Copper(II)-Tannic Acid@Cu with In Situ Grown Gold Nanoparticles as a Bifunctional Matrix for Facile Construction of Label-Free and Ultrasensitive Electrochemical cTnI Immunosensor.

Sensitive detection of cardiac troponin I (cTnI) is of great significance in the diagnosis of a fatal acute myocardial infarction. A redox-active nanocomposite of copper(II)-tannic acid@Cu (CuTA@Cu) was herein prepared on the surface of a glassy carbon electrode by electrochemical deposition of metallic copper combined with a metal stripping strategy. Then, HAuCl4 was in situ reduced to gold nanoparticles (AuNPs) by strong reductive catechol groups in the TA ligand. The AuNPs/CuTA@Cu composite was further utilized as a bifunctional matrix for the immobilization of the cTnI antibody (anti-cTnI), producing an electrochemical immunosensor. Electrochemical tests show that the immunoreaction between anti-cTnI and target cTnI can cause a significant reduction of the electrochemical signal of CuTA@Cu. It can be attributed to the insulating characteristic of the immunocomplex and its barrier effect to the electrolyte ion diffusion. From the signal changes of CuTA@Cu, cTnI can be analyzed in a wide range from 10 fg mL-1 to 10 ng mL-1, with an ultralow detection limit of 0.65 fg mL-1. The spiked recovery assays show that the immunosensor is reliable for cTnI determination in human serum samples, demonstrating its promising application in the early clinical diagnosis of myocardial infarction.

A novel nomogram model for lung adenocarcinoma subtypes based on RNA-modification regulatory genes.

Background: In non-small cell lung cancer (NSCLC), lung adenocarcinoma (LUAD) is the most common subtype. RNA modification has become the frontier and hotspot of current tumor research. Results: In this study, 109 genes that regulate RNA modifications were identified according to The Cancer Genome Atlas (TCGA). A differential gene expression analysis identified 46 differentially expressed RNA modification regulatory genes (DERRGs). LUAD samples were stratified into two distinct clusters based on the expression of these DERRGs. A significant correlation was observed between these clusters and patient survival rates, as well as clinical features. Furthermore, a four-DERRG signature (EIF3B, HNRNPC, IGF2BP1, and METTL3) developed using LASSO regression. According to the calculated risk scores from this signature, LUAD patients were categorized into high-risk and low-risk groups. Patients in the low-risk group exhibited a more favorable prognosis. A prognostic nomogram was crafted, integrating the four-DERRGs signature with clinical parameters. The nomogram was revealed that OS, age, clinical stage, immune cell infiltration, and immune checkpoint molecule expression were significantly linked to the OS of LUAD. GSEA analysis found that the DERRGs were primarily regulated immune pathways. Conclusions: This study developed four DERRGs signatures and formulated a nomogram model for precise prognosis estimation in LUAD patients. The study's insights are instrumental for advancing diagnosis, prognosis, and therapeutic strategies for LUAD.

Practice and Effect Evaluation of Preoperative Management Strategy for Patients Undergoing Day Arthroscopic Surgery Based on Evidence-Based Concept.

Objective: To improve the preoperative management of patients undergoing day arthroscopy. Methods: Based on the evidence-based concept, the preoperative management strategy of daytime arthroscopic surgery was practiced, and the implementation effect of patients undergoing daytime arthroscopic surgery before and after evidence application was compared. The evidence-based strategies adopted in this study include: informed consent,seven aspects: anesthesia assessment, health education, prohibition of drinking and eating, preoperative medication guidance, surgical arrangement, and emergency management. Results: After applying evidence-based strategies, significant improvements were observed in various quality indicators, including reduced waiting time, reduced surgical cancellations, increased patient satisfaction, and reduced unplanned hospitalization rates. Conclusion: Based on the evidence-based concept, the practice of preoperative management strategies for patients undergoing daytime arthroscopic surgery optimizes the daytime surgical process from the strict implementation of patient health education and evaluation, as well as the establishment of compliant and reasonable surgical schedules and emergency mechanisms. This improves the preoperative management behavior of daytime surgery, enhances the patient's medical experience, and enhances the quality of daytime surgical care, providing a good basis for management and decision-making.

