从睾丸cDNA表达文库筛选胰腺癌肿瘤抗原

目的利用胰腺癌患者血清筛选睾丸cDNA噬菌体表达文库，寻找胰腺癌特异性抗原，尤其是肿瘤-睾丸抗原。方法采用血清学筛选重组cDNA表达文库（SEREX）技术筛选睾丸cDNA噬菌体表达文库。对获得的阳性克隆片段进行测序、鉴定和同源性比较。检测40例胰腺导管腺癌患者和40名健康对照血清中抗阳性克隆的抗体。结果筛选共获得107个阳性克隆，代表14个不同抗原基因，其中有13个基因与基因库中已知基因有很高的同源性，这些基因为activin受体AⅡ、LOC92912、KLHL12、IFI16和CAGE等。另有1个阳性克隆HS1在基因库和SEREX数据库中均未发现有同源基因。胰腺癌患者组HS1和HS2克隆抗体阳性率显著高于健康对照组（分别15％与0％比较,Χ^2=4．50，P〈0．05；22．5％与5％比较，Χ^2=5．16，P〈0．05）。结论HS1可能是1个新的肿瘤一睾丸抗原基因。有必要进一步研究这些阳性克隆的生物学功能和临床价值。     

鼻咽癌相关抗原的血清学筛选与鉴定

目的有关鼻咽癌肿瘤抗原的研究还仍然集中于研究EB病毒与鼻咽癌的相互关系上,而有关鼻咽癌细胞本身的肿瘤抗原的研究到目前为止还几乎是空白.采用SEREX技术筛选与鉴定鼻咽癌相关肿瘤抗原.方法从鼻咽癌肿瘤组织中提取mRNA;构建鼻咽癌cDNA表达文库;*"利用SEREX技术筛选鼻咽癌cDNA表达文库;对阳性克隆进行测序、生物信息学分析.结果①构建的鼻咽癌cDNA表达文库大小为1.9×108个重组子.②筛选出10个阳性克隆.③测序:有8个阳性结果.④生物信息学分析:除了一些已知的抗原如MAGE外,发现一个热休克蛋白基因家族的新成员,其在染色体水平上与HSPCB有84%的同源性,在氨基酸水平上有97%的同源性.结论结果显示热休克蛋白新成员可能为鼻咽癌的一个新的肿瘤标记物和鼻咽癌疫苗的候选靶分子.     

人蜕膜组织cDNA表达文库的构建及免疫血清筛选

目的：构建人蜕膜组织的cDNA表达文库,筛选蜕膜组织中与胚胎发生有关的抗原。方法：首先取人流后的蜕膜组织,提取mRNA,构建人蜕膜组织的cDNA表达文库;其次用人蜕膜组织免疫小鼠后获得的异种免疫血清对此表达文库进行血清学筛选,获得阳性克隆,最后对阳性克隆进行测序。结果：获得了人蜕膜组织cDNA表达文库,并对其进行初步筛选,目前暂未获得阳性克隆。结论：本实验构建了人蜕膜组织cDNA表达文库．并对其进行了初步的免疫血清筛选、为今后进一步筛选文库、寻找蜕膜组织中与胚胎发生有关的基因奠定了基础。     

