Computed Tomography-Based Intratumor Heterogeneity Predicts Response to Immunotherapy Plus Chemotherapy in Esophageal Squamous Cell Carcinoma.

RATIONALE AND OBJECTIVES: This study explored the intratumor heterogeneity (ITH) of esophageal squamous cell carcinoma (ESCC) using computed tomography (CT) and investigated the value of CT-based ITH in predicting the response to immune checkpoint inhibitor (ICI) plus chemotherapy in patients with ESCC. MATERIALS AND METHODS: This retrospective study included 416 patients with ESCC who received ICI plus chemotherapy at two independent hospitals between January 2019 and July 2022. Multiparametric CT features were extracted from ESCC lesions and screened using hierarchical clustering and dimensionality reduction algorithms. Logistic regression and machine learning models based on selected features were developed to predict treatment response and validated in separate datasets. ITH was quantified using the score calculated by the best-performing model and visualized through feature clustering and feature contribution heatmaps. A gene set enrichment analysis (GSEA) was performed to identify the biological pathways underlying the CT-based ITH. RESULTS: The extreme gradient boosting model based on CT-derived ITH had higher discriminative power, with areas under the receiver operating characteristic curve of 0.864 (95% confidence interval [CI]: 0.774-0.954) and 0.796 (95% CI: 0.698-0.893) in the internal and external validation sets. The CT-based ITH pattern differed significantly between responding and non-responding patients. The GSEA indicated that CT-based ITH was associated with immunity-, keratinization-, and epidermal cell differentiation-related pathways. CONCLUSION: CT-based ITH is an effective biomarker for identifying patients with ESCC who could benefit from ICI plus chemotherapy. Immunity-, keratinization-, and epidermal cell differentiation-related pathways may influence the patient's response to ICI plus chemotherapy.

The evolution of land policies in China from 1980 to 2019: a policy-text based analysis.

Land resources are indispensable for the development of a country, especially for a country such as China with a very high population relative to its land area. Since reforms were introduced and liberalization was adopted, many policies have been promulgated for the protection and utilization of land resources. In the present study, more than 2000 land policies that have been formulated and promulgated in China from 1980 to 2019 were examined and statistically analyzed to qualitatively and quantitatively introduce the evolution of these policies in China. This paper mainly studies land policy of China from three perspectives as follows: policy intensity, policy tools, and policy targets, and three principal observations emerged from these investigations. First, during the past 40 years, the quantity of land policies introduced showed an increasing trend, while policy intensity became increasingly stabilized. Second, compared with regulatory policies and stimulative policies, it seems that China prefers to adopt guiding policies to manage land resources. Third, the main purpose of land policies issued during 1980-2019 is land market regulation, with subordinate policies for land resources protection.

Predicting response to immunotherapy plus chemotherapy in patients with esophageal squamous cell carcinoma using non-invasive Radiomic biomarkers.

OBJECTIVES: To develop and validate a radiomics model for evaluating treatment response to immune-checkpoint inhibitor plus chemotherapy (ICI + CT) in patients with advanced esophageal squamous cell carcinoma (ESCC). METHODS: A total of 64 patients with advance ESCC receiving first-line ICI + CT at two centers between January 2019 and June 2020 were enrolled in this study. Both 2D ROIs and 3D ROIs were segmented. ComBat correction was applied to minimize the potential bias on the results due to different scan protocols. A total of 788 features were extracted and radiomics models were built on corrected/uncorrected 2D and 3D features by using 5-fold cross-validation. The performance of the radiomics models was assessed by its discrimination, calibration and clinical usefulness with independent validation. RESULTS: Five features and support vector machine algorithm were selected to build the 2D uncorrected, 2D corrected, 3D uncorrected and 3D corrected radiomics models. The 2D radiomics models significantly outperformed the 3D radiomics models in both primary and validation cohorts. When ComBat correction was used, the performance of 2D models was better (p = 0.0059) in the training cohort, and significantly better (p < 0.0001) in the validation cohort. The 2D corrected radiomics model yielded the optimal performance and was used to build the nomogram. The calibration curve of the radiomics model demonstrated good agreement between prediction and observation and the decision curve analysis confirmed the clinical utility. CONCLUSIONS: The easy-to-use 2D corrected radiomics model could facilitate noninvasive preselection of ESCC patients who would benefit from ICI + CT.

