LAMP法检测儿童粪便中肠道腺病毒的建立

目的通过应用环介导等温扩增(LAMP)建立一种快速、简便的可视化检测儿童肠道腺病毒核酸的方法。方法登录GenBank网站下载腺病毒40和41基因型的六邻体序列,根据该区段的保守序列设计4条LAMP引物。将反应体系加入EP管,在恒温条件(65℃)扩增60min后,再以凝胶电泳和钙黄绿素染色评价LAMP法的特异性和灵敏度,并和PCR法的结果相比较。最后,同时用LAMP和PCR法对40份可疑临床样本进行检测,应用卡方检验进行统计学分析。结果 LAMP法能准确地使腺病毒40和41型得到扩增,酶切结果也正确,显示了LAMP法较好的特异性。LAMP的灵敏度比PCR高约10倍。经过对40份临床样本的检测,LAMP与PCR均没有发生非特异扩增,LAMP法和PCR法的一致性为92.5%(P0.05)且LAMP法没有漏检阳性样本。另外,使用钙黄绿素染色的方法更好地显示了产物检测的可视性和简便性。结论该研究初步建立了快速、简便、可视化检测肠道腺病毒的LAMP技术,在基层医疗机构有一定的实用价值。     

Rapid detection of Streptococcus pneumoniae by real-time fluorescence loop-mediated isothermal amplification

Background and aim of study

A significant human pathogenic bacterium, Streptococcus pneumoniae was recognized as a major cause of pneumonia, and is the subject of many humoral immunity studies. Diagnosis is generally made based on clinical suspicion along with a positive culture from a sample from virtually any place in the body. But the testing time is too long. This study is to establish a rapid diagnostic method to identification of Streptococcus pneumoniae.

Methods

Our laboratory has recently developed a new platform called real-amp, which combines loop-mediated isothermal amplification (LAMP) with a portable tube scanner real-time isothermal instrument for the rapid detection of Streptococcus pneumonia. Two pairs of amplification primers required for this method were derived from a conserved DNA sequence unique to the Streptococcus pneumoniae. The amplification was carried out at 63 degree Celsius using SYBR Green for 60 minutes with the tube scanner set to collect fluorescence signals. Clinical samples of Streptococcus pneumoniae and other bacteria were used to determine the sensitivity and specificity of the primers by comparing with traditional culture method.

Results

The new set of primers consistently detected in laboratory-maintained isolates of Streptococcus pneumoniae from our hospital. The new primers also proved to be more sensitive than the published species-specific primers specifically developed for the LAMP method in detecting Streptococcus pneumoniae.

Conclusions

This study demonstrates that the Streptococcus pneumoniae LAMP primers developed here have the ability to accurately detect Streptococcus pneumoniae infections by real-time fluorescence LAMP.     

Structure-based classification of EGFR mutations in operable pre-invasive and invasive non-small cell lung cancer: a cross-sectional study

BackgroundIt has been reported that the structure-based approach for defining functional groups of epidermal growth factor receptor (EGFR) mutations predicts the efficacy of EGFR inhibitors better than the traditional exon-based approach in the advanced stage. However, less is known about this structure-based classification of EGFR mutations in operable early-stage lung adenocarcinoma.MethodsNon-small cell lung cancer (NSCLC) patients with pathological stage I–III or adenocarcinoma in situ (AIS) who had EGFR mutations identified in next-generation sequencing (NGS) testing were recruited. Both exon-based and structure-based groupings of EGFR mutations were compared between the AIS and stage I–III patients using Fisher’s exact test.ResultsIn total 1,012 patients including 66 AIS and 946 stage I–III patients were analyzed in the study. A total of 1185 EGFR mutations were identified in the 1,012 NSCLC patients, of whom 84.39% harbored a single EGFR mutation and 15.61% harbored complex EGFR mutations. As expected, L858R was more common than 19del in our population (39.33% vs. 35.67%). Interestingly, concurrent L858R and 19del mutations were identified in 9 patients (0.89%), and all these patients were diagnosed with multiple primary lung cancer. A higher percentage of atypical EGFR mutations was identified in the AIS cohort than in the stage I–III NSCLC cohort (33.33% vs. 21.66%, P=0.03). According to the structure-based classification of EGFR mutations, 86.07%, 7.11%, 5.04%, and 1.78% of the EGFR mutations were classified as classical-like, P-loop and α C-helix compressing (PACC), exon 20 insertions (Ex20ins), and T790M-like mutations, respectively. The composition of EGFR mutations was different between patients <65 and ≥65 years (P=0.0267) but similar between patients with AIS and stage I–III NSCLC (P=0.1436). However, a higher percentage of Ex20ins occurred in younger (<65 years) patients, nonsmoking patients, and patients with AIS (6.7% vs. 2.5%, P=0.003; 5.8% vs. 0.8%, P=0.0107; and 10.6% vs. 4.7%, P=0.0423, respectively).ConclusionsThis large cross-sectional study delineated the structure-based classification of EGFR mutations in patients with operable NSCLC. While the traditional exon-based EGFR grouping showed difference between AIS and stage I–III NSCLC cohort, no difference was identified in the structural approach. Which approach had better prediction of targeted therapy efficacy in adjuvant settings warrants further investigation.     

