Gene expression patterns can be used as prognostic biomarkers in various types of cancers. We aimed to identify a gene expression pattern for individual distant metastatic risk assessment in patients ...with locoregionally advanced nasopharyngeal carcinoma.
In this multicentre, retrospective, cohort analysis, we included 937 patients with locoregionally advanced nasopharyngeal carcinoma from three Chinese hospitals: the Sun Yat-sen University Cancer Center (Guangzhou, China), the Affiliated Hospital of Guilin Medical University (Guilin, China), and the First People's Hospital of Foshan (Foshan, China). Using microarray analysis, we profiled mRNA gene expression between 24 paired locoregionally advanced nasopharyngeal carcinoma tumours from patients at Sun Yat-sen University Cancer Center with or without distant metastasis after radical treatment. Differentially expressed genes were examined using digital expression profiling in a training cohort (Guangzhou training cohort; n=410) to build a gene classifier using a penalised regression model. We validated the prognostic accuracy of this gene classifier in an internal validation cohort (Guangzhou internal validation cohort, n=204) and two external independent cohorts (Guilin cohort, n=165; Foshan cohort, n=158). The primary endpoint was distant metastasis-free survival. Secondary endpoints were disease-free survival and overall survival.
We identified 137 differentially expressed genes between metastatic and non-metastatic locoregionally advanced nasopharyngeal carcinoma tissues. A distant metastasis gene signature for locoregionally advanced nasopharyngeal carcinoma (DMGN) that consisted of 13 genes was generated to classify patients into high-risk and low-risk groups in the training cohort. Patients with high-risk scores in the training cohort had shorter distant metastasis-free survival (hazard ratio HR 4·93, 95% CI 2·99–8·16; p<0·0001), disease-free survival (HR 3·51, 2·43–5·07; p<0·0001), and overall survival (HR 3·22, 2·18–4·76; p<0·0001) than patients with low-risk scores. The prognostic accuracy of DMGN was validated in the internal and external cohorts. Furthermore, among patients with low-risk scores in the combined training and internal cohorts, concurrent chemotherapy improved distant metastasis-free survival compared with those patients who did not receive concurrent chemotherapy (HR 0·40, 95% CI 0·19–0·83; p=0·011), whereas patients with high-risk scores did not benefit from concurrent chemotherapy (HR 1·03, 0·71–1·50; p=0·876). This was also validated in the two external cohorts combined. We developed a nomogram based on the DMGN and other variables that predicted an individual's risk of distant metastasis, which was strengthened by adding Epstein–Barr virus DNA status.
The DMGN is a reliable prognostic tool for distant metastasis in patients with locoregionally advanced nasopharyngeal carcinoma and might be able to predict which patients benefit from concurrent chemotherapy. It has the potential to guide treatment decisions for patients at different risk of distant metastasis.
The National Natural Science Foundation of China, the National Science & Technology Pillar Program during the Twelfth Five-year Plan Period, the Natural Science Foundation of Guang Dong Province, the National Key Research and Development Program of China, the Innovation Team Development Plan of the Ministry of Education, the Health & Medical Collaborative Innovation Project of Guangzhou City, China, and the Program of Introducing Talents of Discipline to Universities.
The American Joint Committee on Cancer (AJCC) staging system is inadequate for an accurate prognosis in nasopharyngeal carcinoma (NPC). Thus, new biomarkers are under intense investigation. Here, we ...investigated whether the density of TILs could predict prognosis in NPC. First, we used 1490 cases of nasopharyngeal carcinoma samples from two independent cohorts to evaluate the density and distribution of tumor‐infiltrating lymphocytes (TILs). Second, in one cohort, we assessed associations between TILs and clinical outcomes in 593 randomly selected samples (defined as the training set) and validated findings in the remaining 593 samples (defined as the validation set). Furthermore, we confirmed the prognostic value of TILs in a second independent cohort of 304 cases (defined as the independent set). Based on multivariable Cox regression analysis, we also established an effective prognostic nomogram including TILs to improve accuracy in predicting disease‐free survival (DFS) for patients with nondisseminated NPC. We found that high TILs in the training set were significantly associated with favorable DFS hazard ratio (HR) 0.41, 95% confidence interval (CI) 0.28–0.58, p < 0.001, overall survival (OS, HR 0.42, 95% CI 0.27–0.64, p < 0.001), distant metastasis‐free survival (DMFS, HR 0.37, 95% CI 0.23–0.58, p < 0.001) and local‐regional recurrent free survival (LRRFS, HR 0.43, 95% CI 0.25–0.73, p = 0.002). Multivariate analysis showed that TILs are an independent prognostic indicator for DFS in all cohorts. In summary, this study indicated that TILs may reflect the immunological heterogeneity of NPC and could represent a new prognostic biomarker.
