Multimodal neuroimaging fusion unravel structural-functional-neurotransmitter change in Parkinson's disease with impulse control disorders.

BACKGROUND: Impulse control disorders (ICD) in Parkinson's disease (PD) is highly multifactorial in etiology and has intricate neural mechanisms. Our multimodal neuroimaging study aimed to investigate the specific patterns of structure-function-neurotransmitter interactions underlying ICD. METHODS: Thirty PD patients with ICD (PD-ICD), 30 without ICD (PD-NICD) and 32 healthy controls (HCs) were recruited. Gyrification and perivascular spaces (PVS) were computed to capture the alternations of cortical surface morphology and glymphatic function. Seed-based functional connectivity (FC) were performed to identify the corresponding functional changes. Further, JuSpace toolbox were employed for cross-modal correlations to evaluate whether the spatial patterns of functional alterations in ICD patients were associated with specific neurotransmitter system. RESULTS: Compared to PD-NICD, PD-ICD patients showed hypogyrification and enlarged PVS volume fraction in the left orbitofrontal gyrus (OFG), as well as decreased FC between interhemispheric OFG. The interhemispheric OFG connectivity reduction was associated with spatial distribution of µ-opioid pathway (r = -0.186, p = 0.029, false discovery rate corrected). ICD severity was positively associated with the PVS volume fraction of left OFG (r = 0.422, p = 0.032). Furthermore, gyrification index (LGI) and percent PVS (pPVS) in OFG and their combined indicator showed good performance in differentiating PD-ICD from PD-NICD. CONCLUSIONS: Our findings indicated that the co-altered structure-function-neurotransmitter interactions of OFG might be involved in the pathogenesis of ICD.

Transcutaneous auricular vagus nerve stimulation improves anxiety symptoms and cortical activity during verbal fluency task in Parkinson's disease with anxiety.

OBJECTIVE: To investigate the effect of 20/4Hz transcutaneous auricular vagus nerve stimulation (taVNS) on anxiety symptoms in Parkinson's disease (PD) and the potential neural mechanism. METHODS: In the current randomized, double-blind, sham-controlled trial, 30 PD patients with anxiety (PD-A), 30 PD patients without anxiety (PD-nA), and 30 healthy controls (HCs) were enrolled. PD-A patients were randomly (1:1) allotted to real taVNS stimulation group (RS) or sham stimulation group (SS) to explore the efficacy of a two-week treatment of taVNS to promote anxiety recovery. Simultaneously, all participants were measured activation in the bilateral prefrontal cortex during verbal fluency task (VFT) using functional near-infrared spectroscopy. RESULTS: PD-A patients showed significantly decreased oxyhemoglobin in the left triangle part of the inferior frontal gyrus (IFG) during VFT, which was negatively related to the severity of anxiety symptoms. After two-week treatment of taVNS, the interaction of group and time had significant effect on HAMA scores (F = 18.476, p < 0.001, η2 = 0.398). In RS group, compared with baseline, HAMA scores decreased significantly in the post-treatment and follow-up condition (both p < 0.001). Meanwhile, in RS group, HAMA scores were lower than those in SS group in the post-treatment and follow-up condition (p = 0.006, <0.001, respectively). Furthermore, the 20/4Hz taVNS remarkably ameliorated anxiety symptoms in PD patients, directly correlated with the increased activation of the left triangle part of the IFG during VFT in RS group. CONCLUSION: Our results depicted that taVNS could ameliorate the anxiety symptoms of PD-A patients and regulated the function of the left triangle part of the IFG.

Lower serum insulin-like growth factor 2 level in patients with bipolar disorder is associated with the severity of manic symptoms during manic episodes.