Efficacy of dacomitinib in patients with non-small cell lung cancer carrying complex EGFR mutations: a real-world study

BackgroundDacomitinib is a first-line treatment for patients with non-small cell lung cancer (NSCLC) harboring common epidermal growth factor receptor (EGFR) mutations; however, clinical evidence of its activity on NSCLC with complex EGFR mutations is limited.MethodsPatients harboring complex (common mutations co-existing with uncommon mutations), or common (comparison cohort) EGFR mutations, who were treated with dacomitinib, were retrospectively evaluated in the Chinese National Cancer Center and the China PLA hospital between August 2019 and August 2021.ResultsIn total, 72 patients with NSCLC harboring complex (C+U group, n=18) or common (C group, n=54) EGFR mutations and being treated with dacomitinib were enrolled. In the C+U group, 16 cases (88.9%) harbored L858R mutations co-existing with uncommon mutations located from exon 18 to exon 25 of EGFR (mostly E709X), and two cases harbored exon 19 deletion co-existing with G724S or K754E. Among the 15 evaluable patients, the objective response rate (ORR) was 40% (6/15), and the disease control rate (DCR) was 73.3% (11/15). The median progression-free survival (PFS) was 7.5 months [95% confidence interval (CI), 4.4–10.6 months]. Except for the application line of dacomitinib (P=0.039), no significant statistical differences were found in other characteristics and adverse events between the two groups. The Kaplan-Meier method revealed no significant differences in PFS (P=0.889) and overall survival (OS) (P=0.703). However, the stratified analysis found worse PFS in the C+U group than that observed in the C group when receiving 1^st and ≥3^rd line dacomitinib treatment, while its OS was worse than that of group C when receiving ≥3^rd line treatment. Furthermore, in a multivariate analysis, complex mutation status was an independent prognostic factor for OS (P=0.038) in the entire cohort.ConclusionsThis study indicated a worse response and prognosis of patients with NSCLC harboring complex EGFR mutations than those harboring common EGFR mutations when treated with dacomitinib. Further studies and data are needed to confirm this conclusion.     

环介导等温扩增技术检测方法的研究进展

环介导等温扩增技术是一种可用于临床诊断的快速检测方法,可用于检测病毒、细菌、寄生虫等多种病原体。经不断完善,现已在临床检测过程中发挥重要作用。可用显色试剂对反应结果实现可视化,即结果的即时显示,无需后续验证。但是,无论是荧光染料还是显色指示剂,对反应过程都会产生或多或少的影响,目前研究表明羟基萘酚蓝是最好的显色试剂。同时,人们发现电化学技术也可以应用于此,使检测反应更快,操作更加简单。生物传感器可以对多种反应产物进行同步检测,而微流控芯片技术更是方便携带,有望成为实时检测技术的理想载体。     

