Characterisation of the Cinnamomumparthenoxylon (Jack) Meisn (Lauraceae) transcriptome using Illumina paired-end sequencing and EST-SSR markers development for population genetics.

Cinnamomumparthenoxylon is an endemic and endangered species with significant economic and ecological value in Vietnam. A better understanding of the genetic architecture of the species will be useful when planning management and conservation. We aimed to characterize the transcriptome of C.parthenoxylon, develop novel molecular markers, and assess the genetic variability of the species. First, transcriptome sequencing of five trees (C.parthenoxylon) based on root, leaf, and stem tissues was performed for functional annotation analysis and development of novel molecular markers. The transcriptomes of C.parthenoxylon were analyzed via an Illumina HiSeqTM 4000 sequencing system. A total of 27,363,199 bases were generated for C.parthenoxylon. De novo assembly indicated that a total of 160,435 unigenes were generated (average length = 548.954 bp). The 51,691 unigenes were compared against different databases, i.e. COG, GO, KEGG, KOG, Pfam, Swiss-Prot, and NR for functional annotation. Furthermore, a total of 12,849 EST-SSRs were identified. Of the 134 primer pairs, 54 were randomly selected for testing, with 15 successfully amplified across nine populations of C.parthenoxylon. We uncovered medium levels of genetic diversity (PIC = 0.52, Na = 3.29, Ne = 2.18, P = 94.07%, Ho = 0.56 and He = 0.47) within the studied populations. The molecular variance was 10% among populations and low genetic differentiation (Fst = 0.06) indicated low gene flow (Nm = 2.16). A reduction in the population size of C.parthenoxylon was detected using BOTTLENECK (VP population). The structure analysis suggested two optimal genetic clusters related to gene flow among the populations. Analysis of molecular variance (AMOVA) revealed higher genetic variation within populations (90%) than among populations (10%). The UPGMA approach and DAPC divided the nine populations into three main clusters. Our findings revealed a significant fraction of the transcriptome sequences and these newlydeveloped novel EST-SSR markers are a very efficient tool for germplasm evaluation, genetic diversity and molecular marker-assisted selection in C.parthenoxylon. This study provides comprehensive genetic resources for the breeding and conservation of different varieties of C.parthenoxylon.

Predictive ecological niche model for Cinnamomumparthenoxylon (Jack) Meisn. (Lauraceae) from Last Glacial Maximum to future in Vietnam.

Cinnamomumparthenoxylon (Jack) Meisn. is a tree in genus Cinnamomum that has been facing global threats due to forest degradation and habitat fragmentation. Many recent studies aim to describe habitats and assess population and species genetic diversity for species conservation by expanding afforestation models for this species. Understanding their current and future potential distribution plays a major role in guiding conservation efforts. Using five modern machine-learning algorithms available on Google Earth Engine helped us evaluate suitable habitats for the species. The results revealed that Random Forest (RF) had the highest accuracy for model comparison, outperforming Support Vector Machine (SVM), Classification and Regression Trees (CART), Gradient Boosting Decision Tree (GBDT) and Maximum Entropy (MaxEnt). The results also showed that the extremely suitable ecological areas for the species are mostly distributed in northern Vietnam, followed by the North Central Coast and the Central Highlands. Elevation, Temperature Annual Range and Mean Diurnal Range were the three most important parameters affecting the potential distribution of C.parthenoxylon. Evaluation of the impact of climate on its distribution under different climate scenarios in the past (Last Glacial Maximum and Mid-Holocene), in the present (Worldclim) and in the future (using four climate change scenarios: ACCESS, MIROC6, EC-Earth3-Veg and MRI-ESM2-0) revealed that of C.parthenoxylon would likely expand to the northeast, while a large area of central Vietnam will gradually lose its adaptive capacity by 2100.

Microplastics contamination in water supply system and treatment processes.

Due to global demand, millions of tons of plastics have been widely consumed, resulting in the widespread entry of vast amounts of microplastic particles into the environment. The presence of microplastics (MPs) in water supplies, including bottled water, has undergone systematic review, assessing the potential impacts of MPs on humans through exposure assessment. The main challenges associated with current technologies lie in their ability to effectively treat and completely remove MPs from drinking and supply water. While the risks posed by MPs upon entering the human body have not yet been fully revealed, there is a predicted certainty of negative impacts. This review encompasses a range of current technologies, spanning from basic to advanced treatments and varying in scale. However, given the frequent detection of MPs in drinking and bottled water, it becomes imperative to implement comprehensive management strategies to address this issue effectively. Consequently, integrating current technologies with management options such as life-cycle assessment, circular economy principles, and machine learning is crucial to eliminating this pervasive problem.