PER-CRISPR/Cas14a system-based electrochemical biosensor for the detection of ctDNA EGFR L858R.

The detection of epidermal growth factor receptor (EGFR) mutation L858R in circulating tumor DNA (ctDNA) is beneficial for the clinical diagnosis and personalized therapy of non-small cell lung cancer (NSCLC). Herein, for the first time, the combination of the primer exchange reaction (PER) and clustered regularly interspaced short palindromic repeats (CRISPR) and its associated nucleases (Cas) 14a was used in electrochemical biosensor construction for the detection of ctDNA EGFR L858R. EGFR L858R, as the target, induced the isothermal amplification of the PER reaction, and then the CRISPR/Cas14a system was activated; subsequently, the substrate ssDNA-MB was cleaved and the electron on the surface of the gold electrode transferred, resulting in the fluctuation of the electrochemical redox signal on the electrode surface, whereas the electrochemical signal will be stable when EGFR L858R is absent. Therefore, the concentration of EGFR L858R can be quantified by electrochemical signal analysis. The low detection limit is 0.34 fM and the dynamic detection range is from 1 fM to 1 µM in this work. The PER-CRISPR/Cas14a electrochemical biosensor greatly improved the analytical sensitivity. In addition, this platform also exhibited excellent specificity, reproducibility, stability and good recovery. This study provides an efficient and novel strategy for the detection of ctDNA EGFR L858R, which has great potential for application in the diagnosis and treatment of NSCLC.

STARD12/14 are diagnostic and prognostic biomarkers of lung adenocarcinoma associated with epigenetic regulation, immune infiltration and ferroptosis.

Background: Metabolic reprogramming plays an important role in tumor progression and antitumor immunity. START domain-containing proteins (STARDs) are responsible for lipid metabolism. However, the underlying functions of STARDs in lung adenocarcinoma (LUAD) have not been clarified yet. Methods: Oncomine, UALCAN, TCGA and CPTAC were used to explore the expression landscape and clinicopathological characteristics of STARDs in LUAD. Diagnostic and prognostic values were assessed by Kaplan-Meier Plotter, Cox regression analysis, and ROC curve. GeneMANIA, GO, KEGG and GSEA were applied for exploring the potential biological functions. Epigenetic process, including mutation and m6A modification were analyzed by cBioPortal and TCGA. TIMER, TISIDB and TCGA cohort provided an immune signature. The correlation between STARDs expression and ferroptosis was analyzed by TCGA. Finally, the STARDs expression were confirmed by RT-qPCR and western blot. Results: STARD5/10/14 were overexpressed in LUAD compared with normal, while STARD4/7/8/11/12/13 were relatively low. STARD5/12/14 levels were positively related to clinical and lymph node stage. Survival analysis showed high STARD12 expression was associated with favorable overall survival, disease special survival as well as disease free survival, while STARD14 showed the opposite. GSEA analysis found STARD12 and STARD14 were associated with glycolysis, oxidative phosphorylation and tumor related signaling pathways. STARD12 co-expressed genes participated in cell cycle and DNA replication, and STARD14 were enriched in ECM-receptor interaction. Both STARD12 and STARD14 were corelated with epigenetic regulation, especially TP53 mutation and m6A modification. STARD12 expression was positively correlated with TMB level. The level of STARD12 was significantly associated with the abundance of infiltrating immune cells, including B cells, CD8+T cells, macrophages, dendritic cells, and chemokine, receptor, MHC, immunostimulatory related genes. STARD14 was negatively associated with the infiltration of CD8+T cells, while positively with CCL28 and immune checkpoints, including CTLA4 as well as PD-L2. In addition, STARD12/14 could regulate the ferroptosis related genes. Conclusion: STARD12 and STARD14 were expected to be potential biomarkers for LUAD, which were associated with epigenetic regulation, immune infiltration and ferroptosis.