环介导等温扩增技术及其在食源性吸虫感染检测中的应用

随着人们生活水平的提高,土源性寄生虫病的感染率和发病率日趋下降,与之相反,食源性寄生虫病更加引人关注,而食源性吸虫病是其中的重要组成。据WHO统计,目前已知能感染人体的食源性吸虫有100多种,感染者超过4000万人,还有10%的人口面临感染食源性吸虫的风险。目前对于腔道内寄生的食源性吸虫通常采用病原学诊断方法,但对于深部组织内寄生的食源性吸虫则缺乏有效的诊断方法。环介导等温扩增(LAMP)技术是新近发展起来的一种方法,它具有简便快速、灵敏特异、经济实用、且易于普及等优点,有替代传统PCR之势。但国内对LAMP法的了解和应用不多,基于其在食源性吸虫感染方面的诊断价值,本文予以介绍,以供借鉴。     

EGFR immunoexpression,RAS immunoexpression and their effects on survival in lung adenocarcinoma cases

Background

The impacts of epidermal growth factor receptor (EGFR) immunoexpression and RAS immunoexpression on the survival and prognosis of lung adenocarcinoma patients are debated in the literature.

Methods

Twenty-six patients, who underwent pulmonary resections between 2002 and 2007 in our clinic, and whose pathologic examinations yielded adenocarcinoma, were included in the study. EGFR and RAS expression levels were examined by immunohistochemical methods. The results were compared with the survival, stage of the disease, nodal involvement, lymphovascular invasion, and pleural invasion. Nonparametric bivariate analyses were used for statistical analyses.

Results

A significant link between EGFR immunoexpression and survival has been identified while RAS immunoexpression and survival have been proven to be irrelevant. Neither EGFR, nor RAS has displayed a significant link with the stage of the disease, nodal involvement, lymphovascular invasion, or pleural invasion.

Conclusions

Positive EGFR immunoexpression affects survival negatively, while RAS immunoexpression has no effect on survival in lung adenocarcinoma patients.     

田鼠巴贝虫感染FTA-环介导等温扩增检测技术的建立

目的 建立敏感、特异、高效的检测田鼠巴贝虫 FTA-环介导等温扩增技术(LAMP)。方法 根据GenBank公布的田鼠巴贝虫基因序列,就细胞色素B基因保守序列设计了多套LAMP引物,利用LAMP RealTime Turbidimeter LA-320 仪筛选最佳引物、反应条件,建立LAMP检测方法,从保存有血液样本的FTA卡片中提取田鼠巴贝虫 DNA进行LAMP,观察检测效果。结果 设计的4组引物做LAMP试验,其中以引物1,最佳反应温度为64 ℃,恒温下作用,敏感性高,可在1 h内完成,其检出限量为0.687 fg/μL,而PCR的最低检测量为0.687 pg/μL,与间日疟原虫、恶性疟原虫、卵形疟原虫、三日疟原虫、诺氏疟原虫、冈地弓形虫、利什曼原虫、冈比亚锥虫均无交叉反应。以建立的FTA-LAMP法检测20份PCR检测阳性的田鼠巴贝虫感染者血样,其阳性符合率100%。结论 成功建立了检测田鼠巴贝虫特异性细胞色素B基因的FTA-LAMP方法,该法特异性强、灵敏度高、简便、快速、适于现场应用。     