Strong expression of Id-1 in metastatic lymph nodes from esophageal squamous cell carcinoma is associated with better clinical outcome.

BACKGROUND: Over-expression of inhibitor of differentiation or DNA binding 1 (Id-1) is associated with poor prognosis in esophageal squamous cell carcinoma (ESCC). However, some biomarkers discordant expression in metastasis has been reported previously. We aimed to confirm possible differential expression and prognostic value of Id-1 in paired metastatic lymph node (PMLN). METHODS: Expression of Id-1 in primary tumors (PT) and paired regional metastatic lymph nodes of ESCC were evaluated with immunohistochemical (IHC) analysis. Statistical analysis of Kaplan-Meier method was performed to test the prognostic significance of Id-1 expression. RESULTS: The expression of Id-1 was down-regulated in metastatic lymph nodes compared with primary esophageal tumors (P<0.001). Patients with 1 to 2 lymph nodes involved had significantly higher Id-1 expression in metastatic lymph nodes (P=0.028). The similar association was observed between a ratio of involved to examined lymph nodes ≤0.2 and high level Id-1 expression in lymphatic metastases (P=0.011). Better overall survival with statistical significance was observed in patients with higher level Id-1 expression in metastatic lymph nodes (P=0.015). The results of Id-1 expression in metastatic lymph node and paired PT was to predict prognosis effective in out cohort (P=0.035). CONCLUSIONS: The level of Id-1 protein expression was down-regulated from PT to metastatic lymph node. It was contrary to previous studies that strong expression of Id-1 in metastatic lymph nodes was associated with better clinical outcomes in patients with stage T3N1-3M0 ESCC.

Electroacupuncture exerts neuroprotective effects on ischemia/reperfusion injury in JNK knockout mice: the underlying mechanism.

Simple regulation of c-Jun N-terminal kinase (JNK) or p38 mitogen-activated protein kinase (MAPK) pathways is not enough to trigger cell apoptosis. However, activation of the stress activated pathway (JNK/p38 MAPK) together with inhibition of the growth factor activated extracellular signal-regulated kinase (ERK) pathway can promote cell apoptosis. We hypothesized that inhibition of the JNK or p38 pro-apoptotic pathway and activating the ERK pathway could be the mechanism of anti-apoptosis following cerebral ischemia/reperfusion injury. To investigate the mechanism of the protective effect of electroacupuncture on cerebral ischemia/reperfusion injury in JNK knockout mice, mouse models of cerebral ischemia/reperfusion injury were established by Longa's method. Electroacupuncture was conducted at acupoints Chize (LU5), Hegu (LI4), Sanyinjiao (SP6) and Zusanli (ST36) 1.5 hours after ischemia/reperfusion injury for 20 minutes, once a day. The neurological function was evaluated using neurological deficit scores. The expression of phospho-extracellular signal-regulated kinase (p-ERK) and phospho-p38 (p-p38) in JNK knockout mice was detected using double-labeling immunofluorescence and western blot assay. The mRNA expression of ERK and p38 was measured by quantitative real-time polymerase chain reaction. Electroacupuncture improved neurological function, increased the immunoreactivity and relative expression of p-ERK and reduced that of p-p38 in the cerebral cortex and hippocampus on the injured side. Electroacupuncture increased mRNA expression of ERK, but decreased that of p38 in the cerebral cortex and hippocampus on the injured side. In conclusion, electroacupuncture upregulated the protective ERK pathway and inhibited the pro-apoptotic p38 pathway, thereby exerting a neuroprotective effect and improving the neurological function in JNK knockout mice.

A pair of cyclopeptide epimers from the seeds of Celosia argentea.

Two cyclopeptides, celogentin L (1) and its epimer lyciumin A (2) were firstly isolated from Celosia argentea L.. The planar structures of the two compounds were fully determined by spectroscopic data, including 1D-, 2D-NMR, and HR-ESI/MS. The absolute configurations of amino acid components were assigned via chiral-phase HPLC analyses after acid hydrolysis. Furthermore, the configuration of C-N linkage at the glycine Cα was elucidated by extensive analyses of 2D-NMR and comparison of the experimental and calculated electronic circular dichroism (ECD) spectra. Cytotoxicity of the two compounds against human alveolar epithelial A549, hepatocellular carcinoma HepG2, and cervical cancer Hela cell lines was assayed. Although both of them were inactive in these cells, the present findings add new facets for the chemistry of Celosia argentea.

