Osteoclasts are essential for maintaining healthy bone.Pathological elevation of os-teoclastogenesis or osteoclast activity can cause osteoporosis and increase the risk of bone fracture.However,a few options are avail...Osteoclasts are essential for maintaining healthy bone.Pathological elevation of os-teoclastogenesis or osteoclast activity can cause osteoporosis and increase the risk of bone fracture.However,a few options are available for directly measuring osteoclast activity in vivo to test interventions that may affect osteoclasts.Here,we describe an in vivo method to measure osteoclast-mediated bone loss targeted at normal mouse calvaria.The method employs a novel procedure for measuring osteoclast resorption pits using micro-computed tomography.The potential utility of this mouse calvaria model to assess therapies targeting osteoclasts was validated using zoledronic acid,which is a nitrogen-containing bisphosphonate drug used to treat osteoporosis.展开更多
Lateral flow immunoassays(LFIAs)are low-cost,rapid,and easy to use for pointof-care testing(POCT),but the majority of the available LFIA tests are indicative,rather than quantitative,and their sensitivity in antigen t...Lateral flow immunoassays(LFIAs)are low-cost,rapid,and easy to use for pointof-care testing(POCT),but the majority of the available LFIA tests are indicative,rather than quantitative,and their sensitivity in antigen tests are usually limited at the nanogram range,which is primarily due to the passive capillary fluidics through nitrocellulose membranes,often associated with non-specific bindings and high background noise.To overcome this challenge,we report a Beads-on-a-Tip design by replacing nitrocellulose membranes with a pipette tip loaded with magnetic beads.The beads are pre-conjugated with capture antibodies that support a typical sandwich immunoassay.This design enriches the low-abundant antigen proteins and allows an active washing process to significantly reduce non-specific bindings.To further improve the detection sensitivity,we employed upconversion nanoparticles(UCNPs)as luminescent reporters and SARS-CoV-2 spike(S)antigen as a model analyte to benchmark the performance of this design against our previously reported methods.We found that the key to enhance the immunocomplex formation and signal-to-noise ratio lay in optimizing incubation time and the UCNP-to-bead ratio.We therefore successfully demonstrated that the new method can achieve a very large dynamic range from 500 fg/mL to 10μg/mL,across over 7 digits,and a limit of detection of 706 fg/mL,nearly another order of magnitude lower than the best reported LFIA using UCNPs in COVID-19 spike antigen detection.Our system offers a promising solution for ultra-sensitive and quantitative POCT diagnostics.展开更多
The sustainable sourcing of novel bioactive compounds from natural sources is crucial to the success of the pharmaceutical,food,and cosmetics industries.Iris bucharica Foster(syn.Juno bucharica(Foster)Vved.)is a promi...The sustainable sourcing of novel bioactive compounds from natural sources is crucial to the success of the pharmaceutical,food,and cosmetics industries.Iris bucharica Foster(syn.Juno bucharica(Foster)Vved.)is a promising source of novel bioactive molecules,particularly phenolic compounds,which are renowned for their antioxidant properties.In this study,we developed a reliable HPLC-UV-DAD method to identify and quantify phenolic compounds in the leaves and bulbs of I.bucharica,establishing the first set of quality control markers for this species.A total of 21 phenolic compounds were identified in the leaves,with flavonoids isoorientin,guaijaverin,hyperoside,and cosmosiin,the isoflavonoid biochanin A,and the simple phenolic ferulic acid being the most prominent.In comparison,14 compounds were identified in the bulbs,primarily isoflavonoids,including tectoridin and germanaism B,and flavonoid apigenin.The leaves extracts exhibited significant antioxidant activity,whereas the anti-allergic and anti-inflammatory effects were mild.These findings highlight I.bucharica as a sustainable source of bioactive compounds with potential industrial applications.However,further studies are needed to evaluate bioavailability and in vivo efficacy,as well as to optimise extraction methods to realize its industrial potential fully.展开更多
Detecting biomarkers in body fluids by optical lateral flow immune assay(LFIA) technology provides rapid access to disease information for early diagnosis.LFIA is based on an antigen-antibody reaction and is rapidly b...Detecting biomarkers in body fluids by optical lateral flow immune assay(LFIA) technology provides rapid access to disease information for early diagnosis.LFIA is based on an antigen-antibody reaction and is rapidly becoming the preferred choice of physicians and patients for point-of-care testing due to its simplicity,cost-effectiveness,and rapid detection.Observing the optical signal change from the colloidal gold of the traditional LFIA strip has been widely applied for various biomarkers detection in body fluids.Despite the significant progress,rapid real-time detection of color changes in the colloidal gold by the naked eye still faces many limitations,such as large errors and the inability to quantify and accurately detect.New optical LFIA strip technology has emerged in recent years to extend its application scenarios for achieving quantitative detection such as fluorescence,afterglow,and chemiluminescence.Herein,we summarized the development of optical LFIA technology from single to hyphenated optical signals for biomarkers detection in body fluids from invasive and non-invasive sources.Moreover,the challenge and outlook of optical LFIA strip technology are highlighted to inspire the designing of next-generation diagnostic platforms.展开更多
