载血卟啉单甲醚高分子纳米粒用于光声显像引导下声动力治疗:实验研究

目的 制备一种载血卟啉单甲醚（HMME）的多功能分子探针,评估其体外光声成像及声动力治疗（SDT）效果。方法 采用双乳化法制备载HMME的聚乳酸-乙醇酸（PLGA）纳米粒（HMME@PLGA）,观察纳米粒的基本表征和体外增强光声显像情况,通过流式细胞仪检测人乳腺癌MDA-MB-231细胞与HMME@PLGA共孵育后在超声辐照下产生活性氧的能力,并在细胞水平验证其SDT效果。结果 所制备的HMME@PLGA纳米粒大小均一,形态规则,平均粒径为（333.67±17.50）nm,表面电位为（-10.57±1.98）mV,包封率为75.62%,载药量为2.90%,浓度为1 mg/ml的纳米粒与人乳腺癌MDA-MB-231细胞共孵育24 h的细胞存活率为87.21%。随浓度增加,纳米粒光声信号逐渐增强。经超声辐照,HMME@PLGA纳米粒能使人乳腺癌MDA-MB-231细胞产生大量活性氧,并产生明显细胞毒性作用,使细胞大量死亡,Calcein-AM/PI染色呈红色荧光;通过吖啶橙染色观测到细胞溶酶体结构消失。结论 本研究成功制备了包裹HMME的高分子纳米粒,可实现体外光声成像引导下的SDT。     

靶向癌胚抗原载药纳米超声造影剂体外抑制卵巢癌细胞生长

目的 制备靶向癌胚抗原（CEA）载药相变型PLGA纳米粒（Ab-PTX-NPs）,探讨该纳米粒的体外寻靶及抑制肿瘤细胞生长的效能。方法 乳化溶剂挥发法联合碳二亚胺法制备靶向CEA载紫杉醇纳米粒（Ab-PTX-NPs）,采用马尔文粒径分析仪检测其粒径大小。采用高效液相色谱法检测紫杉醇包封率及载药量,恒温摇床透析法检测该纳米粒体外释药特征。激光共聚焦显微镜及流式细胞术观察该纳米粒体外寻靶情况,CCK-8试剂检测卵巢癌细胞存活率。结果 制备的Ab-PTX-NPs粒径为（397.70±99.95）nm,紫杉醇包封率及载药量分别为（67.26±4.15）%和（6.31±0.39）%。抗CEA单克隆抗体与纳米粒连接率为（92.74±5.75）%。共聚焦显微镜下观察到靶向组卵巢癌SKOV3细胞周围见较多造影剂黏附,流式细胞术测得靶向组细胞平均荧光强度明显高于其余各组（P<0.05）,CCK-8试剂法测得靶向组在24 h时,细胞存活率高于纯药组（P<0.05）,而低于无靶组（P<0.05）,当48 h时,靶向组与纯药组细胞存活率差异无统计学意义（P>0.05）。结论 成功制备靶向CEA载药相变型PLGA纳米粒,该纳米粒经低功率聚焦超声治疗仪（LIFU）致相变后可明显增强超声显影,且载药量高,释药快,靶向能力强。     

超声介导正己酰羧甲基壳聚糖载药纳米液滴对卵巢癌细胞的抑制作用

目的 评价采用正己酰羧甲基壳聚糖制备相变型载阿霉素（DOX）纳米液滴的体外超声显像效果及抑制卵巢癌细胞生长的作用。方法 将羧甲基壳聚糖与正己酸酐发生酰化反应合成正己酰羧甲基壳聚糖,采用声振乳化法制备正己酰羧甲基壳聚糖载药纳米液滴,表征其表面形态、粒径、电位;评价其体外超声显像效果,以紫外分光光度计测定药物包封率,采用CCK-8法检测卵巢癌细胞存活率,并进行统计学分析。结果 成功制备正己酰羧甲基壳聚糖载药纳米液滴,镜下为大小均匀的球状物,平均粒径为（458.33±43.50）nm,包封率为（52.06±10.14）%。纳米液滴可增强超声显影效果。超声联合载药纳米液滴组的卵巢癌细胞存活率[（62.54±3.60）%]低于DOX组[（75.55±7.21）%]、载药纳米液滴组[（76.18±4.94）%]和超声组[（89.90±0.83）%],差异有统计学意义（P均<0.05）。结论 超声联合正己酰羧甲基壳聚糖载药纳米液滴可有效抑制卵巢癌细胞生长,在临床诊疗中具有潜在应用价值。     

