非靶向代谢组学方法探究母体羊水中先天性心脏病差异代谢物及相关代谢通路

摘要:目的?通过非靶向代谢组学方法探究先天性心脏病(congenital heart disease，CHD)差异代谢物和代谢通路。方法?采用病例-对照研究，以78例CHD胎儿母体羊水样本作为CHD组，126例非CHD胎儿母体羊水样本作为对照组。应用气相色谱飞行时间质谱(GC-TOF-MS)检测2组代谢物。采用正交偏最小二乘判别分析(OPLS-DA)进行建模，通过多维和单维统计分析寻找差异代谢物。KEGG数据库进行代谢通路富集分析，R语言绘制ROC曲线。结果?CHD组与对照组间存在明显分离趋势，2组间共发现43种差异表达的代谢物(P<0.05)，主要涉及谷氨酸和精氨酸等多条代谢通路。苏氨酸和核糖醇在室间隔缺损(VSD)及法洛四联症(TOF)之间存在差异(P值分别为0.018和0.037)。苯丙氨酸检测VSD敏感性最高(100%)，谷氨酸/焦谷氨酸比值检测VSD特异性最高(97.5%)；尿酸和天冬酰胺检测TOF敏感性最高(100%)，谷氨酸/焦谷氨酸比值检测TOF特异性最高(82.5%)。结论?羊水代谢组学分析证实CHD存在多种代谢物的变化，且多条代谢通路被扰动。     

非靶向代谢组学方法筛选羊水中室间隔缺损自然闭合差异代谢物

摘要：目的 通过基于气相色谱飞行时间质谱（ＧＣ ＴＯＦ／ ＭＳ）的非靶向代谢组学方法探讨羊水中室间隔缺损（ＶＳＤ）自然闭合 差异代谢物。方法 选取 ２０１８ 年 ５ 月至 ９ 月在南京医科大学附属妇产医院临床遗传实验室进行产前诊断的羊水样本 １４ 例， 其中产前胎儿超声心动图检测存在 ＶＳＤ 且产后随访 ＶＳＤ 自然闭合胎儿母体羊水样本 ５ 例，对照组为产前诊断为 ＶＳＤ 但产后 随访未自然闭合胎儿母体羊水样本 ９ 例，采用 ＧＣ ＴＯＦ／ ＭＳ 技术检测所有样本的差异代谢物。通过主成分分析（ＰＣＡ）和正交 偏最小二乘法判别分析（ＯＰＬＳ ＤＡ）比较 ＶＳＤ 自然闭合组和未自然闭合组代谢谱差异，通过多维和单维统计分析寻找差异代 谢物。结果 ＰＣＡ 和 ＯＰＬＳ ＤＡ 模型均显示 ＶＳＤ 自然闭合组和未自然闭合组间无明显分离趋势。单维和多维分析均显示鼠 李糖和苯甲酸在 ＶＳＤ 自然闭合组较未自然闭合组显著降低。结论 本研究发现了 ＶＳＤ 自然闭合差异代谢物，为预测 ＶＳＤ 能 否自然闭合提供了潜在的标志物，也为 ＶＳＤ 自然闭合病理机制的探索提供更多线索。     

异位妊娠特征性代谢组学的研究

目的:采用超高效液相色谱/飞行时间质谱(UPLC/Q-TOF MS)联用技术,探索异位妊娠的特征性代谢物,为建立异位妊娠特征性代谢物组诊断模型提供依据,以期早期诊断异位妊娠。方法:本研究初步检测了9例异位妊娠,15例流产及20例正常妊娠患者的血清标本。利用UPLC-Q/TOF MS联用分析技术,比较正常妊娠、流产、异位妊娠的血液全组分代谢组学差异,用SIMCA-P11.0软件进行主成分分析和偏最小二乘法-显著性分析方法对各组代谢指纹数据进行分析,筛选出异位妊娠可能的特异性代谢物。结果:将异位妊娠和正常妊娠,异位妊娠和流产进行疾病聚类分析,发现了13个有可能成为异位妊娠的特征性代谢产物。结论:通过比对异位妊娠与宫内妊娠(包括正常宫内妊娠及宫内妊娠流产)的代谢差异,发现其可能的特征性代谢产物,对于探索异位妊娠的早期诊断有重要意义。     

大鼠实验性胃溃疡的代谢组学研究

目的:探讨大鼠实验性胃溃疡发生与自愈过程中代谢组学的变化及其机制.方法:Wistar雄性大鼠完全随机分为正常组与不同时间模型组;采用乙酸灼烧法制备胃溃疡模型;运用气相色谱-质谱(GC-MS)获得大鼠胃黏膜代谢物谱,进行主成分分析(PCA)及偏最小二乘法判别分析(PLS-DA)进行模式识别.结果:与正常组比较,不同时间模型组胃黏膜代谢物谱发生明显变化;PCA得分图各组区分良好;PLS-DA获得与分组密切相关的代谢标志物包括某些有机酸、氨基酸、脂肪酸与胆固醇;表明大鼠胃溃疡的发生与自愈过程中出现能量代谢及物质代谢异常.结论:从代谢组学的角度揭示了大鼠实验性胃溃疡代谢物谱的动态变化,较全面地阐释了胃溃疡的发生及自愈机制.     