Two New Benzoquinone Derivatives from Vietnamese Knema globularia Stems.

The chemical investigation of the stems of Knema globularia led to the isolation of two new benzoquinones derivatives, embenones A and B (1 and 2), along with three known compounds (3-5). The structures of the isolated compounds were determined using spectroscopic techniques, including HRESIMS, 1D and 2D NMR, in conjunction with comparison to existing literature data. Compounds 1 and 2 represent new carbon skeletons in nature. Furthermore, all isolated compounds were evaluated for their α-glucosidase inhibitory activity, with compounds 1-3 exhibiting superior potency relative to the positive control (acarbose, IC50 331 µM). Their IC50 values ranged from 1.40 to 96.1 µM.

Non-Heme Iron Enzymes Catalyze Heterobicyclic and Spirocyclic Isoquinolone Core Formation in Piperazine Alkaloid Biosynthesis.

We report the discovery and biosynthesis of new piperazine alkaloids-arizonamides, and their derived compounds-arizolidines, featuring heterobicyclic and spirocyclic isoquinolone skeletons, respectively. Their biosynthetic pathway involves two crucial non-heme iron enzymes, ParF and ParG, for core skeleton construction. ParF has a dual function facilitating 2,3-alkene formation of helvamide, as a substrate for ParG, and oxidative cleavage of piperazine. Notably, ParG exhibits catalytic versatility in multiple oxidative reactions, including cyclization and ring reconstruction. A key amino acid residue Phe67 was characterized to control the formation of the constrained arizonamide B backbone by ParG.

Tuning Molecular Motion Enhances Intrinsic Fluorescence in Peptide Amphiphile Nanofibers.

Peptide amphiphiles (PAs) are highly tunable molecules that were recently found to exhibit aggregation-induced emission (AIE) when they self-assemble into nanofibers. Here, we leverage decades of molecular design and self-assembly study of PAs to strategically tune their molecular motion within nanofibers to enhance AIE, making them a highly useful platform for applications such as sensing, bioimaging, or materials property characterization. Since AIE increases when aggregated molecules are rigidly and closely packed, we altered the four most closely packed amino acids nearest to the hydrophobic core by varying the order and composition of glycine, alanine, and valine pairs. Of the six PA designs studied, C16VVAAK2 had the highest quantum yield at 0.17, which is a more than 10-fold increase from other PA designs including the very similar C16AAVVK2, highlighting the importance of precise amino acid placement to anchor rigidity closest to the core. We also altered temperature to increase AIE. C16VVAAK2 exhibited an additional 4-fold increase in maximum fluorescence intensity when the temperature was raised from 5 to 65 °C. As the temperature increased, the secondary structure transitioned from ß-sheet to random coil, indicating that further packing an already aligned molecular system makes it even more readily able to transfer energy between the electron-rich amides. This work both unveils a highly fluorescent AIE PA system design and sheds insights into the molecular orientation and packing design traits that can significantly enhance AIE in self-assembling systems.

An Electrochemical Aptasensor for the Detection of Freshwater Cyanobacteria.

Aphanizomenon is a genus of cyanobacteria that is filamentous and nitrogen-fixing and inhabits aquatic environments. This genus is known as one of the major producers of cyanotoxins that can affect water quality after the bloom period. In this study, an electrochemical aptasensor is demonstrated using a specific aptamer to detect Aphanizomenon sp. ULC602 for the rapid and sensitive detection of this bacterium. The principal operation of the generated aptasensor is based on the conformational change in the aptamer attached to the electrode surface in the presence of the target bacterium, resulting in a decrease in the current peak, which is measured by square-wave voltammetry (SWV). This aptasensor has a limit of detection (LOD) of OD750~0.3, with an extension to OD750~1.2 and a sensitivity of 456.8 µA·OD750-1·cm-2 without interference from other cyanobacteria. This is the first aptasensor studied that provides rapid detection to monitor the spread of this bacterium quickly in a targeted manner.

Pertussis seasonal variation in Northern Vietnam: the evidence from a tertiary hospital.

