Monitoring response to neoadjuvant therapy for breast cancer in all treatment phases using an ultrasound deep learning model.

Purpose: The aim of this study was to investigate the value of a deep learning model (DLM) based on breast tumor ultrasound image segmentation in predicting pathological response to neoadjuvant chemotherapy (NAC) in breast cancer. Methods: The dataset contains a total of 1393 ultrasound images of 913 patients from Renmin Hospital of Wuhan University, of which 956 ultrasound images of 856 patients were used as the training set, and 437 ultrasound images of 57 patients underwent NAC were used as the test set. A U-Net-based end-to-end DLM was developed for automatically tumor segmentation and area calculation. The predictive abilities of the DLM, manual segmentation model (MSM), and two traditional ultrasound measurement methods (longest axis model [LAM] and dual-axis model [DAM]) for pathological complete response (pCR) were compared using changes in tumor size ratios to develop receiver operating characteristic curves. Results: The average intersection over union value of the DLM was 0.856. The early-stage ultrasound-predicted area under curve (AUC) values of pCR were not significantly different from those of the intermediate and late stages (p< 0.05). The AUCs for MSM, DLM, LAM and DAM were 0.840, 0.756, 0.778 and 0.796, respectively. There was no significant difference in AUC values of the predictive ability of the four models. Conclusion: Ultrasonography was predictive of pCR in the early stages of NAC. DLM have a similar predictive value to conventional ultrasound for pCR, with an add benefit in effectively improving workflow.

Effect of surgery-first approach on quality of life and mental health of orthognathic patients: A systematic review and meta-analysis.

Objectives: This study intends to explore the effects of the surgery-first approach (SFA) on quality of life and mental health of patients who undergo orthognathic surgery compared to the conventional three-stage approach (CTA). Data: The analysis included eight studies with a total of 307 patients, of which one was randomized controlled trial (RCT), one was clinical controlled trial (CCT), and six were non-randomized studies of interventions (NRSIs). Sources: Electronic databases such as Medline, Embase, Scopus, and Web of Science were searched for eligible trials up to April 2023. Study selection: RCTs, CCTs, and NRSIs, which compared the quality of life or mental health of orthognathic patients treated with SFA and CTA, were included in this study. The meta-analysis showed that the standardized mean differences (SMD) of Oral Health Impact Profiles-14 (OHIP-14) scores and the Orthognathic Quality of Life Questionnaire (OQLQ) between SFA and CTA were -1.58 (P = 0.05) and -2.99 (P < 0.00001) at the termination of the first-stage treatment, which altered to -0.94 (P = 0.54) and 0.09 (P = 0.65) after total treatment. Two studies applied the Psychosocial Impact of Dental Aesthetics Questionnaire (PIDAQ) and the Beck Depression Inventory (BDI-II) to examine mental health, resulting in a trend similar to the former scales. Conclusion: In contrast to the conventional procedure, orthognathic treatment with SFA can instantly enhance the quality of life at the end of the first-stage treatment but has similar effects after the overall treatment. Moreover, SFA has a positive impact on psychological conditions. Clinical significance: This study first systematically reviewed the effect of SFA on patients' mental well-being. According to our findings, it is better to select SFA if possible. Otherwise, the patient's psychological condition should be monitored appropriately throughout decompensation for better well-being both physically and mentally.

WTAP Mediates NUPR1 Regulation of LCN2 Through m6A Modification to Influence Ferroptosis, Thereby Promoting Breast Cancer Proliferation, Migration and Invasion.

