Background Runzaoling prescription (RZL) has been used clinically for many years, and has shown potential therapeutic effects in previous studies for the treatment of Sjögren's Syndrome (SS). Even so, the specific active components and underlying mechanisms by which RZL exerts its anti-SS effects remain unclear. Materials and methods Ultra-high performance liquid chromatography coupled with Q-Exactive Orbitrap tandem mass spectrometry (UHPLC-Q-Exactive Orbitrap MS/MS) was employed to systematically identify the chemical constituents of RZL in vitro, as well as those components that enter the bloodstream in mice. The main active components, core genes and related signaling pathways of RZL for SS were screened by network pharmacology. Furthermore, the affinity between important components and their primary targets was predicted using molecular docking technology. Finally, the animal model of spontaneous SS (NOD mice) was established to evaluate the efficacy of RZL. Results and Conclusions 728 components were identified in RZL, and 49 prototype components as well as 7 metabolites were identified from the blood of mice. Network pharmacological analysis showed that RZL targeted key proteins like ICAM1, TNF, TLR4, and EGFR through active components such as isorhapontigenin, hypaphorine and cyanidin, modulating pathways like NF-κB, PI3K/AkT, MAPK, HIF-1 and TNF to exhibit the anti-inflammatory effects for treating SS. Molecular docking revealed that key active components of RZL effectively interact with proteins in the NF-κB pathway. Animal studies indicated that RZL alleviated pathological damage, enhanced saliva production and submandibular gland index in SS model mice. Additionally, it may regulate the TLR4/MyD88/NF-κB signaling pathway and effectively inhibit the inflammatory response.
Hydrazine is a well-known hazardous environmental pollutant and toxic compound.Therefore, there is an urgent demand for an effective analytical monitoring method to safeguard the public from unforeseen exposure to harmful contaminants. Functional surface-enhanced Raman scattering (SERS) has recently emerged as a highly advantageous technique for sensing trace biochemical substances, owing to its unique capacity for fingerprint-like molecular identification, making it suitable for hydrazine detection. Herein, we used UV-nanoimprint lithography (UV-NIL) combined with glancing angle deposition of electron beam evaporation (GLAD) to prepare T-shaped PVP/Au nanocylinder arrays with gaps. T-shaped hollow Au nanocylinder arrays with gaps could be achieved by dissolving PVP nanocylinder with 60°C hot water. 4-Mercaptobenzaldehyde incorporated on the T-shaped hollow Au nanocylinder arrays (T-shaped hollow Au nanocylinder arrays@4-MBA), reacted with hydrazine to form Raman active benzaldehyde hydrazone based on the Schiff base reaction, enabling sensitive and selective SERS detection of hydrazine. The limit of detection (LOD) of hydrazine was as low as 4.1 × 10-9 mol/L. In addition, T-shaped hollow Au nanocylinder arrays@4-MBA platform prepared by this method can maintain high repeatability and stability after six month of fabrication. Therefore, the T-shaped hollow Au nanocylinder arrays functionalized with 4-MBA provides an efficient platform for rapid, sensitive, stable, and highly selective detection of hydrazine.
