抗绿脓杆菌单抗制备及对小鼠绿脓杆菌感染的保护性作用

目的制备抗绿脓杆菌单克隆抗体,观察其对绿脓杆菌感染的烧伤小鼠及正常小鼠的保护作用。方法应用NS-1细胞和经绿脓杆菌免疫后的BALB/C小鼠脾细胞制备杂交瘤细胞株,收集其分泌的抗体,观察其对绿脓杆菌感染的Ⅲ°烧伤小鼠及正常小鼠的免疫保护作用。结果该单克隆抗体可预防绿脓杆菌感染。结论我们的研究结果为抗绿脓杆菌单克隆抗体在临床上应用的可行性提供了实验室基础。     

丙种球蛋白和富含IgM的免疫球蛋白中抗绿脓杆菌抗体含量及其保护性质的比较研究

近年来,绿脓杆菌引起的传染病发病率在不断增长.临床医生认为,这种感染很难治疗,这是因为病原体对抗生素产生耐药性以及常常因免疫保护力低下而发生。有资料表明,超免疫血浆及特异性人和动物的免疫球蛋白含有各种绿脓杆菌抗原的抗体,并由绿脓杆菌引起动物感染的试验中证明它们的保护作用。也已证明市售的丙种球蛋白具有被动保护小鼠免于致死感染。然而,人们知道,市售的丙种球蛋白中基本上为 IgG,而正常的抗细菌抗体则主要与 IgM 组分相关。通过正常人血清组分抗体分布研究表明,抗绿脓杆菌抗体主要浓缩于 IgM 组分。     

黄芪多糖抗呼吸道绿脓杆菌感染的实验研究

目的:探讨黄芪多糖拮抗绿脓杆菌在呼吸道黏附,阻断感染作用及机制,从而证明黄芪多糖是补益类中药黄芪中起以上作用的主要成分。方法:复制慢性支气管炎(慢支)感染绿脓杆菌小鼠模型。从120只模型小鼠中随机选择20只进行细菌定量培养,观察感染后35 d内小鼠气管带菌情况。另100只小鼠随机再均分为5组。低、中、高剂量治疗组分别以0.26、0.78和1.56 mg黄芪多糖灌胃,阳性对照组以慢支固本冲剂1 ml灌胃,阴性对照组以1 ml蒸馏水灌胃,各组均连用10 d。从各组随机取10只小鼠,先取其气管组织作细菌定量培养,观察小鼠气管带菌数;再取其气管标本进行病理切片,用光学显微镜观察气管黏膜上皮的病理变化。将各组另10只小鼠进行活体气管和支气管原位固定,用扫描电镜观察其超微结构及绿脓杆菌黏附数。结果:1各治疗组细菌培养菌落形成单位数明显低于阴性对照组(P均<0.01),高剂量治疗组与阳性对照组差异无显著性(P>0.05)。2光镜观察:各治疗组气管黏膜的病理变化轻于阴性对照组;高剂量治疗组和阳性对照组病变基本消失。3电镜观察:治疗组纤毛损伤、黏附的绿脓杆菌数明显少于阴性对照组,与阳性对照组相似。结论:黄芪多糖对慢支病变小鼠气管黏膜上皮有修复作用,能阻断绿脓杆菌在呼吸道的黏附,从而起到抗绿脓杆菌感染的作用;黄芪多糖是补益类中药黄芪中抗细菌黏附的有效成分。     

特重度烧伤患者绿脓杆菌感染的临床特点与护理

目的 探讨特重烧伤患者绿脓杆菌感染的特点与护理治疗方法。方法 回顾分析本院64例特重烧伤患者绿脓杆菌感染的救治与护理过程。结果 本组44例特重烧伤合并绿脓杆菌感染患者，除8例因经济原因无法救治外，4例死于各种并发症。其成功救治除了临床上对绿脓杆菌感染的早期预防和高敏感性抗生素的使用外，护理中严格控制交叉感染、注重日常护理常规以及红外线辐射床等辅助措施也起重要作用。结论 特重烧伤患者绿脓杆菌感染的救治重在预防交叉感染、及时应用有效抗生素和加强常规护理措施。     

抗脂多糖单克隆抗体的制备及功能研究

摘 要 目的：制备广谱结合活性抗脂多糖单克隆抗体,并探讨其保护活性。 方法：用噬菌体展示技术筛选出模拟脂多糖保守结构的多肽;将模拟肽与蓝载体交联后免疫BALB/c小鼠并制备单克隆抗体;间接ELISA与免疫荧光检测其结合活性;小鼠体内、外实验检测其保护活性。结果：经过三次融合,筛选到一株具广谱结合活性的抗脂多糖单克隆抗体SMU-3A8;间接ELISA与免疫荧光检测显示其可以和4种商品化脂多糖及6种G-菌发生交叉反应;体外实验显示SMU-3A8可以部分抑制脂多糖刺激小鼠单核巨噬细胞系RAW264.7产生NO;体内实验表明其可以延长致死量鼠伤寒杆菌感染BALB/c小鼠的生存时间、提高生存率。结论：在本实验中,我们成功制备了一株具广谱结合活性,对鼠伤寒菌感染小鼠模型有保护作用的抗脂多糖单克隆抗体;为脂多糖相关研究提供了一个有力工具。 关键词 脂多糖 单克隆抗体 噬菌体展示 模拟肽     