基于液相色谱质谱联用系统的急性缺血性脑梗死患者血清代谢轮廓分析

目的通过代谢组学分析平台分析大动脉粥样硬化型急性缺血性脑梗死患者血清代谢轮廓变化,建立疾病区分模型并寻找特征代谢物,探讨其可能的代谢机制。方法用代谢组学分析平台对25例大动脉粥样硬化型急性缺血性脑梗死患者和23例健康志愿者的血清样本进行分析,用基于模式识别的多元统计学分析方法对实验数据进行处理并对差异性代谢物的变化趋势进行分析。结果成功构建了血清代谢轮廓的主成分分析模型(PCA)及正交偏最小二乘判别分析模型(OPLS-DA),从模型中筛选出60个具有显著差异的特征离子,在35个被鉴定的内源性代谢物中,缺血性脑梗死组与健康人对照组比较,25个代谢物含量明显升高,剩余的10个代谢物呈下降趋势。结论通过对该类型缺血性脑梗死患者血清代谢轮廓的分析,找到了与疾病相关的一组特征代谢物。     

预测急性心肌梗死患者转归的血清代谢物谱研究

目的采用液-质联用代谢组学研究平台寻找能在急性心肌梗死(AMI)患者入院时即预测患者转归的特异性代谢物。方法运用超高效液相色谱串联质谱(UPLC-MS/MS)代谢组学研究平台分析48例心绞痛患者、24例AMI好转者、24例AMI死亡者及48名体检健康者(正常对照组)代谢物数据,对代谢轮廓数据的变异性进行分析,构建疾病区分模型。采用受试者工作特征(ROC)曲线评估特征代谢物的临床诊断效能。结果成功构建了"正常对照组-心绞痛组-AMI好转组-AMI死亡组"的主成分分析(PCA)模型(R2X=40.2%,Q2=13.9%)及正交偏最小二乘判别分析(OPLS-DA)模型(R2X=79.0%,R2Y=83.2%,Q2=65.0%)。验证结果显示模型的预测准确度达100%。筛选并鉴定出10种可用于AMI预后判断的代谢物离子,其中4-羟基-6-二十二烷酮、N-乙酰-白三烯、溶血磷脂酰胆碱(LPC)[18:2(9Z,12Z)]、LPC(18:0)、LPC(P-16:0)、LPC[P-18:1(9Z)]、LPC[20:1(11Z)]、LPC[20:2(11Z,14Z)]在AMI死亡患者的血清中水平较低,L-苏氨酸、3-甲基-2-丁烯-1-硫醇水平较高,与AMI好转患者比较差异均有统计学意义(P0.05)。结论采用代谢组学分析方法找到的特征代谢物具有很好的预测AMI患者预后的能力,可作为潜在的疾病转归标志物进一步研究。     

代谢组学方法分析重症谵妄患者的血清代谢物

目的基于磁共振代谢组学方法分析发生谵妄的重症患者血清代谢物标志物。 方法选择2016年2月至2017年9月皖南医学院弋矶山医院重症医学科（ICU）发生谵妄的患者18例（谵妄组）,同时选择匹配的未发生谵妄的18例患者作为对照（非谵妄组）。采用磁共振（¹H磁共振）方法检测2组患者血清代谢组学,主成分分析（PCA）法进行多变量数据分析,偏最小二乘法-判别分析（PLS-DA）法进行模型验证,正交偏最小二乘法-判别分析（OPLS-DA）筛选及鉴定2组患者间存在显著差异的代谢产物。 结果与非谵妄组患者相比,谵妄组患者的异亮氨酸、亮氨酸、缬氨酸、丙氨酸和乙酰乙酸的峰值明显增高,而柠檬酸的峰值则明显降低。 结论ICU患者发生谵妄时,机体氨基酸代谢、三羧酸循环和酮体代谢异常,基于磁共振的代谢组学技术有助于发现谵妄患者差异标志代谢物。     