SYT7 is a key player in increasing exosome secretion and promoting angiogenesis in non-small-cell lung cancer.

Lung cancer is the leading cause of cancer-related mortality, and non-small cell lung cancer (NSCLC) accounts for approximately 85% of all lung cancer cases. Our previous study confirmed that synaptotagmin 7 (SYT7) promoted NSCLC metastasis in vivo and in vitro. Studies have shown that SYT7 is an important regulatory molecule of exocytosis in various cells. However, the characteristics of SYT7 across cancers and the function of SYT7 in tumor exosome secretion remain unclear. In this study, we conducted systematic pancancer analyses of SYT7, namely, analyses of expression patterns, diagnostic and prognostic values, genetic alterations, methylation, immune infiltration, and potential biological pathways. Furthermore, we demonstrated that SYT7 increased the secretion of exosomes from A549 and H1299 cells, promoting the migration, proliferation, and tube formation of human umbilical vein endothelial cells (HUVECs). Notably, SYT7 promoted angiogenesis by transferring exosomes containing the molecule centrosomal protein of 55 kDa (CEP55) protein to HUVECs. The CEP55 protein levels was downregulated in STAT1 inhibitor-treating SYT7-overexpresion NSCLC cells. We further found that SYT7 activated the mTOR signaling pathway through the downstream molecule CEP55, thereby promoting the invasion and metastasis of NSCLC cells. SYT7 promoted exosome secretion by NSCLC cells through upregulating syntaxin-1a and syntaxin-3. In vivo, SYT7 promoted the tumorigenesis, angiogenesis and metastasis of A549 cells through the exosome pathway. Our study is of great importance for understanding the mechanism of tumor exosome secretion and the role of exosomes in tumor progression.

Analysis and Recognition of Voluntary Facial Expression Mimicry Based on Depressed Patients.

Many clinical studies have shown that facial expression recognition and cognitive function are impaired in depressed patients. Different from spontaneous facial expression mimicry (SFEM), 164 subjects (82 in a case group and 82 in a control group) participated in our voluntary facial expression mimicry (VFEM) experiment using expressions of neutrality, anger, disgust, fear, happiness, sadness and surprise. Our research is as follows. First, we collected a large amount of subject data for VFEM. Second, we extracted the geometric features of subject facial expression images for VFEM and used Spearman correlation analysis, a random forest, and logistic regression-based recursive feature elimination (LR-RFE) to perform feature selection. The features selected revealed the difference between the case group and the control group. Third, we combined geometric features with the original images and improved the advanced deep learning facial expression recognition (FER) algorithms in different systems. We propose the E-ViT and E-ResNet based on VFEM. The accuracies and F1 scores were higher than those of the baseline models, respectively. Our research proved that it is effective to use feature selection to screen geometric features and combine them with a deep learning model for depression facial expression recognition.

Compare the clinical value of two minimally invasive approaches to locating radial nerve in the posterior humeral approach.

PURPOSE: To compare the clinical value between locating radial nerve (RN) guided by Color Doppler ultrasonography and posterior antebrachial cutaneous nerve (PACN) in the posterior humeral approach. METHODS: The five fresh adult cadavers (ten upper arms) were selected to compare the two methods of locating the RN in the posterior humeral approach (guided by ultrasound and PACN) by measuring the operation time, the length of incision, and the area of subcutaneous free. And the comparison between the two groups was statistically analyzed by paired t-test. RESULTS: The results of this study demonstrated that the length of incision and the area of subcutaneous free in the ultrasound group were smaller than that in the PACN group (P < 0.05), while the operation time was just the opposite (P < 0.05). However, after excluding the time of ultrasound location, the operation time in the ultrasound group was shorter than that in the PANC group, and the difference was statistically significant (P < 0.05). CONCLUSION: The RN can be quickly and safely exposed by both methods. The ultrasound approach requires a long learning curve, but is more minimally invasive and can help determine whether the intraoperative nerve is compressed by the plate. And the PACN method requires a longer incision and a wider area of subcutaneous free, while specialized equipment and professional training for surgeons are not required. In a word, these two methods have advantages and disadvantages, so they should be selected based on the exact situation.

