视神经组织工程重建中基因治疗工具的作用

背景:目前治疗视神经疾病的方法主要有细胞移植治疗,基因治疗,药物治疗,手术干预等.其中,基因治疗研究逐步受到重视,并有望成为某些视神经疾病的可供选择的治疗方法之一.目的:从各种基因载体应用、神经营养因子、组织工程修复以及临床应用等方面,全面了解视神经损伤后基因治疗修复的方法.方法:以"视神经修复(optic nerve regeneration),基因治疗(gene therapy),组织工程(tissue-engineering)"为检索词,应用计算机检索维普数据库,Pubmed数据库,Elsevier数据库相关文章.纳入与视神经损伤后基因治疗修复密切相关的文献,排除重复性研究.结果与结论:共检索到268篇文献,排除无关重复的文献,保留29篇文献进行综述.目前视神经损伤后,越来越多的研究采用非病毒及病毒载体从基因角度感染视网膜节细胞,其中病毒载体包括慢病毒载体,腺病毒载体,腺相关病毒载体,当前大部分研究是采用重组的腺相关病毒载体.通过这些基因载体工具的应用,能增强视网膜节细胞的存活,促进受损视神经轴突的再生.     

腺病毒介导的骨形态发生蛋白基因增强在骨组织工程中的应用

背景:将目的基因转导到靶细胞中需要载体工具,载体可协助基因进入细胞及表达蛋白,载体的选择是转基因成败的关键。目的:总结腺病毒介导的骨形态发生蛋白基因增强用于诱导成骨和修复骨缺损的研究现状。方法:应用计算机检索PubMed数据库及中国期刊网全文数据库1989-01/2012-01有关腺病毒介导骨形态发生蛋白基因转染的诱导成骨作用,骨形态发生蛋白基因修饰的组织工程化骨修复骨缺损的文章,英文检索词为"adenovirus,bone morphogenetic protein,gene transfection,bone tissue engineering",中文检索词为"腺病毒,骨形态发生蛋白,基因转染,骨组织工程"。排除重复性及非中英文语种研究,共保留24篇文献进行综述。结果与结论:腺病毒介导的骨形态发生蛋白基因增强的组织工程利用基因工程技术将编码特定功能因子的基因转移到种子细胞上,使其持续产生生长因子。转基因后的细胞停留支架材料表面缓慢释放生长因子,促进成骨前体细胞增殖分化,也可在骨缺损处诱导成骨前体细胞向成骨细胞定向分化,从而修复骨缺损。     

帕金森病基因治疗的研究与应用

目的:随着基因技术的发展和对帕金森病发病机制研究的深入,帕金森病被认为是中枢神经系统疾病中最适合基因治疗的疾病之一,总结基因治疗帕金森病的主要策略的应用研究。资料来源:应用计算机检索Medline1997-01/2004-08与帕金森病的基因治疗相关文献,检索词“Parkinsondisease,genetherapy,stemcells”,并限定文献语种为英文。资料选择:共收集到93篇相关文献,对这些文献进行筛选,选择帕金森病基因治疗的实验性文章,其中相似的研究以近3年的文章优先。资料提炼:在72篇帕金森病基因治疗相关文献中排除相似的研究,选择18篇文献,在21篇有关干细胞应用于帕金森病治疗的文献中选择1篇与基因工程联合应用有关的文献。资料综合:将筛选到的19篇文献按导入编码多巴胺合成过程中所需的酶类基因(5篇),导入抗凋亡蛋白基因及编码神经营养因子的基因(9篇),调整基底核环路的功能活性(4篇),以及与干细胞联合应用(1篇)来归类。结论:介导基因递送的载体中以重组的病毒载体为主,导入上述相关基因对帕金森动物模型行为均有改善,多基因联合治疗及与干细胞的联合治疗是帕金森病的基因治疗的趋势。     

