，高倍视野下计数内皮细胞及黏附的微气泡的数目，通过计算微气泡与内皮细胞的比值对两者之间的结合作用进行定量分析。 结果 无论是正常内皮细胞，或损伤内皮细胞，仅见少量的对照组微气泡的黏附作用；而镜下可见大量携VCAM-1单抗的白蛋白微气泡黏附在损伤内皮细胞表面，黏附数目显著高于黏附于正常内皮细胞表面的数目。 结论 携VCAM-1单抗的靶向性声学造影剂能够特异性结合在损伤内皮细胞表面，开拓了超声成像技术检测血管内皮损伤、评价血管内皮功能新的研究领域。     

含氟烷人血白蛋白靶向超声造影剂的制备研究

目的为靶向超声造影剂的制备进行基础实验研究。方法将含氟烷人血白蛋白超声造影剂与ICAM-1抗体混合,加入0.1%戊二醛,4℃孵育2h,促使抗体与微泡外壳充分交联。普通光镜评估靶向微泡的大小、形态。免疫荧光法进行靶向微泡的鉴定。结果普通光镜下可见携ICAM-1抗体的靶向微泡大小、形态与普通白蛋白微泡对照无明显改变。荧光显微镜证实微泡与ICAM-1抗体整合成功。结论戊二醛交联法适用于白蛋白靶向超声造影剂的制备,方法简便,成本低,重复性好。     

靶向性声学造影剂与不同程度损伤血管内皮细胞相互作用的实验研究

目的观察携VCAM-1单克隆抗体的白蛋白声学造影剂与不同程度损伤血管内皮细胞的相互作用,并探讨这种靶向性声学微气泡对内皮细胞的黏附作用与内皮细胞受损程度之间的关系。方法以不同浓度肿瘤坏死因子-α(tumor necrosis factorα,TNF-α)作为干预因子培养6组不同程度损伤的人脐静脉血管内皮细胞,采用免疫细胞化学染色与病理图文分析法检测各组血管内皮细胞表面VCAM-1的表达水平;采用SPDP交联法制备携VCAM-1单克隆抗体的靶向性白蛋白微气泡,倒置显微镜下观察其与各组损伤内皮细胞的结合作用,并于高倍视野下(200×)计数内皮细胞及黏附的微气泡的数目,计算微气泡与内皮细胞的比值作为微气泡的黏附率;各组损伤内皮的免疫微气泡的黏附率与其表面VCAM-1的表达水平进行相关性分析。结果随着TNF-α干预浓度的增高,内皮细胞表面VCAM-1的表达水平逐渐增高,且各组之间具有显著性差异;镜下观察可见损伤内皮细胞免疫微气泡黏附作用亦随着干预浓度的增高而增强,且微气泡对损伤内皮细胞的黏附率与细胞表面VCAM-1的表达水平呈正相关。结论携VCAM-1单克隆抗体的靶向性声学造影剂不仅能够与损伤血管内皮细胞特异性结合,其黏附率亦能够反映内皮细胞损伤的程度。     

抗VEGF抗体介导靶向超声造影剂的体外实验研究

目的以抗血管内皮因子（VEGF）抗体为配体研制能与血管内皮细胞特异性结合的靶向脂质体超声造影剂并检测其体外寻靶能力。 方法以静电吸附法将抗VEGF抗体连接到脂质体造影剂微泡的表面；体外培养ECV304人血管内皮细胞，用免疫荧光法检测靶向造影剂与其体外结合能力，以普通造影剂为对照组。 结果所制备的靶向超声造影剂与普通微泡无显著差异；免疫荧光实验结果显示靶向造影剂能在体外与血管内皮细胞特异性结合。 结论携抗VEGF抗体的脂质体靶向造影剂能通过静电吸附法成功制备，且在体外能与血管内皮细胞特异性结合。     

超声成像方式和造影剂剂量对造影剂微气泡后散射强度的影响

目的：采用声学密度法，观察实验动物犬在改变造影剂剂量和仪器设置条件时心腔内不同类型的造影剂切散射强度的差异，探讨超声成像方式和造影剂剂量对造影剂微气泡后散射强度的影响。方法：取主动脉根部短轴切面，将仪器分别设置为基波、谐波1．8／3．6和谐波2．1／4．2状态，经周围静脉注射不同剂量的声振白蛋白造影剂、levovist和含氟碳造影剂，记录不同仪器设置状态和不同造影剂剂量时的后散射二维图像，并与右室压力曲线同步。结果：仪器设置条件的改变对声振白蛋白造影剂微气泡后散射强度的影响不大，却对levovist和含氟碳造影剂后散射强度的影响差异具有显著性；声振白蛋白造影剂、levovist和含氟碳造影剂舒张末期和收缩末期后散射强度均与造影剂剂量成反比，而舒张末期和收缩末期后散射强度之差不受造影剂剂量改变的影响。结论：不同类型的造影剂微气泡在超声仪器设置条件改变时后散射强度变化不一，与微气泡的大小和与之相匹配的共振频率有关。提示运用声学密度法，测量微气泡后散射强度舒张末期和收缩末期之差反映心腔内压力高低的临床应用中，应考虑到超声成像方式对微气泡后散射强度的影响，特定的造影剂应和特定的二次谐波频率相匹配。     