Supramolecular assemblies (SMAs) formed during traditional decoction are increasingly recognized as an important physical-state contributor to the efficacy of traditional Chinese medicine (TCM), but whether these assemblies form reproducibly in Chinese herbal formula granules—and how excipients and thermal history modulate their behavior—remains poorly characterized. We compared the herb pair Magnolia officinalis–Glycyrrhiza uralensis prepared as a traditional co-decoction (MG) and as reconstituted formula granules (MGFG). SMAs were enriched by centrifugation followed by dialysis to yield MG-SMAs and MGFG-SMAs (0, 30, and 60 min re-decoction). An excipient-containing model was generated by adding dextrin, and the corresponding assembly-enriched colloidal fraction was operationally designated MGE-SMAs. We systematically compared assembly morphology and solution-phase colloidal properties using scanning electron microscopy (SEM) and dynamic light scattering (DLS) and probed the assembly microenvironment using Fourier-transform infrared spectroscopy (FT-IR) and ultraviolet–visible spectroscopy (UV–vis). Chemical profiling was performed by Ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry/mass spectrometry (UPLC-Q-TOF-MS/MS) combined with multivariate analyses, and representative constituents were quantified by High-performance liquid chromatography (HPLC). Anti-Staphylococcus aureus activity was evaluated by growth-inhibition curves (OD600), MIC/MBC assays, and Live/Dead staining. The MG decoction presented a highly heterogeneous dispersion (1040.97 ± 91.00 nm; PDI 0.98 ± 0.02), whereas its centrifugation- and dialysis-derived colloidal fraction, MG-SMAs, exhibited a narrower nanoscale particle-size distribution (229.43 ± 3.27 nm; PDI 0.15 ± 0.07) with a similarly weakly negative ζ-potential and reduced conductivity, consistent with enrichment of a defined colloidal fraction. Addition of dextrin and reconstitution from granules increased dispersity (PDI 0.56–0.71) and reduced |ζ| (≈ −7 mV), indicating excipient- and processing-induced heterogeneity. Re-decoction was associated with time-dependent changes in the colloidal properties of MGFG-derived fractions. Compared with unheated MGFG-SMAs, MGFG-SMAs-30 exhibited a lower mean hydrodynamic particle size and PDI, while extending re-decoction to 60 min produced no further reduction in these parameters. UPLC-Q-TOF-MS/MS revealed broadly overlapping component coverage but a preparation-dependent redistribution of metabolites, and HPLC confirmed selective enrichment and depletion trends among marker compounds. Functionally, MG and MG-SMAs showed the same MIC against S. aureus (0.8 mg/mL), whereas excipient-containing systems required higher inhibitory concentrations; among granule-derived samples, MGFG-SMAs-30 produced the strongest inhibition. Although the excipients increased the concentration required for antibacterial inhibition, reheating partially improved the activity of the granule-derived preparations, suggesting that both composition and assembly state may influence bioactivity. This study established an analytical workflow integrating colloidal characterization with high-resolution chemical profiling to investigate how excipients and thermal history affect the formation and properties of assembly-enriched colloidal fractions derived from formula granules.
Introduction Gastric diseases have become a global health challenge due to infectious agents and unhealthy lifestyles, with a notable trend of rising incidence among younger populations in recent years, including children and adolescents who increasingly suffer from gastric discomfort. Biling Weitong Granules, an improved formulation derived from the classical prescriptions "Jinlingzi Powder" and "Zuojin Pill", embodies the therapeutic principles of "unblocking and descending theory" and "qi-blood harmony theory" in spleen-stomach disease treatment. This compound demonstrates efficacy in regulating qi flow, activating blood circulation, harmonizing the stomach, and alleviating pain, making it clinically applicable for various acute and chronic gastric disorders. However, its precise pharmacological mechanisms remain incompletely elucidated. Methods The chemical components of Biling Weitong Granules were identified using UHPLC-Q-Orbitrap-MS technology. Subsequently, an acute gastritis mouse model was established by absolute ethanol induction, with histopathological damage to gastric tissues evaluated through H&E staining. An integrated analysis combining network pharmacology, metabolomics, and transcriptomics was then conducted to investigate the mechanism underlying the anti-acute gastritis effects of Biling Weitong Granules. The expression levels of PI3K, AKT, P50, P65, IKKα, IκBα and their phosphorylated forms were detected by Western blotting. Results This study identified a total of 125 compounds in Biling Weitong Granules through comprehensive analysis. Network pharmacology analysis revealed that the therapeutic effects against acute gastritis may be mediated through key pathways including "Apoptosis", "TNF signaling pathway", "JAK-STAT signaling pathway", and "PI3K-Akt signaling pathway". Furthermore, integrated metabolomics and transcriptomics analyses demonstrated that Biling Weitong Granules exert their therapeutic effects by regulating metabolic abnormalities of glutathione and other amino acids, consequently modulating critical cellular processes such as oxidative stress, inflammatory responses, and apoptosis. Western blot analysis revealed that the phosphorylation levels of p-PI3K, p-AKT, p-P65, p-P50, p-IKKα, and p-IκBα were reduced after treatment with Biling Weitong Granules. Conclusion Biling Weitong Granules (BLWTG) exert therapeutic effects against acute gastritis (AG) by modulating metabolic abnormalities of glutathione and other amino acids, thereby influencing multiple signaling pathways including "Apoptosis", "TNF signaling pathway", "JAK-STAT signaling pathway", and "PI3K-Akt signaling pathway". These findings reveal the complex mechanistic basis underlying the anti-AG effects of this traditional Chinese medicine.