LncRNA NONRATT021972 Was Associated with Neuropathic Pain Scoring in Patients with Type 2 Diabetes.

BACKGROUND: Long noncoding RNAs were involved in the processes of diabetes. Our study was aimed to explore clinical potential of LncRNA NONRATT021972 in diabetic neuropathic pain and investigate detailed mechanisms. METHODS: 154 patients with type 2 diabetes were enrolled as experimental group paired with control. Patients without diabetes but neuropathy were enrolled to explore exclusive role of LncRNA NONRATT021972 in neuropathy. Real-time PCR and ELISA were performed to examine expression of LncRNA and TNF-α in flood. Neuropathic pain scores were calculated with data from NPQ. Streptozotocin was used for SD adult male rats to establish diabetes for NONRATT021972 siRNA or saline treatment. Neuropathic pain behaviors and expression of TNF-α were assessed. RESULT: Patients with type 2 diabetes had a significantly higher concentration of LncRNA NONRATT021972 in blood and more severe symptoms of neuropathic pain. LncRNA NONRATT021972 was positively associated with neuropathic pain scores of type 2 diabetes. TNF-α level increased in patients with type 2 diabetes. Animal experiment showed that LncRNA NONRATT021972 siRNA attenuated inflammation via decreasing TNF-α and alleviated neuropathic pain. CONCLUSION: LncRNA NONRATT021972 increased in type 2 diabetes and was positively associated with neuropathic pain scoring in type 2 diabetes. LncRNA NONRATT021972 exacerbated neuropathic pain via TNF-α related pathways.

Enantioselective Recognition for Many Different Kinds of Chiral Guests by One Chiral Receptor Based on Tetraphenylethylene Cyclohexylbisurea.

A neutral chiral receptor based on TPE cyclohexylbisurea was synthesized and could discriminate the enantiomers of many different kinds of chiral reagents, including chiral acidic compounds, basic compounds, amino acids, and even neutral alcohols. The (1)H NMR spectra disclosed that the ability of chiral recognition could be ascribed to the multiple hydrogen bonds and CH-π interactions between the TPE urea receptor and the enantiomer of the chiral guest, which led to the selective aggregation of the receptor with one of the two enantiomers. This result exhibited a great potential in enantiomer discernment and high-throughput analysis of enantiomer composition of these chiral analytes by one chiral AIE molecule.

Comparison of pain relief between patient-controlled epidural analgesia and patient-controlled intravenous analgesia for patients undergoing spinal fusion surgeries.

INTRODUCTION: This meta-analysis aimed to compare the postoperative analgesic effects of patient-controlled epidural analgesia (PCEA) and patient-controlled intravenous analgesia (PCIA) for patients undergoing spinal fusion surgeries. METHOD: Relevant articles were identified using computerized and manual search strategies. Statistical analyses were undertaken by the CMA 2.0 statistical software. RESULTS: Nine cohort studies with a total of 436 patients undergoing spinal fusion surgeries were incorporated in the present meta-analysis. There were significant differences between the PCEA and PCIA groups in the visual analogue scale score of patients undergoing spinal fusion [standardized mean difference = 0.27, 95 % confidence interval (95 % CI) = 0.070-0.470, P = 0.008]. However, no obvious difference was observed in the rate of side effects between the PCIA and PCEA groups (side effects: odds ratio = 0.957, 95 % CI = 0.536-1.708, P = 0.882). CONCLUSION: Our findings suggested that PCEA may be more effective in relieving pain than PCIA for patients undergoing spinal fusion surgeries.

The ABO blood group predicts survival in esophageal squamous cell carcinoma in patients who ever smoked: a retrospective study from China.

The association between clinical characteristics, prognosis, and the ABO blood group of esophageal squamous cell carcinoma (ESCC) is rarely discussed. The aims of the current study were to investigate the correlation of the ABO blood group with the clinicopathological characteristics in a consecutive cohort of patients with ESCC and to assess whether the ABO blood group was associated with prognosis. A total of 511 patients with locoregional ESCC who underwent curative treatment were retrospectively analyzed at a single institution between January 2007 and December 2008. The relationship between the ABO blood group and clinicopathological variables was assessed by chi-squared analysis. The Kaplan-Meier method was used to estimate the 5-year overall survival (OS). The Cox proportional hazards model was used in univariate and multivariate analyses of OS. There were no significant differences in the clinicopathological characteristics among patients with different ABO blood groups. The 5-year OS rates were 50.0 % for patients with blood type A, 45.4 % for type B, 50.8 % for type O, and 60.7 % for type AB. In a subgroup analysis of 321 patients who ever smoked, the B/O group had a poorer OS compared with the A/AB group (p = 0.0245). Multivariate analysis revealed an unfavorable and independent impact of the B/O group on patient survival with ESCC who ever smoked (p = 0.011). Findings suggest the B/O blood type as a predictor of mortality in ESCC patients who ever smoked. Future studies conducted prospectively are warranted to confirm this work and to better understand the underlying biological mechanisms.