Background:Pistacia chinensis Bunge has been traditionally used to manage various conditions,including asthma,pain,inflammation,hepatoprotection,and diabetes.The study was conducted to investigate the antioxidant and ...Background:Pistacia chinensis Bunge has been traditionally used to manage various conditions,including asthma,pain,inflammation,hepatoprotection,and diabetes.The study was conducted to investigate the antioxidant and anti-lipoxygenase(LOX)properties of the isolated compound 2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-4H-chromen-4-one from Pistacia chinensis.Methods:LOX assay and antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl(DPPH)assay were performed.Molecular docking studies were conducted using a molecular operating environment.Results:The LOX assay revealed significant inhibitory effects at 0.2µM concentration,with an IC50 value of 37.80µM.The antioxidant effect demonstrated dose-dependency across 5 to 100µg/mL concentrations,reaching 93.09%at 100µg/mL,comparable to ascorbic acid’s 95.43%effect.Molecular docking studies highlighted strong interactions with the lipoxygenase enzyme,presenting an excellent docking score of-10.98 kcal/mol.Conclusion:These findings provide valuable insights into Pistacia chinensis’chemical components and biological effects,reinforcing its traditional medicinal applications.展开更多
Succinylcholine(SC)is a widely used depolarizing muscle relaxant,but improper use can lead to arrhythmias and,in severe cases,pose a life-threatening risk.Additionally,some criminals exploit SC for illicit activities....Succinylcholine(SC)is a widely used depolarizing muscle relaxant,but improper use can lead to arrhythmias and,in severe cases,pose a life-threatening risk.Additionally,some criminals exploit SC for illicit activities.Therefore,rapid SC detection is paramount for clinical practice and public safety.Currently,however,limited methods are available for the rapid detection of SC.A fluorescent indicator displacement assay sensor based on molecular recognition of an amide naphthotube was developed.This sensor enabled the rapid fluorescent detection of SC through competitive binding between SC and methylene blue with the amide naphthotube.The sensor exhibited exceptional sensitivity with a detection limit as low as 1.1μmol/L and a detection range of 1.1~60μmol/L,coupled with outstanding selectivity and robust stability.Furthermore,this sensor accurately determined SC levels in biological samples such as serum.In summary,this research provides a new solution for the rapid and accurate sensing of SC in complex matrices and offers new insights for the swift identification and detection of toxins.展开更多
Micro RNA-133a(mi RNA-133a) and cardiac troponin I(c Tn I) are different-type crucial biomarkers of acute myocardial infarction(AMI), whose levels are great significance for AMI diagnosis and treatment. Herein,a novel...Micro RNA-133a(mi RNA-133a) and cardiac troponin I(c Tn I) are different-type crucial biomarkers of acute myocardial infarction(AMI), whose levels are great significance for AMI diagnosis and treatment. Herein,a novel photoelectrochemical-electrochemical(PEC-EC) dual-mode biosensing platform for dual-target assays of mi RNA-133a and c Tn I was developed. In which, a PEC-EC dual-mode sensing platform for mi RNA-133a was constructed based on the changes of the photocurrent inhibition effect and the electrochemical signal of Fc on the Fc-hairpin DNA probe(Fc-HP)/Zn Cd S-quantum dots(QDs)/ITO electrode. Furthermore, under magnetic separation and the specific interaction between c Tn I and its aptamer, the N-doped porous carbon-Zn O polyhedra(NPC-Zn O)-hemin-capture DNA probe hybrid(NH-CP) was obtained and introduced to the Fc-HP/Zn Cd S-QDs/ITO electrode via hybridization between NH-CP and Fc-HP. The hemin molecules encapsulated in NH-CP could effectively induce the photocurrent-polarity-switching of the FcHP/Zn Cd S-QDs/ITO electrode and generate a new electrochemical signal originating from hemin. Thus,c Tn I was assayed sensitively and selectively by the PEC-EC dual-mode biosensing platform. Here, Fc and hemin not only serve as the electrochemical indicators, but also respectively inhibit the photocurrent and switch the photocurrent polarity of Zn Cd S-QDs. Furthermore, the proposed biosensing platform could be easily expanded to the detection of other multiplex-type biomarkers via the change of the sequences of the related DNA probes, implying its significant potential in clinical diagnosis and biological analysis.展开更多