Objective: Accumulating evidence has indicated that neurodevelopmental defects may underlie the pathophysiology of bipolar disorder (BD). Insulin-like growth factors (IGFs) are a family of neurotrophic factors that are essential for the survival and development of neurons. The present study aims to investigate whether IGF-2 signaling is implicated in the pathophysiological processes of BD. Method: 50 healthy controls and 78 patients with BD, including 23 patients who diagnosed acute depressive episode and 55 patients who diagnosed acute manic episode, were recruited in this study. The 17-item Hamilton Depression Rating Scale (HAMD-17) and the Young Mania Rating Scale (YMRS) were used to assess the severity of the depressive and manic symptoms, respectively. The serum IGF-2 level was determined by an enzyme-linked immunosorbent assay (ELISA). The Kolmogorov-Smirnov and Mann-Whitney U tests were used for between-group comparisons and spearman analysis was used to analyze correlations. Results: Patients with BD had lower serum IGF-2 levels (66.08 ± 21.22 ng/ml) when compared to healthy controls (88.72 ± 31.55 ng/ml). BD patients were divided into manic episode and depressive episode subgroups. We found that serum IGF-2 levels were reduced in both the mania and depression subgroups (mania: 67.19 ± 21.52 ng/ml, depression: 63.43 ± 20.67 ng/ml; P < 0.001), while no significant difference was observed between two groups (P > 0.05). Spearman correlation analyses revealed that the levels of serum IGF-2 were negatively correlated with the YMRS scores in BD patients (r = -0.522, P < 0.001). Furthermore, IGF-2 was found to be an independent contributor to the severity of symptoms in patients with manic episodes (B = -0.610, t = -5.299, P < 0.001). Conclusion: Lower serum IGF-2 levels were found in BD patients and correlated with the severity of the manic symptoms in these patients during manic episodes. These results suggest that reduced IGF-2 levels might be involved in the pathophysiology of BD, and serum IGF-2 could be a peripheral biomarker for the evaluation of the severity of manic symptoms in BD patients.

Corrigendum: Lower serum insulin-like growth factor 2 level in patients with bipolar disorder is associated with the severity of manic symptoms during manic episodes.

[This corrects the article DOI: 10.3389/fpsyt.2024.1354999.].

Enhanced in vivo antiviral activity against pseudorabies virus through transforming gallic acid into graphene quantum dots with stimulation of interferon-related immune responses.

With the urgent need for antiviral agents, antiviral materials with high biocompatibility and antiviral effects have attracted a lot of attention. In this study, gallic acid, a natural polyphenolic compound, was transformed into biocompatible graphene quantum dots (GAGQDs) which exhibit enhanced antiviral activity against pseudorabies virus (PRV). The as-prepared GAGQDs inhibit PRV proliferation with a 104-fold reduction in viral titers. Investigation of the antiviral mechanism revealed that GAGQDs inhibit the adsorption, invasion and replication of PRV infection. Treatment with GAGQDs regulates the expression levels of interferon-related antiviral proteins, including mitochondrial antiviral-signaling protein (MAVS), signal transducer and activator of transcription 1 (STAT1) and 2',5'-oligoadenylate synthetase 1 (OAS1), suggesting that GAGQDs can stimulate innate antiviral immune responses, resulting in enhanced antiviral effects. More importantly, GAGQD treatments alleviate clinical symptoms and reduce mortality in PRV-infected mice. Our results reveal the enhanced therapeutic effects of GAGQDs against PRV infection in vitro and in vivo, suggesting the potential of GAGQDs as a promising novel antiviral agent.

Integrative transcriptomic and metabolomic analysis in mice reveals the mechanism by which ginseng stem-leaf saponins enhance mucosal immunity induced by a porcine epidemic diarrhea virus vaccination.

Porcine epidemic diarrhea virus (PEDV) is a main cause of severe enteric disease in piglets, leading to millions of dollars lost annually in the global pig industry. Parenteral vaccination is limited in generating sufficient mucosal immunity, which is crucial for early defense against PEDV. Here, we orally administered ginseng stem-leaf saponins (GSLS) to mice before parenteral vaccination and found that GSLS significantly enhanced the phagocytosis of dendritic cells, promoted the activities of CD4+ T cells and increased PEDV-specific IgA antibodies in the intestinal mucosa. Transcriptomic results showed that the altered genes following GSLS treatment were mostly related to the immune response and metabolism. In addition, integrated analysis of the transcriptome and metabolome revealed that the mechanism by which GSLS enhances mucosal immunity may be associated with progesterone-related pathways. Further studies are needed to explore the detailed molecular mechanisms.

Early Oral Administration of Ginseng Stem-Leaf Saponins Enhances the Peyer's Patch-Dependent Maternal IgA Antibody Response to a PEDV Inactivated Vaccine in Mice, with Gut Microbiota Involvement.

