微电场通过非整合素途径活化粘着斑激酶促进滋养细胞迁移/侵袭研究

目的探讨生理性微电场促进体外培养的人胎盘滋养细胞迁移/侵袭功能是否与滋养细胞表面整合素(integrin)表达有关。方法用150 m V/mm的直流微电场刺激滋养细胞,时间分别为5、10和15 h,测定其迁移情况并观察细胞形态变化。Western blot检测刺激前后1 h内粘着斑激酶FAK活化情况和细胞表面integrinα1、integrinα5、integrinαV和integrinα1蛋白表达水平。结果在含有10%胎牛血清的培养基中,150 m V/mm电场刺激下滋养细胞向负极定向迁移,迁移速度和距离较对照组明显增加(P=0.021),胞体拉长,垂直于电场方向排列;胞内FAKTyr397位点于刺激后5、10、30、60 min内迅速活化并逐渐加强(P0.05);刺激前后滋养细胞表面integrinα1、integrinα5、integrinαV、和integrinα1蛋白表达水平无明显改变(P0.05)。结论生理性直流微电场可能通过非整合素途径活化FAK从而促进滋养细胞迁移/侵袭功能,但其详细机制仍需进一步研究。     

蜕膜巨噬细胞调控滋养细胞侵袭力的研究

目的 探讨蜕膜巨噬细胞对滋养细胞侵袭力的调控。方法 取20 ~ 35岁女性正常早孕（6 ~ 7⁺⁶周）行人工流产的蜕膜组织15 ~ 20 g。将蜕膜组织充分清洗后使用Collagenase和Dnase消化2次,每次30 min,细胞悬液过滤后通过CD14⁺免疫磁珠分选,并行免疫荧光鉴定。流式细胞仪分析巨噬细胞M1型（CD80⁺和CD86⁺）和M2型（CD163⁺和CD206⁺）。获得的巨噬细胞置于6孔板培养,24 h后收集细胞上清。巨噬细胞上清与滋养细胞系HTR-8/SVneo共培养24 h,实时定量PCR检测HTR-8/SVneo的水通道蛋白1（AQP 1）mRNA相对表达量变化,以及基质金属蛋白酶-2（MMP-2）和MMP-9 mRNA相对表达量的变化。应用实时无标记细胞分析技术（RTCA）检测人早孕蜕膜巨噬细胞上清对人滋养层细胞系HTR-8/SVneo侵袭力的影响。结果 成功建立人类早孕蜕膜巨噬细胞体外培养体系。流式细胞检测显示,人类正常早孕蜕膜组织中巨噬细胞以M2型为主,约占全部巨噬细胞的87%。巨噬细胞上清使HTR-8/SVneo的AQP 1和MMP-9 mRNA相对表达量升高（P均<0.001）。RTCA结果显示巨噬细胞上清明显增强HTR-8/SVneo的侵袭力（P<0.001）。结论 体外培养条件下,蜕膜巨噬细胞可能通过介导AQP1表达升高而增强滋养细胞的侵袭力。     

微电场对胎盘滋养细胞迁移/侵袭相关信号通路活性的影响

目的探讨滋养细胞迁移/侵袭相关促分裂原活化蛋白激酶/细胞外信号调节蛋白激酶(MAPK/ERK),磷酸肌醇-3激酶/丝氨酸苏氨酸蛋白激酶(PI3K/Akt)和蛋白酪氨酸激酶/信号转导和转录活化因子3(JAK/STAT3)信号传导通路在微电场刺激下的活化水平。方法将胎盘滋养细胞用场强为150mV/mm的直流微电场加以刺激,通过蛋白质印迹法(Western blot)测定电刺激前和后各时间点滋养细胞MAPK/ERK、PI3K/Akt和JAK/STAT3信号传导通路相关活性分子的活化水平。滋养细胞在加电前分别用MAPK抑制剂PD980599(100μmol/L)和Akt抑制剂LY294002(20μmol/L)处理1h,在上述相应含抑制剂的培养基中用场强为150mV/mm直流微电场刺激5h,观察抑制剂对微电场刺激细胞行为的影响。结果 Western blot结果显示,滋养细胞在150mV/mm微电场作用下,p42/44MAPK Thr202/Thr204位点、Akt Ser473位点于5min开始活化,10~60min内逐渐加强,STAT3S727位点在微电场作用下上述各时间点无明显磷酸化水平改变。相应信号通路抑制剂处理后滋养细胞对微电场刺激的应答反应均受到明显抑制。结论微电场对滋养细胞迁移/侵袭功能的影响与MAPK/ERK、PI3K/Akt信号通路的活化有关。     