BACKGROUND: Pertussis is a highly contagious and dangerous respiratory disease that threatens children's health in many countries, including Vietnam, despite vaccine coverage. From 2015 to 2018, Vietnam experienced an increasing number of pertussis patients. Therefore, this study aimed to investigate the trend and examine the seasonal variations of pertussis in North Vietnam. METHODS: Data were collected from medical records of all under-5-year-old inpatients admitted to the National Children's Hospital in Hanoi, Vietnam (VNCH) 2015-2018. A descriptive analysis was performed to describe the distribution of incident cases by year and season. Linear multivariable regression was conducted to investigate the association between the incidence of cases and seasonality adjusted by age and vaccination status. RESULTS: We identified 1063 laboratory-confirmed patients during 2015-2018, including 247 (23.2%) severe patients. The number of pertussis patients admitted to VNCH per 1000 hospitalizations was 3.2 in 2015, compared to 1.9, 3.1, and 2.1 in 2016, 2017, and 2018, respectively. Outbreaks occurred biennially; however, there was no significant difference in the number of severe patients over this period. Most cases occurred in the hot season (509 patients, or nearly half of the study population). With the adjustment of the vaccination rate and average age, the risk of pertussis-associated hospitalization in the mild season and the hot season was 21% (95% CI [0.12; 0.3]) and 15% (95% CI [0.05; 0.25]) higher than that in the warm season, respectively. The rate of hospitalizations was high in the mild season (28.9%) and the warm season (30.8%), nearly twice as much as that in the hot season; nevertheless, the death rate was only striking high in the mild season, about 5-6 times as much as those in the other seasons. CONCLUSION: The pertussis incidence in Northern Vietnam varied between seasons, peaking in the hot season (April-July). However, severe patients and deaths increased in the mild season (December-March). Interventions, for example, communication activities on pertussis and vaccination, are of immense importance in lowering the prevalence of pertussis. In addition, early diagnoses and early warnings performed by health professionals should be encouraged.

Gustavson syndrome is caused by an in-frame deletion in RBMX associated with potentially disturbed SH3 domain interactions.

RNA binding motif protein X-linked (RBMX) encodes the heterogeneous nuclear ribonucleoprotein G (hnRNP G) that regulates splicing, sister chromatid cohesion and genome stability. RBMX knock down experiments in various model organisms highlight the gene's importance for brain development. Deletion of the RGG/RG motif in hnRNP G has previously been associated with Shashi syndrome, however involvement of other hnRNP G domains in intellectual disability remain unknown. In the current study, we present the underlying genetic and molecular cause of Gustavson syndrome. Gustavson syndrome was first reported in 1993 in a large Swedish five-generation family presented with profound X-linked intellectual disability and an early death. Extensive genomic analyses of the family revealed hemizygosity for a novel in-frame deletion in RBMX in affected individuals (NM_002139.4; c.484_486del, p.(Pro162del)). Carrier females were asymptomatic and presented with skewed X-chromosome inactivation, indicating silencing of the pathogenic allele. Affected individuals presented minor phenotypic overlap with Shashi syndrome, indicating a different disease-causing mechanism. Investigation of the variant effect in a neuronal cell line (SH-SY5Y) revealed differentially expressed genes enriched for transcription factors involved in RNA polymerase II transcription. Prediction tools and a fluorescence polarization assay imply a novel SH3-binding motif of hnRNP G, and potentially a reduced affinity to SH3 domains caused by the deletion. In conclusion, we present a novel in-frame deletion in RBMX segregating with Gustavson syndrome, leading to disturbed RNA polymerase II transcription, and potentially reduced SH3 binding. The results indicate that disruption of different protein domains affects the severity of RBMX-associated intellectual disabilities.

Streamlining apartment provider evaluation: A spherical fuzzy multi-criteria decision-making model.