What's new?
Doctors typically use tumor stage to help determine cancer prognosis, but for nasopharyngeal cancer, it is not precise enough. These authors turned to the immune system for prognostic clues. They looked at the density and distribution of tumor‐infiltrating lymphocytes (TILs) in NPC patients from China. TILs turned out to be a strong independent predictor of disease‐free survival: greater numbers of TILs, they found, meant better outcomes. Once a standardized method for evaluating TILs can be developed, this metric could be extremely valuable for predicting disease progression in NPC patients.
Objective
Oral lichen planus (OLP) is a T cell‐mediated inflammatory disease with uncertain etiology. Exosomes are cell‐derived vesicles containing biological cargo, being associated with the ...development of multiple inflammatory diseases. The present study aims to investigate the role of T cell‐derived exosomes in the pathogenesis of OLP.
Methods
Exosomal marker CD63 was detected in OLP lesions by immunohistochemistry. Twenty‐three cytokines in T cell‐derived exosomes were assessed using luminex xMAP‐based assay. After co‐incubating with exosomes, the apoptosis of keratinocytes and the proliferation of Jurkat cells were assessed via flow cytometry and cell counting kit‐8 assay, respectively.
Results
CD63 was highly expressed in the lymphocyte infiltrated areas of OLP lesions. OLP T cell‐derived exosomes contained upregulated interleukin‐7, ‐10, ‐12, ‐17 and downregulated interleukin‐1β, ‐5, and interferon‐γ. Both exosomes from OLP patients and controls induced the apoptosis of keratinocytes and altered their morphology. Moreover, healthy control–derived exosomes markedly inhibited the proliferation of Jurkat cells, whereas OLP‐derived exosomes exhibited no inhibitory effect.
Conclusions
OLP T cell‐derived exosomes have an aberrant cytokine profile and could trigger the apoptosis of keratinocytes in vitro, which may be involved in the pathogenesis of OLP.
Herein, a Janus three-dimensional (3D) DNA nanomachine was constructed for the simultaneous and sensitive fluorescence detection and imaging of dual microRNAs (miRNAs) in cancer cells, which could ...effectively eliminate signal interference in a homogeneous nanoparticle-based 3D DNA nanostructure caused by the proximity of the two different signal probes to achieve accurate co-location in the same position of living cancer cells. In this system, the Janus nanoparticles were synthesized as the carrier for immobilizing two different oligonucleotides on two different functionalized hemispheres of the nanoparticles to form a Janus 3D DNA nanostructure, which could convert trace amounts of miRNA-21 and miRNA-155 targets into massive FAM and Cy5-labeled duplexes to induce two remarkable fluorescence emissions by the catalytic hairpin assembly (CHA) and 3D DNA walker cascade nucleic acid amplification strategy, realizing sensitive detection and imaging of miRNA targets in cancer cells. Impressively, in comparison with current miRNA imaging methods based on nanoparticle assemblies, the proposed strategy could efficiently eliminate "false positive" results obtained in single type miRNA detection and distinctly increase the immobilization concentration of two different signal probes using Janus nanoparticles as the carrier to further enhance fluorescence intensity, resulting in accurate co-location in the same position of living cells. Meanwhile, the proposed fluorescence imaging technology makes it possible to visualize low concentrations of miRNAs with tiny change associated with some cancers, which could significantly improve the accuracy and precision compared to those of the conventional fluorescence
in situ
hybridization (FISH) approach. Therefore, it could serve as persuasive evidence for supplying accurate information to better understand biological processes and investigate mechanisms of various biomolecules and subcellular organelles, resulting in the further validation of their function in tumor proliferation and differentiation. This strategy provided an innovative approach to design new generations of nanomachines with ultimate applications in bioanalysis and clinical diagnoses.
A Janus three-dimensional DNA nanomachine was constructed for the simultaneous and sensitive fluorescent detection and imaging of dual microRNAs in the cancer cells.