种子细胞在颌骨缺损修复中的研究与应用

背景：颌骨缺损时形态的修复不仅要考虑形态也要考虑咬合关系能否建立。目的：对近年来骨组织工程中种子细胞应用于颌骨缺损修复治疗的研究现状做一综述。方法：以Bone tissue engineering,seeded cells,Osteogenic Differentiation为英文检索词,以骨组织工程,种子细胞,成骨分化,骨缺损修复为中文检索词,检索1994年1月至2011年4月PubMed数据库和CNKI数据库相关文献。阅读标题和摘要进行初筛,排除因研究目的与此文无关及内容重复性文献,共保留58篇文献进行综述。结果与结论：综合分析各种种子细胞应用于骨组织工程的研究现状,认为骨髓间充质干细胞研究最多,而脐血间充质干细胞和脐带间充质干细胞的研究因其能克服骨髓间充质干细胞应用的诸多缺点故也较为热门。胚胎干细胞存在伦理学上的困境,若能够有所突破,仍有望成为重要的颌骨缺损时骨组织工程的种子细胞之一。基因修饰的种子细胞能增强骨组织工程种子细胞的增殖能力和定向成骨分化的能力。但基因工程技术仍有待提高,其安全性和有效性仍有待商榷。     

慢病毒载体的构建及优化

目的:针对慢病毒载体的基因组装、转导及基因表达的优化论述,及不同种属的慢病毒载体的最低活性加以说明,同时对慢病毒遗传体系的产生、转导效率和生物安全性进行总结。资料来源:应用计算机检索Pubmed以及美国SCI和Medline数据库2000-01/2004-12有关慢病毒载体的论文,检索词“lentivirusvectors,immunodeficiency”,并限定文章语言种类为English。同时应用计算机检索中国期刊网数据库1995-01/2005-12有关慢病毒载体构建的文章,限定文章语言种类为汉语,检索词“慢病毒载体”。资料选择:对检索到的相关信息及文章进行整理,选取针对性强的文章。纳入标准:①临床研究性文章。②基础研究文章。排除标准:①其他病毒载体的结构文献以及重复性资料。②诸多病毒实验应用性文献。③重复同一研究。资料提炼:共收集到198篇关于慢病毒结构原理的相关文献,其中包括英文文献150篇,中文文献48篇,有21篇符合纳入标准。资料综合:介绍不同源慢病毒载体的构建和慢病毒的基因位点。影响基因转移的一个主要因素是细胞的基因表达。人类转基因载体的一个重要特点是可以调节转基因的表达。调节转基因信号的另一个途径是运用可切割的前病毒产物。转基因载体是建立在逆转录病毒之上,主要包括肿瘤逆转录病毒和慢病毒,这些病毒载体系统是哺乳动物细胞外源基因传递、整合和表达的有效途径。结论:蛋白酶、rev.慢病毒蛋白和VSV/G糖蛋白均具有细胞毒性和抑制细胞的作用,当连续表达时要求使用诱导表达系统。     

帕金森病基因治疗的研究与应用

目的：随着基因技术的发展和对帕金森病发病机制研究的深入，帕金森病被认为是中枢神经系统疾病中最适合基因治疗的疾病之一，总结基因治疗帕金森病的主要策略的应用研究。 资料来源：应用计算机检索Medline1997-01／2004-08与帕金森病的基因治疗相关文献，检索词“Parkinson disease，genetherapy．stemcells”，并限定文献语种为英文。 资料选择：共收集到93篇相关文献，对这些文献进行筛选，选择帕金森病基因治疗的实验性文章，其中相似的研究以近3年的文章优先。 资料提炼：在72篇帕金森病基因治疗相关文献中排除相似的研究，选择18篇文献，在21篇有关干细胞应用于帕金森病治疗的文献中选择1篇与基因工程联合应用有关的文献。 资料综合：将筛选到的19篇文献按导入编码多巴胺合成过程中所需的酶类基因（5篇），导入抗凋亡蛋白基因及编码神经营养因子的基因（9篇），调整基底核环路的功能活性（4篇），以及与干细胞联合应用（1篇）来归类。 结论：介导基因递送的载体中以重组的病毒载体为主，导入上述相关基因对帕金森动物模型行为均有改善，多基因联合治疗及与干细胞的联合治疗是帕金森病的基因治疗的趋势。     