Analysis on heterogeneity of hepatocellular carcinoma immune cells and a molecular risk model by integration of scRNA-seq and bulk RNA-seq.

Background: Studies have shown that hepatocellular carcinoma (HCC) heterogeneity is a main cause leading to failure of treatment. Technology of single-cell sequencing (scRNA) could more accurately reveal the essential characteristics of tumor genetics. Methods: From the Gene Expression Omnibus (GEO) database, HCC scRNA-seq data were extracted. The FindCluster function was applied to analyze cell clusters. Autophagy-related genes were acquired from the MSigDB database. The ConsensusClusterPlus package was used to identify molecular subtypes. A prognostic risk model was built with the Least Absolute Shrinkage and Selection Operator (LASSO)-Cox algorithm. A nomogram including a prognostic risk model and multiple clinicopathological factors was constructed. Results: Eleven cell clusters labeled as various cell types by immune cell markers were obtained from the combined scRNA-seq GSE149614 dataset. ssGSEA revealed that autophagy-related pathways were more enriched in malignant tumors. Two autophagy-related clusters (C1 and C2) were identified, in which C1 predicted a better survival, enhanced immune infiltration, and a higher immunotherapy response. LASSO-Cox regression established an eight-gene signature. Next, the HCCDB18, GSA14520, and GSE76427 datasets confirmed a strong risk prediction ability of the signature. Moreover, the low-risk group had enhanced immune infiltration and higher immunotherapy response. A nomogram which consisted of RiskScore and clinical features had better prediction ability. Conclusion: To precisely assess the prognostic risk, an eight-gene prognostic stratification signature was developed based on the heterogeneity of HCC immune cells.

FOXP family DNA methylation correlates with immune infiltration and prognostic value in NSCLC.

Background: Forkhead box P (FOXP) family was introduced as a double-edged sword in tumorigenesis and influenced immunotherapy response by modulating host immunity. This study aimed to summarize the involvement of the FOXP family in non-small cell lung cancer (NSCLC). Methods: The UALCAN, Gene Expression Profiling Interactive Analysis (GEPIA), and Reverse transcription-quantitative polymerase chain reaction (RT‒qPCR) were used to analyse the expression levels of the FOXP family in NSCLC. The prognostic impact was evaluated using Kaplan-Meier Plotter. MethSurv, UALCAN, and cBioPortal were applied to analyse the DNA methylation and mutation status of the FOXP family respectively. COEXPEDIA, STRING, and GeneMANIA were used to explore the interaction mechanism. Finally, TISIDB was used to investigate all of the immune-related characteristics regulated by the FOXP family. Results: The expression levels of FOXP1/3/4 were dysregulated in NSCLC tissues than that in normal tissues. Groups with low expression levels of FOXP1/4 and high expression levels of FOXP2/3 were associated with poor prognosis in NSCLC. The transcriptional levels of FOXP2/3/4 were correlated with DNA methylation in NSCLC. FOXP1/3/4 DNA methylation were correlated with prognosis. Pathway enrichment analysis indicated the FOXP family was mainly related to immune-related pathways. After DNA methylation, the correlations between FOXP family and immune factors were opposite to that before alteration in NSCLC. Conclusion: This study elucidated FOXP family could serve as vital diagnostic and prognostic biomarkers in NSCLC. Our study highlighted novel potential functions of FOXP family DNA methylation in regulation of immune-related signatures in NSCLC.

Junctional adhesion molecule-like protein promotes tumor progression via the Wnt/ß-catenin signaling pathway in lung adenocarcinoma.