多发性骨髓瘤相关抗原的免疫学筛选及生物信息学分析

本研究采用“cDNA表达文库的血清学分析（ serological analysis of cDNA expression library, SEREX ）”技术筛选骨髓瘤HMy2细胞cDNA表达文库。将得到的30个阳性克隆全部进行测序,并进行BLAST同源序列比对分析。结果表明：获得已知基因6条,骨髓瘤相关肿瘤抗原新基因12条。经部分序列EST拼接,已知基因6条如环指蛋白167、KLF10因子、TPT1蛋白、p02蛋白、cDNA FLJ46859 fis、DNMT1甲基转移酶等,它们与其他肿瘤的形成、发展与预后有一定的关系。利用生物信息学对新基因结构和功能初步分析和预测显示,其中MMSA-3、MMSA-8和MMSA-11有完整的编码区,编码的蛋白长度分剐为215、160和122个氨基酸;除MMSA-1可能定位于性染色体,其余均可能定位于常染色体;MMSA-4和控制转录的肿瘤蛋白高度相似,MMSA-5可能参与细胞的吞噬作用,MMSA-7可能使NF-κB失活,MMSA-12可能为淋巴细胞的胞质蛋白。CrELISA初步分析表明,MMSA-3和MMSA-7等新基因特异性较高。结论：本项研究所筛选和鉴定的肿瘤抗原可用于多发性骨髓瘤的早期免疫学诊断、残留病灶监测、判断预后及肿瘤疫苗制备等多种研究。     

非洲爪蟾肝脏肿瘤cDNA表达文库异种同源抗原的筛选

目的:通过重组cDNA文库的血清学筛选(SEREX)技术获得来自非洲爪蟾(Xenopus laevis)肝脏肿瘤文库中具有潜在治疗价值的异种同源抗原的cDNA序列.方法:用人肝癌细胞株SMMC-7721免疫家兔后收集的抗血清作为筛选血清,采用SEREX技术筛选Xenopus laevis肝脏肿瘤cDNA表达文库.结果:通过两轮筛选获得19个阳性克隆,通过DNA序列测定和分子生物学技术确定这19个克隆编码16个蛋白.其中Transgelin 2、RAS guanyl releasing protein 1和Chromobox protein homolog 6可能与肝细胞肝癌有关.结论:用异种免疫血清筛选异种同源肿瘤抗原拓宽了SEREX 技术的应用范围,鉴定Transgelin 2、RAS guanyl releasing protein 1和Chromobox protein homolog 6可能为异种同源抗原.     

与人体肥胖相关的神经肽Y基因克隆及序列分析

目的 克隆与肥胖相关的人神经肽Y(NPY)基因,并鉴定该克隆基因序列的正确性.方法 根据GeneBank中收录的NPY基因序列,设计该基因上下游序列,经过PCR反应合成NPY的cDNA.然后与pET28a+载体重组,筛选阳性克隆和DNA序列分析鉴定,测序后与GeneBank中NPY基因进行同源性比较和序列分析.结果 PCR扩增出一个100 bp左右的DNA片段,与载体重组后和DNA序列分析鉴定,DNA序列分析显示克隆的DNA片段是人NPY基因.且所克隆的基因共编码36个氨基酸,分子量为4.2 kD,与GeneBank中NPY基因序列同源性达100%.结论 克隆的人NPY基因与Genebank中NPY序列完全一致,为进一步应用分子生物学技术深人研究NPY与人体肥胖发生、发展及转化等相关作用机制及应用该基因进行其基因蛋白表达奠定了基础.     

与人体肥胖相关的神经肽Y受体Y1基因的克隆及序列分析

目的 克隆与肥胖相关的人神经肽Y受体Y1(NPYlR)基因,并鉴定该克隆基因序列的正确性.方法 从人的脂肪组织中提取总RNA并进行反转录,以此为模板用PCR扩增NPY1R基因的cDNA,然后与pET 28a+载体重组,筛选阳性克隆和DNA序列分析鉴定,测序后与GeneBank中NPY1R基因进行同源性比较和序列分析.结果 PER扩增出一个1100 bp左右的DNA片段,与载体重组后DNA序列分析鉴定,DNA序列分析显示克隆的DNA片段是人NPY1R基因,且所克隆的基因共编码384个氨基酸,分子量为44 KD,与GeneBank中NPY1R基因序列同源性达100%.结论 克隆的人NPY1R基因与GeneBank中NPY1R序列完全一致,为进一步应用分子生物学技术深入研究NPY1R与人体肥胖发生、发展及转化等相关作用机制及应用该基因进行其基因蛋白表达奠定了基础.     