骨组织工程中的细胞外基质材料及其种子细胞

背景:目前,骨组织工程在骨再造及骨缺损修复领域方面发展迅速,其中细胞外基质材料和种子细胞的选择应用是决定骨组织工程成功的前提和基础.目的:对骨组织工程中的细胞外基质材料和种子细胞的研究现状进行综述,并分析了这些细胞外基质材料和种子细胞的优缺点及今后尚待解决的问题和发展趋势.方法:应用计算机检索Pubmed数据库和CNKI数据库中1994-01/2009-12关于骨组织工程的文章,在标题中和摘要中以"骨组织工程,种子细胞,细胞外基质材料"或"bone tissue engineering,seeded cell,extral cellular matrix materials"为检索词进行检索.选择文章内容与骨组织工程中的种子细胞和细胞外基质材料有关者,同一领域文献则选择近期发表或发表在权威杂志文章.初检得到246篇文献,根据纳入标准选择相关的30篇文献进行综述.结果与结论:细胞外基质材料尚不够完善,还存在这样或那样的缺陷.如基质材料的降解率与成骨速度不协调;基质材料在体内所引发的免疫反应等;种子细胞的来源及培养体系尚未完全确定;种子细胞与细胞外基质材料的相互作用关系,尤其是如何增加种子细胞在基质材料表面的生物活性及分化方向方面,仍需进一步深入研究.     

慢病毒载体的构建及优化

目的：针对慢病毒载体的基因组装、转导及基因表达的优化论述。及不同种属的慢病毒载体的最低活性加以说明，同时对慢病毒遗传体系的产生、转导效率和生物安全性进行总结。资料来源：应用计算机检索Pubmed以及美国ScI和Medline数据库2000-01／2004-12有关慢病毒载体的论文，检索词“lentivirus vectors，immunodeficiency”，并限定文章语言种类为English。同时应用计算机检索中国期刊网数据库1995-01／2005-12有关慢病毒载体构建的文章，限定文章语言种类为汉语，检索词“慢病毒载体”。资料选择：对检索到的相关信息及文章进行整理。选取针对性强的文章。纳入标准：①临床研究性文章。②基础研究文章。排除标准：①其他病毒载体的结构文献以及重复性资料。②诸多病毒实验应用性文献。③重复同一研究。资料提炼：共收集到198篇关于慢病毒结构原理的相关文献，其中包括英文文献150篇，中文文献48篇，有21篇符合纳入标准。资料综合：介绍不同源慢病毒载体的构建和慢病毒的基因位点。影响基因转移的一个主要因素是细胞的基因表达。人类转基因载体的一个重要特点是可以调节转基因的表达。调节转基因信号的另一个途径是运用可切割的前病毒产物。转基因载体是建立在逆转录病毒之上，主要包括肿瘤逆转录病毒和慢病毒，这些病毒载体系统是哺乳动物细胞外源基因传递、整合和表达的有效途径。结论：蛋白酶、rev．慢病毒蛋白和VSV／G糖蛋白均具有细胞毒性和抑制细胞的作用，当连续表达时要求使用诱导表达系统。     