LncRNA SNHG8在胎盘植入的表达及其对滋养细胞侵袭及迁移的影响

目的 探讨lncRNA SNHG8在胎盘植入（placenta accreta,PA）中的表达及对滋养细胞侵袭迁移的影响。方法 采用q RT-PCR检测PA组及对照组各30例胎盘组织中lncRNA SNHG8的表达,同时与30例PA患者产前超声评分做相关性分析。采用Transwell和划痕实验检测干扰lncRNA SNHG8后对人绒毛膜滋养层细胞（HTR8/SVneo cells）侵袭和迁移的影响,同时,Western blot检测MMP-2和MMP-9的表达情况;StarBase软件预测lncRNA SNHG8下游靶点,并在两组胎盘组织中检测其表达;双荧光素酶报告实验检测lncRNA SNHG8与miR-542-3p的靶向关系。结果 与对照组比较,lncRNA SNHG8在胎盘植入组胎盘组织中表达上调（P <0.05）,并与产前超声评分呈正相关。干扰lncRNA SNHG8后抑制了滋养细胞的侵袭迁移（P <0.05）,MMP-2及MMP-9蛋白表达也明显下降（P <0.05）。生物学预测miR-542-3p存在与lncRNA SNHG8的结合位点,miR-542-3...     

mir-181a对滋养细胞HTR-8/SVneo功能的影响

目的 MicroRNAs在子痫前期的发生发展过程中发挥了重要作用,本实验通过观察mir-181a对人滋养细胞HTR-8/SVneo凋亡功能的影响,进一步探讨mir-181a在子痫前期发病过程中的功能.方法 采用瞬时转染技术对人滋养细胞HTR-8/SVneo分别进行过表达与抑制mir-181a,应用RT-PCR检测mir-181a mRNA的表达.应用荧光显微镜检测细胞凋亡的变化.结果 RT-PCR显示过表达组和抑制组分别与阴性对照组和空白对照组相比均有明显差异（P＜0.05）,说明转染有效.细胞凋亡结果显示与阴性对照组和空白对照组相比,过表达mir-181a实验组细胞凋亡能力增加,抑制组细胞凋亡能力降低,差异有统计学意义（P＜0.05）;阴性对照组和空白对照组比较无统计学意义（P＞0.05）.结论 mir-181a能促进人滋养细胞HTR-8/SVneo的凋亡,有望作为子痫前期的治疗靶点.     

体外血管内皮生长因子单克隆抗体对C6胶质瘤细胞增殖、侵袭和迁移力的影响

目的:在体外观察血管内皮生长因子(VEGF)单克隆抗体对C6胶质瘤细胞增殖、侵袭和迁移力的影响。方法:采用氯化钴(Co Cl2)模拟缺氧环境,予以0、0.1、1、10μg/m L的VEGF抗体,对常氧或缺氧条件下对C6胶质瘤细胞进行处理。通过MTT实验检测细胞增殖,Transwell实验检测细胞的侵袭和迁移能力,Western blotting检测HIF-1α蛋白、FAK/Pyk2总蛋白及磷酸化FAK-Tyr397/Pyk2-Tyr402表达水平变化。结果:200μmol/L Co Cl2明显抑制C6细胞增殖(P<0.05),10μg/m L VEGF抗体可明显抑制C6细胞增殖(P<0.05)。与常氧相比,缺氧可使C6细胞的迁移、侵袭能力增强(P<0.05)。常氧和缺氧下,VEGF抗体的干预均增强C6细胞的迁移、侵袭能力(P<0.05),且相对于常氧,缺氧可进一步增强VEGF抗体促C6细胞迁移、侵袭的作用(P<0.05)。Co Cl2干预可增加C6细胞中HIF-1α的含量;在常氧和缺氧情况下,只有1μg/m L VEGF抗体可降低FAK的总蛋白量(P<0.05);在缺氧情况下,VEGF抗体可明显增高Pyk2-Tyr402位点的磷酸化水平。结论:在缺氧情况下,VEGF抗体的干预可使Pyk2-Tyr402位点的磷酸化水平增高,引起C6细胞的侵袭迁移能力进一步增强。     

