高分辨率熔解曲线分析法检测非小细胞肺癌KRAS和BRAF基因突变

目的探讨高分辨率熔解曲线分析(HRM)法检测非小细胞肺癌(NSCLC)中KRAS和BRAF基因突变用于临床检测的可行性。方法用HRM法检测64例NSCLC患者KRAS基因第2外显子和BRAF基因第15外显子的突变情况,用直接测序法对结果进行验证。结果 HRM法检测结果表明有9例NSCLC患者发生KRAS基因突变(14.06%)、4例发生BRAF基因突变(6.25%),直接测序法证实两法的结果完全一致;共检测出4种KRAS基因突变类型,G12C(GGT>TGT)的突变率最高(44.4%),BRAF基因突变型均为V600E。结论用HRM法检测临床样本KRAS和BRAF基因突变,具有操作简便、结果准确、成本低的优点,适用于临床检测。     

非小细胞肺癌患者肿瘤组织中EGFR和KRAS基因突变的 分子病理检测分析

目的:探讨非小细胞肺癌患者肿瘤组织中EGFR和KRAS基因各亚型突变情况。方法:应用直接测序方法检测非小细胞肺癌石蜡组织中1 273例EGFR基因和1 062例KRAS基因突变情况。结果:非小细胞肺癌肿瘤组织中EGFR基因总突变率为36.68%(467/1 273),外显子18、19、20和21的突变率分别为1.02%(13/1 273)、18.93%(241/1 273)、2.59%(33/1 273)和15.95%(203/1 273);EGFR基因各外显子之间双重突变共17例(1.34%),其中18外显子与20外显子双重突变3例(0.24%),19外显子与20外显子双重突变7例(0.55%),19外显子与21外显子双重突变4例(0.31%)和20外显子与21外显子双重突变3例(0.24%);EGFR基因各外显子内双重突变共2例(2.18%),均为21外显子双重突变。KRAS基因总突变率为3.01%(32/1 062),外显子2的密码子5、12、13和25的突变率分别为0.09%(1/1 062)、2.64%(28/1 062)、0.18%(2/1 062)和0.09%(1/1 062),外显子3密码子61的突变率为0.09%(1/1 062)。结论:非小细胞肺癌患者中EGFR基因存在较高的突变率,尤其为19和21外显子突变,其基因突变亚型分类能指导EGFR-TKI的肿瘤靶向治疗,KRAS基因突变率虽低但不容忽视,其基因突变预示着EGFR-TKI原发耐药。     

高分辨率熔解曲线分析在血友病B F9基因突变检测中的应用

目的 利用高分辨率熔解曲线(HRM)技术建立简便有效的血友病B(HB)基因诊断的方法.方法 收集2005年1月至2010年6月就诊于上海瑞金医院的55例HB患者的外周血标本,抽提外周血基因组DNA.采用PCR扩增结合测序的方法对40例HB患者进行F9基因突变检测,利用其中21例带有F9基因1～7号外显子突变的HB患者标本建立相应的HRM突变筛查方法.采用F9基因1～7号外显子的PCR-HRM突变筛查方法及8号外显子的DNA直接测序方法,对15例未知F9基因突变的HB患者进行基因诊断.结果 通过PCR扩增结合测序,40例HB患者均检测到F9基因突变.21例带有F9基因1～7号外显子突变的HB患者标本中,19例(90％)患者的突变可通过PCR-HRM技术进行检测.通过F9基因1～7号外显子PCR-HRM突变筛查及8号外显子的DNA直接测序,15例未知F9基因突变的HB患者均检测到F9基因突变.55例HB患者中共检测到34种F9基因突变.结论 HB基因诊断的新方法,即PCR-HRM筛查F9基因1～7号外显子突变结合8号外显子的DNA测序法操作简便、结果可靠.     

2种方法检测非小细胞肺癌EGFR基因突变的比较

目的比较测序法和Taqman实时荧光PCR法检测表皮生长因子受体(EGFR)基因19、21外显子基因突变的一致性,为寻找适宜临床应用的EGFR基因突变检测方法提供实验依据。方法收集60例非小细胞肺癌患者肿瘤组织,应用测序法和Taqman实时荧光PCR法分别检测EGFR基因19、21外显子基因突变,比较两种方法的有效性。结果两法检测EGFR基因19、21外显子突变结果符合率分别为96.7%(58/60)和98.3%(59/60),差异无统计学意义(P>0.05)。结论 2种方法均可用于EGFR基因19、21外显子突变检测,测序法更适合指导临床分子靶向治疗。     

非小细胞肺癌838例EGFR、KRAS基因突变状态及临床意义

目的探讨非小细胞肺癌(NSCKC)中EGFR、KRAS基因突变与患者临床病理特征的相关性。方法采用ARMS法和Sanger测序法检测838例NSCLC样本中EGFR和KRAS基因的突变情况,同时收集患者的临床病理资料,并分析之间的相关性。结果 EGFR、KRAS突变率分别为42.24%、10.98%。EGFR突变以21外显子L858R和19外显子Del突变为主,占所有突变的87.85%; EGFR突变在女性患者中的突变率明显高于男性(P0.01),在腺癌中的突变率明显高于非腺癌(P0.01),在不吸烟患者中的突变率明显高于吸烟患者(P0.01)。KRAS突变以外显子2突变为主,占所有突变的92.39%,其中12密码子突变占86.96%; KRAS突变在男性患者中的突变率明显高于女性(P0.01),在腺癌中的突变率明显高于非腺癌(P0.01),在吸烟患者中的突变率明显高于不吸烟者(P0.01)。EGFR、KRAS突变与患者年龄、有无淋巴结转移无相关性(P0.05)。结论潍坊地区NSCLC患者中EGFR突变以21外显子L858R和19外显子Del突变为主; KRAS突变以外显子2第12密码子突变为主; EGFR和KRAS基因突变与患者性别、组织学类型及吸烟密切相关。     