非小细胞肺癌患者EGFR、ALK基因突变及临床病理特征与预后的相关性研究

目的探讨非小细胞肺癌(NSCLC)患者EGFR、ALK基因突变状态及病理特征,分析二者与患者预后情况相关性。方法收集2015年1月至2018年1月苏州大学附属第一医院病理确诊的NSCLC病例262例,记录患者临床病理特征及转归,采用突变增阻滞系统(ARMS)-Taqman探针法及RT-PCR检测患者肿瘤样本的EGFR和ALK基因外显子突变情况,采用Kaplan-Meier法和Cox风险回归考察不同基因突变情况的预后、临床病理特征与临床结局的相关性。结果262例NSCLC患者中,168例为野生型,ALK突变12例,突变率4.6%;EGFR突变82例,突变率31.30%,其中18号外显子突变2例,19号外显子突变37例,20号外显子突变9例,21号外显子突变31例;各组间年龄、病理类型比较差异有统计学意义,P<0.05;Wild type组中位OS时间11.6月,ALK组中位OS时间15.0月,EGFR组中位OS时间36.8月,EGFR组OS明显优于ALK组OS时间及Wild type组OS时间,P<0.05;EGFR19号外显子突变组、21号外显子突变组OS生存时间显著高于18号外显子突变组和20号外显子突变组,P<0.05;EGFR阴性(RR=15.751,95%CI:9.252~26.816)、鳞癌(RR=18.344,95%CI:6.187~54.388)为OS的危险因素。结论EGFR突变状态、病理类型对非小细胞肺癌预后具有良好的预测价值,鳞癌和EGFR阴性患者生存期短。EGFR基因少见突变(18号外显子、20号外显子)患者的中位OS时间更短。     

Genetic modifiers of EGFR dependence in non-small cell lung cancer

Lung adenocarcinomas harboring activating mutations in the epidermal growth factor receptor (EGFR) represent a common molecular subset of non-small cell lung cancer (NSCLC) cases. EGFR mutations predict sensitivity to EGFR tyrosine kinase inhibitors (TKIs) and thus represent a dependency in NSCLCs harboring these alterations, but the genetic basis of EGFR dependence is not fully understood. Here, we applied an unbiased, ORF-based screen to identify genetic modifiers of EGFR dependence in EGFR-mutant NSCLC cells. This approach identified 18 kinase and kinase-related genes whose overexpression can substitute for EGFR in EGFR-dependent PC9 cells, and these genes include seven of nine Src family kinase genes, FGFR1, FGFR2, ITK, NTRK1, NTRK2, MOS, MST1R, and RAF1. A subset of these genes can complement loss of EGFR activity across multiple EGFR-dependent models. Unbiased gene-expression profiling of cells overexpressing EGFR bypass genes, together with targeted validation studies, reveals EGFR-independent activation of the MEK-ERK and phosphoinositide 3-kinase (PI3K)-AKT pathways. Combined inhibition of PI3K-mTOR and MEK restores EGFR dependence in cells expressing each of the 18 EGFR bypass genes. Together, these data uncover a broad spectrum of kinases capable of overcoming dependence on EGFR and underscore their convergence on the PI3K-AKT and MEK-ERK signaling axes in sustaining EGFR-independent survival.The term “oncogene addiction” has been used to describe the phenomenon whereby tumor cells exhibit singular reliance on an oncogene or oncogenic pathway for their survival, despite the accumulation of multiple genetic lesions (1). In non-small cell lung cancer (NSCLC), this principle is perhaps best exemplified with the finding that epidermal growth factor receptor (EGFR) mutations predict response to EGFR tyrosine kinase inhibitors (TKIs) gefitinib and erlotinib, and thus represent a dependency in the subset of tumors harboring these alterations (2–6). However, though EGFR-mutant NSCLCs typically respond dramatically to EGFR TKIs, clinical responses are not universal, even within this genetically defined cohort, with the rate of objective response estimated to be ∼71% (5, 6). Furthermore, the overwhelming majority of patients who initially respond to EGFR inhibitors ultimately develop resistance to therapy (7). A deeper understanding of the genetic underpinnings of EGFR addiction, and how EGFR-mutant cells can overcome reliance on EGFR, may improve clinical outcomes.Here, we have applied an unbiased screening approach to identify genetic modifiers of EGFR dependence in NSCLC. Mounting evidence supports the existence of several genetic modifiers of EGFR dependence in EGFR-mutant NSCLC, which can reduce the degree to which these tumors rely on EGFR and thereby contribute to EGFR TKI resistance (8). Examples include amplification of the MET receptor tyrosine kinase (RTK) (9), activation of the NF-κB pathway (8), amplification of the HER2 (ERBB2) RTK (10), amplification of the CRKL gene (11), and activation of the AXL kinase (12). Notably, MET bypass can be reciprocally achieved via EGFR activation in MET-dependent cells (13), and analogous examples of reciprocal kinase switching have been reported in other kinase-driven cancer models (14, 15). These and other findings suggest that compensatory kinase switching may be a more general way in which oncogene-dependent cancers overcome reliance on their primary driver kinase (14, 16), but the full-range of kinases capable of mediating EGFR bypass has not been systematically studied.Recent advances in large-scale functional genetic libraries have made it possible to query a wide range of genetic perturbations for their ability to modulate specific cellular phenotypes in mammalian systems (17, 18). Using the model of EGFR-mutant, erlotinib-sensitive NSCLC cells, we have performed a systematic ORF-based screen to identify kinase and kinase-related genes whose overexpression can complement loss of EGFR activity in an EGFR-dependent context. Our findings indicate broad potential for EGFR substitution in the setting of EGFR dependence, with compensatory mechanisms commonly conferring EGFR-independent activation of the PI3K-AKT and MEK-ERK signaling pathways. Importantly, this approach has recovered known mechanisms of erlotinib resistance as well as identified novel mediators of EGFR bypass in EGFR-mutant NSCLC. These data support the idea that the EGFR-dependent state can be redundantly driven by diverse genetic inputs that commonly converge on shared downstream signaling nodes.     