载10-羟基喜树碱液态氟碳纳米粒制备及其表征、体外增强超声显像效果观察

目的 研制一种包裹液态氟碳（PFP）的新型载药纳米级超声造影剂,考察其基本特性并观察其体外增强超声显像效果。方法 采用旋转蒸发和探头超声法制备载10-羟基喜树碱（10-HCPT）的液态氟碳（PFP）纳米粒,在光学显微镜和透射电镜下观察纳米粒形态,采用马尔文仪测量纳米粒粒径和电荷。采用紫外分光光度计测定纳米粒包封率,并应用低强度聚焦超声（LIFU）辐照载药液态氟碳纳米粒溶液,观察纳米粒相变情况及其增强超声显像效果。结果 制备的载药脂质纳米粒外观为乳白色混悬液,在油镜及透射电镜下观察,载药纳米粒形态规则,分布均一,平均粒径约（500.82±25.97）nm,表面平均电位为（-47.77±3.09）mV;在体外经LIFU辐照后,可见纳米粒发生液气相变形成微泡,在基波模式和谐波模式下,均可显著增强超声显影。结论 成功研制了包裹液态氟碳的载药脂质纳米粒,可望成为一种新型的多功能超声造影剂。     

载单宁酸铁和紫杉醇相变型纳米粒用于体外超声显像及治疗视网膜母细胞瘤

目的 制备可用于早期诊断和治疗视网膜母细胞瘤（Rb）的纳米粒，体外评价其超声显像和治疗效果。方法 制备载单宁酸铁（Fe^ⅢTA）和紫杉醇（PTX）并包裹全氟戊烷（PFP）的聚乳酸-羟基乙酸共聚物（PLGA）纳米粒，检测其粒径、电位、电镜、吸光度、包封率、载药量等相关表征，评估808激光（1 W/cm²，5 min）辐照前后温度变化和超声信号改变，光镜下观察纳米粒相变情况。体外检测Fe^ⅢTA/PLGA/PFP纳米粒的生物安全性。设立空白对照组、激光组、Fe^ⅢTA/PLGA/PTX/PFP（FPTP）组、Fe^ⅢTA/PLGA/PTX/PFP+激光（FPTP+激光）组及不同浓度（0、0.250、0.500、1.000 g/L）FPTP+激光组，检测激光辐照5 min后各纳米粒组对肿瘤细胞的杀伤效果。结果 成功制备出Fe^ⅢTA/PLGA/PTX/PFP纳米粒，粒径（155.8±55.68）nm，电位（-27.3±5.14）mV，透射电子显微镜下见黑色Fe^ⅢTA外壳包裹核心灰白球形结构，PTX包封率（70.89±8.03）%，载药率（9.61±0.63）%。体外激光辐照FPTP纳米粒后温度上升与浓度呈正相关，B模式和造影模式超声信号也随浓度增加而增强。FPTP+激光组对Y79细胞的杀伤作用比空白对照组、激光组、FPTP组更强，并随浓度增加而增强。结论 成功制备出载单宁酸铁和紫杉醇的相变型纳米粒，具有良好的光热效应和超声显像效果，对Y79细胞有强杀伤作用，有望用于Rb实现诊疗一体。     

"适配子-PLGA纳米粒"靶向超声/光声相变造影剂的制备

目的 研制一种适配子偶联的包裹纳米金棒和液态氟碳的PLGA纳米粒。方法 采用双乳化法制备包裹纳米金棒及全氟己烷(PFH)的PLGA纳米粒(GNP);用碳二亚胺法将适配子与PLGA纳米粒连接获得"适配子-PLGA纳米粒"靶向相变造影剂(AP-GNP)。光镜、激光共聚焦显微镜及电镜下观察及检测AP-GNP基本性质;激光共聚焦显微镜下及流式细胞术观察及检测适配子与纳米粒连接情况;以激光仪辐照AP-GNP观察其光致相变情况;观察AP-GNP与乳腺癌细胞MCF-7特异性结合情况。结果 成功制备适配子修饰的包裹纳米金棒和PFH的PLGA纳米粒, 其平均粒径为(472.43±25.82)nm, 适配子与纳米粒的连接率为达97.98%;激光辐照AP-GNP后, 光致相变效果明显;AP-GNP粘附于MCF-7细胞周围, 未修饰适配子GNP未见明显特异性结合。结论 成功制备偶联的包裹纳米金棒和PFH的PLGA纳米粒, 其光致相变效果明显, 靶向性能良好。     