BACKGROUND: Lung adenocarcinoma (LUAD) is a heavy social burden worldwide. Because the mechanisms involved in LUAD remain unclear, the prognosis of LUAD remains poor. Consequently, it is urgent to investigate the potential mechanisms of LUAD. Junctional adhesion molecule-like protein (JAML), is recognized as a tumorigenesis molecule in gastric cancer. However, the role of JAML in LUAD is still unclear. Here we aimed to evaluate the role of JAML in LUAD. METHODS: qRT-PCR, Western blotting and immunohistochemistry were conducted to investigate the expression of JAML in LUAD tissues. JAML was knocked down and overexpressed in LUAD cells using transient transfection by siRNA and plasmids or stable transfection by lentivirus. Proliferation potential of LUAD cells were detected by Cell Counting Kit-8, EdU incorporation and Colony formation assay. Migration and invasion abilities of LUAD cells were determined by wound healing, transwell migration and invasion assays. Cell cycle and cell apoptosis were detected by flow cytometry. The effects of JAML in vivo were studied in xenograft tumor models. Western blotting was used to explore the molecular mechanisms of JAML function. In addition, rescue experiments were performed to verify the possible mechanisms. RESULTS: JAML expression was elevated in LUAD tissues compared with peritumor tissues, and this upregulation was positively related to pT and pTNM. Furthermore, both in vitro and in vivo, JAML silencing markedly repressed malignant behaviors of LUAD cells and vice versa. Knockdown of JAML also mediated cell cycle arrest at G0/G1 phase and promoted apoptosis in LUAD cells. Mechanistically, silencing JAML repressed the process of epithelial-mesenchymal transition by inactivating the Wnt/ß-catenin pathway in LUAD cells. Effects of JAML can be rescued by Wnt/ß-catenin pathway activator in A549 cells. CONCLUSIONS: Our data reveal the oncogenic role of JAML in LUAD, indicating that JAML may be a predictive biomarker and novel therapeutic target for LUAD.

Autophagy Induced by BCL2-Related ceRNA Network Participates in the Occurrence of COPD.

Purpose: Chronic obstructive pulmonary disease (COPD) is a predominant cause of mortality worldwide. Autophagy, which depends on a lysosomal degradation pathway, plays an essential role in the occurrence of COPD. The aim of our study was to identify the potential function of autophagy and construct a BCL2-related competing endogenous RNA (ceRNA) network that induces autophagy in COPD. Methods: Blood sample data from GSE31568, GSE24709, and GSE61741 were collected from the Gene Expression Omnibus (GEO) database. Differentially expressed miRNAs in COPD and controls were identified via GEO2R. Transcription factors were obtained from FunRich. DIANA, miRDB, miRTarBase, and TargetScan were used to predict target genes of miRNAs. Autophagy genes were collected from the Human Autophagy Database (HADb). The GSE151052 dataset was used to identify autophagy-related differentially expressed genes in tissues. Functional enrichment and protein-protein interaction (PPI) network analyses were conducted via Metascape and the STRING network. Spearman correlation analysis was used to analyze the relationship between autophagy-related differentially expressed genes and lung function. The BCL2-related ceRNA network was modeled by Cytoscape. Results: We obtained 41 differentially expressed miRNAs and 10 significantly different transcription factors. We identified 19 autophagy-related differentially expressed genes that were significantly different (P<0.05) in tissue samples. The most significant enrichment in Metascape was an autophagy item, which further confirmed autophagy participation in the occurrence of COPD. PPI network analysis found four genes (BCL2, BECN1, MAPK8, and ITPR1), among which BCL2 was correlated with both FEV1/FVC and FEV1 prediction. Finally, the BCL2-related ceRNA network was constructed to clarify the interaction of RNAs and occurrence of autophagy, including 18 miRNAs and 65 lncRNAs. Conclusion: We identified 19 autophagy-related differentially expressed genes that participated in COPD; among them, BCL2 was correlated with lung function, and a BCL2-related ceRNA network was constructed, which further revealed the potential mechanism of autophagy involvement in COPD.