逆转录环介导等温扩增技术在诺如病毒基因检测中的应用

目的建立逆转录环介导等温扩增技术(RT-LAMP)快速检测诺如病毒的方法。方法采用逆转录环介导等温扩增技术(RT-LAMP)扩增诺如克病毒特异性基因,并与RT-PCR方法比较。结果与RT-PCR技术相比,RT-LAMP法更加简便快速,能够在等温条件下进行,特异性好,具有更高的灵敏性。结论RT-LAMP方法具有灵敏度高,特异性强,操作简便快速,不需要复杂仪器设备等优点,为临床检测诺如病毒快速简便的新方法。     

Correlation of EGFR G873R mutation with prognosis of docetaxel in non-small cell lung cancer

BackgroundClinical features of epidermal growth factor receptor (EGFR) mutations have been commonly recognized in variant cancers. The role of EGFR mutations in non-small cell lung cancer (NSCLC) has spurred research and drug development efforts. However, there are still mutations that have not been widely reported, and their influences on NSCLC have not been fully elucidated; EGFR G873R mutation is just one of them. The aim of this study was to investigate the correlation between EGFR G873R mutation and the prognosis of chemotherapy in NSCLC.MethodsA total of 54 patients with NSCLC were enrolled in this study. Immunohistochemical staining was used to detect the expression of EGFR. A DNA extraction kit (GeneRead DNA FFPE Kit) was used to extract total DNA from resected cancer tissues. Genomic DNA targets were amplified by polymerase chain reaction (PCR), and then the amplicons were purified and sequenced. Statistical methods were performed to detect the relationship between EGFR G873R mutation and various clinicopathological features and the effect of EGFR G873R mutation on the prognosis of chemotherapy.ResultsEGFR G873R mutation did not show statistical significance, with EGFR high expression identified in 30 cases (P>0.05). Patients with EGFR G873R mutation had a significantly favorable prognosis of docetaxel (P=0.032), and for patients treated with docetaxel, EGFR G873R mutation was significantly correlated with better 5-year disease-free survival (DFS; P=0.026) and overall survival (OS; P=0.026). However, there was no statistical significance found between EGFR G873R mutation and the prognosis of vinorelbine (P>0.05), and for patients treated with vinorelbine, EGFR G873R mutation had no statistical significance with 5-year DFS (P>0.05) and OS (P>0.05).ConclusionsEGFR G873R mutation was remarkably correlated with the prognosis of docetaxel in NSCLC, which indicates that EGFR G873R may be employed as a promising biomarker to identify individuals with better prognosis of docetaxel and as an antitumor target for NSCLC treatment.     