一个具有诊断价值的弓形虫基因的发现

目的用弓形虫成囊株感染的小鼠血清筛选弓形虫RH株速殖子cDNA文库,以寻找诊断弓形虫病的抗原基因。方法收集成囊株(Prugniaud株)慢性感染的小鼠血清,经滤膜吸附法去除大肠杆菌抗体,筛选弓形虫RH株速殖子cDNA文库。对3次复筛得到的阳性克隆进行插入片段的核苷酸序列测定,结果送GenBank进行同源性分析。根据序列测定结果,选取其中1个具有完整开放阅读框架的基因进行分析。结果获得4个持续阳性反应克隆。测序和同源性分析显示,WT1为弓形虫P30基因;WT4为弓形虫N-乙酰磷酸丙糖转移酶基因;WT9与已知的基因无同源性。WT12克隆基因与一未知的弓形虫基因有很高的同源性,且具有1个513bp大小的完整开放阅读框架,其蛋白质结构和功能预测可能是1个跨膜信号蛋白。结论筛选得到的1个阳性克隆有望成为早期诊断抗原基因,值得进一步研究。     

与人体肥胖相关的神经肽Y受体Y5基因的克隆及序列分析

目的 克隆与肥胖相关的人神经肽Y受体Y5(NPY5R)基因,并鉴定该克隆基因序列的正确性.方法 从人的脂肪组织中提取总RNA并进行反转录,以此为模板用PCR扩增NPY5R基因的cDNA,然后与pET28a+载体重组,筛选阳性克隆和DNA序列分析鉴定,测序后与CeneBank中NPY5R基因进行同源性比较和序列分析.结果 PCR扩增出-个1 300 bp左右的DNA片段,与载体重组和DNA序列分析鉴定,DNA序列分析显示克隆的DNA片段是人NPY5R基因,且所克隆的基因共编码445个氨基酸,分子量为50KD,与CeneBank中NPY5R基因序列同源性达100%.结论 克隆的人NPY5R基因与CeneBank中NPY5R序列完全一致,通过对其相关生物学信息的明确分析,为进一步应用分子生物学技术深入研究NPY5R与人体肥胖发生、发展及转化等相关作用机制及应用该基因进行其基因蛋白表达奠定了基础.     

日本血吸虫微管蛋白基因的获得和分析

目的从日本血吸虫(schistosoma japonicum，Sj)成虫cDNA文库中获得并分析日本血吸虫新的表达基因，为日本血吸虫病的防治提供药物靶标或侯选疫苗。方法构建日本血吸虫成虫cDNA文库，随机挑取重组阳性克隆进行测序，对部分序列进行步移法测序获取全长cDNA，并进行生物信息学分析和登录。结果获得了1个日本血吸虫新基因，全长1439bp，编码443个氨基酸，与肝片形吸虫微管蛋白基因具有79％的同源性。结论表达序列标签法、步移法测序和生物信息学技术三者相结合，有利于发现日本血吸虫新基因。     

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.     

Structure and function of "metalloantibiotics"

Although most antibiotics do not need metal ions for their biological activities, there are a number of antibiotics that require metal ions to function properly, such as bleomycin (BLM), streptonigrin (SN), and bacitracin. The coordinated metal ions in these antibiotics play an important role in maintaining proper structure and/or function of these antibiotics. Removal of the metal ions from these antibiotics can cause changes in structure and/or function of these antibiotics. Similar to the case of "metalloproteins," these antibiotics are dubbed "metalloantibiotics" which are the title subjects of this review. Metalloantibiotics can interact with several different kinds of biomolecules, including DNA, RNA, proteins, receptors, and lipids, rendering their unique and specific bioactivities. In addition to the microbial-originated metalloantibiotics, many metalloantibiotic derivatives and metal complexes of synthetic ligands also show antibacterial, antiviral, and anti-neoplastic activities which are also briefly discussed to provide a broad sense of the term "metalloantibiotics."