Potassium (K+) is an essential macronutrient for plant growth and development. Transporters from the KT/HAK/KUP family play crucial roles in K+ homeostasis and cell growth in various plant species. ...However, their physiological roles in maize are still unknown. In this study, we cloned ZmHAK5 and ZmHAK1 and investigated their functions in maize (Zea mays L.). In situ hybridization showed that ZmHAK5 was mainly expressed in roots, especially in the epidermis, cortex, and vascular bundle. ZmHAK5 was characterized as a high‐affinity K+ transporter. Loss of function of ZmHAK5 led to defective K+ uptake in maize, under low K+ conditions, whereas ZmHAK5‐overexpressing plants showed increased K+ uptake activity and improved growth. ZmHAK1 was upregulated under low K+ stress, as revealed by RT‐qPCR. ZmHAK1 mediated K+ uptake when heterologously expressed in yeast, but its transport activity was weaker than that of ZmHAK5. Overexpression of ZmHAK1 in maize significantly affected K+ distribution in shoots, leading to chlorosis in older leaves. These findings indicate that ZmHAK5 and ZmHAK1 play distinct roles in K+ homeostasis in maize, functioning in K+ uptake and K+ distribution, respectively. Genetic manipulation of ZmHAK5 may represent a feasible way to improve K+ utilization efficiency in maize.
Transporters from the KT/HAK/KUP family play crucial roles in K+ homeostasis and cell growth in plants. Here we demonstrate that K+ transporters ZmHAK5 and ZmHAK1 play distinct roles in K+ homeostasis in maize, functioning in K+ uptake and K+ distribution, respectively.
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•CS-EDTA-HTlc composite was successfully synthesized for efficient removal of Cu(II).•CS-EDTA-HTlc composite can prevent the adsorbent from aggregation and loss.•The removal of Cu(II) ...followed the pseudo-second-order kinetic model.•The adsorption data fitted well with the Freundlich model.•Mechanisms included complexation and isomorphic substitution.
Adsorption is recognized as one of the most promising methods for heavy metal removal from contaminated water. However, the adsorption efficiency often decreases because of easy aggregation and an inevitable certain loss of adsorbents during the adsorption process. Herein, a highly efficient chitosan (CS) assisted ethylenediaminetetraacetate-intercalated Mg-Al hydrotalcite-like compound (EDTA-HTlc) composite absorbent (CS-EDTA-HTlc) was synthesized by using a simple ion-exchange/chemical crosslinking process to remove Cu(II), used as a template heavy metal, from an aqueous solution. Characterization by FESEM, PXRD, FTIR, TG-DTG, BET specific surface area, and pore volume revealed that CS-EDTA-HTlc had a high specific surface area and many different functional groups (such as carboxyl, hydroxyl, and amino) with strong complexing ability. The adsorption capacity of CS-EDTA-HTlc for Cu(II) was investigated by varying experimental conditions such as adsorbent dosage, initial solution pH, adsorption time, initial Cu(II) concentration, and temperature. The adsorption equilibrium was reached within 120 min. The Cu(II) removal efficiency came 98% with the initial Cu(II) concentration of 200 mg·L-1 and the adsorbent dosage of 1.0 g·L-1. The adsorption kinetics and isotherms data of Cu(II) on CS-EDTA-HTlc were well described by the pseudo-second-order kinetic equation and the Freundlich model, respectively. The adsorption was a spontaneous exothermic process, and the possible adsorption mechanisms analyzed by the XPS and PXRD spectra involved the complexation of Cu(II) ions by carboxyl/hydroxyl/amino groups and the isomorphic substitution of Mg(II) in brucite-like layers. The results suggest that CS-EDTA-HTlc has potential applications in natural Cu(II) contaminated wastewater treatment.
Electrochemiluminescence (ECL), as a sensitive and controllable assay, offers a considerable opportunity for multiple types of biomarkers detection. However, constructing such a biosensor remains a ...significant challenge. Herein, an ultrasensitive and versatile ECL biosensor was constructed to detect multiple types of biomarkers from breast cancer by taking the strategies of nonenzymatic catalytic hairpin assembly (CHA) and hybridization chain reaction (HCR) amplification, as well as aptamer-triggered emitter release. Concretely, with the appearance of target 1 microRNA-21 (miRNA-21), abundant double-stranded DNA (dsDNA) polymers were generated on this biosensing surface via amplification circuits of CHA and HCR, which could be intercalated into substantial (Ru(bpy)2dppzCl2) as ECL indicators to obtain an obvious enhancement of ECL signal for target 1 detection with a detection limit (0.1 fM). Furthermore, in the presence of target 2 human mucin 1 (MUC1) protein, the ECL signal had a distinct decrease, because aptamer recognition induced the release of Ru(bpy)2dppzCl2 from the sensing surface, thus, achieving a sensitive detection for MUC1 with a detection limit (2.4 fg·mL–1). Simultaneously, this sensing platform was applied to monitor the biomarkers from MDA-MB-231 breast cancer cells, suggesting that this method was applicable to detect real samples. Therefore, this platform is an applicable and versatile implement for the determination of multiple types of biomarkers to improve diagnostic accuracy and efficiency.