Spider-web-inspired membrane reinforced with sulfhydryl-functionalized cellulose nanocrystals for oil/water separation.

The cellulose nanocrystals (CNC) has attracted widespread attention in reinforced materials. However, the application of CNC in electrospinning has been limited due to its self-polymerization. Herein, a cobweb-like nanofibrous membrane was fabricated by electrospinning the polyacrylonitrile (PAN) and sulfydryl-functionalized CNC (SC). The SC content could reach to 48 wt% after the thiolation modification. The membrane with ultrafine fibers and interlaced nets possessed outstanding porosity (91.7%) and underwater superoleophobicity. An ultrahigh permeation flux of 1244 L·m-2·h-1 with a separation efficiency of >99.9% was achieved driven by gravity. The mechanical properties also enhanced significantly with the increase of SC. When the addition amount of SC was 48 wt%, the maximum tensile stress was 2.9 MPa, which was 3.4 times than that of the PAN membrane. The antifouling performance and chemical stability endowed the SC(48)/PAN membrane with intriguing reusability, thus making it exhibit enormous potential in oil/water separation.

Serum ITIH4 in coronary heart disease: a potential anti-inflammatory biomarker related to stenosis degree and risk of major adverse cardiovascular events.

Aim: This study aimed to investigate the correlation of ITIH4 with inflammatory cytokines, stenosis degrees and prognosis in coronary heart disease (CHD) patients. Methods: Serum ITIH4 levels of 300 CHD patients and 30 controls, together with levels of TNF-α, IL-6, IL-8 and IL-17A of CHD patients, were determined using ELISA. Results: Serum ITIH4 was reduced in CHD patients versus controls (p < 0.001). ITIH4 was negatively linked with TNF-α, IL-6, IL-8, IL-17A, C-reactive protein, serum creatinine and Gensini score in CHD patients (all p < 0.050). ITIH4 quartile level negatively correlated with the cumulative major adverse cardiovascular event rate (p = 0.041). Conclusion: Serum ITIH4 may serve as an anti-inflammatory biomarker that negatively associates with stenosis degree and major adverse cardiovascular event risk in CHD patients.

What is this article about? This study aimed to find the clinical value of measuring the protein ITIH4 in patients with coronary heart disease (CHD). What was done? A total of 300 CHD patients and 30 non-CHD people (with chest pain or suspected CHD symptoms) were enrolled in this study. Blood ITIH4 levels of all people were detected. What were the results? Blood ITIH4 levels were lower in CHD patients compared with non-CHD people. In CHD patients, a high level of ITIH4 was associated with low inflammation, reduced vessel narrowness and a good prognosis. What do the results mean? Blood ITIH4 serves as an anti-inflammatory biomarker, whose high level represents better conditions in CHD patients.

Wheat-associated Pseudomonas taiwanensis WRS8 reduces cadmium uptake by increasing root surface cadmium adsorption and decreasing cadmium uptake and transport related gene expression in wheat.