Ferroptosis is involved in various pathophysiological diseases, including triple-negative breast cancer (TNBC). Targeting ferroptosis is considered as a novel anti-TNBC strategy. Nevertheless, the regulatory mechanism of ferroptosis during TNBC progression is unclear. Here, the role of WTAP in ferroptosis during TNBC progression  was investigated. The clinicopathological significance of WTAP, NUPR1 and LCN2 was analyzed by Kaplan-Meier method. Cell viability was assessed using MTT assay. Transwell assay was employed to analyze cell migration and invasion. GSH/GSSG and Fe2+ levels in TNBC cells were analyzed using kits. m6A level was examined using m6A dot blot assay. NUPR1 mRNA stability was analyzed using RNA degradation assay. RIP was performed to analyze the interaction between eIF3a and NURP1. Herein, our results revealed that WTAP, NUPR1 and LCN2 expressions were significantly elevated in TNBC. NUPR1 silencing inhibited TNBC cell proliferation, migration and invasion by inducing ferroptosis. NUPR1 positively regulated LCN2 expression in TNBC cells, and LCN2 knockdown induced ferroptosis to suppress TNBC cell malignant behaviors. Our molecular study further revealed that WTAP promoted NUPR1 expression in an m6A-EIF3A mediated manner. And, as expected, WTAP knockdown promoted ferroptosis to suppress TNBC cell malignant behaviors, which were abrogated by NUPR1 overexpression. WTAP upregulated LCN2 by regulation of NUPR1 m6A modification, thereby suppressing ferroptosis to contribute to accelerate TNBC progression. Our study revealed the cancer-promoting effect of WTAP, NUPR1 and LCN2 in TNBC and clarified the relevant mechanism, providing a theoretical basis for developing novel diagnostic and therapeutic strategies for TNBC.

Ordered heterogeneity in dual-ligand MOF to enable high electrochemiluminescence efficiency for bioassay with DNA triangular prism as signal switch.

Herein, the microRNA-141 electrochemiluminescence (ECL) bioassay was developed using the dual-ligand metal-organic framework (d-MOF) with ordered heterogeneity, which simultaneously contained the luminophore ligands (1,1,2,2-tetra(4-carboxylbiphenyl)ethylene, denoted as TCBPE) and the coreactant ligands (1,4-diazabicyclo[2.2.2]octane, denoted as DN2H2). The resultant d-MOF revealed significantly enhanced ECL intensity without any exogenous coreactants, which was 3.53 times higher in comparison with that of single-ligand MOF (only TCBPE as ligands) even with the addition of exogenous DN2H2. Thanks to the ordered heterogeneity in d-MOF, the intramolecular rotation of TCBPE was restricted via oriented coordination and the spatial location of DN2H2 was reasonably arranged due to the framework structure, which could not only enhance the excitation efficiency but also improve the electron-transfer efficiency based on the synergistic enhancement effect between structures and compositions in micro/nano confined space. Based on this, the proposed biosensor employed a novel DNA triangular prism (DNA TP) as signal switch to detect microRNA-141, achieving the low detection limit at the level of 22.9 aM and a broad linear ranging from 100 aM to 100 pM. The precise design of the ordered d-MOFs by co-assembling the luminophore and coreactant ligands holds a promise strategy to achieve ECL MOFs and construct the ECL biosensors in diagnostic analysis.

iCVM: An Interpretable Deep Learning Model for CVM Assessment Under Label Uncertainty.

The Cervical Vertebral Maturation (CVM) method aims to determine the craniofacial skeletal maturational stage, which is crucial for orthodontic and orthopedic treatment. In this paper, we explore the potential of deep learning for automatic CVM assessment. In particular, we propose a convolutional neural network named iCVM. Based on the residual network, it is specialized for the challenges unique to the task of CVM assessment. 1) To combat overfitting due to limited data size, multiple dropout layers are utilized. 2) To address the inevitable label ambiguity between adjacent maturational stages, we introduce the concept of label distribution learning in the loss function. Besides, we attempt to analyze the regions important for the prediction of the model by using the Grad-CAM technique. The learned strategy shows surprisingly high consistency with the clinical criteria. This indicates that the decisions made by our model are well interpretable, which is critical in evaluation of growth and development in orthodontics. Moreover, to drive future research in the field, we release a new dataset named CVM-900 along with the paper. It contains the cervical part of 900 lateral cephalograms collected from orthodontic patients of different ages and genders. Experimental results show that the proposed approach achieves superior performance on CVM-900 in terms of various evaluation metrics.

