查找使用阿拉丁产品的科研文献

Exploring T7 RNA polymerase-assisted CRISPR/Cas13a amplification for the detection of BNP via electrochemiluminescence sensing platform

Zaiyong Zhang,Jinglong Li,Chunlin Chen,Yuwei Tong,Dehui Liu,Cuizhi Li,Huan Lu,Li Huang,Wanling Feng,Xiaoting Sun

Brain natriuretic peptide (BNP) is considered to be an important biomarker of heart failure (HF) attracting attention. However, its low concentration and short half-life in blood lead to a low-sensitivity detection of BNP, which is a challenge that has to be overcome. In this work, we propose a highly specific, highly sensitive T7 RNA polymerase-assisted clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a system to detect BNP via an electrochemiluminescence (ECL) sensing platform and incorporate Exo III-hairpin and dumbbell-shaped hybridization chain reaction (HCR) technologies. In this detection scheme, the ECL sensing platform possesses low background signal and high sensitivity. Firstly, the T7 promoter-initiated T7 RNA polymerase acts as a signal amplification technique to generate large amounts of RNAs that can activate CRISPR/Cas13a activity. Secondly, CRISPR/Cas13a is able to trans -cleave the surrounding trigger strand to produce DNA1. Thirdly, DNA1 is involved in the co-amplification reaction of Exo III and hairpin DNA, which subsequently triggers a dumbbell-shaped HCR technology. Eventually, a large number of Ru (II) molecules are inserted into the interstitial space of the dumbbell-shaped HCR to generate a strong ECL signal. The CRISPR/Cas13a possesses outstanding specificity for a single base and increased sensitivity. The tightly conformed dumbbell-shaped HCR provides higher sensitivity than the traditional linear HCR amplification technique. Ultimately, the clever combination of several amplification reactions enables the limit of detection (LOD) as low as 3.2 fg/mL. It showed promise for clinical sample testing, with recovery rates ranging from 98.4% to 103% in 5% human serum samples. This detection method offered a valuable tool for early HF detection, emphasizing the synergy of amplification strategies and specificity conferred by CRISPR/Cas13a technology.

Improved LFIAs for highly sensitive detection of BNP at point-of-care

Yan Gong,Jie Hu,Jane Ru Choi,Minli You,Yamin Zheng,Bo Xu,Ting Wen,Feng Xu

Defect-rich CoOOH nanorings: A biocompatible and cost-efficient material for clinical diagnosis of children heart failure

Yi Wang,Aiping Tu,Xinwen Zhang,Mengyao Li,Laicheng Liu,Yongfei Li,Jiangling Wu,Pu Zhang

Nanomaterials have been widely used in the detection of biomarkers, but only a few of them can be really applied in clinical disease diagnosis in hospital. Herein, we fabricate a kind of defect-rich CoOOH nanorings (NRs) and employ them as probes to improve the clinical diagnosis performance for children heart failure (HF). This diagnosis platform is dependent on ultrasensitive and specific detection of the “gold standard biomarker” of HF, B-type natriuretic peptide (BNP), which is realized based on the prevention of fluorescence resonance energy transfer. The detection range for BNP is 30–800 fg/mL with a limit of detection as low as 30 fg/mL. For diagnosis of a large number of clinical cases of children who behave sign or symptom of HF (respiratory distress, edema, or tachycardia), a very close correlation ( r  = 0.990) is obtained between this method and commercially available chemiluminescence immunoassay (CLIA, Siemens) that commonly adopted by hospital. It is worth noting that the present method has better diagnostic performance for children HF than CLIA including specificity (84.6% vs 61.5%), accuracy (82.2% vs 66.7%) and positive predictive value (40.0% vs 20.0%). Moreover, this method is more cost-effective than CLIA for BNP detection. The defect-rich, biocompatible, and low-cost CoOOH nanomaterial is promising to be applied in early clinical diagnosis of HF in hospital.

An Immunocolorimetric Sensing System for Highly Sensitive and High-Throughput Detection of BNP with Carbon-Gold Nanocomposites Amplification

Xin Liu,Ying Gan,Fengheng Li,Yong Qiu,Yuxiang Pan,Hao Wan,Ping Wang

Conventional immunocolorimetric sensing of biomolecules continues to be challenged with low sensitivity although its wide application as a diagnostic tool in medicine and biotechnology. Herein, we present a multifunctional immunocolorimetric sensing system for sensitive and high-throughput detection of B-type natriuretic peptide (BNP) with carbon-gold nanocomposite (CGNs) amplification. Using a “green” strategy, monodisperse carbon nanospheres (CNs) were successfully synthesized by glucose carbonization. A simple and efficient hydrothermal method was developed to assemble abundant gold nanoparticles (AuNPs) onto the surfaces of CNs. The resulting CGNs were characterized and utilized for biomarker detection with superior properties of easy manufacturing, good biocompatibility, satisfactory chemical stability, and high loading capacity for biomolecules. As a proof of concept, the as-prepared CGNs were conjugated with horseradish peroxidase-labeled antibody against BNP ([email protected]) functioning as the carrier, signal amplifier, and detector for the sensitive detection of BNP. Under optimal conditions, the established[email protected]immunoprobe remarkably enhanced the detection performance of BNP, achieving signal amplification of more than 9 times compared to the conventional method. Based on our self-developed bionic electronic eye (e-Eye) and[email protected]immunoprobe, the multifunctional sensing system displayed a wide dynamic linear range of 3.9–500 ng/mL and a LOD of 0.640 ng/mL for BNP detection with high specificity, good accuracy and reproducibility. This portable sensing system with enhanced performance demonstrates great potential for BNP detection in point of care applications, and offers a universal and reliable platform for in vitro biomarker detection.Keywords:bionic e-Eye;carbon-gold nanocomposites;immunocolorimetric sensor;BNP