Metal-resistant bacteria can reduce Cd accumulation in plants, but mechanisms underlying this effect are poorly understood. In this study, a highly effective Cd-resistant WRS8 strain was obtained from the rhizoshere soil of Triticum aestivum L. Yangmai-13 and identified as Pseudomonas taiwanensis based on 16S rRNA gene sequence analysis. Strain WRS8 was investigated for its effects on Cd availability and wheat tissue Cd contents and the related mechanisms using a hydroponic culture experiment. In strain WRS8-inoculated solution, the Cd concentration reduced and the pH and cell-adsorbed Cd increased with time. Strain WRS8 increased the wheat root and above-ground tissue dry weights by 11-36% compared to the controls. In strain WRS8-inoculated wheat plants, the Cd contents of the roots and above-ground tissues decreased by 78-85% and 88-94% and the Cd bioconcentration and translocation factors decreased by 78-85% and 46-58% at days 3 and 10, respectively, compared with the controls. The root surface-adsorbed Cd contents increased by 99-121% in the WRS8 strain-inoculated wheat plants at days 3 and 10 compared to the controls. Furthermore, strain WRS8 colonized the wheat root surfaces and interiors and reduced the expression levels of the LCT1 and HMA2 genes involved in Cd accumulation and transport in wheat roots by 46% and 30%, respectively, compared to the controls. In the Cd-contaminated soils, strain WRS8 significantly reduced the available Cd content by 20-24% and increased the pH compared to the controls. These findings showed the important role of strain WRS8 in reducing solution and soil Cd availability and suggested that strain WRS8 reduced the wheat tissue Cd accumulation by increasing root surface Cd adsorption and decreasing wheat root Cd uptake and transport-related gene expression and may provide a new and effective wheat rhizobacteria-enhanced approach for reducing wheat Cd uptake in Cd-polluted environments.

Homogeneous Electrochemiluminescence Biosensor for the Detection of RNase A Activity and Its Inhibitor.

Ribonuclease A (RNase A) is increasingly considered as a biomarker for tumor diagnosis, and it is of great significance to develop an ultrasensitive, cost-effective assay for RNase A detection. Electrochemiluminescence (ECL) technology has distinctive advantages in the development of biosensors for diverse targets. However, most of the ECL biosensors require the complex process of electrode modification, which is laborious and time consuming. In this work, an immobilization-free homogeneous ECL assay was developed for the highly sensitive detection of RNase A activity for the first time. On the basis of the fact that RNase A can specifically hydrolyze RNA at the site of ribonucleotide uracil (rU), a rU-containing chimeric DNA probe is designed and labeled with Ru(bpy)32+ (act as ECL indicator). The chimeric DNA probe hardly diffuses to the surface of negatively charged indium tin oxide (ITO) electrode due to the strong electrostatic repulsion between the negatively charged DNA and ITO electrode, resulting in a weak ECL signal detected. When the RNase A is present, the chimeric DNA probe is hydrolyzed into small fragments, which contains little negative charge and can diffuse easily to the ITO electrode surface due to the decreased electrostatic repulsion. In this case, an enhanced ECL signal can be detected. Under the optimal conditions, there is a linear relationship between the ECL signal and the concentration of RNase A in the range of 0.001-0.10 ng/mL, and the detection limit is 0.2 pg/mL. In addition, the proposed ECL sensing system is also applied to detect the RNase A inhibitor, taking As3+ as an example. The proposed homogeneous ECL sensing system provides a new approach for the highly sensitive and convenient detection of RNase A as well as other ribonucleases only by redesigning a responding chimeric DNA probe.

High-performance non-enzymatic glucose sensor by hierarchical flower-like nickel(II)-based MOF/carbon nanotubes composite.

A hierarchical three-dimensional flower-like nickel(II)-terephthalic acid (Ni(TPA)) metal-organic framework (MOF) was synthesized through a simple solvothermal method. Then the single-walled carbon nanotubes (SWCNT) were doped with the synthesized Ni(TPA) MOF by the ultrasonic method to improve the chemical stability and the electrochemical activity of the MOF. To explore the potential application of the nanocomposite, the Ni(TPA)-SWCNT modified glassy carbon electrode was prepared and used as a non-enzymatic electrochemical sensor for glucose. The results show that the nanocomposite demonstrates remarkably higher electrochemical response as well as stronger electrocatalytic activity toward glucose oxidation, in comparison with the single-component of Ni(TPA) and SWCNT. The Ni(TPA)-SWCNT modified electrode exhibits outstanding performance including excellent selectivity, wide linear range from 20 µM to 4.4 mM, low detection limit of 4.6 µM, and fast response (<5 s) for glucose detection. For glucose analysis in real serum samples, the developed sensor reveals a good agreement with the automatic biochemical analyzer, with the deviation rate of 0-6.7% and correlation coefficient (r) of 0.9940 (n = 20, P < 0.0001), indicating high accuracy and great potential of the sensor for practical application in fast analysis of glucose.