Monomer emission and aggregate emission of TPE derivatives in the presence of γ-cyclodextrin.

It was found for the first time that neutral amphiphilc tetraphenylethylene (TPE) derivatives showed an enhanced monomer emission and a decreased aggregate emission when they were included in the cavity of γ-cyclodextrin. This result provided a new insight into the aggregation-induced emission (AIE) effect.

Video-assisted thoracic surgery versus open thoracotomy for non-small-cell lung cancer: a propensity score analysis based on a multi-institutional registry.

OBJECTIVES: Comparative long-term survival and oncological outcomes for patients with non-small-cell lung cancer (NSCLC) who undergo video-assisted thoracic surgery (VATS) or conventional open lobectomy remain uncertain. We conducted a multi-institutional propensity-matched study to stratify potential differences in these outcomes. METHODS: We established a multi-institutional registry for 4312 patients with NSCLC who underwent lobectomy between 2001 and 2008 from eight institutions in the People's Republic of China. Age, gender, histological type and tumour staging were entered into a non-parsimonious multivariable logistic regression model to assess long-term survival outcomes. The predicted probability derived from the logistic equation was used as the propensity score for each individual. Based on similar propensity scores, we matched 1458 of the 1700 patients who underwent VATS lobectomy with 1458 of the 2612 patients who underwent open lobectomy and compared their long-term survival outcomes. RESULTS: The mean age of the 2916 matched patients was 59 (standard deviation = 11) years. After propensity-matching, VATS and open lobectomy patients were similar in regards to important prognostic variables. Three prognostic factors were independently associated with improved survival in the multivariate analysis: age < 60 (P < 0.001), female gender (P = 0.013) and pathological staging (P < 0.001). Patients who underwent VATS vs open lobectomy had similar long-term survival (P = 0.07). CONCLUSIONS: The current propensity score analysis suggests that well-matched patients with NSCLC who underwent standardized VATS lobectomy had similar long-term survival outcomes when compared with those who underwent open lobectomy.

Comparison of the prognostic values of various nutritional parameters in patients with esophageal squamous cell carcinoma from Southern China.

BACKGROUND: Nutritional evaluation is important for patients with esophageal cancer, but the impact of undernutrition on outcome of those patients is not well elucidated. Our aim is to assess the impact of baseline nutritional status on overall survival (OS) in Chinese patients with esophageal squamous cell carcinoma (ESCC) and to detect a most appropriate indicator for nutritional evaluation. METHODS: 502 patients from Southern China diagnosed as ESCC in Sun Yat-Sen University Cancer Center were included. A series of nutritional indicators were introduced to evaluate the baseline nutritional status. Kaplan-Meier method was used to estimate the 5-year OS and the log-rank test was used to determine the survival differences. Cox proportional hazards model was used in the univariate and multivariate analyses of OS. RESULTS: With a median follow up time of 30 months, the median OS for the entire patient group was 37.3 months with the 5-year OS rate of 43.0%. Only performance status, AJCC 6th stage and body mass index (BMI) were the independent prognostic factors in multivariate analysis of OS. The median OS for patients with BMI less than 18.5, patients with BMI within 18.5-24.9 and patients with BMI more than 24.9 were 19.2, 43.2 and 51.6 months, respectively, with the 5-year OS rates of 25.2%, 46.1% and 48.1% (P<0.001). Patients with BMI <18.5 tended to present with a more advanced stage disease and a poorer tumor grade. CONCLUSIONS: Baseline nutritional status is predictive of OS in Chinese patients with ESCC. BMI is a steady indicator for nutritional evaluation and a sensitive prognostic parameter for ESCC patients. Treatment optimization in ESCC patients with low BMI should integrate the modalities and individual nutritional support.