Direct Quantification of S100β and BNP in Whole Blood via a GHNP-Enhanced SERS Plasmonic Sensor for Ultra-Early Ischemic Stroke Diagnosis and Clinical Stratification

Mengyue Wang,Jinyang Wu,Chengcheng Wang,Dongyu Liu,Ying Wang,Peng Zhao

Ischemic stroke (IS) demands ultra-early diagnosis to improve outcomes, yet conventional imaging modalities often fail to capture the critical window for intervention due to inherent temporal delays. Here, we report a surface-enhanced Raman scattering (SERS)-based plasmonic nanosensor that leverages gold hexagonal nanoparticles (GHNPs) as high-efficiency plasmonic amplifiers to enable simultaneous quantification of two key IS biomarkers-S100β and brain natriuretic peptide (BNP)-directly in whole blood. By engineering a sandwich immunoassay on a silver nanoparticle-modified conductive substrate, the GHNPs generate dense plasmonic “hotspots” that substantially amplify Raman signals, allowing for sensitive and stable detection without sample pretreatment. The sensor was validated in a rat middle cerebral artery occlusion (MCAO) model, capturing dynamic biomarker elevations within hours post-ischemia, and in clinical blood samples from IS patients, where biomarker levels were significantly elevated compared to healthy controls. The platform achieved detection limits as low as 7.18 fg/mL for S100β and 8.98 fg/mL for BNP, markedly outperforming conventional ELISA. By enabling rapid, multiplexed, and highly accurate quantification of these two complementary biomarkers, our SERS platform directly addresses the diagnostic gap between biochemical onset and clinical imaging, offering a powerful tool for ultra-early IS diagnosis and clinical stratification with strong potential for translational application.

High content screening identifies licoisoflavone A as a bioactive compound of Tongmaiyangxin Pills to restrain cardiomyocyte hypertrophy via activating Sirt3

Rui Guo,Ningning Liu,Hao Liu,Junhua Zhang,Han Zhang,Yingchao Wang,Mirko Baruscotti,Lu Zhao,Yi Wang

Background : Cardiac hypertrophy is a prominent feature of heart remodeling, which may eventually lead to heart failure. Tongmaiyangxin (TMYX) pills are a clinically used botanical drug for treating multiple cardiovascular diseases including chronic heart failure. The aim of the current study was to identify the bioactive compounds in Tongmaiyangxin pills that attenuate cardiomyocytes hypertrophy, and to investigate the underlying mechanism of action. Methods and Results : The anti-hypertrophy effect of TMYX was validated in isoproterenol-induced cardiac hypertrophy model in C57BL/6 mice. After TMYX treatment for 2 weeks, the heart ejection fraction and fractional shortening of the mice model was increased by approximately 20% and 15%, respectively, ( p  < 0.05). Besides, TMYX dose-dependently reduced the cross section area of cardiomyocytes in the angiotensin-II induced hypertrophy H9c2 model ( p  < 0.01). Combining high content screening and liquid chromatography mass spectrometry, four compounds with anti-cardiac hypertrophy effects were identified from TMYX, which includes emodin, licoisoflavone A, licoricone and glyasperin A. Licoisoflavone A is one of the compounds with most significant protective effect and we continued to investigate the mechanism. Primary cultures of neonatal rat cardiomyocytes were treated with a hypertrophic agonist phenylephrine (PE) in the presence or absence of licoisoflavone A. After 48 h of treatment, cells were harvested and mitochondrial acetylation was analyzed by western blotting and Image analysis. Interestingly, the results suggested that the anti-hypertrophic effects of licoisoflavone A depend on the activation of the deacetylase Sirt3 ( p  < 0.01). Finally, we showed that licoisoflavone A-treatment was able to decrease relative ANF and BNP levels in the hypertrophic cardiac cells ( p  < 0.01), but not in cells co-treated with Sirt3 inhibitors (3-TYP) ( p  > 0.05). Conclusion : TMYX exerts its anti-hypertrophy effect possibly through upregulating Sirt3 expression. Four compounds were identified from TMYX which may be responsible for the anti-hypertrophy effect. Among these compounds, licoisoflavone A was demonstrated to block the hypertrophic response of cardiomyocytes, which required its positive regulation on the expression of Sirt3. These results suggested that licoisoflavone A is a potential Sirt3 activator with therapeutic effect on cardiac hypertrophy.