Uncommon EGFR mutations in lung carcinoma: features and treatment outcomes in a retrospective French cohort

BackgroundThe best management for rare epidermal growth factor receptor (EGFR) mutations in advanced non-small cell lung carcinoma (NSCLC) remains uncertain. The literature indicates that response to usual treatment could differ in certain subgroups such as exon 20 insertion/duplication (E20ID), other single uncommon mutation (OSUM), and EGFR complex mutation (ECM).MethodsIn this observational, regional, multi-center, retrospective study, we gathered data on uncommon EGFR mutations in NSCLC from 2007 to 2021. We analyzed patient characteristics, prognostic factors and treatment outcomes [objective response rate (ORR), disease control rate (DCR), progression free survival (PFS) and overall survival (OS)].ResultsAmong 119 patients with an uncommon EGFR mutant, 34 harbored E20ID, 23 ECM, and 62 OSUM. There were significantly more non-smokers in E20ID. Female gender and performance status <2 were associated with a better prognosis. Among the 97 metastatic patients with available data for 1st line treatment, median estimated OS was 21 months (95% CI: 18–31 months), with better non-significant OS for ECM. Median estimated PFS was 7 months (95% CI: 4–9 months). We found significant differences in ORR, DCR and PFS favoring 1st line chemotherapy for E20ID, whereas the outcomes for OSUM and ECM were more favorable for tyrosine kinase inhibitor (TKI) (mainly 2nd and 3rd generation).ConclusionsThere were variations in treatment outcomes among subgroups in our cohort. Exon 20 insertions showed better ORR and PFS with 1st line chemotherapy compared to TKI. Conversely, other rare EGFR mutations including ECM had better ORR and PFS with TKI than chemotherapy. There was no significant difference in OS among treatment groups overall or within rare mutation subgroups.     

Correlation between epidermal growth factor receptor mutations and nuclear expression of female hormone receptors in non-small cell lung cancer: a meta-analysis

Background

Compared with male, female non-small cell lung cancer (NSCLC) patients have better response when treated with epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKIs), suggesting a potential association between female hormones and EGFR mutation. However, the results provided by previous studies were inconclusive and controversial. We sought to examine the link between the expression of nuclear female hormone receptors and EGFR mutations in NSCLC.

Methods

Electronic databases were used to search the relevant articles. The involved hormone receptors included estrogen receptor (ER) and progesterone receptor (PR). The primary endpoint was the occurrence of ER/PR expression and EGFR mutation in NSCLC patients.

Results

Five studies fulfilled the criteria and were included in our analysis. Patients with high ER-β expression had higher positive EGFR mutation than low ER-β patients (44.2% vs. 23.7%), and there was a significant difference between the two groups [odds radio (OR) 3.44, 95% confidence interval (CI): 2.40-4.93, Z=6.72, P<0.001]. However, there is no significant correlation between EGFR mutations and ER-α (when included ER-α3, OR 1.20, 95% CI: 0.62-2.33, Z=0.55, P=0.58; and when included ER-α4, OR 1.18, 95% CI: 0.62-2.25, Z=0.51, P=0.61) or PR (OR 1.29, 95% CI: 0.40-4.10, Z=0.43, P=0.67). No significant publication bias was observed.

Conclusions

High nuclear expression of ER-β, but not ER-α or PR is correlated with EGFR mutations in NSCLC. The underlying mechanism and potential translational relevance warrant further investigation.     

The co-mutation of EGFR and tumor-related genes leads to a worse prognosis and a higher level of tumor mutational burden in Chinese non-small cell lung cancer patients

BackgroundLung cancer is the leading cause of cancer mortality in China. The clinicopathologic features and genetic profile of Chinese lung cancer patients need to be investigated. This study evaluated the gene mutation profile, analyzed the frequency of concurrent genes in epidermal growth factor receptor (EGFR)-mutant non-small cell lung cancer (NSCLC) patients, and determined its prognostic value.MethodsWe collected the clinical data from 151 initially diagnosed patients NSCLC. Tumor samples underwent targeted next-generation sequencing (NGS). Progression-free survival (PFS) and overall survival (OS) were analyzed using the Kaplan-Meier method.ResultsAmong the 151 participants, the mutational profile revealed alterations in 29 genes, where TP53 (37.09%) and EGFR (30.46%) exhibited the highest mutation rates. Mutations in the EGFR gene were most prevalent (40%) in adenocarcinoma (LUAD) and were only present in 8.8% of participants with squamous cell carcinoma (LUSC). The most frequently mutated genes in LUAD patients were TP53 (47%), followed by KRAS (11.7%). In all 39 participants with EGFR mutations, TP53, KRAS, PIK3CA, APC, and FBXW7 were also mutated. Those with only EGFR mutation appeared to have a better prognosis; however, the difference was not statistically significant. Tumor mutational burden (TMB) was roughly significantly increased in patients who harbored EGFR and other mutant driver genes, compared with only EGFR mutant patients. The TMB value was significantly higher in those with P53 mutation than in P53 wild-type patients.ConclusionsWe described the genetic profiles of NSCLC and compared the difference in genetic profiles between LUAD and LUSC. The concomitant genetic alterations might be a poor prognostic factor for patients with EGFR mutation, and TMB might be prognostically related to the co-mutations of EGFR and other genes.     