An in situ assembly strategy for the construction of a sensitive and reusable electrochemical aptasensor.

The 5'-end thiolated aptamer is attached on the electrode surface through hybridization with immobilized assistant strands. Then the signal enhancing platform of gold nanoparticles and an electroactive Cu2+-l-cysteine tag are in situ tethered on the 5'-end of the aptamer, through which, a sensitive and reusable aptasensor is facilely constructed.

Graphene oxide directed in-situ synthesis of Prussian blue for non-enzymatic sensing of hydrogen peroxide released from macrophages.

A novel electrochemical non-enzymatic hydrogen peroxide (H2O2) sensor has been developed based on Prussian blue (PB) and electrochemically reduced graphene oxide (ERGO). The GO was covalently modified on glassy carbon electrode (GCE), and utilized as a directing platform for in-situ synthesis of electroactive PB. Then the GO was electrochemically treated to reduction form to improve the effective surface area and electroactivity of the sensing interface. The fabrication process was characterized by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX) and atomic force microscopy (AFM). The results showed that the rich oxygen containing groups play a crucial role for the successful synthesis of PB, and the obtained PB layer on the covalently immobilized GO has good stability. Electrochemical sensing assay showed that the modified electrode had tremendous electrocatalytic property for the reduction of H2O2. The steady-state current response increased linearly with H2O2 concentrations from 5µM to 1mM with a fast response time (less than 3s). The detection limit was estimated to be 0.8µM. When the sensor was applied for determination of H2O2 released from living cells of macrophages, satisfactory results were achieved.

Immobilization free electrochemical biosensor for folate receptor in cancer cells based on terminal protection.

The determination of folate receptor (FR) that over expressed in vast quantity of cancerous cells frequently is significant for the clinical diagnosis and treatment of cancers. Many DNA-based electrochemical biosensors have been developed for FR detection with high selectivity and sensitivity, but most of them need complicated immobilization of DNA on the electrode surface firstly, which is tedious and therefore results in the poor reproducibility. In this study, a simple, sensitive, and selective electrochemical FR biosensor in cancer cells has been proposed, which combines the advantages of the convenient immobilization-free homogeneous indium tin oxide (ITO)-based electrochemical detection strategy and the high selectivity of the terminal protection of small molecule linked DNA. The small molecule of folic acid (FA) and an electroactive molecule of ferrocence (Fc) were tethered to 3'- and 5'-end of an arbitrary single-stranded DNA (ssDNA), respectively, forming the FA-ssDNA-Fc complex. In the absence of the target FR, the FA-ssDNA-Fc was degraded by exonuclease I (Exo I) from 3'-end and produced a free Fc, diffusing freely to the ITO electrode surface and resulting in strong electrochemical signal. When the target FR was present, the FA-ssDNA-Fc was bound to FR through specific interaction with FA anchored at the 3'-end, effectively protecting the ssDNA strand from hydrolysis by Exo I. The FR-FA-ssDNA-Fc could not diffuse easily to the negatively charged ITO electrode surface due to the electrostatic repulsion between the DNA strand and the negatively charged ITO electrode, so electrochemical signal reduced. The decreased electrochemical signal has a linear relationship with the logarithm of FR concentration in range of 10fM to 10nM with a detection limit of 3.8fM (S/N=3). The proposed biosensor has been applied to detect FR in HeLa cancer cells, and the decreased electrochemical signal has a linear relationship with the logarithm of cell concentration ranging from 100-10000cell/mL. Compared with the traditional heterogeneous electrochemical FR biosensors, the proposed biosensor owns the merits of the simplicity and high specificity, presenting the great potential application in the area of early diagnosis of cancers.