抗体偶联白蛋白声学微气泡的方法学研究

目的 制备抗体偶联的白蛋白声学微气泡造影剂，并探讨偶联反应的条件。 方法 采用N-succinimidyl 3-（2-pyridyldithio）propionate，SPDP）法将人IgG抗体与白蛋白声学微气泡以不同的质量比进行共价偶联，玻片凝集法与免疫荧光染色法检验抗体与白蛋白声学微气泡造影剂的交联并确定最佳反应比例。 结果 SPDP法制备的免疫微气泡的玻片凝集反应以及免疫荧光染色均呈阳性，IgG抗体与白蛋白声学微气泡造影剂质量比为1：5时免疫荧光反应强度最高。 结论 应用SPDP法能够将抗体有效地结合到白蛋白声学微气泡造影剂表面，为进一步靶向性声学微气泡造影剂的制备奠定了基础。     

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.     

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。     

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。     

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。     

免疫脂质体微泡造影剂的制备及体外靶向研究

目的制备抗人肝细胞癌免疫脂质体微泡造影剂,并探讨其体外靶向作用.方法采用静电吸附法将抗人肝细胞癌单克隆抗体HAb18结合到本试验室研制的脂膜氟烷微泡造影剂表面,制备免疫脂质体微泡造影剂;采用玻片凝集实验、荧光免疫染色实验证明抗体与脂质体微泡的结合;通过花环形成及花环形成阻断实验研究免疫脂质体微泡的靶向作用.结果静电吸附法可使单抗HAb18牢固地结合到脂质体微泡上;免疫脂质体微泡围绕靶细胞形成花环,花环形成率达90%以上.结论采用静电吸附法制备的免疫脂质体微泡造影剂在体外能与人肝癌细胞高效特异结合.     

携IL-8单抗靶向超声造影剂与血管内皮细胞相互作用的实验研究

目的 观察自制携带IL-8单抗靶向超声造影剂与损伤血管内皮细胞的相互作用,探讨IL-8在粥样斑块形成过程中的作用和评价血管内皮功能的新方法.方法 采用交联剂将抗人IL-8单克隆抗体偶联到SonoVue微泡表面,制备靶向性超声微泡;倒置显微镜下分别观察SonoVue微泡、靶向性微泡与正常内皮细胞、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泡数,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 仅有极少量对照组SonoVue微泡黏附于正常及损伤内皮细胞;而镜下可见携IL-8单抗的SonoVue微泡黏附于内皮细胞,黏附于损伤内皮细胞表面的微泡数显著高于正常内皮细胞.结论 携IL-8单抗的靶向性超声造影剂能够特异性结合在损伤细胞表面,拓展了超声成像技术检测血管内皮损伤、评价血管内皮功能的新领域.     

靶向VEGFR-3超声造影剂的制备及其体外寻靶实验研究

目的 制备一种特异性靶向淋巴管内皮细胞的高分子超声造影剂,并考察其体外寻靶能力.方法 采用双乳化法制备高分子造影剂羟基端乳酸-羟基乙酸共聚物(PLGA-COOH);并用碳二亚胺法将造影剂与抗体偶联制备出靶向高分子造影剂.检测该造影剂一般特性及体外寻靶能力,并与普通高分子造影剂做比较.结果 高分子造影剂平均粒径771.2 nm.体外寻靶试验显示,该靶向高分子造影剂较多并牢固的聚集到淋巴管内皮细胞表面;普通高分子造影剂对照组未见造影剂和细胞的结合.结论 成功制备出结合VEGFR-3抗体的高分子超声造影剂.该造影剂在体外对淋巴管内皮细胞有较强的特异性亲和力.     

抗体微矩阵玻片经硅酸钠处理后对红细胞血型抗体吸附能力的研究

目的探讨经硅酸钠修饰后用作抗体微矩阵载体的玻璃基片对血型抗体的吸附能力。方法用硅酸钠溶液在玻片上形成一层膜,再酸化以形成硅酸,然后用红细胞血型抗体包被后,以可与其发生特异性抗原抗体免疫反应的对应红细胞作反应指示剂,检测经硅酸钠处理后的玻片对血型抗体的吸附能力,以及血型抗体的免疫活性。结果红细胞血型抗体可以牢固的结合在硅酸钠处理过的玻片上,并且可以与其对应的红细胞抗原发生免疫学反应。结论经硅酸钠处理的玻片对血型抗体有很高的结合效力,同时抗体仍保持很高的免疫活性,因而可充当蛋白质抗体微矩阵基片。     

靶向包载经PEG修饰的重组腺病毒PLGA造影剂的制备及其体外寻靶实验

目的采用共价结合的方法制备靶向包载经PEG修饰的重组腺病毒PLGA-COOH造影剂,并考察其体外寻靶能力。方法采用碳二亚胺法将包载经PEG修饰的重组腺病毒PLGA-COOH造影剂与抗体共价结合,制备靶向高分子超声造影剂,检测该造影剂一般特性及体外寻靶能力。结果体外寻靶实验显示,该靶向超声造影剂与普通包载经PEG修饰的重组腺病毒PLGA COOH造影剂对照能聚集并牢固结合到人脐静脉血管内皮细胞表面。结论成功制备出靶向包载经PEG修饰的重组腺病毒PLGA-COOH造影剂,该靶向造影剂在体外具有较强的特异性寻靶结合能力。     

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.     

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.