Construction and validation of an eight pyroptosis-related lncRNA risk model for breast cancer.

OBJECTIVE: We developed a risk model based on pyroptosis-related long non-coding RNAs (lncRNAs) and assessed its prognostic value and clinical significance in breast cancer (BRCA). METHODS: BRCA RNA sequencing data with corresponding clinical information were retrieved from The Cancer Genome Atlas (TCGA) database. Univariate Cox regression analysis was used to examine correlations between prognosis of BRCA patients and the expression levels of pyroptosis-related lncRNAs. A prognostic model was developed and validated by identifying the correlation of risk scores with tumor immune infiltration and immune cell function through immune response analysis. Functional analyses of focal dysfunction-related lncRNA were also carried out. Lastly, single sample gene set enrichment analysis (ssGSEA) was conducted to determine the differences in immune responses between the low- and high-risk groups. RESULTS: We divided the TCGA-BRCA dataset into 3 clusters by consensus clustering, and identified 11 pyroptosis-related lncRNAs that are differentially expressed between tumors and normal tissues. In addition, we determined if PD-L1 expression is associated with clustering and gene expression. The list was further narrowed down to eight pyroptosis-related lncRNAs and their regression coefficients were obtained through LASSO regression analysis. The relative proportion of 22 different immune cells in the BRCA microenvironment was determined using the CIBERSORT algorithm to explore the indicative effects of risk score on the tumor microenvironment (TME). We found that the resting mast cells, M0, and M2 macrophages were positively correlated with the risk scores. CONCLUSION: The potential role of pyroptosis-related lncRNAs in BRCA prognosis may be exploited as a treatment target for patients with BRCA.

Development of Hollow Electrochemiluminescent Nanocubes Combined with a Multisite-Anchored DNA Nanomachine for Mycotoxin Detection.

Polycyclic aromatic hydrocarbons (PAHs) are regarded as promising electrochemiluminescent (ECL) emitters owing to their high quantum efficiency and inexpensive production. Despite the fact that the ECL properties of the pure PAH microcrystal (such as rubrene microcrystals, Rub MCs) have gained extensive attention, it is a challenge in controlling the morphology and size to reduce the inner filter effect. Herein, an advanced ECL emitter of palladium nanoparticle-functionalized hollow PAH-metal nanocubes was prepared by an in situ redox deposition method (the resultant nanocomposites were abbreviated as Pd-Rub-Ag@Au nanocubes). Specifically, the rubrene-decorated Ag@Au nanocubes (Rub-Ag@Au nanocubes) were prepared using the Ag@Au nanocubes as a template and a rubrene cation radical (Rub•+) as a reductant, and then Pd nanoparticles (Pd NPs) were in situ reduced on the surface of Rub-Ag@Au nanocubes. Impressively, compared with the Rub MCs, Pd-Rub-Ag@Au nanocubes showed uniform size and significantly enhanced ECL efficiency and intensity in the aqueous media. As a proof-of-concept, the Pd-Rub-Ag@Au nanocube-based ECL biosensing platform combined with a multisite-anchored DNA nanomachine was constructed for ochratoxin A (OTA, a type of mycotoxin) detection. The DNA nanomachine covered with high-density recognizing sequences could operate toehold-mediated strand displacement amplification on the sensing platform and promote the movement efficiency and velocity greatly. Due to the advanced performance of Pd-Rub-Ag@Au nanocubes and high recognition efficiency of the DNA nanomachine, the proposed biosensor for OTA detection can achieve a detection limit of 4.7 fg/mL ranging from 0.01 to 100 pg/mL, which offers an ingenious method for the further application of PAHs.

Swing Arm Location-Controllable DNA Walker for Electrochemiluminescence Biosensing.