Background Kongxian Dan (KXD), a classic traditional Chinese prescription for treatingexcessive mucus secretion, which possesses broad therapeutic potential. This study aims to identify its active components against asthma and elucidate the underlying mechanisms. Methods Active components of KXD absorbed into mouse serum were identified using LC-MS/MS. Potential targets and related signaling pathways were predicted via network pharmacology. An OVA-induced mouse asthma model as well as human bronchial epithelial BEAS-2B cells were established to evaluate the therapeutic effects. Inflammatory cells, cytokines and oxidative stress markers in BALF and lung tissue were measured. Histopathology was performed to assess airway inflammation, mucus secretion, remodeling, and hepatorenal toxicity. The expression of mucin (MUC5AC) and TMEM16A was detected via immunofluorescence, and related key proteins were analyzed by western blot. Results Sixteen active components derived from KXD were identified. Network pharmacology analysis suggested a critical association with calcium signaling pathways. In vivo and in vitro experiments showed that KXD significantly alleviated airway inflammation, reduced inflammatory cell infiltration and cytokine levels, and suppressed decreased oxidative stress. It also attenuated mucus hypersecretion and airway remodeling without hepatorenal toxicity. Mechanistically, KXD reduced intracellular calcium concentration by downregulating PLC, IP3R, and TMEM16A expression. Conclusion KXD mitigates asthma-related airway inflammation and excessive mucus secretion, potentially through the inhibition of the PLC/IP3R/Ca²⁺/TMEM16A signaling pathway.
Cancer is a serious threat to human health, and early diagnosis and treatment are the key to increasing the survival rate and improving the prognosis of cancer patients. Nucleic acid aptamer sensors demonstrate considerable potential for cancer applications owing to their exceptional specificity and strong binding affinity. This paper reviews the optimization of nucleic acid aptamer screening processes, the discovery of commonly used nucleic acid aptamers, and the recent research progress on nucleic acid aptamer sensors in cancer diagnosis and therapy. Furthermore, this review emphasizes the promising applications of nucleic acid aptamer sensors in multi-target high-sensitivity detection, real-time dynamic monitoring, and personalized precision medicine. This review systematically summarizes current advances in nucleic acid aptamer sensors, emphasizing their pivotal role in advancing non-invasive, intelligent, and precise cancer diagnostics and therapeutics, while proposing novel strategies for early intervention and efficient cancer management.
Tetrastigma hemsleyanum Diels & Gilg (TH) is a herbaceous vine in the family Vitaceae with known immune-enhancing properties. However, the bioactive components and therapeutic mechanisms of TH remain unclear. Therefore, this study aimed to establish a comprehensive approach integrating pharmacodynamic validation, serum pharmacochemistry, and network pharmacology to determine the immunomodulatory effects of TH and screen for its bioactive components. Firstly, the immunomodulatory effects of TH on mice were analyzed and confirmed by monitoring body weight changes, organ indices, serum results, and TCR sequencing. Secondly, the prototype components absorbed into blood in plasma after administration of TH ethanol extract (THE) were analyzed and identified by UPLC-Q-Exactive Orbitrap MS, and ten prototype components were characterized, mainly phenolic acids and flavonoids. Finally, the obtained plasma components were input into the SwissTarget Prediction tool for drug target prediction, and disease targets were obtained from the Genecard and DrugBank databases. Subsequently, a protein-protein interaction (PPI) network was constructed to screen and determine core targets such as AKT1,STAT3,HIF1A. By identifying the plasma components of THE and their predicted core targets, this study provides a pharmacological basis for further investigation into the immunomodulatory mechanisms of TH.