聚合物基因载体的设计与优化

目的:随着基因治疗研究的不断深入,选择更安全和更有效的基因转移载体已成为基因治疗成功的关键。为此本文就聚合物基因载体的设计与优化进行综述。资料来源:应用计算机检索PUBMED1990-01/2005-12期间的相关文章,检索词为“genetherapy,non-viralvectors,cationicpolymer/DNAcomplex”,并限定文章语言种类为English。同时计算机检索万方中国全文期刊数据库1990-01/2005-12期间的相关文章,检索词为“基因治疗,非病毒载体,阳离子聚合物/基因复合物”,并限定文章语言种类为中文。资料选择:对资料进行初审,并查看每篇文献后的引文。纳入标准:文章所述内容应与聚合物基因载体的设计及优化研究相关。排除标准:重复研究或Meta分析类文章。资料提炼:共收集到60篇相关文献,28篇文献符合纳入标准,排除的32篇文献为内容陈旧或重复。符合纳入标准的28篇文献中,26篇涉及基因转移体系,2篇涉及Polyplex转染过程中的问题以及聚合物载体的优化。资料综合:本文首先简要介绍了两类基因转移体系:病毒载体转移体系和非病毒转移体系,以及非病毒转移体系的分类。然后详细介绍了聚合物基因载体的发展现状,列举了几类近年设计的新型阳离子聚合物载体,指出其优点和不足、结构与功能的关系,以及优化的方法。最后介绍了以阳离子聚合物作为基因载体进行细胞转染的机制,并探讨了如何对聚合物基因载体进行优化。结论:在非病毒转移体系中,聚合物基因载体由于其设计灵活及具有很好的生物相容性等优点而在人类基因治疗中具有很大的潜力。然而低转染效率限制了其在临床上的应用。随着对聚合物基因转移机制的进一步理解,聚合物基因转移体系将成为人类基因治疗的有力工具。     

骨组织工程种子细胞的研究近况

目的:分析比较各种种子细胞在骨组织工程中作用,归纳较理想的种子细胞的条件,为骨缺损患者修复与恢复功能选择最适宜的种子细胞提供科学依据。资料来源:应用计算机检索Medline1990-01/2006-01有关骨组织工种、种子细胞、骨缺损修复方面的文献,检索词为“bone,tissueengineering,seedcell,bonedefect,bonerepair”并限定文章语言种类为英文。中文文献应用计算机从中国期刊全文数据库检索。资料选择:对资料进行初审,纳入标准:选取有关骨组织工程种子细胞实验或临床研究文献。排除标准:①非随机对照实验、重复实验。②综述、讲座、笔谈等非原著性文献。资料提炼:共收集了384篇有关骨组织工程种子细胞实验或临床研究方面文献,其中23篇符合纳入标准,排除361篇。对23篇文章中提及的①骨种子细胞:成骨细胞,骨髓基质细胞,间充质干细胞,骨髓间充质干细胞。②骨外种子细胞:骨外间充质干细胞,胚胎干细胞。③转基因干细胞等种子细胞进行综合分析。资料综合:较理想的种子细胞的条件应该是:①取材方便,对供体损伤小。②在体外培养体系中具有较强的增殖传代能力,生物活性好,衰亡率低,能在较短时间内得到数量较多并易定向分化为成骨细胞。③植入机体后能适应受区环境,保持高质量成骨活性,且远期效果良好。④生物毒性低,组织相容性好,免疫活性低,致癌性低等。结论:骨组织工程中对种子细胞的研究已进入临床研究阶段,并取得可喜的进展,其中以间充质干细胞最好,它取材容易,安全性好,是很有发展前景的种子细胞。     

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.     

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.     

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.     

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.     

Modified micro-superior percutaneously-assisted total hip: early experiences & case reports

Because of the extensile nature and familiarity of the standard posterior-lateral approach to the hip, a family of "micro-posterior" approaches has been developed. This family includes the Percutaneously-Assisted Total Hip (PATH) approach, the Supercapsular (SuperCap) approach and a newer hybrid approach, the Supercapsular Percutaneously-Assisted Total Hip (SuperPATH) approach. Such approaches should ideally provide a continuum for the surgeon: from a "micro" (external rotator sparing) posterior approach, to a "mini" (external rotator sacrificing) posterior approach, to a standard posterior approach. This could keep a surgeon within his comfort zone during the learning curve of the procedure, while leaving options for complicated reconstructions for the more practiced micro-posterior surgeons. This paper details one author's experiences utilizing this combined approach, as well as permutations of this entire micro-posterior family of approaches as applied to more complex hip reconstructions.     

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.     

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.