载印度墨水相变型光声/超声双模态造影剂的制备及体外显影

目的 制备一种新型的光声/超声双模态造影剂,观察其体外光致相变作用以及光声/超声显影效果。方法 采用多步乳化法 合成载印度墨水及全氟己烷(PFH)的高分子微球(i-PFH-PLGA),检测其基本理化性质、光致相变作用及体外增强光声及超声成像的显像效果。结果 制备的i-PFH-PLGA呈球型,平均粒径为(542.1±68.91)nm。经激光辐照后,显微镜下可观察到较多的i-PFH-PLGA纳米粒发生液气相转变产生微泡;体外光声成像实验显示,i-PFH-PLGA纳米粒可检测到明显的光声信号,且光声信号的强度随纳米粒浓度的增加而增强;体外超声成像实验显示,经激光照射后,i-PFH-PLGA纳米粒组回声强度较辐照前明显增强(P<0.05),而单纯的液态氟碳和印度墨水辐照前后回声强度未见明显改变。结论 成功制备出的载印度墨水和液态氟碳的双模态造影剂可用于体外光声/超声成像研究。     

基于类芬顿反应构建磁性纳米粒治疗卵巢癌:体外实验

目的 构建具有类芬顿反应性能的磁性靶向纳米粒,评价其体外光声成像效果及联合催化反应下抑制卵巢癌细胞的作用。方法 以双乳化法制备包裹四氧化三铁（Fe₃O₄）及葡萄糖氧化酶（GOx）的磁性聚乳酸羟基乙酸共聚物（PLGA）纳米粒Fe₃O₄@PLGA-GOx,观察其基本特性、体外稳定性及释药效果;采用酶标仪监测其联合催化活性,以共聚焦显微镜观察细胞摄取纳米粒能力;以荧光显微镜和流式细胞术观察纳米粒在联合催化作用下产生细胞毒性活性氧的能力;用细胞计数试剂盒（CCK-8）检测磁性Fe₃O₄@PLGA-GOx体外杀伤人卵巢癌SKOV3细胞的能力,并评价其体外光声成像效果。结果 成功制备出磁性纳米粒Fe₃O₄@PLGA-GOx,平均粒径（290.81±108.52）nm,表面电位（－18.41±5.90）mV;Fe₃O₄及GOx的包封率分别为（91.34±2.35）%和（26.53±6.72）%。外部磁场作用下,细胞对纳米粒摄取增加;随纳米粒浓度增加,光声信号增强。Fe₃O₄@PLGA-GOx可介导类芬顿反应,产生活性氧并发挥细胞毒性作用杀伤肿瘤细胞。结论 成功制备的具有类芬顿反应性能磁性纳米粒在光声成像引导下具有体外催化治疗肿瘤细胞作用。     

声动力学治疗对MDA-MB-231细胞增殖及侵袭能力的影响

目的 探讨超声辐照血卟啉单甲醚和声诺维对乳腺癌细胞株MDA-MB-231增殖及侵袭能力的影响。方法 将对数生长期的MDA-MB-231细胞制成细胞悬液,加入HMME和SonoVue后用超声进行辐照。分3组：对照组,不接受治疗;辐照组1：输出声强0.3 W/cm²,辐照时间60 s;辐照组2：输出声强0.6 W/cm²,辐照时间30 s。采用MTS法绘制细胞生长曲线,平板克隆方法观察细胞形成克隆的能力,并用Transwell小室法检测细胞侵袭能力。结果 对照组的生长曲线陡直,而辐照组的曲线平缓。接种2周后对照组出现肉眼可见的细胞克隆,而辐照组未出现。结晶紫染色后观察,辐照组仅可见由少许细胞聚集成的细胞团。接种后24 h、48 h观察,对照组、辐照组1、辐照组2穿越小室的细胞数分别为42.00±13.52、3.52±1.26、2.75±0.96（P<0.05）及104.25±12.61、36.25±3.40、28.75±4.99（P<0.05）。对照组穿越小室的细胞数显著高于辐照组（P<0.05）。结论 超声辐照联合HMME及SonoVue能有效抑制MB-231细胞增殖,降低细胞形成克隆的能力,并显著降低细胞的侵袭能力。     