In the context of the thriving real estate market in developing countries like Vietnam, understanding consumer preferences and effectively addressing them through a comprehensive multi-criteria decision-making (MCDM) framework is paramount for real estate providers. This study presents a two-stage MCDM model that integrates the Delphi technique and the Technique for Order Preference by Similarity to Ideal Solution (TOPSIS) based on Spherical Fuzzy Sets (SFSs). Initially, the SF-Delphi technique validates critical criteria influencing customers' apartment selection in Vietnam. Secondly, the SF-TOPSIS method evaluates the top ten apartment providers. To ensure robustness and validity, a comparative analysis compares the results with those from the Intuitionistic Fuzzy TOPSIS (IF-TOPSIS) and Fuzzy TOPSIS (F-TOPSIS) methods. Subsequently, five rank correlation coefficients (Spearman, Kendall, Goodman-Kruskal, Weighted rank measure of correlation, Weighted Similarity) are used to assess the relationships between various TOPSIS techniques applied to apartment suppliers in Vietnam. The correlation coefficients demonstrate strong agreement among the TOPSIS methods, with the smallest coefficient being 0.7778, surpassing the threshold of 0.7. This high level of consistency confirms the efficacy of the proposed TOPSIS approach with different Fuzzy Sets in reliably evaluating customers' preferences for apartment suppliers. Notably, the legal aspect's prominence underscores its critical role in shaping customer choices, emphasizing the significance of considering legal factors in the context of apartment supply and demand in Vietnam. Furthermore, using SFSs makes this approach particularly suited to capture consumer perceptions within the dynamic and uncertain business environment characterized by volatility, uncertainty, complexity, and ambiguity (VUCA).

Pilot and full scale applications of floating treatment wetlands for treating diffuse pollution.

Floating treatment wetlands (FTW) are nature-based solutions for the purification of open water systems such as rivers, ponds, and lakes polluted by diffuse sources as untreated or partially treated domestic wastewater and agricultural run-off. Compared with other physicochemical and biological technologies, FTW is a technology with low-cost, simple configuration, easy to operate; has a relatively high efficiency, and is energy-saving, and aesthetic. Water remediation in FTWs is supported by plant uptake and the growth of a biofilm on the water plant roots, so the selection of the macrophyte species is critical, not only to pollutant removal but also to the local ecosystem integrity, especially for full-scale implementation. The key factors such as buoyant frame/raft, plant growth support media, water depth, seasonal variation, and temperature have a considerable role in the design, operation, maintenance, and pollutant treatment performance of FTW. Harvesting is a necessary process to maintain efficient operation by limiting the re-pollution of plants in the decay phase. Furthermore, the harvested plant biomass can serve as a green source for the recovery of energy and value-added products.

Assessment of availability, readiness, and challenges for scaling-up hypertension management services at primary healthcare facilities, Central Highland region, Vietnam, 2020.

INTRODUCTION: Vietnam aims for 95% of commune health stations (CHSs) to have functional hypertension management programs by 2025. However, limited resources may impede the Central Highland region health system from achieving this goal. We assessed the availability and readiness of hypertension management services at CHSs in the Central Highland region and identified challenges to facilitate evidence-based planning. METHODS: We used a mixed-methods cross-sectional design to assess hypertension management services using WHO's service availability and readiness assessment (SARA) tools in all 579 CHSs in the region, combined with twenty in-depth interviews of hypertension program focal points at communal, district, and provincial levels in all four provinces. We descriptively analyzed quantitative data and thematically analyzed qualitative data. RESULTS: Hypertension management services were available at 65% of CHSs, and the readiness of the services was 62%. The urban areas had higher availability and readiness indices in most domains (basic amenities, basic equipment, and essential medicines) compared to rural areas, except for staff and training. The qualitative results showed a lack of trained staff and ambiguity in national hypertension treatment guidelines, insufficient essential medicines supply mechanism, and low priority and funding limitations for the hypertension program. CONCLUSION: The overall availability and readiness for hypertension diagnosis and management service at CHSs in the Central Highland region were low, reflecting inadequate capacity of the primary healthcare facilities. Some measures to strengthen hypertension programs in the region might include increased financial support, ensuring a sufficient supply of basic medicines, and providing more specific treatment guidelines.

Experience With the Routine Use of Electronic Patient-Reported Outcome Measures for Patients With Lung Cancer.