[Performance evaluation of small snail control projects in Hubei Province in 2009 and 2010].

OBJECTIVE: To evaluate the performance of small Oncomelania snail control projects in Hubei Province in 2009 and 2010, so as to explore the best snail control project in different areas. METHODS: The expense-effect, expense-efficiency and cost-utility analyses were carried out to analyze the small snail control projects with reference to the relevant schistosomiasis japonica prevention and control index system. RESULTS: The coverage rate of snail control was 40.15%, the decrease rate of the snail areas was 32.86%, the unit cost for the snail control was 0.39 Yuan/m2, the cost of reducing 1% of the snail area was 67.34 ten thousand Yuan, the total benefit was 15 554.14 ten thousand Yuan, the ratio of cost to benefit was 7.03, the net benefit was 13 341.44 ten thousand Yuan, and the ratio of cost to net benefit was 6.03, and the investment ratio was 1 : 2.11 in the 183 small snail control projects. CONCLUSION: There is an obvious and respectable short-period effect of the small snail control projects.

Expression and prognostic relevance of tumor carcinoembryonic antigen in stage IB non-small cell lung cancer.

BACKGROUND: High serum carcinoembryonic antigen (CEA) levels have been reported to be associated with poor prognosis in non-small cell lung cancer (NSCLC), while the prognostic role of tumor CEA expression remains to be defined. The present study investigated the expression of tumor CEA in stage IB NSCLC, and correlated it with clinicopathological features and prognosis. PATIENTS AND METHODS: Immunohistochemistry for tumor CEA was assessed in the specimens of 183 patients with stage IB NSCLC. Receiver-operating characteristic (ROC) curve analysis was used to determine the cut-off score for tumor positivity. RESULTS: High CEA expression was detected more frequently in adenocarcinomas (72.2%) and other NSCLCs (69.0%) than in squamous cell carcinomas (25.4%, P<0.001). Both univariate and multivariate analysis indicated that tumor CEA was an independent prognostic factor for overall and disease-free survival (P<0.05). CONCLUSIONS: Elevated expression of tumor CEA may be an adverse prognostic indicator in stages IB NSCLC.

HtrA1 expression associated with the occurrence and development of esophageal cancer.

BACKGROUND: The purposes of this study were to measure both the mRNA and protein expression levels of high-temperature requirement serine peptidase 1 (HtrA1) in human esophageal cancer tissues and their adjacent, comparatively normal esophageal tissues. METHODS: The expression levels of HtrA1 mRNA and protein in both tissue types were measured by semi-quantitative RT-PCR (reverse transcription-polymerase chain reaction) and Western blotting. The clinical and pathological correlation between HtrA1 expression levels and the occurrence and development of esophageal cancer was analyzed. RESULTS: The expression levels of HtrA1 mRNA and protein in esophageal carcinoma were significantly lower than the levels expressed in their adjacent normal esophageal tissue (p < 0.05). The more highly undifferentiated esophageal tumor cells expressed lower HtrA1 mRNA and protein expression levels (p < 0.05). Patients with tumors in early pathological stages (I-II) had significantly higher HtrA1 mRNA and protein expression levels than did patients with tumors in mid-to-late pathological stages (III-IV) (p < 0.05). Patients with positive lymph node metastasis had significantly lower HtrA1 mRNA and protein expression levels than did patients with lymph node-negative disease (p < 0.05). CONCLUSIONS: HtrA1 expression is associated with the occurrence and development of esophageal cancer.

Twist expression associated with the epithelial-mesenchymal transition in gastric cancer.

This study aimed to investigate the expression of Twist in gastric cancer tissues and its correlation between Twist and the epithelial-mesenchymal transition (EMT). By means of RT-PCR and Western blot, the mRNA and protein expressions of Twist, E-cadherin, and Vimentin in 61 gastric cancer tissues and adjacent normal tissues were detected. The positive rates of Twist, E-cadherin, and Vimentin mRNA expression in gastric cancer tissues were 73.9. 40.6, and 60.9 %, respectively; compared to the expression of these genes in adjacent normal tissues (2.9, 75.4, and 27.5 %), the differences were significant (p < 0.05). The E-cadherin protein expression level in gastric cancer tissues was significantly lower than that in the adjacent normal tissues (p < 0.05). After the transfection of Twist siRNA into the MKN45 cells, the protein expression of Twist was significantly reduced (p < 0.05), the protein expression of E-cadherin was significantly increased, and the number of cells that passed through the Transwell chamber was significantly lower than that in the non-transfected control group as well as the transfected control group (p < 0.05). Twist may be associated with the epithelial-mesenchymal transition in gastric cancer and the tumorigenesis, invasion, and metastasis of gastric cancer.