Neonatal piglets during the first week of life are highly susceptible to porcine epidemic diarrhoea virus (PEDV) infection, with mortality rates reaching 80-100%. Passive lactogenic immunity remains the most effective way to protect neonates from infection. Although safe, inactivated vaccines provide little or no passive protection. Here, we administered ginseng stem-leaf saponins (GSLS) to mice before parenteral immunization with an inactivated PEDV vaccine to investigate the effect of GSLS on the gut-mammary gland (MG)-secretory IgA axis. Early oral GSLS administration potently increased PEDV-specific IgA plasma cell generation in the intestine, facilitated intestinal IgA plasma cell migration to the MG by enhancing the chemokine receptor (CCR)10-chemokine ligand (CCL)28 interaction, and ultimately promoted specific IgA secretion into milk, which was dependent on Peyer's patches (PPs). Additionally, GSLS improved the gut microbiota composition, especially increasing probiotic abundance, and these microflora members promoted the GSLS-enhanced gut-MG-secretory IgA axis response and were regulated by PPs. In summary, our findings highlight the potential of GSLS as an oral adjuvant for PEDV-inactivated vaccines and provide an attractive vaccination strategy for lactogenic immunity induction in sows. Further studies are required to evaluate the mucosal immune enhancement efficacy of GSLS in pigs.

Dumbbell-like upconversion nanoparticles synthesized by controlled epitaxial growth for light-heat-color tri-modal sensing of carcinoembryonic antigen.

Accurate quantitative analysis of tumor markers in a wide linear range has important practical significance towards complex clinical samples in cancer identification and monitoring of tumor development stages, but remains challenging. Herein, three-layer dumbbell-like upconversion nanoparticles NaErF4:Tm@NaYF4@NaNdF4 (labeled as UCNPs) combined with G-quadruplex (G4) DNAzyme are reported for tri-modal sensing of carcinoembryonic antigen (CEA) in a wide range using upconversion luminescence (UCL), photothermal and catalysis signal readouts. Initially, dumbbell-like UCNPs were controlled synthesized by a three-dimensional epitaxial growth strategy through tuning the concentration of Nd precursors. After surface functionalization, G4zyme-UCNPs-cDNA/Apt-MB was subsequently fabricated by biotin-streptavidin interaction and DNA hybridization. Quantitative detection of CEA was achieved by competitive interaction and magnetic separation, and the intensities of tri-modal signals (light, heat and catalysis-based chrominance) of dissociative probes are linearly related to the concentration of CEA. The results showed that the tri-modal sensing method exhibited a wide linear range (0.005-2000 ng/mL) and low limit of detection (LOD) across three models: the luminescence model (0.005-50 ng/mL, LOD = 0.910 pg/mL), the catalysis model (10-1000 ng/mL, LOD = 0.387 ng/mL), and the temperature model (50-2000 ng/mL, LOD = 1.114 ng/mL). These findings suggest that the tri-modal sensing platform is suitable for use in the analysis of a wide range of complex and diverse clinical samples.

Immune Enhancement of Nanoparticle-Encapsulated Ginseng Stem-Leaf Saponins on Porcine Epidemic Diarrhea Virus Vaccine in Mice.

Porcine epidemic diarrhea virus (PEDV) causes severe enteric disease in pigs, particularly neonatal piglets. Current vaccines do not provide complete protection against PEDV. Ginseng stem-leaf saponins (GSLS), a promising oral adjuvant candidate, can improve intestinal immune responses in poultry and mice. However, its low stability limits further use. Poly lactic-co-glycolic acid (PLGA), a biocompatible and biodegradable nanoparticle, has been widely used in biomedicine for stable and targeted drug delivery. In this study, we developed GSLS-PLGA nanoparticles (GSLS-NPs) and evaluated the mucosal adjuvant efficacy in vitro and in vivo. GSLS-NPs significantly enhanced antigen internalization and pro-inflammatory cytokine secretion by DC2.4 cells. Mice orally administered GSLS-NPs before intramuscular inoculation generated CD11b+CD8α- and CD11b-CD103+ dendritic cells in the spleen and draining mesenteric lymph nodes, respectively, which are the types mainly responsible for antigen presentation. Additionally, enhanced neutralizing and non-neutralizing antibody responses and expanded activities of specific effector and memory CD4+ and CD8+ T cells were also observed in mice immunized with PEDV vaccines plus GSLS-NPs compared to mice receiving the vaccines alone. Furthermore, GSLS-NPs showed a good safety profile and presented great advantages over GSLS aqueous solution. Collectively, our results highlight the potential of GSLS-NPs as a mucosal adjuvant and provide an attractive vaccination strategy for combatting PEDV. Further study is required to evaluate the efficacy of this mucosal adjuvant in swine.

HPLC-DAD fingerprints combined with chemometric techniques for the authentication of plucking seasons of Laoshan green tea.