Bungarus snake envenomation is an acute and potentially fatal condition often mistaken for bites from non-venomous snakes. As a result, it is crucial to diagnose this condition rapidly and accurately. This study presents the development of a rapid test strip for detecting Bungarus venom, utilizing AIENPs conjugated with the venom as a probe. The methodologies employed encompass antibody preparation, purification, specificity testing, and the optimization of conditions. The blocking agent should be 2 mg, the conjugation time 3 minutes, and the minimum amount of AIENPs 1 mg. The results demonstrate that the polyclonal antibody is particular, with the test strip capable of detecting as low as 100 ng/mL of Bungarus venom and distinguishing it from other common Chinese snake venoms. The simulation testing of real samples demonstrated a 98% positive rate and a 0% false-positive rate. Although clinical samples were not tested, the strip is sensitive, accurate, and simple, offering significant potential for diagnosing Bungarus snake bites and providing new insights for the development of other test strips.Plain Language summary:In many remote areas of the world, the lack of effective diagnostic measures for vulnerable populations affected by snakebite poisoning, as well as poor medical facilities in these areas, seriously affect the timely treatment of snakebite patients. For different types of snakebites, the appropriate antivenom is required to achieve the best treatment effect. Therefore, an accurate diagnosis is essential to select the appropriate antivenom and ensure the effectiveness of treatment. The B. multicinctus is an extremely deadly venomous snake that poses a great medical challenge. In response to this situation, this study developed a single antibody test strip for the rapid detection of B. multicinctus venom based on AIE technology, using AIENPs conjugated with krait venom as a labeled probe, which can quickly, accurately, and sensitively distinguish B. multicinctus venom from other snake venoms. This test is not only simple and cost-effective, but also suitable for promotion in remote areas, and will effectively reduce the mortality rate of patients bitten by B. multicinctus.
Synthetic cathinones represent a major and rapidly evolving subclass of new psychoactive substances. Due to their potent central nervous system-stimulating effects and psychoactive properties, these compounds have become a primary focus of forensic science and toxicology laboratories worldwide. The abuse or acute overdose of these substances often leads to extremely severe health consequences, posing a significant threat to global public health and safety. With the continuous tightening of legislation, the molecular structures of synthetic cathinones have undergone rapid mutation. This necessitates the development of more advanced and sensitive analytical strategies to achieve accurate detection and identification of target compounds within complex biological samples. This review systematically summarizes the recent advances in the forensic analysis of synthetic cathinones over the past five years. It provides an in-depth discussion of biological sample pretreatment techniques, including the latest trends towards miniaturization and green chemistry principles. Regarding instrumental analysis, this review comprehensively evaluates analytical methods for synthetic cathinones and delves into the challenges of "structural elucidation" for novel analogs and positional isomers. Finally, by incorporating recent typical cases of poisoning and polydrug abuse, this review offers a detailed reference and technical support for addressing the threats associated with synthetic cathinone abuse.
The aggregation-induced electrochemiluminescence (AIECL) of phenanthroimidazole-functionalized tetraphenylethene (pTPI) was reported in this work. The electrochemiluminescence (ECL) intensity of pTPI was significantly enhanced with the increase of water content in DMF/water mixture. The strong emission in the aggregated state could be attributed to the restriction of intramolecular motions, which effectively suppressed the nonradiative relaxation pathway. The ECL behavior of pTPI in N,N-dimethylformamide (DMF) was investigated by transient ECL test, where the annihilation reaction was found to follow the "S-path". To achieve the application of pTPI in aqueous solution, pTPI nanowires (pTPI NWs) were synthesized via nanoprecipitation method. With triethylamine (TEA) as a coreactant, pTPI NWs demonstrated bright anodic ECL emission, with the relative ECL efficiency reaching 10.6% compared with that of Ru(bpy)(2+)(3). This work paved a new avenue for development of highly efficient ECL luminophores, and the developed pTPI NWs showed promising application prospects in the fields of biosensing and light-emitting devices.
Laccase is a member of the blue multicopper oxidase family,which catalyzes the oxidation of phenolic compounds in the presence of oxygen.As a versatile biocatalyst with broad substrate specificity,laccase holds promise for various applications such as biosensing,environmental remediation,and green catalysis.However,the practical deployment of native laccase is often hindered by the limitations of high production costs and poor operational stability.In the catalytic process of natural laccase,the redox couple between monovalent and divalent copper(Cu2+/Cu+)plays a key role in transferring electrons from the reducing substrate to oxygen molecules.Inspired by this mechanism,layered potassium birnessite(KBir)with remarkable laccase-mimicking activity was synthesized in this work.By utilizing the redox characteristics of the interlayer manganese couple(Mn4+/Mn3+),active center and catalytic function of natural laccase were successfully mimicked.Steady-state kinetic analysis confirmed that KBir had excellent catalytic efficiency,along with good stability under various conditions(Temperature,pH,inorganic salts,and organic solvents),making it a promising alternative to natural laccase.Based on this,KBir was successfully applied to detection of quercetin(QUE),demonstrating great potential in the field of biosensing.This work provided new insights into the rational design of advanced laccase-mimicking enzymes and highlighted their broad application prospects.