声学密度法分析压力对微气泡后散射强度的影响

目的 探讨声学密度法在犬心腔内声振白蛋白造影剂后散射时间—强度曲线随心动周期的变化规律，分析收缩期压力和造影剂剂量的改变对其后散射强度的影响。方法 取主动脉根部短轴切面，经多巴酚丁胺逐步升压后，周围静脉注射不同剂量声振白蛋白造影剂，记录不同收缩期压力和不同造影剂剂量时的后散射二维图像，并与右室压力曲线同步。结果 心腔内微气泡后散射时间—强度曲线随心动周期发生可逆性的周期性变化，收缩末期后散射强度达到最低，舒张末期恢复原强度；舒张末期和收缩末期后散射强度之差随收缩期压力的升高逐渐加大，与收缩期压力高低相关(r＝0．64，P＜0．0001)，与造影剂剂量无关(P＝0．57)。结论 压力作用致微气泡后散射强度的减弱主要与微气泡的暂时被压缩有关，利用声学密度法，通过观察心腔内微气泡后散射时间—强度曲线的变化，测量微气泡舒张末期和收缩末期后散射强度之差可定量估测心腔内压力的高低。     

自然组织谐波成像与基波成像在心脏声学造影中的对比观察

组织谐波成像是新近开展的一种超声成像技术，其探头接受声波的频率是发射频率的两倍（分别为１．７５ＭＨｚ和３．５ＭＨｚ），我们将该技术应用于心脏声学造影检查，观察其对造影剂微气泡的显像效果，并与传统的基波成像技术相比较，发现前者右心显影更强，密度更高，散在造影剂微气泡颗粒更粗，更亮，并能清晰显示左心少量的穿肺的造影剂，这是传统方法难于探及的，但要达到心肌显影，还需改进造影剂的制备，开发出新的心肌声学造影剂。     

压力对心腔内造影剂微气泡后向散射强度的影响

目的与方法：本研究应用声学密度法在体外搏动心脏模型中定量分析压力对声振白蛋白微气泡后向散射强度的影响,并观察正常人体心腔内双氧水微气泡后向散射强度在心动周期中的变化规律。结果：Ａｌｂｕｎｅｘ声振白蛋白微气泡在心腔内压力的作用下,其大小可发生可逆性变化,而这种变化可相应地引起后向散射强度可逆的周期性变化。舒张末期和收缩末期后向散射强度之差与造影剂浓度无关,而与收缩末期压力高低密切相关。在初步的人体研究中,我们也观察到了同样的心腔内造影剂后向散射强度的变化规律。结论：有可能通过应用声学密度法观察心腔内造影剂微气泡后向散射强度变化、无创性估计心腔内压力。     

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.     

Dissociable correlates of two classes of retrieval processing in prefrontal cortex

Although substantial evidence suggests that the prefrontal cortex (PFC) implements processes that are critical for accurate episodic memory judgments, the specific roles of different PFC subregions remain unclear. Here, we used event-related functional magnetic resonance imaging to distinguish between prefrontal activity related to operations that (1) influence processing of retrieval cues based on current task demands, or (2) are involved in monitoring the outputs of retrieval. Fourteen participants studied auditory words spoken by a male or female speaker and completed memory tests in which the stimuli were unstudied foil words and studied words spoken by either the same speaker at study, or the alternate speaker. On "general" test trials, participants were to determine whether each word was studied, regardless of the voice of the speaker, whereas on "specific" test trials, participants were to additionally distinguish between studied words that were spoken in the same voice or a different voice at study. Thus, on specific test trials, participants were explicitly required to attend to voice information in order to evaluate each test item. Anterior (right BA 10), dorsolateral prefrontal (right BA 46), and inferior frontal (bilateral BA 47/12) regions were more active during specific than during general trials. Activation in anterior and dorsolateral PFC was enhanced during specific test trials even in response to unstudied items, suggesting that activation in these regions was related to the differential processing of retrieval cues in the two tasks. In contrast, differences between specific and general test trials in inferior frontal regions (bilateral BA 47/12) were seen only for studied items, suggesting a role for these regions in post-retrieval monitoring processes. Results from this study are consistent with the idea that different PFC subregions implement distinct, but complementary processes that collectively support accurate episodic memory judgments.     

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.     

Guns and Knives in New Mexico: Patterns of Penetrating Trauma, 1978–1993

Objective: To identify patterns of nonfatal and fatal penetrating trauma among children and adults in New Mexico using ED and medical examiner data.
Methods: The authors retrospectively sampled in 5-year intervals all victims of penetrating trauma who presented to either the state Level-1 trauma center or the state medical examiner from a 16-year period (1978–1993). Rates of nonfatal and fatal firearm and stabbing injury were compared for children and adults.
Results: Rates of nonfatal injury were similar (firearm, 34.3 per 100,000 person-years; stabbing, 35.1). However, rates of fatal injury were significantly different (firearm, 21.9; stabbing, 2.7; relative risk: 8.2; 95% confidence interval: 5.4, 12.5). From 1978 to 1993, nonfatal injury rates increased for children (p = 0.0043) and adults (p < 0.0001), while fatal penetrating injury remained constant. The increase in nonfatal injury in children resulted from increased firearm injury rates. In adults, both stabbing and firearm nonfatal injury rates increased.
Conclusions: Nonfatal injury data suggest that nonfatal violence has increased; fatal injury data suggest that violent death rates have remained constant. Injury patterns vary by age, mechanism of trauma, and data source. These results suggest that ED and medical examiner data differ and that both are needed to guide injury prevention programs.