The role of ramucirumab with docetaxel in epidermal growth factor receptor mutant and wild-type non-small cell lung cancer

BackgroundRamucirumab paired with docetaxel extends progression free survival and overall survival in non-small cell lung cancer (NSCLC) following progression on platinum therapy. There is some data that epidermal growth factor receptor (EGFR) mutant disease would respond better to vascular endothelial growth factor receptor (VEGFR) therapy than EGFR wild type disease.MethodsThis retrospective, single-institution cohort study reports outcomes of patients who received docetaxel with or without ramucirumab according to EGFR status. Clinical data including age, performance status, metastatic burden and prior treatment history was obtained and reported with time on treatment and overall survival as primary endpoints. Data analysis was performed for three cohorts: EGFR mutant disease receiving docetaxel and ramucirumab (EGFR-doce/ram), EGFR mutant disease receiving docetaxel alone (EGFR-doce) and EGFR wild type disease receiving docetaxel and ramucirumab (WT-doce/ram).ResultsPatients in the EGFR-doce/ram cohort had a median time on docetaxel of 1.4 months (95% CI: 0.72–5.2 months) and of 0.8 months (95% CI: 0.2–6.5 months) on ramucirumab. Patients in the EGFR-doce cohort were on docetaxel for a median 1.4 months (95% CI: 0.9–2.4 months). Patients in the WT-doce/ram cohort had a median time on docetaxel of 2.3 months (95% CI: 1.6–4.1 months) and on ramucirumab of 1.4 months (95% CI: 0.8–3.2 months). There was no significant difference between time on ramucirumab or docetaxel between the cohorts. Overall survival for the three cohorts was noted to be 6.7 months (95% CI: 2.5–16.2 months) for the EGFR-doce/ram cohort, 4.9 months (95% CI: 4.2–12.5 months) for the EGFR-doce cohort and 6.6 months (95% CI: 4.3–12.8 months) for the WT-doce/ram cohort. There was no significant difference in overall survival between the cohorts.ConclusionsOur data did not support the initial hypothesis that patients with EGFR mutant disease would do better with the addition of ramucirumab. Our study was limited by small sample size, retrospective nature and inability to control for confounders including prior bevacizumab or immune checkpoint inhibitor (ICI) exposure. This study offers real-world estimates to clinicians and patients about the length of time they can expect to derive benefit from the combination of ramucirumab and docetaxel.     

Correlation between familial cancer history and epidermal growth factor receptor mutations in Taiwanese never smokers with non-small cell lung cancer: a case-control study

Background

Lung cancer is a leading cause of cancer deaths in the world. Cigarette smoking remains a prominent risk factor, but lung cancer incidence has been increasing in never smokers. Genetic abnormalities including epidermal growth factor receptor (EGFR) mutations predominate in never smoking lung cancer patients. Furthermore, familial aggregations of patients with these mutations reflect heritable susceptibility to lung cancer. The correlation between familial cancer history and EGFR mutations in never smokers with lung cancer requires investigation.

Methods

This was a retrospective case-control study that evaluated the prevalence of EGFR mutations in lung cancer patients with familial cancer history. Never smokers with lung cancer treated at a hospital in Taiwan between April 2012 and May 2014 were evaluated. Inclusion criteria were never smokers with non-small cell lung cancer (NSCLC). Exclusion criteria involved patients without records of familial cancer history or tumor genotype.