PURPOSE: The Christie NHS Foundation Trust launched their electronic patient-reported outcome measures (ePROMs) service in January 2019 in the routine clinical setting. The lung cancer questionnaires consist of 14 symptom items, adapted from the Common Terminology Criteria for Adverse Events (version 5.0) and the EuroQol EQ-5D-5L quality-of-life (QoL) tool. Patients with lung cancer are invited to complete questionnaires assessing their symptoms and QoL using an online platform. METHODS: The ePROM responses and clinical, pathologic, and treatment data for patients who completed the questionnaires between January 2019 and December 2020 were extracted from electronic medical records. The symptom and QoL scores of patients who completed baseline pretreatment ePROMs and also those who completed ePROMs pre- and postpalliative lung systemic anticancer therapy (SACT) or radical thoracic radiotherapy were evaluated. Pretreatment questionnaires were analyzed according to age, Eastern Cooperative Oncology Group performance status (ECOG PS), and Adult Comorbidity Evaluation-27 (ACE-27) comorbidity score. RESULTS: One thousand four hundred eighty patients with lung cancer were included. There were no statistically significant differences in symptoms and QoL scores between age groups. Cough (P = .006) and EQ-5D-5L mobility scores (P = .006) were significantly worse for patients with an ECOG PS of 0-1. Dyspnea (P = .035), hemoptysis (P = .023), nausea (P = .041), mobility (P = .004), and self-care (P = .0420) were significantly worse for those with higher ACE-27 scores (2-3 v 0-1). Palliative SACT was associated with a significant improvement in cough (P < .001) and hemoptysis (P = .025), but significantly negatively affected mobility (P = .013). Patients receiving radical thoracic radiotherapy reported a significant improvement in hemoptysis (P = .042) but worse pain (P = .002) and fatigue (P = .01). Other changes in symptom and QoL scores were not significant. CONCLUSION: The symptoms and QoL reported at baseline and before and after both palliative SACT and radical thoracic radiotherapy are clinically relevant and meaningful. We have demonstrated that routine implementation of ePROMs into clinical practice is feasible and can inform clinical practice and future research.

Roles of microalgae-based biofertilizer in sustainability of green agriculture and food-water-energy security nexus.

For years, agrochemical fertilizers have been used in agriculture for crop production. However, intensive utilization of chemical fertilizers is not an ecological and environmental choice since they are destroying soil health and causing an emerging threat to agricultural production on a global scale. Under the circumstances of the increasing utilization of chemical fertilizers, cultivating microalgae to produce biofertilizers would be a wise solution since desired environmental targets will be obtained including (1) replacing chemical fertilizer while improving crop yields and soil health; (2) reducing the harvest of non-renewable elements from limited natural resources for chemical fertilizers production, and (3) mitigating negative influences of climate change through CO2 capture through microalgae cultivation. Recent improvements in microalgae-derived-biofertilizer-applied agriculture will be summarized in this review article. At last, the recent challenges of applying biofertilizers will be discussed as well as the perspective regarding the concept of circular bio-economy and sustainable development goals (SDGs).

The Complete Chloroplast Genome of An Ophiorrhiza baviensis Drake Species Reveals Its Molecular Structure, Comparative, and Phylogenetic Relationships.

Ophiorrhiza baviensis Drake, a flowering medical plant in the Rubiaceae, exists uncertainly within the Ophiorrhiza genus' evolutionary relationships. For the first time, the whole chloroplast (cp) genome of an O. baviensis Drake species was sequenced and annotated. Our findings demonstrate that the complete cp genome of O. baviensis is 154,770 bp in size, encoding a total of 128 genes, including 87 protein-coding genes, 8 rRNAs, and 33 tRNAs. A total of 59 SSRs were screened in the studied cp genome, along with six highly variable loci, which can be applied to generate significant molecular markers for the Ophiorrhiza genus. The comparative analysis of the O. baviensis cp genome with two published others of the Ophiorrhiza genus revealed a high similarity; however, there were some notable gene rearrangements in the O. densa plastome. The maximum likelihood phylogenetic trees were constructed based on the concatenation of the rps16 gene and the trnL-trnF intergenic spacer sequence, indicating a close relationship between the studied O. baviensis and other Ophiorrhiza. This study will provide a theoretical molecular basis for identifying O. baviensis Drake, as well as species of the Ophiorrhiza genus, and contribute to shedding light on the chloroplast genome evolution of Rubiaceae.

The Biosynthetic Gene Cluster of Mushroom-Derived Antrocin Encodes Two Dual-Functional Haloacid Dehalogenase-like Terpene Cyclases.