Here, we described a novel swing arm location-controllable DNA walker based on the DNA tetrahedral nanostructures (DTNs) for nucleic acid detection using the polycyclic aromatic hydrocarbon (PAH) microcrystals (TAPE-Pe MCs) consisting of the nonplanar molecular tetrakis(4-aminophenyl)ethene (TAPE) and planar molecular perylene (Pe) as electrochemiluminescence (ECL) luminophores. Specifically, the swing arm strands and track strands were fixed simultaneously on the DTNs to obtain the location-controllable DNA walker, which possessed an improved reaction efficiency compared to that of a fixed swing arm-based DNA walker due to the quantitative and orderly swing arm on the DTNs. On the other hand, the Pe microcrystals doped by TAPE molecules could decrease the π-π stacking of Pe molecules for the ECL efficiency enhancement, achieving a blue-shifted and intense ECL emission. Therefore, we defined this enhanced and blue-shifted ECL phenomenon as "inhibition of conjugation-driven ECL (IC-ECL)". To prove these principles, a location-controllable DNA walker-based ECL biosensor was developed with microRNA let-7a as target molecules. The ECL biosensor achieved a low detection limit of 4.92 fM within a wide linear range from 10 fM to 100 nM. This approach offers a new insight for ECL efficiency increase and location-controllable strategies with improved reaction efficiency, demonstrating potential in diagnostic analysis.

A synergistic promotion strategy remarkably accelerated electrochemiluminescence of SnO2 QDs for MicroRNA detection using 3D DNA walker amplification.

Developing low-cost and efficient methods to enhance the electrochemiluminescence (ECL) intensity of luminophores is highly desirable and challenging. Herein, we develop a synergistic promotion strategy based on three types of co-reaction accelerators to achieve an efficient SnO2 quantum dots (SnO2 QDs)-based ternary ECL system. Specifically, the MnO2 nanoflowers (MnO2 NFs), Ag nanoparticles (Ag NPs) and hemin/G-quadruplex were rationally selected as co-reaction accelerators. Owing to the synergistic effect, the deft integration of three types of co-reaction accelerators enabled better structural stability, more exposed catalytic active sites, and faster charge transfer, thus more effectively facilitating the reduction of co-reactant (S2O82-) compared with that of the single co-reaction accelerator. To demonstrate the practical utility of this principle, an "on-off-super on" ECL biosensor was constructed in combination with a 3D DNA walker, which showed a superior linear range (10 aM-100 pM) and a low detection limit (2.9 aM) for the highly-sensitive miRNA-21 detection. In general, this work firstly reported that three types of co-reaction accelerators were deftly integrated to remarkably amplify the ECL emission of SnO2 QDs, and provided brand-new perspectives for research on the ingenious design of the structure and component of highly efficient co-reaction accelerators.

Potential Role of Integrin α5ß1/Focal Adhesion Kinase (FAK) and Actin Cytoskeleton in the Mechanotransduction and Response of Human Gingival Fibroblasts Cultured on a 3-Dimension Lactide-Co-Glycolide (3D PLGA) Scaffold.

BACKGROUND The stability of orthodontic treatment is thought to be significantly affected by the compression and retraction of gingival tissues, but the underlying molecular mechanism is not fully elucidated. The objectives of our study were to explore the effects of mechanical force on the ECM-integrin-cytoskeleton linkage response in human gingival fibroblasts (HGFs) cultured on 3-dimension (3D) lactide-co-glycolide (PLGA) biological scaffold and to further study the mechanotransduction pathways that could be involved. MATERIAL AND METHODS A compressive force of 25 g/m² was applied to the HGFs-PLGA 3D co-cultured model. Rhodamine-phalloidin staining was used to evaluate the filamentous actin (F-actin) cytoskeleton. The expression level of type I collagen (COL-1) and the activation of the integrin alpha5ß1/focal adhesion kinase (FAK) signaling pathway were determined by using real-time PCR and Western blotting analysis. The impacts of the applied force on the expression levels of FAK, phosphorylated focal adhesion kinase (p-FAK), and COL-1 were also measured in cells treated with integrin alpha5ß1 inhibitor (Ac-PHSCN-NH 2, ATN-161). RESULTS Mechanical force increased the expression of integrin alpha5ß1, FAK (p-FAK), and COL-1 in HGFs, and induced the formation of stress fibers. Blocking integrin alpha5ß1 reduced the expression of FAK (p-FAK), while the expression of COL-1 was not fully inhibited. CONCLUSIONS The integrin alpha5ß1/FAK signaling pathway and actin cytoskeleton appear to be involved in the mechanotransduction of HGFs. There could be other mechanisms involved in the promotion effect of mechanical force on collagen synthesis in addition to the integrin alpha5ß1 pathway.