The effects of RKIP gene expression on the biological characteristics of human triple-negative breast cancer cells in vitro.

The purpose of this study was to investigate the effect of Raf kinase inhibitor protein (RKIP) on the growth, proliferation, invasion and metastasis of triple-negative breast cancer (TNBC) cells to provide experimental evidence for developing future therapies against human TNBC. The pcDNA3.1-RKIP eukaryotic expression vector was constructed and transfected into the TNBC cell line MDA-MB-231. The alterations of the biological characteristics of RKIP-transfected MDA-MB-231 cells were analyzed using the following approaches: a growth curve, a 3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide (MTT) assay, bromodeoxyuridine (BrdU) staining and a cell migration assay. The effects of the RKIP gene on MMP-1 and MMP-2 expression were also examined. The pcDNA3.1 empty vector-transfected and mock-transfected MDA-MB-231 cells were used as control groups. Compared with the empty vector-transfected and mock-transfected cells, the cell growth of RKIP-transfected MDA-MB-231 cells was significantly reduced. The empty vector-transfected group was not significantly different compared with the mock-transfected MDA-MB-231 cells. The results of the MTT and BrdU assays demonstrated that the proliferation of pcDNA3.1-RKIP-transfected cells was significantly reduced compared to the control cells (P < 0.05). The result of the cell migration assay suggested that the cross-membrane migration rate of the pcDNA3.1-RKIP-transfected cells was significantly lower than that of the control MDA-MB-231 cells (P < 0.05). We also demonstrated that RKIP may inhibit MMP-1 and MMP-2 expression in MDA-MB-231 cells. The RKIP gene may play a role in inhibiting cellular proliferation. The RKIP gene may also have some inhibitory effects on the invasiveness and metastatic capability of human TNBC cells.

A practical molecular assay to predict survival in resected non-squamous, non-small-cell lung cancer: development and international validation studies.

BACKGROUND: The frequent recurrence of early-stage non-small-cell lung cancer (NSCLC) is generally attributable to metastatic disease undetected at complete resection. Management of such patients depends on prognostic staging to identify the individuals most likely to have occult disease. We aimed to develop and validate a practical, reliable assay that improves risk stratification compared with conventional staging. METHODS: A 14-gene expression assay that uses quantitative PCR, runs on formalin-fixed paraffin-embedded tissue samples, and differentiates patients with heterogeneous statistical prognoses was developed in a cohort of 361 patients with non-squamous NSCLC resected at the University of California, San Francisco. The assay was then independently validated by the Kaiser Permanente Division of Research in a masked cohort of 433 patients with stage I non-squamous NSCLC resected at Kaiser Permanente Northern California hospitals, and on a cohort of 1006 patients with stage I-III non-squamous NSCLC resected in several leading Chinese cancer centres that are part of the China Clinical Trials Consortium (CCTC). FINDINGS: Kaplan-Meier analysis of the Kaiser validation cohort showed 5 year overall survival of 71·4% (95% CI 60·5-80·0) in low-risk, 58·3% (48·9-66·6) in intermediate-risk, and 49·2% (42·2-55·8) in high-risk patients (p(trend)=0·0003). Similar analysis of the CCTC cohort indicated 5 year overall survivals of 74·1% (66·0-80·6) in low-risk, 57·4% (48·3-65·5) in intermediate-risk, and 44·6% (40·2-48·9) in high-risk patients (p(trend)<0·0001). Multivariate analysis in both cohorts indicated that no standard clinical risk factors could account for, or provide, the prognostic information derived from tumour gene expression. The assay improved prognostic accuracy beyond National Comprehensive Cancer Network criteria for stage I high-risk tumours (p<0·0001), and differentiated low-risk, intermediate-risk, and high-risk patients within all disease stages. INTERPRETATION: Our practical, quantitative-PCR-based assay reliably identified patients with early-stage non-squamous NSCLC at high risk for mortality after surgical resection. FUNDING: UCSF Thoracic Oncology Laboratory and Pinpoint Genomics.