Porous Hydrogel-Encapsulated Photonic Barcodes for Multiplex Detection of Cardiovascular Biomarkers

JingJing Ji,Wenbin Lu,Yi Zhu,Hong Jin,Yuyu Yao,Huidan Zhang,Yuanjin Zhao

Early detection of cardiac troponin I (cTnI), B-type natriuretic peptide (BNP), and myoglobin (Myo) is essential for the diagnosis of acute myocardial infarction (AMI) and heart failure (HF). We designed a porous hydrogel-encapsulated photonic crystal (PhC) barcode-based suspension array for multiple cardiovascular marker detection. The hybrid hydrogel was composed of polyethylene glycol diacrylate (PEGDA) and gelatin, resulting in a porous and hydrophilic scaffold which ensured stability of the PhC in aqueous solutions. The encapsulated PhC barcodes had stable diffraction peaks for the corresponding markers. Using a sandwich format, the proposed suspension array was used for simultaneous multiplex detection of cardiovascular biomarkers in a single tube. The immunoassay results we tested on cTnI, BNP, and Myo could be assayed in the ranges of 0.01 to 1000 ng/mL, 0.1 to 10 000 pg/mL, and 1 to 10 000 ng/mL with limits of detection of 0.009 ng/mL, 0.084 pg/mL, and 0.68 ng/mL at 3σ, respectively. This method also showed acceptable accuracy and repeated detection, and the results were consistent with the results of conventional clinical methods for detecting actual clinical samples. Therefore, suspension arrays based on hydrogel-encapsulated PhC barcodes are highly promising for AMI diagnosis.

Black ginseng: a novel medicine for treating heart failure

Dou Peiyuan,Liu Linlin,Jin Mozhu,Huang Jing,Lekhooa Rose Makhotso,Ran Xiaoku,Yan Xiaohui

Introduction: Black ginseng (BG) was processed by “steaming and drying” (generally nine times) repeatedly to produce “rare saponins” and secondary ginsenosides. Both ginseng (GS) and red ginseng (RG) were commonly used in treating heart failure (HF), and the latter was confirmed to be more potent, implying the presence of rare ginsenosides that contribute positively to the treatment of heart failure. Previous research indicated that rare ginsenosides are more abundant in BG than in RG. Consequently, this study aims to investigate the effects of BG and its components on HF to elucidate the active substances and their underlying mechanisms in the treatment of HF.Methods: The effects of BG and its fractions (water-eluted fraction (WEF), total saponin fraction (TSF), and alcohol-eluted fraction (AEF)) on rats with isoproterenol (ISO)-induced HF were explored, and steroids belonging to the hypothalamic–pituitary–adrenal (HPA) and hypothalamic–pituitary–gonadal (HPG) axes were determined quantitatively using the ultra-performance liquid chromatography–triple quadrupole tandem mass spectrometry (UPLC-QqQ-MS/MS) method. In addition, 16S rDNA sequencing was performed on the gut microbiota, followed by GC–MS analysis of short-chain fatty acids (SCFAs), and the biochemical indexes related to energy metabolism and the serum cyclic nucleotide system were also analyzed by ELISA.Results: Based on a thorough evaluation of energy metabolism and the endocrine system, it was observed that the effects of BG components on the hypothalamic–pituitary–thyroid (HPT) and HPA axes were more pronounced. Notably, the treatment efficacy of the low dose of the total saponin fraction (TSFL), water decoction (WD), and high dose of the polysaccharide fraction (PSFH) was superior based on pharmacodynamic indicators such as brain natriuretic peptide (BNP), creatine kinase (CK), and estradiol (E2)/T). Furthermore, the WD and BG components exhibited significant effects on androgens (T and androstenedione (A4)). The TSFL group exerts an anti-inflammatory effect by regulating Lactobacillus/Erysipelotrichales. The WD, PSFH, and TSFL may impact inflammatory cytokines through the gut microbiota (Lactobacillus/Erysipelotrichales) and their metabolites (acetate and butyrate), exerting an anti-inflammatory effect.Discussion: The BG and all its split components demonstrated varying levels of efficacy in alleviating HF, and TSF and PSF exhibited a significant protective effect on HF. The main active components in TSF were revealed to be ginsenosides Rk1, Rk3, 20-(S)-Rg3, and 20-(S)-Rh2 by the H9C2 cell experiment. The decoction of BG and its components exhibited a potent impact on androgen hormones, with an elevation trend. This phenomenon may be attributed to the activation of the eNOS-NO pathway through androgen regulation, thereby contributing to its anti-HF activities. The WD, PSFH, and TSFL may exert anti-inflammatory effects through the intestinal flora (Lactobacillaceae/Erysipelotrichaceae) and its metabolites (acetic acid and butyric acid), which affect the inflammatory factors. The different mechanisms of action of each component of HF also reflect the significance and necessity of the overall role of traditional Chinese medicine (TCM). Our research was the first to report that the E2/T is related to HF and can be used as an indicator to evaluate HF.