DNA Structure Transition-Induced Affinity Switch for Biosensing Based on the Strong Electrochemiluminescence Platform from Organic Microcrystals.

Here we described an effective recognition strategy using the target-triggered DNA structure transition as an affinity switch for nucleic acid detection based on the strong electrochemiluminescence (ECL) platform of 9,10-diphenylanthracene (DPA) doped perylene (Pe) microcrystals (DPA@Pe MCs). Specifically, the target-triggered rolling-circle amplification (RCA) could generate a long, single-stranded DNA with repeated G-quadruplex units, which would hinder the access of quenching probes due to the steric hindrance effects offered by the DNA structure transition. Using this effective recognition strategy, an ECL biosensor with ultrasensitive and accurate characteristics was proposed to detect microRNA-21, which showed an excellent linear response from 10 aM to 1 pM with the detection limit down to 4.14 aM. The DNA structure transition-induced affinity switch strategy offered a potential applications in clinical diagnosis analysis.

Mechanical force promotes the proliferation and extracellular matrix synthesis of human gingival fibroblasts cultured on 3D PLGA scaffolds via TGF­ß expression.

Human gingival fibroblasts (HGFs) are responsible for connective tissue repair and scarring, and are exposed to mechanical forces under physiological and pathological conditions. The exact mechanisms underlying gingival tissue reconstruction under mechanical forces remain unclear. The present study aimfed to investigate the effects of mechanical forces on the proliferation and extracellular matrix synthesis in HGFs by establishing a 3­dimensional (3D) HGF culture model using poly(lactide­co­glycolide) (PLGA) scaffolds. HGFs were cultured in 3D PLGA scaffolds and a mechanical force of 0, 5, 15, 25 or 35 g/cm2 was applied to HGFs for 24 h. A mechanical force of 25 g/cm2 induced the highest proliferation rate, and thus was selected for subsequent experiments. Cell viability was determined using the MTT assay at 0, 24, 48 and 72 h. The expression levels of type I collagen (COL­1) and matrix metallopeptidase (MMP)­1 were examined by reverse transcription­quantitative polymerase chain reaction and ELISA, and transforming growth factor (TGF)­ß expression was evaluated by ELISA. The application of mechanical force on HGFs cultured on the 3D PLGA scaffolds resulted in a significant increase in cell proliferation and COL­1 expression, as well as a decrease in MMP­1 expression. A TGF­ß1 inhibitor was also applied, which attenuated the effects of mechanical force on HGF proliferation, and COL­1 and MMP­1 expression, thus suggesting that TGF­ß signaling pathways may mediate the mechanical force­induced alterations observed in HGFs. In conclusion, these findings helped to clarify the mechanisms underlying mechanical force­induced HGF proliferation and ECM synthesis, which may promote the development of targeted therapeutics to treat various diseases, including gingival atrophy caused by orthodontic treatment.

Inhibiting ROS-TFE3-dependent autophagy enhances the therapeutic response to metformin in breast cancer.