心肌特异性导向肽靶向诊疗造影剂增强心肌分子显影及其在大鼠体内的分布

目的 制备心肌特异性靶向肽（PCM）修饰的载药纳米诊疗探针,探讨其心肌靶向显影能力及其在大鼠体内的分布情况。方法 采用双乳化法及碳二亚胺法制备PCM靶向载卡维地洛纳米诊疗探针（PCM/C-PFPs）,检测其基本性质、大鼠体内显影及体内分布情况。结果 PCM/C-PFPs纳米探针呈球形,平均粒径为（415.00±6.24） nm,电位为（-20.27±1.55） mV;包封率为（78.23±3.45）%;C-PFPs与多肽PCM连接率为（98.98±1.02）%。加热至约60℃时,该探针发生明显相变,并产生大量微气泡。声诺维组和PCM/C-PFPs+低频聚焦超声（LIFU）组大鼠心脏超声显影效果明显,但声诺维组显影仅维持数分钟,而PCM/C-PFPs+LIFU组心脏超声信号可持续至1 h,且心脏显影的声强度高于其他各组（P均<0.05）。PCM/C-PFPs+LIFU组大鼠心脏切片中可见大量聚集的红色探针,明显多于C-PFPs+LIFU组和单纯PCM/C-PFPs组,且多于该组其他组织（肝、肾、肺、脾）。结论 本研究成功制备的心肌靶向诊疗造影剂,具有良好的靶向心脏超声显影和大鼠体内分布特性。     

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.     

Interprofessional approaches to creating safe,high quality health care

Three supplementary perspectives are presented arguing that interprofessional collaboration is both necessary and desirable. Nonetheless, there are often too many serious intra-professional barriers and obstacles to interprofessional collaboration to make it successful. Some of these barriers, it is argued and illustrated, are found in the multiple ways in which professional identity is tacitly acquired and embodied in the practitioners' habitual, everyday practice. The paper then explores ways in which reflection, especially Second order reflection, can help to elucidate and overcome these obstacles, as well as increasing professional adaptability and competence.     

Systematic Reviews Published in the January 2011 Issue of The Cochrane Library

ABSTRACT

The Cochrane Library of Systematic Reviews is published quarterly as a DVD and monthly online. The January 2011 issue (first quarterly DVD for 2011) contains 4515 complete reviews, 1985 protocols for reviews in production, and 13,521 one-page summaries of systematic reviews published in the general medical literature. In addition, there are citations of 641,000 randomized controlled trials, and 14,018 cited papers in the Cochrane methodology register. The health technology assessment database contains over 9300 citations. One hundred and seven new reviews have been published in the last 3 months, of which five have potential relevance for practitioners in pain and palliative medicine.     

Prise en charge des entorses externes de cheville : étude clinique préliminaire sur l’efficacité du dispositif Myolux™

Ankle sprains are the most common injury of the musculoskeletal system and are associated with significant societal and economic impacts. It has been proven that classical therapeutic strategies may not be effective in preventing recurrent injuries: the recurrence rates reported in the literature can reach 73%. In order to provide an effective rehabilitation solution, a destabilizing orthosis was developed. This device is equipped with a mechanical articulator reproducing the subtalar mechanics and placed under the heel. In this paper, we present the main results of a preliminary clinical study conducted between 2004 and 2007. All subjects included in this study were treated with the abovementioned orthosis during 10 rehabilitation sessions of 30 minutes each. Data show a relatively low recurrence rate of 12% for the overall population. Moreover, it's of primary importance to note that this satisfactory ratio is largely reduced (3% of recurrence rate) for the 29 patients who performed one training session per month after the 10th initial rehabilitation sessions. Hence, the destabilizing orthosis appears to be an effective solution to prevent recurrent ankle sprains. However, joint protection requires long-term and regular training sessions. This result has motivated the development of a similar device allowing patients to perform training sessions at home. Finally, data obtained in this study are promising awaiting the final results of the comparative, multicentric and independent clinical trials currently managed by the Hospices Civils de Lyon.     