Cardiac troponin I (cTnI) exhibits high specificity and sensitivity for myocardial injury, serving crucial biomarker for the early diagnosis of high-risk cardiovascular diseases (CVD), such as acute myocardial infarction (AMI). Recently, to meet the demand for early screening of high-risk CVD, various nanomaterial-based analytical methods with high-performance have been developed for the precise detection of low-abundance cTnI blood samples. This review summarized recent research advancements over the past five years, focusing on physicochemical properties of different nanomaterials to enhance the performance of electrochemical and optical detection methods for cTnI. It also discussed the limitations and prospects of as-developed methods in clinical applications, providing a detailed outline for the development of new methods for precise early screening of high risk CVD.
The ultra-low abundance of tumor markers and interference from complex matrices in liquid biopsy represent the primary bottlenecks for early cancer screening,and single amplification strategyis can bardly achieve a good balance among sensitivity,specificity,and reaction speed.In recent years,clustered regularly interspaced short palindromic repeats(CRISPR)and CRISPR-associated proteins(Cas)system-assisted molecular self-assembly strategy was used in detection of tumor biomarker.Leveraging the precise recognition and efficient cleavage activity of CRISPR-Cas system,this synergistic strategy effectively overcomes the challenges of background leakage and sluggish kinetics associated with traditional enzyme-free self-assembly.By constructing a cascade signal amplification loop,CRISPR-Cas system-assisted molecular self-assembly strategies enable the ultrasensitive and highly specific detection of trace markers such as circular tumor DNA(ctDNA)and micro RNA(miRNA).Herein,recent research progress on CRISPR-Cas system-assisted molecular self-assembly strategies for tumor marker detection was reviewed.The applications of this dual signal amplification mechanism in the field of physical sensing were introduced.The deep integration of the CRISPR-Cas system-assisted molecular self-assembly strategies with artificial intelligence,solid-state reagent storage,and microfluidic technology constituted a pivotal direction for advancing its application in clinical point-of-care testing.
The abuse of drugs,including narcotics and psychotropic substances,antibiotics,and additives,has become a significant threat to global public health security and social stability.Therefore,the development of rapid,efficient,portable,and on-site screening technologies suitable for resource-limited environments is of great importance for the early detection,effective regulation,and monitoring of trends in drug abuse.Lateral flow immunoassay(LFIA),leveraging its operational simplicity,rapid response,cost-effectiveness,and suitability for non-laboratory settings,has emerged as a pivotal platform for point-of-care testing(POCT)of abuse drugs.Among these,nanotags,serving as signal carriers,play a key role in enabling high-performance LFIA by conjugating antibodies to output detection signals.This review began with a brief introduction to LFIA technology,systematically summarized recent research progress in LFIA recognition mechanisms,novel nanolabels,and signal transduction strategies for drug abuse detection,and discussed the remaining challenges and future trends in this field.
Poor targeting specificity and low intracellular bioavailability have become key factors restricting the improvement of clinical therapeutic efficacy in tumor treatment.The tumor microenvironment(TME)exhibits characteristics including acidity,hypoxia,overexpression of specific enzymes,and redox imbalance,which provide natural targets for tumor tissue-specific response behavior of nanodrugs.Moreover,the functional homeostasis of subcellular organelles such as mitochondria,lysosomes,endoplasmic reticulum,and nucleus is a core determinant regulating the fate of tumor cells.This paper focuses on the signal-driven cascade targeting strategy,which takes TME characteristic signals as the primary trigger to help nanodrugs overcome in vivo physiological barriers and achieve specific responses in tumor tissues.Subsequently,cascade reactions expose organelle-targeting moieties,and secondary precise localization is realized by virtue of the microenvironmental differences between subcellular organelles and cytosol,thereby achieving specific intervention in subcellular organelle functions.This study systematically summarizes the design principles of cascade targeting driven by acidic,enzymatic,redox,and hypoxic signals,with a focus on the application pathways of core mechanisms(e.g.,charge reversal,structural conformation change,and prodrug activation)in subcellular organelle precision therapy,as well as the synergistic potential of this strategy to ameliorate tumor therapy resistance and remodel the tumor immune microenvironment.This work aims to provide a theoretical reference for the development of intelligent nanodrugs and precise tumor therapy.