Autophagy modulation is a potential therapeutic strategy for breast cancer, and a previous study indicated that metformin exhibits significant anti-carcinogenic activity. However, the ability of metformin to induce autophagy and its role in breast cancer cell death remains unclear. In this study, we exposed MCF-7 cells to different concentrations of metformin (2.5, 5, and 10 mM) for 48 h, and metformin-induced significant apoptosis in the MCF-7 cells. The expression levels of CL-PARP (poly(ADP-ribose) polymerase 1) and the ratio of BAX to BCL-2 were significantly increased. In addition to apoptosis, we showed that metformin increased autophagic flux in MCF-7 cells, as evidenced by the upregulation of LC3-II and downregulation of P62/SQSTM1. Moreover, pharmacological or genetic blocking of autophagy increased metformin-induced apoptosis, indicating a cytoprotective role of autophagy in metformin-treated MCF-7 cells. Mechanistically, metformin-induced TFE3(Ser321) dephosphorylation activated TFE3 nuclear translocation and increased of TFE3 reporter activity, which contributed to lysosomal biogenesis and the expression of autophagy-related genes and, subsequently, initiated autophagy in MCF-7 cells. Importantly, we found that metformin triggered the generation of reactive oxygen species (ROS) in MCF-7 cells. Furthermore, N-acetyl-l-cysteine (NAC), a ROS scavenger, abrogated the effects of metformin on TFE3-dependent autophagy. Notably, TFE3 expression positively correlated with breast cancer development and poor prognosis in patients. Taken together, these data demonstrate that blocking ROS-TFE3-dependent autophagy to enhance the activity of metformin warrants further attention as a treatment strategy for breast cancer.

Hemin as electrochemically regenerable co-reaction accelerator for construction of an ultrasensitive PTCA-based electrochemiluminescent aptasensor.

In this work, hemin was firstly used as electrochemically regenerable co-reaction accelerator for signal amplification to develop an ultrasensitive aptasensor for Aflatoxin M1 (AFM1) detection. Initially, the perylenetetracarboxylic acid (PTCA) was directly employed as luminophore to construct the ECL sensing nano-platform by combining Au nanoparticles (Au NPs) for immobilizing thiol-terminated hairpin probe (H1). Then with the help of hairpin H2, H3, the AFM1-catalyzed hairpin assembly (CHA) was executed to produce the H1-H3 duplex, which could further initiate the hybridization chain reaction (HCR) to generate dendritic DNA polymers consisting of G-rich sequence for capturing large quantities of hemin on the electrode surface. Herein, hemin as electrochemically regenerable co-reaction accelerator could interact with the co-reactant (S2O82-) to obviously improve the luminous efficiency of the PTCA. Therefore, a strong and stable ECL signal was achieved by the employment of hemin as electrochemically regenerable co-reaction accelerator. The proposed aptasensor determined AFM1 down to 0.09pgmL-1 within a linear range of 0.4pgmL-1 to 400ngmL-1. With excellent sensitivity and stability, the strategy provided an efficient and simple method for the trace detection of biomolecules in clinical analysis.

Evaluation of surgical anti-adhesion products to reduce postsurgical intra-abdominal adhesion formation in a rat model.

BACKGROUND: Adhesions frequently occur after abdominal surgery. Many anti-adhesion products have been used in clinic. However, the evidences are short for surgeons to reasonably choose the suitable anti-adhesion produces in clinical practice. This study provided such evidence by comparing the efficiency of five products to prevent abdominal adhesion formation in a rat model. METHODS: Fifty-six Sprague-Dawley rats were randomly divided into seven groups: sham-operation group, adhesion group, and five product groups (n = 8). The abdomens of rats were opened. The injuries were created on abdominal wall and cecum in the adhesion and product groups. The wounds on abdominal wall and cecum of rats in the adhesion group were not treated before the abdomens were closed. The wounds on abdominal wall and cecum of rats in the product groups were covered with anti-adhesion product: polylactic acid (PLA) film, Seprafilm®, medical polyethylene glycol berberine liquid (PEG), medical sodium hyaluronate gel (HA), or medical chitosan (Chitosan). Fourteen days after surgery, the adhesions were evaluated by incidence, severity, adhesion area on abdominal wall and adhesion breaking strength. RESULTS: The application of PLA film and Seprafilm® significantly reduced the incidence, severity, adhesion area and breaking strength of cecum-abdomen adhesion (P<0.05). HA, PEG and Chitosan failed to significantly reduce the cecum-abdomen adhesion (P>0.05). The statistical significances in the incidence and severity of abdomen-adipose adhesion between adhesion group and the product groups were not achieved. However, Seprafilm® was more effective to reduce abdomen-adipose adhesion than PLA film. Furthermore, it was found that the products tested in this study did not effectively reduce cecum-adipose adhesion. The application of PEG could result in abdomen-small intestine adhesion. CONCLUSION: Based on the results of this study, the preference order of anti-adhesion products used to reduce postsurgical intra-abdominal adhesion formation is Seprafilm > PLA >> HA > Chitosan > PEG.