HEV保护性抗体的制备与功能研究

目的研究基于戊型肝炎病毒中和表位(p179)的小鼠单克隆抗体的HEV中和保护作用。方法构建含HEV中和表位p179基因的真核表达质粒venus-179,以尾静脉注射方法对BALB/c小鼠实施基因免疫,ELISA法检测免疫后小鼠血清中抗体应答水平,通过单克隆抗体的方法制备HEV抗体,通过C3A细胞株人HEV细胞感染模型,采用RT-PCR法检测中和试验后保护性抗体的中和活性。结果成功制备了6株针对戊型肝炎病毒中和表位(p179)的抗体,其中抗体株2G8F9能有效中和HEV病毒,证实其产生的特异性抗体能够有效阻断HEV感染易感细胞C3A,具有中和HEV的活性。结论戊型肝炎病毒中和表位(p179)的基因疫苗venus-179免疫小鼠制备的单克隆抗体具有中和作用的免疫应答。     

抗人脑钠肽单克隆抗体的制备及其鉴定

目的:制备抗人脑钠肽单克隆抗体并对其进行初步鉴定。方法:实验于2004-12/2006-02在南京医科大学第一附属医院心血管病研究所进行。利用碳化二亚胺法将合成的人脑钠肽与牛血清白蛋白偶联制备完全抗原,反复免疫Balb/c小鼠,应用杂交瘤技术将免疫小鼠的脾细胞与Balb/c小鼠骨髓瘤细胞(SP2/0)在PEG下融合,HAT选择性培养,间接酶联免疫吸附方法对细胞培养上清检测、筛选,选择筛选结果脑钠肽抗体阳性的细胞株连续进行3次亚克隆,建立稳定分泌脑钠肽单克隆抗体的杂交瘤细胞株。扩大培养阳性单克隆细胞株后,腹腔注射小鼠,制备腹水。对腹水亲和层析纯化,所得的单克隆抗体行初步鉴定。结果:免疫小鼠血清呈脑钠肽抗体阳性,经筛选得到3株稳定分泌脑钠肽单克隆抗体的杂交瘤细胞株,纯化的单克隆抗体经鉴定为IgG型,蛋白质印迹分析证实单克隆抗体可特异识别脑钠肽,具备较高的特异性及敏感性。结论:成功制备了3株抗脑钠肽特异性的单克隆抗体,可用于脑钠肽免疫检测方法的建立。     

抗人脑钠肽单克隆抗体的制备及其鉴定

目的：制备抗人脑钠肽单克隆抗体并对其进行初步鉴定。 方法：实验于2004-12／2006-02在南京医科大学第一附属医院心血管病研究所进行。利用碳化二亚胺法将合成的人脑钠肽与牛血清白蛋白偶联制备完全抗原，反复免疫Balb／c小鼠，应用杂交瘤技术将免疫小鼠的脾细胞与Balb／c小鼠骨髓瘤细胞（SP2／0）在PEG下融合，HAT选择性培养，间接酶联免疫吸附方法对细胞培养上清检测、筛选，选择筛选结果脑钠肽抗体阳性的细胞株连续进行3次亚克隆，建立稳定分泌脑钠肽单克隆抗体的杂交瘤细胞株。扩大培养阳性单克隆细胞株后，腹腔注射小鼠，制备腹水。对腹水亲和层析纯化，所得的单克隆抗体行初步鉴定。 结果：免疫小鼠血清呈脑钠肽抗体阳性，经筛选得到3株稳定分泌脑钠肽单克隆抗体的杂交瘤细胞株，纯化的单克隆抗体经鉴定为IgG型，蛋白质印迹分析证实单克隆抗体可特异识别脑钠肽，具备较高的特异性及敏感性。 结论：成功制备了3株抗脑钠肽特异性的单克隆抗体，可用于脑钠肽免疫检测方法的建立。     

抗重组人内皮抑素单克隆抗体的制备

目的 用重组人内皮抑素融合蛋白作为抗原,制备鼠抗人内皮抑索单克隆抗体.方法 采用麦芽糖结合蛋白理组内皮抑素(MBP-endostatin)免疫Balb/c小鼠,取其脾脏与同系骨髓瘤细胞(SP2/0)融合,通过酶联免疫吸附试验(EUSA)筛选阳性克隆.结果 经2次亚克隆后获得一株阳性的杂交瘤细胞.结论 抗人内皮抑素单克隆抗体的制备为肿瘤治疗的基础研究及检测奠定了基础.     

绿脓杆菌感染的抗菌药物治疗

绿脓杆菌(学名:铜绿假单胞菌,PA)是假单胞菌属中最为多见的一种机会致病菌,广泛分布于自然界中,可通过环境污染、交叉感染、内源性感染和医源性感染等途径传播.该菌常殖于正常人呼吸道、皮肤黏膜以及医疗设备特别是吸氧器、湿化器、气管插管、呼吸机活瓣与管道等处,当宿主免疫功能低下、长期使用广谱抗生素、糖皮质激素、肿瘤放疗、化疗、气管切开及接受吸氧、气管插管机械通气、留置导尿管或胃管等因素存在时,极易引起感染.临床上绿脓杆菌常可引起尿路感染、烧伤创面及褥疮感染、败血症、肺部感染、脑膜炎等.     

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.