华南地区172例非小细胞肺癌表皮生长因子受体和KRAS突变分析

目的:完善华南地区非小细胞肺癌(NSCLC)患者表皮生长因子受体(EGFR)和KRAS基因突变谱,探讨其与临床表型的关系。方法:直接测序法对172例NSCLC患者肿瘤组织DNA的EGFR 18-21外显子和KRAS 2号外显子进行突变鉴定并分析。结果:EGFR和KRAS在本群体中突变率分别为32%(55/172)和8.1%(14/172),其中热点突变为EGFR 19号外显子缺失,21号外显子L858R和KRAS 12位密码子点突变。女性、无吸烟史患者与男性、吸烟者相比,EGFR突变率显著增高,而KRAS突变率显著降低。腺癌患者EGFR突变率显著高于非腺癌患者,KRAS突变则无显著差别。结论:华南地区人群EGFR具有丰富的突变谱。完善该地区的突变数据库,确证各种突变对TKI的治疗敏感性的影响,对靶向治疗在NSCLC患者中更深入、广泛开展具有重要的指导作用。     

NSCLC患者肿瘤组织中EGFR和K-Ras基因各亚型的突变情况及意义

目的探讨非小细胞肺癌(NSCLC)患者肿瘤组织中EGFR和KRAS基因突变状况以及临床意义。方法选取2015年2月至2018年1月在我院治疗的NSCLC患者110例,采用荧光PCR检测EGFR和KRAS基因突变状况。结果 110例患者中共检测到EGFR基因突变为33.64%,其中外显子18、19和20突变分别占为27.03%、32.43%、16.22%,外显子18和20共同突变占13.51%,外显子19和20共同突变占10.81%;110例患者中共检测KRAS基因突变为2.73%;未检测到EGFR与KRAS基因双重突变;女性患者EGFR基因突变率为62.79%,明显高于男性患者(P0.05);腺癌患者EGFR基因突变率为42.17%,明显高于非腺癌患者(P0.05);KRAS基因突变与患者年龄、性别、吸烟史、病灶大体类型、肿瘤直径、TNM分期、分化程度以及淋巴结转移无关(P0.05)。结论 NSCLC患者肿瘤组织中EGFR突变率较高,而KRAS基因突变率较低,其中EGFR突变与患者性别、病理类型有关。     

胰腺癌中EGFR、KRAS、BRAF基因突变状况分析及意义

目的:检测胰腺癌组织EGFR、KRAS、BRAF基因突变状况,为胰腺癌EGFR靶向治疗研究奠定基础.方法:提取胰腺癌及胰腺良性病变石蜡组织切片中基因组DNA,PCR扩增EGFR 18、19、21外显子片段,KRAS2、3外显子片段,BRAF15外显子片段,采用直接测序法检测其突变状况.结果:32例胰腺癌患者中有24例患者存在KRAS基因突变,10例良性病变组织均未发现突变,两者差异具有统计学意义(x2=14.57,P=1.35× 10-4),其中22例12密码子突变(G12D14例,G12V8例);2例61密码子突变(Q61L).所有检测样本中未见BRAF突变.共3例EGFR突变,其中包括1例19外显子突变(de1746-750),2例21外显子突变(L858R),10例良性病变组织未见突变.结论:在胰腺癌中KRAS基因突变可能为EGFR通路失调的主要原因,其次是EGFR突变,BRAF突变未见.     

某地区结直肠癌患者KRAS基因及BRAF基因突变状态的检测分析

目的 检测某地区结直肠癌患者KRAS基因和BRAF基因的突变状态及其与年龄、性别的关系.方法 由某地区30例结直肠癌患者石蜡组织中提取DNA,通过直接测序法及荧光定量PCR法检测KRAS基因及BRAF基因突变状态.结果 KRAS基因突变率为43.3%,共发现6种突变类型,主要位于12、13密码子,其中以c.38G>A突变率最高(38.5%).单因素及多因素分析均提示KRAS突变与年龄或性别无相关性.BRAF基因突变率为0.结论 某地区结直肠癌患者KRAS基因突变率高,靶向治疗前进行突变状态检测具有重要临床价值.     

HRM法检测结直肠癌组织KRAS基因密码子12和13点突变

目的 建立HRM法检测大肠癌患者肿瘤组织KRAS(v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog)基因突变的方法 .方法 采用HRM法对含不同比例KRAS基因突变型质粒的系列混合样本进行检测,以评价其灵敏度.应用HRM法检测60份大肠癌患者新鲜肿瘤组织KRAS基因密码子12和13的突变状况,并与直接测序法的结果 进行比较分析.结果 HRM法只需在PCR结束后直接运行高分辨熔解,即可获得检测结果 .HRM法可检出系列混合样本中突变型质粒比例为10%的突变,其检测灵敏度达10%.HRM法从60份大肠癌患者组织标本中,检出17份KRAS基因密码子12或13突变(28.3%);直接测序法检出15份(25.0%)突变,2份未检出KRAS基因突变.HRM法检测的敏感度为100%(15/15),特异度为96%(43/45).结论 HRM法在筛选大肠癌标本的KRAS基因突变类型时,具有操作简便、快速、灵敏,单管避免污染等优点,完全符合临床个体化治疗的要求,值得推广.     

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.     

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.     

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.     

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.     

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."