Inhibiting ROS-STAT3-dependent autophagy enhanced capsaicin-induced apoptosis in human hepatocellular carcinoma cells.

Capsaicin, which is the pungent ingredient of red hot chili peppers, has been reported to possess anticancer activity, including that against hepatocellular carcinoma. However, the precise molecular mechanisms by which capsaicin exerts its anticancer effects remain poorly understood. Herein, we have tested the involvement of autophagy in the capsaicin mechanism of action in human hepatocellular carcinoma. HepG2 cancer cells were treated with different doses of capsaicin (50, 100 and 200µmol/L) for 6, 12, and 24 h. Flow cytometry and Caspase-3 activity assay were performed to determine cell apoptosis. Immunofluorescence was performed to visualize LC3-positive puncta. Western blotting was used to detect the expression of the hallmarks of apoptosis and autophagy. Capsaicin can induce apoptosis in HepG2 cells. The expression levels of CL-PARP and Bcl-2 were significantly increased. In line with the apoptosis, capsaicin can trigger autophagy in HepG2 cells. Capsaicin increased LC3-II and beclin-1 expression and GFP-LC3-positive autophagosomes. Pharmacological or genetic inhibition of autophagy further sensitized HepG2 cells to capsaicin-induced apoptosis. Mechanistically, capsaicin upregulated the Stat3 activity which contributed to autophagy. Importantly, we found that capsaicin triggered reactive oxygen species (ROS) generation in hepatoma cells and that the levels of ROS decreased with N-acetyl-cysteine (NAC), a ROS scavenger. Moreover, NAC abrogated the effects of capsaicin on Stat3-dependent autophagy. In this study, we demonstrated that capsaicin increased the phosphorylation of signal transducer and activator of transcription 3 (p-STAT3)-dependent autophagy through the generation of ROS signaling pathways in human hepatoma. Inhibiting autophagy could enhance capsaicin-induced apoptosis in human hepatocellular carcinoma.

Triple Quenching of a Novel Self-Enhanced Ru(II) Complex by Hemin/G-Quadruplex DNAzymes and Its Potential Application to Quantitative Protein Detection.

Herein, a novel "on-off" electrochemiluminescence (ECL) aptasensor for highly sensitive determination of thrombin has been constructed based on the triple quenching of the effect of hemin/G-quadruplex DNAzymes upon the Ru(II) complex-based ECL system. First, a strong initial ECL signal was achieved by the dual amplification strategies of (i) intramolecular coreaction of a self-enhanced Ru(II)-based molecule (PTCA-PEI-Ru(II)) and (ii) intermolecular coreaction between PTCA-PEI-Ru(II) and nicotinamide adenine dinucleotide (NADH), which was named the signal-on state. Then, a novel triple quenching of the effect of multifunctional hemin/G-quadruplex DNAzymes upon the Ru(II) complex-based ECL system was designed to realize the desirable signal-off state, which was outlined as follows: (i) the hemin/G-quadruplex DNAzymes mimicked NADH oxidase to oxidize NADH and in situ generate the H2O2, consuming the coreactant of NADH; (ii) its active center of hemin could oxidize the excited state PTCA-PEI-Ru(II)* to PTCA-PEI-Ru(III), making the energy and electron transfer quench; (iii) it also acted as horseradish peroxidase (HRP) to catalyze the H2O2 for in situ producing the quencher of O2. Based on triple quenching of the effect of hemin/G-quadruplex DNAzymes, the highly sensitive "on-off" thrombin aptasensor was developed with a wide linear detection range of 1.0 × 10(-14) M to 1.0 × 10(-10) M and a detection limit down to the femtomolar level.