Clinical Trial Updates and Hotline Sessions presented at the European Society of Cardiology Congress 2007

This article provides information and commentaries on trials which were presented at the Hotline and Clinical Trial Update Sessions at the European Society of Cardiology Congress 2007 in Vienna. The key presentations were performed by leading experts in the field with relevant positions in the trials or registries. It is important to note that unpublished reports should be considered as preliminary data, as the analysis may change in the final publications. The comprehensive summaries have been generated from the oral presentation and the webcasts of the European Society of Cardiology and should provide the readers with the most comprehensive information of relevant publications.     

Prospects for the control of neglected tropical diseases by mass drug administration

The prospects for the control of neglected tropical diseases, including soil-transmitted helminthiasis, shistosomiasis, lymphatic filariasis, onchocerciasis and trachoma, through mass drug administration, are exemplified by the elimination of the trachoma as a public-health problem in Morocco. In spite of this and other striking successes, mass drug administration programs are faced with major challenges resulting from suboptimal coverage and lack of efficacy. At current suboptimal coverage rates, programs may need prolongation for an extended period, increasing costs and undermining sustainability. Community participation through health education and information appears to be crucial to improve coverage and to achieve sustainability. Implementation of complementary measures, such as vector control, improved hygiene and environmental sanitation, are important to further control transmission and to prevent re-emergence of the infection and, again, may only be achieved effectively through community-based initiatives. To reduce costs and to relieve pressure on the health system, combining neglected tropical disease programs in areas where diseases coexist and integration with existing control programs for malaria, tuberculosis and HIV/AIDS is advocated. The risk of developing drug resistance is of particular concern in view of the lack of alternative drugs, and reduced treatment efficacy due to emerging resistance is evident for the soil-transmitted helminths and onchocerciasis. Given the risk for the development of drug resistance and the need for a high degree of participation, close attention should be paid to the monitoring of the coverage and efficacy of the different program components.     

Balance and Boundaries in Grief Counseling: An Intervention Framework for Volunteer Training

Volunteers or paraprofessional counselors are commonly used to provide supportive care to the bereaved. These counselors generally are trained in basic listening skills, providing a generic, nonspecific approach to intervention that remains to be proven effective. The present paper outlines a framework that provides paraprofessionals with a broader model for intervention with the bereaved. Attention to boundaries as a helper and balance in the grief recovery are central to the model. Interventions are described that provide the paraprofessional counselor with more options for tailoring their counseling strategy to the individual. These include techniques that are presumed to be more specific to the enhancement of grief recovery.     

A new method for the simultaneous analysis of unmodified and modified urinary nucleosides and nucleobases by high performance liquid chromatography

Details are given of a new, rapid and simple pre-fractionation method and an isocratic high performance liquid chromatography system suitable for parallel analysis of nucleosides and nucleobases from urine and other biological fluids. The quantitative recovery and excellent reproducibility of the method is demonstrated by analysis of representative standard RNA catabolites. The advantage of this new method for application to biological samples is discussed.     

Bacterial meningitis: an update of new treatment options

The outcome of bacterial meningitis critically depends on the rapid initiation of bactericidal antibiotic therapy and adequate management of septic shock. In community-acquired meningitis, the choice of an optimum initial empirical antibiotic regimen depends on the regional resistance patterns. Pathogens resistant to antibacterials prevail in nosocomial bacterial meningitis. Dexamethasone is recommended as adjunctive therapy for community-acquired meningitis in developed countries. In comatose patients, aggressive measures to lower intracranial pressure <20 mmHg (in particular, external ventriculostomy, osmotherapy and temporary hyperventilation) were effective in a case–control study. Although many experimental approaches were protective in animal models, none of them has been proven effective in patients. Antibiotics, which are bactericidal but do not lyse bacteria, and inhibitors of matrix metalloproteinases or complement factor C5 appear the most promising therapeutic options. At present, vaccination is the most efficient method to reduce disease burden. Palmitoylethanolamide appears promising to enhance the resistance of the brain to infections.