Laoshan green teas plucked in summer and autumn were measured by high performance liquid chromatography-diode array detector (HPLC-DAD). After baseline correction, the fingerprints data were resolved by multivariate curve resolution-alternating least squares (MCR-ALS) and a total of 57 components were acquired. Relative concentrations of these components were afterwards applied to distinguish plucking seasons using principal component analysis (PCA), support vector machines (SVM) and partial least squares-discriminant analysis (PLS-DA). For both SVM and PLS-DA models, the total recognition rates of training set, cross-validation and testing set were 100%, 91.3% and 100%, respectively. Besides, three variable selection methods were employed to determine characteristic components for the authentication of summer and autumn teas. Results showed that PLS-DA model based on three characteristic components selected by VIP possesses identical predictive ability as the original model. This study demonstrated that our proposed strategy is competent for the authentication of plucking seasons of Laoshan green tea.

Recent advances in single extracellular vesicle detection methods.

Extracellular vesicles (EVs) are secreted by a variety of cells. They are known for their pertinent role in intercellular communication, and participation in different pathological processes, making them ideal candidate for utilization as a biomarker for diagnosis and treatment of diseases. In contemporary years, the concept of a well-established liquid biopsy technology, and detection and utilization of EVs as a biomarkers have received unprecedented attention. Many rapid and precise EVs detection methods have been proposed, however, majority of them detect EVs in a bulk. As the prevalent heterogeneity of single extracellular vesicle (SEV) plays an important role in the analysis of disease progression, therefore, to prevent information loss, increased attention has been paid to SEV detection with remarkable successes. Technologies like fluorescence labeling, micro imaging and microfluidic chip were successfully employed for EVs detection at SEV level. This review summarizes the recent advances in SEV detection methods, their potential targets, applications as well as concludes future prospects for developing new SEV detection strategies.

Gold nanoclusters-poly(9,9-dioctylfluorenyl-2,7-diyl) dots@zeolitic imidazolate framework-8 (ZIF-8) nanohybrid based probe for ratiometric analysis of dopamine.

Ratiometric analysis of dopamine (DA) in complex biological system is urgently desired. In this work, a novel dual-emission fluorescence probe was fabricated by embedding both gold nanoclusters (AuNCs) and poly(9,9-dioctylfluorenyl-2,7-diyl) (PFO) dots into zeolitic imidazolate framework-8 (ZIF-8) (abbreviated as ZIF-8@AuNCs-PFO) and applied to ratiometric analysis of DA. Remarkably, encapsulating AuNCs and PFO dots into ZIF-8 not only achieved an excellent aggregation induced emission (AIE) enhancement effect on AuNCs (5 times increase), but also brought about an unique DA-triggered asynchronous fluorescence changes of AuNCs and PFO dots. The as-prepared probe exhibited excellent performance toward DA in the concentrations range from 0.01 to 10000 µmol L-1 and good selectivity over interfering substances. The detection limit of DA was as low as 4.8 nmol L-1. Furthermore, good stability and practicability of the probe in human serum samples suggesting its great potential for diagnostic purposes. Moreover, the quenching mechanism of AuNCs was intensively studied and summarized as three synergistic processes: (i) electron transfer between AuNCs and DA; (ii) DA-triggered architecture change of ZIF-8; (iii) fluorescence resonance energy transfer (FRET) between AuNCs and polydopamine (PDA), which offered an important theory for ZIF-based fluorescent probes.

Dual-colored carbon dots-based ratiometric fluorescent sensor for high-precision detection of alkaline phosphatase activity.

Probing the level and activity of alkaline phosphatases (ALP) is of great significance for biomedical research on cellular functions and clinical diagnosis of cancers. Herein, a novel dual-colored carbon dots (CDs)-based ratiometric fluorescent sensor was constructed to accomplish high sensitive and accurate determination of ALP relyed on the difference of fluorescence resonance energy transfer (FRET) between blue light emitted CDs (B-CDs)-MnO2 nanohybrid and yellow light emitted CDs (Y-CDs)-MnO2 nanohybrid. The ratiometric fluorescent sensor enabled sensitive discrimination of ALP against other enzymes in a linear range of 0.1-500 U/L with a limit of detection of 0.02 U/L. The lower error and signal fluctuation, more satisfactory LODs and higher R value (R2 = 0.99552) of the ratiometric sensing platform than single signal detection mode (R2 = 0.85231; R2 = 0.64260) indicated the superiority of the ratiometric fluorescence detection mode.Besides, the excellent analytical performance towards ALP in biological system demonstrated the potential application in clinical diagnosis.

Antiviral activity of Piscidin 1 against pseudorabies virus both in vitro and in vivo.