Nasopharyngeal carcinoma (NPC) is an aggressive malignancy with extremely skewed ethnic and geographic distributions. Increasing evidence indicates that targeting the tumor microenvironment (TME) ...represents a promising therapeutic approach in NPC, highlighting an urgent need to deepen the understanding of the complex NPC TME. Here, we generated single-cell transcriptome profiles for 7581 malignant cells and 40,285 immune cells from fifteen primary NPC tumors and one normal sample. We revealed malignant signatures capturing intratumoral transcriptional heterogeneity and predicting aggressiveness of malignant cells. Diverse immune cell subtypes were identified, including novel subtypes such as CLEC9A
dendritic cells (DCs). We further revealed transcriptional regulators underlying immune cell diversity, and cell-cell interaction analyses highlighted promising immunotherapeutic targets in NPC. Moreover, we established the immune subtype-specific signatures, and demonstrated that the signatures of macrophages, plasmacytoid dendritic cells (pDCs), CLEC9A
DCs, natural killer (NK) cells, and plasma cells were significantly associated with improved survival outcomes in NPC. Taken together, our findings represent a unique resource providing in-depth insights into the cellular heterogeneity of NPC TME and highlight potential biomarkers for anticancer treatment and risk stratification, laying a new foundation for precision therapies in NPC.
In this work, we developed a sensitive and efficient ratiometric electrochemical method for lipopolysaccharide (LPS) detection using Cu-based metal–organic frameworks (Cu-MOFs) as a catalyst and ...target-triggered quadratic cycles for signal amplification. First, in the presence of target LPS, the conformation change of the specifically designed hairpin probes 1 (HP1) triggered the target cyclic-induced polymerization to produce the output DNA with the aid of phi29 DNA polymerase (phi29). Then, the obtained output DNA hybridized with ferrocene-labeled hairpin probes 2 (Fc-HP2, which were immobilized on the electrode) to generate a nicking endonuclease (N.BstNBI) cleavage site. Thus, with N.BstNBI, the original signal molecules of Fc left from the electrode, and the single-stranded capture-probe-modified sensing interface was obtained. At this time, signal probes conducted by Au-nanoparticles-functionalized Cu-MOFs and labeled hairpin probes 3 (HP3/AuNPs/Cu-MOFs) were hybridized with capture probes for hairpin assembly. Herein, AuNPs/Cu-MOFs were not only used as nanocarriers for immobilizing HP3 but also acted as electroactive materials for signal reporting. With the proposed target-triggered quadratic cycles, the cleavage sites of Fc-HP2 were cut, and capture probes were obtained to hybridize with HP3/AuNPs/Cu-MOFs, which caused the signal decrease of Fc. Then Cu-MOFs were closed to the electrode for the signal increase of Cu-MOFs. Furthermore, when glucose was present in the detection solution, AuNPs/Cu-MOFs catalyzed the oxidation of glucose to realize the enzyme-free signal amplification. By measuring the peak currents ratio of the Cu-MOFs and Fc, the proposed aptasenor for LPS detection showed a low detection limit (0.33 fg/mL) and a wide linear range from 1.0 fg/mL to 100 ng/mL with high accuracy and sensitivity. This ratiometric electrochemical approach is expected to be a valuable strategy for detection of other analytes.
Ligand-protected gold nanoclusters (Au NCs) are promising electrochemiluminescence (ECL) emitters because of their striking optical properties and excellent biocompatibility, but free vibration and ...rotation of their ligand result in low ECL efficiency, dramatically limiting their applications. Herein, using the ligand of Au NCs as one of the building units, a Au NC-based metal–organic framework (Au NC-based MOF) was constructed by the coordination-assisted self-assembly strategy, which not only impedes the ligand rotation-induced energy dissipation but also diminishes the self-quenching effect due to the spatial distribution of Au NCs. As a proof of concept, the prepared GSH-Au NCs@ZIF-8 gives rise to a 10-fold enhanced anodic ECL efficiency compared to that of densely aggregated GSH-Au NCs with triethylamine as the coreactant. Based on high ECL efficiency of GSH-Au NCs@ZIF-8, a “signal off” sensing platform was proposed with rutin as a model analyte, achieving a low detection limit of 10 nM. Therefore, the strategy paves an effective and alternative methodology to enhance ECL efficiency of metal NCs, considerably broadening their potential applications in sensing analysis, clinical diagnosis, and light-emitting devices.
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