Accurate identification of single nucleotide variants (SNVs) is essential for diagnosing pathogen drug resistance. However, current isothermal amplification-based assays often suffer from insufficient specificity in SNVs discrimination and limited multiplexing capability. Here, an enzyme-free nucleic acid circuit for SNVs detection based on a four-way junction (4WJ) was developed. By exploiting a toehold-mediated strand exchange (TMSE) mechanism within an"X-shaped"topological framework, the circuit enabled highly sensitive discrimination of single-base mismatches. By using katG S315T mutation associated with isoniazid resistance in M. tuberculosis as a model, the circuit reliably distinguished targets differing by SNVs. When coupled with loop-mediated isothermal amplification (LAMP), the system achieved selective detection of mutant targets down to 1 copy/mu L, generating signals significantly stronger than 5000 copies/mu L wild-type background. Moreover, modular extension of the supporting strands enabled the construction of an OR logic gate for simultaneous detection of isoniazid and rifampicin resistance mutations (rpoB S531L), demonstrating the multiplexing capability of the platform. This strategy operated without the need for precise thermal cycling instrumentation and offered a rapid, simple, and high-fidelity approach, providing a promising method for preliminary screening of pathogen drug resistance in resource-limited settings.
Bacterial infections pose a severe threat to human health and public health security.Existing detection methods suffer from limitations including long time duration,complex operation,and reliance on professional skills.Moreover,the detection systems often lack antibacterial functionality,leading to delays in early warning and intervention for infection,thus creating an urgent demand for rapid detection and efficient bacteriostatic technologies.To address these issues,a dual-functional system integrating colorimetric detection and bacteriostasis based on chlorogenic acid(CGA),a natural herbal extract,was developed in this study.Firstly,the detection medium was screened based on simulated body fluid(SBF),and its key components and CGA concentration were further optimized.Using Escherichia coli(E.coli)and Staphylococcus aureus(S.aureus)as model bacteria,the visual detection range for both strains with CGA reached 104-108 CFU/mL,and the maximum bacteriostatic rate was 100%.Biocompatibility experiments demonstrated that the hemolysis rate of CGA was below 5%within the effective concentration range.After treating human umbilical vein endothelial cells(HUVECs)and mouse fibroblasts(L929 cells)with this system,the cell viability exceeded 80%,preliminarily indicating good biocompatibility.In practical application tests,this system showed a certain ability to identify infections in terms of warning about bacterial contamination of contact lenses and detecting bacterial infections in clinical urine samples.This work provided a green,convenient,and low-cost new strategy for rapid diagnosis,prevention,and control of bacterial infections,holding broad application prospects in the fields such as medical device quality control and daily hygiene protection.
Copper-based metal-organic framework nanofibers (Cu-MOF NFs) were synthesized via a solvothermal method using 1,2,4,5-benzenetetracarboxylic acid (H4BTEC) as ligand for electrochemical detection of metronidazole (MET) residues in the gingival crevicular fluid (GCF) of periodontitis patients. The morphology and crystal structure of the samples were characterized using scanning electron microscopy (SEM) and X-ray diffraction (XRD). The results showed that the Cu-MOF NFs prepared with a reaction time of 60 min exhibited a long-range ordered crystal structure and had the highest specific surface area (121.4 m(2)/g). By optimizing the electrochemical detection parameters, square wave voltammetry (SWV) results showed that the constructed electrochemical sensor exhibited high sensitivity toward MET detection, the detection sensitivity was 228.18 & micro;A/(& micro;mol & centerdot;cm(2)), linear detection range was 1-6 & micro;mol/L, and limit of detection (3 sigma) was 0.333 & micro;mol/L. When applied to detection of MET in real GCF samples, this sensor also showed satisfactory detection accuracy, thus providing preliminary research experience for its practical application in complex oral environment.