BACKGROUND: Swine-origin virus infection spreading widely could cause significant economic loss to porcine industry. Novel antiviral agents need to be developed to control this situation. METHODS: In this study, we evaluated the activities of five broad-spectrum antimicrobial peptides (AMPs) against several important swine-origin pathogenic viruses by TCID50 assay. Plaque reduction assay and cell apoptosis assay were also used to test the activity of the peptides. Protection effect of piscidin against pseudorabies virus (PRV) was also examined in mouse model. RESULTS: Piscidin (piscidin 1), caerin (caerin 1.1) and maculatin (maculatin 1.1) could inhibit PRV by direct interaction with the virus particles in a dose-dependent manner and they could also protect the cells from PRV-induced apoptosis. Among the peptides tested, piscidin showed the strongest activity against PRV. Moreover, in vivo assay showed that piscidin can reduce the mortality of mice infected with PRV. CONCLUSION: In vitro and in vivo experiments indicate that piscidin has antiviral activity against PRV.

Caerin1.1 Suppresses the Growth of Porcine Epidemic Diarrhea Virus In Vitro via Direct Binding to the Virus.

Porcine epidemic diarrhea (PED) has re-emerged in recent years and has already caused huge economic losses to the porcine industry all over the world. Therefore, it is urgent for us to find out efficient ways to prevent and control this disease. In this study, the antiviral activity of a cationic amphibian antimicrobial peptide Caerin1.1 against porcine epidemic diarrhea virus (PEDV) was evaluated by an in vitro system using Vero cells. We found that even at a very low concentration, Caerin1.1 has the ability to destroy the integrity of the virus particles to block the release of the viruses, resulting in a considerable decrease in PEDV infections. In addition, Caerin1.1 showed powerful antiviral activity without interfering with the binding progress between PEDV and the receptor of the cells, therefore, it could be used as a potential antiviral drug or as a microbicide compound for prevention and control of PEDV.

Target-triggered signal-on ratiometric electrochemiluminescence sensing of PSA based on MOF/Au/G-quadruplex.

Signal-amplified ratiometric electrochemiluminescence (Sa-RECL) provides an attractive approach to maximize signal-to-noise ratio through enhancing signals and eliminate interferences. In this work, we prepared a novel metal-organic framework (MOF)/Au/G-quadruplex as both quenchers and enhancers to fabricate a target-triggered ratiometric ECL sensor for high sensitive and accurate detection of prostatic specific antigen (PSA). The ratiometric ECL sensor using the dual-potential-dependent ECL emitters (quantum dots (QDs) and luminol) and MOF/Au/G-quadruplex not only achieved signal self-calibration but also realized cooperative amplification. After the sequential hybridization among of complementary DNA-QDs, PSA aptamer and pDNA-Au-Hemin-MIL-DNAzyme and the further competition of PSA, the pDNA-Au-Hemin-MIL-DNAzyme probe would keep away from the electrode surface, causing a switchover of their ECL signals from "off-on" state to "on-off" state. The ratiometric ECL aptasensor exhibits high-sensitive and accurate analytical performance toward PSA with a linear detection range from 0.5 to 500 ng mL-1 and a detection limit of 0.058 ng mL-1 (S/N = 3). The novel ratiometric ECL biosensor has been successfully applied to determine different serum samples of PSA, indicating its potential application in the clinical diagnosis.

Cellular hnRNP A1 Interacts with Nucleocapsid Protein of Porcine Epidemic Diarrhea Virus and Impairs Viral Replication.

The nucleocapsid (N) protein is a major structural component of porcine epidemic diarrhea virus (PEDV), which is predicted to be a multifunctional protein in viral replication. Heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) is a cellular protein participating in the splicing of pre-mRNA in the nucleus and translation regulation in the cytoplasm. According to our previous proteomic study about PEDV infection in vivo, hnRNP A1 was thought to be a cellular factor influencing PEDV replication. In this report, PEDV N protein was discovered to colocalize with cellular hnRNP A1 in perinuclear region of PEDV infected cells. Co-immunoprecipitation (CO-IP) results clearly demonstrated that PEDV N protein could bind to human hnRNP A1. Replication of PEDV was inhibited by silencing the expression of hnRNP A1 in CCL-81 cells, suggesting the positive effect of hnRNP A1 on PEDV infection.

The construction of recombinant Lactobacillus casei expressing BVDV E2 protein and its immune response in mice.