晚期氧化蛋白产物对妊娠滋养细胞生物学功能的影响

目的观察晚期氧化蛋白产物（AOPP）对妊娠滋养细胞（HTR8/SVneo）功能的影响。 方法体外培养HTR8/SVneo细胞株,用次氯酸（HClO）氧化鼠白蛋白（MSA）体外制备AOPP-MSA,将HTR8/SVneo滋养细胞与不同浓度（50 μg/ml组、100 μg/ml组或200 μg/ml组）的AOPP-MSA共同培养。四甲基偶氮唑蓝（MTT）比色法检测滋养细胞的增殖能力,以平均吸光度（A）值表示;侵袭小室检测滋养细胞侵袭能力,以穿膜细胞数表示;检测上清液中滋养细胞分泌的β-HCG水平评价滋养细胞分化能力;Hoechest33258染色检测滋养细胞凋亡。采用单因素方差分析比较不同浓度AOPP下,滋养细胞的增殖能力、侵袭能力、细胞分泌β-HCG、细胞凋亡数的差异。 结果与对照组比较,AOPP浓度为50 μg/ml时,对HTR8SVneo滋养细胞增殖、侵袭、分化能力及凋亡影响不明显（P>0.05）;AOPP浓度为100 μg/ml和200 μg/ml时,AOPP明显抑制滋养细胞增殖、侵袭和分化能力,并促进滋养细胞凋亡（P<0.05）。 结论AOPP可以抑制滋养细胞的增殖、侵袭、分化能力及诱导滋养细胞凋亡。降低血浆中AOPP水平可能有利于改善滋养细胞生物学功能。     

不同间歇时间磁刺激对星形胶质细胞迁移能力的影响及机制分析

目的 探讨不同间歇时间磁刺激对星形胶质细胞迁移能力的影响及相关机制。 方法 将传代星形胶质细胞分为对照组、1 s间歇组、5 s间歇组和10 s间歇组,分别给予相应间歇时间磁刺激,观察不同间歇时间磁刺激对星形胶质细胞迁移能力的影响;星形胶质细胞在磁刺激作用下,采用PEA-15磷酸化阻滞剂Bis I、细胞外调节蛋白激酶（ERK1/2）阻滞剂U0126处理细胞,采用Transwell实验检测星形胶质细胞迁移能力,采用Western blot技术检测pPEA-15和pERK1/2表达。 结果 1 s间歇时间磁刺激可明显增强星形胶质细胞迁移能力,促进PEA-15及ERK1/2磷酸化,并提高基质金属蛋白酶-9（MMP-9）表达;加入Bis I可降低ERK1/2磷酸化水平及MMP-9表达,减弱磁刺激对星形胶质细胞的促迁移作用;经U0126试剂处理后,发现磁刺激对星形胶质细胞的促迁移作用显著下降。 结论 1 s间歇时间磁刺激能促进星形胶质细胞PEA-15磷酸化,提高ERK1/2磷酸化水平,进而增强下游蛋白MMP-9表达,从而促进星形胶质细胞迁移。     

缺氧通过刺激胸腺基质淋巴细胞生成素分泌促进宫颈癌细胞体外侵袭

目的:探讨缺氧诱导宫颈癌细胞体外侵袭的可能调节机制。方法:用ELISA法分析常氧和缺氧培养对宫颈癌细胞株HeLa和CaSki细胞的胸腺基质淋巴细胞生成素(thymic stromal lymphopoietin,TSLP)分泌水平及TSLP对侵袭相关分子基质金属蛋白酶-2(matrix metalloprotein-2,MMP-2)、MMP-9和白细胞介素-8(interleukin-8,IL-8)表达水平的影响。Matrigel侵袭实验分别检测重组TSLP、缺氧和TSLP中和性抗体对宫颈癌细胞侵袭力的影响。结果:与常氧环境相比,缺氧培养下HeLa、CaSki细胞TSLP分泌水平显著升高,差异有统计学意义(P0.01)。重组TSLP(100ng/mL)体外刺激显著促进HeLa、CaSki细胞的侵袭力(P0.001),并促进其分泌MMP-2、MMP-9和IL-8(P0.01)。缺氧培养可显著促进HeLa、CaSki细胞的侵袭力(P0.001);TSLP中和性抗体(5μg/mL)能抑制HeLa、CaSki细胞侵袭(P0.05),且可明显抑制缺氧对HeLa、CaSki细胞侵袭力的促进作用(P0.05)。结论:缺氧可通过促进宫颈癌细胞分泌TSLP,促进宫颈癌细胞体外侵袭。     