Results

This study included 246 never smokers with lung cancer. The study population mainly involved never smoking women with a mean age of 60 years, and the predominant tumor histology was adenocarcinoma. Lung cancer patients with familial cancer history had an increased prevalence of EGFR mutations compared to patients without family history [odds ratio (OR): 5.9; 95% confidence interval (CI): 3.3-10.6; P<0.001]. Specifically, 57 out of 85 cancer patients (67%) with familial cancer history had these mutations, while 41 out of 161 patients (25%) without family history harbored mutations. Subgroup analysis also revealed that patients with familial lung cancer history had stronger association with EGFR mutations (OR: 7.5; 95% CI: 3.4-16.3; P<0.001) compared to patients with family history of non-pulmonary cancers (OR: 5.0; 95% CI: 2.5-10.0; P<0.001).

Conclusions

The study demonstrated an increased prevalence of EGFR mutations in Taiwanese never smoking lung cancer patients with familial cancer history. Moreover, a sizable proportion of never smoking cancer patients harbored these mutations. These observations have implications for the treatment of lung cancer in never smokers.     

Chromosomal imbalances in lung adenocarcinomas with or without mutations in the epidermal growth factor receptor gene

Background and objective: Epidermal growth factor receptor (EGFR) mutations are common in lung adenocarcinomas of Asian patients, implying a good response to treatment with the EGFR tyrosine kinase inhibitors, gefitinib and erlotinib. However, the distinct chromosomal imbalances between lung adenocarcinomas with and those without EGFR mutations have not been fully elucidated. Methods: Seventy‐seven patients of surgically resected lung adenocarcinoma were analysed for the EGFR exon 19 deletion and the L858R mutation, using mutant‐enriched PCR, and for chromosomal imbalance alterations using comparative genomic hybridization. Results: EGFR mutations were detected in 42 (54.5%) patients, including 22 with the exon 19 deletion and 20 with the L858R mutation. The mean number of chromosomal arms with imbalance alterations was significantly higher in tumours with EGFR mutations than in those lacking these two mutations. The minimal regions with gain on 1q23–q31, 6p12–p21.1 and 7q11.2, and loss on 3p21, 8p22–p23, 9q33, 10q25 and 13q13, differed significantly between lung adenocarcinomas with or without EGFR mutations. However, neither EGFR mutations, nor any of the common chromosomal imbalance alterations alone, exhibited significant associations with tumour stage or disease‐specific survival of the patients. Conclusions: These results indicate that imbalance alterations at several chromosomal regions occur significantly more frequently in lung adenocarcinomas with EGFR mutations than in those without such mutations. Tumour growth‐related genes in these chromosomal regions should be further investigated to improve our understanding of the common genetic alterations in lung adenocarcinomas with EGFR mutations.     

RT-LAMP与LAMP法检测结核分枝杆菌的效果比较

目的 比较RT-LAMP、LAMP、L-J法检测MTB的效果,为结核病的快速诊断提供依据。方法 用常见的非结核分枝杆菌与其他常见的呼吸道感染病菌(金黄色葡萄球菌、铜绿假单胞菌、肺炎克雷伯杆菌)检测特异性,并用酶切实验对结核分枝杆菌特异性产物进行酶切鉴定;用RT-LAMP、LAMP、L-J法检测100名结核病患者和22名来自无结核病患者痰标本中的结核分枝杆菌测试灵敏性;对浓度为1 ng/μL H37Rv标准株进行对倍比稀释用于RT-LAMP极限检测实验。结果 RT-LAMP、LAMP、L-J方法的阳性率分别为100%,92%和88%;RT-LAMP和L-J, RT-LAMP和LAMP差异均有统计学意义;RT-LAMP的灵敏度比LAMP高10倍;RT-LAMP不仅可以识别活菌,并且能够重复检测MTB的单一拷贝。结论 RT-LAMP检测效果优于LAMP及L-J,适于基层医院结核病诊断的推广和应用。