血清学三联筛查联合羊水穿刺在18、21-三体综合征产前诊断中的应用

目的探究血清学三联[甲胎蛋白(AFP)、β-人绒毛膜促性腺激素(β-HCG)及雌三醇(μE3)]筛查联合羊水穿刺在18、21-三体综合征产前诊断中的应用。方法对2015年1月至2017年12月于该院产检的26 158例孕中期孕妇进行血清学三联筛查,对染色体非整倍体病高风险孕妇进一步进行羊水穿刺检查,并随访至其妊娠结束。结果26 158例孕妇中,18-三体综合征和21-三体综合征的筛查阳性率分别为0.58%和7.15%,总阳性率为7.73%(2 022例)。对高风险孕妇进一步行羊水穿刺检查,有1 921例孕妇接受羊水穿刺做胎儿染色体检查,其中21-三体综合征22例(1.15%),18-三体综合征13例(0.68%)。2 022例高风险孕妇不良妊娠结局发生率为7.47%,24 136例低风险孕妇不良妊娠结局发生率为2.18%,高风险孕妇不良妊娠结局发生率明显高于低风险孕妇(P0.05)。高龄组孕妇21-三体综合征阳性筛查率为28.28%,18-三体综合征阳性筛查率为3.18%;低龄组孕妇21-三体综合征阳性筛查率为6.36%,18-三体综合征阳性筛查率为0.48%。高龄组孕妇筛查高风险率明显高于低龄组(P0.05)。结论血清学三联筛查联合羊水穿刺可以检查胎儿染色体结构及数目是否异常,对18、21-三体综合征的预防具有较高的应用价值,对不良妊娠结局具有一定的指导作用,对预防缺陷新生儿出生具有重要的意义。     

乙型病毒性肝炎、原发性肝细胞癌患者血清代谢组学研究

目的研究乙型病毒性肝炎、原发性肝细胞癌(HCC)患者及健康体检者代谢物变化。方法收集49例HCC、29例乙型肝炎DNA阴性、26例乙型肝炎DNA阳性患者血清作为病例组,收集46例健康体检者血清作为对照组,采用AvanceⅡ-600MHz核磁共振仪检测血清中小分子代谢产物,通过聚类分析筛选出各组的特征代谢物。结果与对照组比较,乙型肝炎DNA阴性组患者血清中有12种代谢物质差异有统计学意义(P0.05),乙型肝炎DNA阳性组患者血清中有15种代谢物质差异有统计学意义(P0.05);肝癌组患者血清中有21中代谢物质差异有统计学意义(P0.05),其中苯乙尿酸水平上升,其他代谢物下降。与DNA阳性组比较,乙型肝炎DNA阴性组患者血清中有14种代谢物质差异有统计学意义(P0.05)。结论核磁共振氢谱可有效筛查出2型肝炎、HCC患者之间有显著差异的血清代谢物,在无创诊断HCC方面具有广阔的应用前景。     

基于GC-MS的慢性髓系白血病代谢轮廓分析

目的:应用基于气相色谱-质谱联用(GC-MS)的代谢组学方法对慢性髓系白血病(CML)患者血浆进行代谢轮廓分析,寻找与疾病相关的潜在标志物,并初步探讨其涉及的相关代谢通路。方法:选取2015年2月至2015年4月在苏州大学附属第一医院初次确诊的26例CML患者,及26例同期异基因造血干细胞供者(健康对照),另取26例经过一代酪氨酸激酶抑制剂(TKI)治疗后获得最佳疗效的CML患者(治疗后对照)。应用气相色谱质谱联用技术(GC-MS)方法对三组血浆样本进行代谢物无靶标检测,采集代谢物指纹图谱,结合模式识别分析方法及t检验筛选差异代谢物,通过受试者工作曲线(receiver operating characteristic curve,ROC)分析,评价差异代谢物临床效能,并进行Met PA代谢通路分析。结果:健康人及初诊CML患者血浆代谢表型明显不同。与健康对照组比较,CML组有6个血浆代谢物表现出差异,肉豆蔻酸、甘油丰度均较健康对照组低,而肌醇、半乳糖、乳酸、甘氨酸丰度均较健康对照组高(VIP1,P0.05,AUC0.7),经过TKI药物治疗后获得最佳疗效的CML患者上述差异血浆代谢物均有向正常人水平恢复的趋势。以上CML患者血浆代谢紊乱通路主要与半乳糖代谢、丙酮酸代谢、甘油酯代谢、甘氨酸丝氨酸苏氨酸代谢相关(影响值0.10)。结论:CML患者血浆中代谢物水平出现显著变化,代谢组学结合模式识别分析方法可能成为CML辅助诊断的新方法。     

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.