Bovine viral diarrhea virus (BVDV) is the etiological agent of BVD causes substantial economic losses and endemic in world-wide cattle population. Mucosal immunity plays an important role in protection against BVDV infection and Lactobacillus casei is believed as an excellent live vaccine vector for expressing foreign genes. In this study, we have constructed a novel recombinant L. casei/pELX1-E2 strain expressing the most immunogenic E2 antigen of BVDV; using growth phage dependent surface expression system pELX1. The expression of E2 protein was verified by SDS-PAGE, Western blotting, and Immunofluorescence microscopic analysis. The immune responses triggered by the E2 producing recombinant L. casei were evaluated in BALB/c mice revealed that oral and intranasal (IN) administration of the recombinant strain was able to induce a significantly higher level of specific anti-E2 mucosal IgA and serum IgG as well as the greater level of cellular response by IFN-γ and IL-12 than those of intramuscular (IM) and control groups of mice. However, IN inoculation was found the most potent route of immunization. The ability of the recombinant strain to induce serum neutralizing antibody against BVDV and reduced viral load after viral challenge indicated better protection of BVDV infection. Therefore, this recombinant L. casei expressing E2 could be a safe and promising mucosal vaccine candidate against BVD.

Comparative Proteome Analysis of Porcine Jejunum Tissues in Response to a Virulent Strain of Porcine Epidemic Diarrhea Virus and Its Attenuated Strain.

Porcine epidemic diarrhea virus (PEDV), a predominant cause of acute enteric infection, leads to severe dehydrating diarrhea and mortality in piglets all over the world. A virulent PEDV YN13 strain, isolated in our laboratory, was attenuated to yield an attenuated PEDV strain YN144. To better understand the pathogenesis mechanism and the virus-host interaction during infection with both PEDV YN13 and YN144 strains, a comparative proteomic analysis was carried out to investigate the proteomic changes produced in the primary target organ, using isobaric tags for relative and absolute quantitation (iTRAQ) labeling, followed by liquid chromatography tandem-mass spectrometry (LC-MS/MS). A total of 269 and 301 differently expressed proteins (DEPs) were identified in the jejunum tissues of the piglets inoculated with YN13 and YN144, respectively. Bioinformatics analysis revealed that these proteins were involved in stress responses, signal transduction, and the immune system. All of these involved interferon-stimulated genes (ISGs) which were up-regulated in jejunums by both of the PEDV-infected groups. Based on the comparative analysis, we proposed that different changes induced by YN13 and YN144 in heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1), eukaryotic initiation factor 4G1 (eIF4G1), and some members in the heat shock protein (HSP) family, may be responsible for differences in their pathogenicity.

Signal-Amplified Near-Infrared Ratiometric Electrochemiluminescence Aptasensor Based on Multiple Quenching and Enhancement Effect of Graphene/Gold Nanorods/G-Quadruplex.

Dual-signaling ratiometric electrochemiluminescence (ECL) technology has attracted particular attention in analytical science due to its precise measurement to normalize variation in environmental changes. Creating new mated ECL report units with two emitting states and improving the detection sensitivity are major challenges for ratiometric ECL measurement. Here, we fabricate an ultrasensitive near-infrared ratiometric ECL aptasensor based on a dual-potential signal amplification strategy triggered by the quencher/enhancer [graphene/hemin/gold nanorods/G-quadruplex-hemin (rGO-H-AuNRs-G4H) composite]. The composite was initially prepared through three consecutive steps: the π-π stacking interaction between hemin and graphene, in-site growth of AuNRs, and surface ligand exchange. Dual ECL quenching of quantum dots (QDs) and multiple signal enhancement of luminol can be achieved simultaneously by the fabrication of the sandwich "thrombin aptamer I (TBA1)-TB-TBA2 (rGO-H-AuNRs-G4H)" mode: (i) the formation of three-dimensional G-quadruplex between aptamer and thrombin not only shortens the distance between the donor (QDs) and receptor (rGO-H and AuNRs) to trigger electrochemiluminescence energy transfer but also provides the place for intercalating hemin; (ii) the hemin intercalated into G4 structure and hemin connected onto rGO together with AuNRs/rGO nanomaterials can achieve the multiple peroxidase-like catalysis of H2O2 to greatly enhance the ECL of luminol. The ratiometric ECL aptasensor self-calibrated by the internal reference (luminol or QDs) exhibits ultrasensitive and accurate analytical performance toward thrombin (TB) with a linear detection range from 100 ng/mL to 0.5 pg/mL and a detection limit of 4.2 fg/mL [defined as signal-to-noise ratio (S/N) = 3].