Highly efficient electrogenerated chemiluminescence quenching of PEI enhanced Ru(bpy)3²âº nanocomposite by hemin and Au@CeO2 nanoparticles.

In this work, a new signal amplified strategy based on the quenching effect of hemin and Au nanoparticles decorated CeO2 nanoparticles (Au@CeO2 NPs) for ultrasensitive detection of thrombin (TB) is reported for the first time. Herein, the poly(ethylenimine) (PEI) enhanced Ru(bpy)3(2+) nanocomposite was implemented by direct chemical polymerization, which could provide the desirable enhanced initial ECL signal. Furthermore, the detection aptamer of thrombin (TBA 2) was immobilized on Au@CeO2 NPs to form TBA 2/Au@CeO2 conjugates. Then, the G-rich DNA of TBA 2 sequence could fold into a G-quadruplex structure to embed hemin to obtain the quenching probe of hemin/TBA 2/Au@CeO2 conjugates. In the presence of target TB, the sandwiched structure could be formed between capture aptamer (TBA 1), TB and hemin/TBA 2/Au@CeO2 conjugates, thereby resulting in a proportional quenching in ECL response with TB, due to the quenching of both hemin and Au@CeO2 NPs. As a result, the signal-off aptasensor showed a wider linear range response from 10(-13) to 10(-8) M with lower detection limit of 0.03 pM.

Ultrasensitive immunoassay based on a pseudobienzyme amplifying system of choline oxidase and luminol-reduced Pt@Au hybrid nanoflowers.

A multi-functional luminol-reduced Pt@Au hybrid flower-like nanocomposite (luminol-Pt@AuNF) which not only acts as an efficient signal probe but also constitutes a pseudobienzyme amplifying system with choline oxidase (ChOx) was firstly synthesized and applied to the construction of a solid-state luminol electrochemiluminescence (ECL) immunosensor for cardiac troponin I (cTnI) detection.

A supersandwich electrochemiluminescence immunosensor based on mimic-intramolecular interaction for sensitive detection of proteins.

An electrochemiluminescence (ECL) immunoassay protocol was developed based on mimic-intramolecular interaction for sensitive detection of prostate specific antigen (PSA). It was constructed by integrating the ECL luminophore (tris(4,4'-dicarboxylicacid-2,2'-bipyridyl)-ruthenium(ii)dichloride (Ru(dcbpy)3(2+))) and coreactant (histidine) into the supersandwich DNA structure. This strategy was more effective in amplifying the ECL signal by shortening the electronic transmission distance, improving the ECL luminous stability and enhancing the ECL luminous efficiency. The ECL matrices denoted as MWCNTs@PDA-AuNPs were fabricated through spontaneous oxidative polymerization of dopamine (DA) on multiwalled carbon nanotubes (MWCNTs) and reducing HAuCl4 to produce gold nanoparticles (AuNPs) by DA simultaneously. Then, the prepared matrices were applied to bind capture antibodies. Moreover, supersandwich Ab2 bioconjugate was designed using a PAMAM dendrimer to immobilize the detection antibody and supersandwich DNA structure. The PAMAM dendrimer, with a plurality of secondary and tertiary amine groups, not only facilitated high-density immobilization of the detection antibody and supersandwich DNA structure, but also greatly amplified the ECL signal of Ru(dcbpy)3(2+). The supersandwich DNA structure contained multiple Ru(dcbpy)3(2+) and histidine, further amplifying the ECL signal. The proposed supersandwich immunosensor showed high sensitivity with a detection limit of 4.2 fg mL(-1) and a wide linear range of 0.01 pg mL(-1)-40.00 ng mL(-1). With the excellent stability, satisfying precision and reproducibility, the proposed immunosensor indicates promising practicability for clinical diagnosis.