CCR3在肾透明细胞癌中的表达及对肾透明细胞癌细胞侵袭、转移能力的影响

目的研究CCR类趋化因子受体3(CCR3)在肾透明细胞癌中的表达及对肾透明细胞癌细胞侵袭、转移能力的影响。方法回顾性选取2019年4月至2020年4月在湖南中医药高等专科学校附属第一医院就诊的肾透明细胞癌组织标本50例,选取50例癌旁组织为对照,进行免疫组化染色。选取人肾透明细胞癌细胞系786-0和人肾透明细胞癌细胞系A498,细胞培养,检测CCR3 mRNA表达水平。构建CCR3沉默慢病毒载体作为CCR3抑制组,对照组中加入一段无义序列,空白组只加生理盐水。细胞迁移、侵袭实验观察细胞迁移、侵袭能力,蛋白质印迹(Western blotting)检测磷酸化细胞外调节蛋白激酶(p-ERK1/2)、基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)水平。结果CCR3在肾透明细胞癌组织中阳性表达比率(48.00%)低于癌旁组织阳性表达(12.00%),差异有统计学意义(P <0.05)。A498人肾透明细胞癌细胞系中CCR3 mRNA表达量高于786-0人肾透明细胞癌细胞系中表达量,差异有统计学意义(P <0.05)。与空白组相比,对照组癌细胞迁移、侵袭数量增加,CCR3抑制组癌细胞迁移、侵袭数量减少(P <0.05);与对照组相比,CCR3抑制组癌细胞迁移、侵袭数量减少,差异有统计学意义(P <0.05)。与空白组相比,对照组p-ERK1/2、MMP-2、MMP-9蛋白表达水平升高,CCR3抑制组p-ERK1/2、MMP-2、MMP-9蛋白表达水平降低(P <0.05);与对照组相比,CCR3抑制组p-ERK1/2、MMP-2、MMP-9蛋白表达水平降低,差异有统计学意义(P <0.05)。结论 CCR3在肾透明细胞癌细胞中显示高表达,抑制CCR3表达通过调控p-ERK1/2、MMP-2、MMP-9蛋白,从而抑制肾透明细胞癌细胞侵袭和迁移。     

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.     

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.     

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.     

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.     

Imaging choices in occult hip fracture

Background: Hip fracture is a common injury, with an incidence rate of > 250,000 per year in the United States. Diagnosis is particularly important due to the high dependence on the integrity of the hip in the daily life of most people. Objectives: In this article we review the literature focused on hip fracture detection and discuss advantages and limitations of each major imaging modality. Discussion: Plain radiographs are usually sufficient for diagnosis as they are at least 90% sensitive for hip fracture. However, in the 3–4% of Emergency Department (ED) patients having hip X-ray studies who harbor an occult hip fracture, the Emergency Physician must choose among several methods, each with intrinsic limitations, for further evaluation. These methods include computed tomography, scintigraphy, and magnetic resonance imaging. Conclusion: We present an evidence-based algorithm for the evaluation of a patient suspected to have an occult hip fracture in the ED. Also outlined are future directions for research to distinguish more effective techniques for identifying occult hip fractures.     

Systematic Reviews Published in the April 2012 Issue of the Cochrane Library

ABSTRACT

The Cochrane Library of Systematic Reviews is published quarterly as a DVD but monthly online. The April 2012 issue (second DVD for 2012) contains 5045 complete reviews, 2182 protocols for reviews in production, and 17,084 short summaries of systematic reviews published in the general medical literature. In addition, there are citations of 674,000 randomized controlled trials, and 15,400 cited papers in the Cochrane methodology register. The health technology assessment database contains just over 11,000 citations. One hundred and seventeen new reviews have been published in the last 3 months of which 12 have potential relevance for practitioners in pain and palliative medicine. The impact factor of the Cochrane Library stands at 6.186. Readers are encouraged to access the full report for any articles of interest as only a brief commentary is provided.     

Platelets,thrombosis, coagulation,and atherosclerosis

When I first got the invitation to join a medical delegation going to Moldova, I thought for a moment that our destination was the fictional country in the old Marx Brothers movie Duck Soup. On further checking, it turns out that entertaining place was called Freedonia. I now know that Moldova is indeed a real country, bordered on the west by Romania and on the other three sides by the Ukraine. It is a proud country, rich with traditions, and its people are warm, giving, eager to learn ways to improve their healthcare system, and deeply appreciative of our attempts to help them in the task.