(-)-Antrocin (1), produced by the medicinal mushroom Antrodia cinnamomea, is a potent antiproliferative compound. The biosynthetic gene cluster of 1 was identified, and the pathway was characterized by heterologous expression. We characterized a haloacid dehalogenase-like terpene cyclase AncC that biosynthesizes the drimane-type sesquiterpene (+)-albicanol (2) from farnesyl pyrophosphate (FPP). Biochemical characterization of AncC, including kinetic studies and mutagenesis, demonstrated the functions of two domains: a terpene cyclase (TC) and a pyrophosphatase (PPase). The TC domain first cyclizes FPP to albicanyl pyrophosphate, and the PPase domain then removes the pyrophosphate to form 2. Intriguingly, AncA (94 % sequence identity to AncC), in the same gene cluster, converts FPP into (R)-trans-γ-monocyclofarnesol instead of 2. Notably, Y283/F375 in the TC domain of AncA serve as a gatekeeper in controlling the formation of a cyclofarnesoid rather than a drimane-type scaffold.

Growth of freshwater cyanobacterium Aphanizomenon sp. ULC602 in different growing and nutrient conditions.

Aphanizomenon sp. ULC602, recently isolated in a Belgian lake, is a filamentous, nitrogen-fixing, freshwater cyanobacterium that is one of the primary producers of cyanotoxins following its bloom formation, causing water contamination. This study aims to evaluate the effects of growing conditions and essential nutrients on the growth of Aphanizomenon sp. ULC602 via its production of chlorophyll-a (Chlo-a). Our results indicated that this bacterium could grow well at temperatures ranging from 18 to 25°C with an optimal pH of 6.0-7.5 under continuous lighting. It grew slowly in the absence of a carbon source or at lower carbon concentrations. The addition of nitrogen from nitrate and urea led to a less than 50% reduction of Chlo-a content compared to the medium lacking nitrogen. The iron bioavailability significantly stimulated the Chlo-a production, but it was saturated by an iron concentration of 0.115 mM. Moreover, a decrease in Chlo-a biomass was observed under sulfur deficiency. The bacterium could not grow well in media containing various phosphorus sources. In conclusion, as the growth and consequent forming bloom of cyanobacteria can be stimulated or inhibited by environmental conditions and eutrophication, our investigation could contribute to further studies to control the blooming of the target bacterium in freshwater.

Ce3+/Ce4+-Doped ZrO2/CuO Nanocomposite for Enhanced Photocatalytic Degradation of Methylene Blue under Visible Light.

In recent years, photocatalysis has been used as an environmentally friendly method for the degradation of organic pigments in water. In this study, Ce3+/Ce4+-doped ZrO2/CuO as a mixed semiconductor oxide was successfully prepared by a one-step hydrothermal method. The Ce3+/Ce4+-doped ZrO2/CuO has shown high degradation efficiency of methylene blue (MB), and the maximum degradation percentage was observed to be 94.5% at 180 min under irradiation visible light. The photocatalytic activity increases significantly by doping Ce3+/Ce4+ in ZrO2/CuO for MB degradation. Ce3+/Ce4+ doping is shown to reduce the (e-/h+) recombination rate and improve the charge transfer, leading to enhanced photocatalytic activity of materials. The materials were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), FTIR, EDS, BET and diffuse reflectance spectroscopy (DRS).

Highly sensitive detection of EGFR L858R mutation at the mRNA level.

The missense mutation EGFR L858R implies increased sensitivity to EGFR tyrosine kinase inhibitor (TKIs) therapy, despite a significant non-response rate. Currently, detection of EGFR L858R mutation is mostly DNA based, therefore, the allele-specific expression level of the mutated gene and its clinical relevance is hidden. Based on the extendable blocking probes and hot-start protocol for reverse transcription, we have developed and validated a novel one-step realtime RT-PCR assay that enables detection of EGFR L858R mutation at the mRNA level. This RNA-based assay was able to detect the EGFR L858R mutation in a 10,000-fold excess of its wildtype counterpart, indicating an analytical sensitivity of 0.01%. In comparison to the reference DNA-based assay, the RNA-based assay further detected the EGFR L858R mutation in significantly additional formalin-fixed paraffin-embedded (FFPE) samples (19.2% vs 15.0%). Interestingly, our data showed that the relative mRNA levels of EGFR L858R mutation varied greatly in tumor tissues (∼4 logs); and the circulating mRNA of EGFR L858R mutation was detectable in plasma of NSCLC patients. This novel RNA-based PCR assay provides a simple and ultrasensitive tool for detection of EGFR L858R mutation at the mRNA level as a new class of biomarkers.