Background:Ethylhexyl triazone(EHT)and diethylhexyl butamido triazone(HEB)both belong to the recently developed class of triazine ultraviolet filters.However,their toxicity profiles remain unclear.Objective:To assess ...Background:Ethylhexyl triazone(EHT)and diethylhexyl butamido triazone(HEB)both belong to the recently developed class of triazine ultraviolet filters.However,their toxicity profiles remain unclear.Objective:To assess the genotoxic and phototoxic effects of EHT and HEB.Methods:The genotoxicity of EHT and HEB was assessed using in vitro bacterial reverse mutation assays,chromosomal aberration assays,and micronucleus assays.Meanwhile,their phototoxicity was evaluated using in vitro 3T3 neutral red uptake(NRU)phototoxicity assays and in vivo skin phototoxicity tests.Results:In the bacterial reverse mutation assay,the number of bacterial colonies was not significantly higher in the EHT and HEB groups than in the solvent control group.Similarly,the chromosomal aberration assay revealed no increase in aberration rates after either EHT or HEB treatment.In the micronucleus assay,the frequency of micronuclei was comparable between the treatment and control groups.Finally,based on the 3T3 NRU phototoxicity assay,both EHT and HEB(photo-irritation factor<2 and mean photo effect value<0.1)were classified as non-phototoxic.The skin phototoxicity test in vivo showed the same results as in vitro.Conclusion:Results from a series of genotoxicity and phototoxicity assays indicate that EHT and HEB possess neither genotoxic nor phototoxic potential.These findings provide experimental evidence supporting the safety of EHT and HEB for topical applications.展开更多
Objective:To explore the mechanism of action of Zangjiangzhi capsule(ZJZC)in treating hyperlipidemia(HLP).Methods:The components of ZJZC were analyzed and identified using ultra-high performance liquid chromatography ...Objective:To explore the mechanism of action of Zangjiangzhi capsule(ZJZC)in treating hyperlipidemia(HLP).Methods:The components of ZJZC were analyzed and identified using ultra-high performance liquid chromatography with Q-Exactive Orbitrap tandem mass spectrometry(UHPLC-Q-Exactive-Orbitrap-MS/MS).Network pharmacology analysis was used to explore the mechanism of action of ZJZC in HLP treatment.The SwissTargetPrediction database was used to predict compound targets,and GeneCards,DisGeNet,OMIM,and DRUGBANK databases were used to identify HLP-related targets.Proteineprotein interaction diagrams were constructed using the STRING database.The targets were subjected to gene ontology and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.The“herbingredient-target”network was visualized using Cytoscape.Preliminary validation was performed using molecular docking and enzyme-linked immunosorbent assay.Results:Ninety compounds were identified in ZJZC,including 34 flavonoids,12 phenols,10 terpenoids,10 alkaloids,8 organic acids,8 anthraquinones,and 9 other compounds.In total,904 targets were identified for these compounds.Among them,158 targets intersected with the HLP target network.Network pharmacology analysis showed that MAPK1,PPAR-a,RXRA,HSP90AA1,PIK3R1,AKT1,PIK3CA,IL6,TNF,and ESR1 are the key targets of action.KEGG enrichment analysis identified 164 pathways.Among these,the AGE-RAGE signaling pathway in diabetic complications,lipid and atherosclerosis pathways,regulation of lipids in adipocytes,and insulin resistance are related to HLP.Molecular docking showed good affinity between the key targets and ingredients.Further,ZJZC treatment in mice resulted in lower expression of MAPK1 protein and increased expression of PPAR-a protein,which have been shown to be strongly associated with HLP.Conclusions:This study showed that ZJZC contains various active ingredients and can modulate multiple targets and pathways associated with HLP,providing evidence at the molecular level for its clinical application in the treatment of HLP.展开更多
Silver ion(Ag^(+))is a highly toxic metal ion,and its monitoring in water or food resources has become extraordinarily necessary within the scope of human health.In the light of the fact of Ag^(+)-induced folding stru...Silver ion(Ag^(+))is a highly toxic metal ion,and its monitoring in water or food resources has become extraordinarily necessary within the scope of human health.In the light of the fact of Ag^(+)-induced folding structure of specific peptides,an unlabeled and highselectivity Ag^(+)assay is presented by means of intrinsic fluorescence of peptides.Under the quenching effect of gold nanoparticles(AuNPs),characteristic fluorescence of peptides could be considerably reduced by rapid modification.Along with the Ag adding,the fluorescence signals of peptide-AuNPs are largely enhanced by the behavior between peptides and Agt.This is basically involving the formation of 4-coordinated complexes,generating the changes of peptides in structure and fluorescence properties.Under this circumstance,the adverse influence of plenty of interfering ions is suppressed,including the toxic Hg^(2+),Pb^(2+).The results highlight that Ag ions could be selectively recognized as low as 2.4 nmol/L with a linear range of 5 to 800 nmol/L.In comparison with other programs,the given approach declares simplicity,sensitivity,and superior selectivity.Furthermore,the biosensor excels in the practical application in water samples(e.g.,lake,tap and drinking water)owing to its non-interference and on-site rapid determination.展开更多
Hair follicle stem cell(HFSC),capable of self-renewal and differentiation in hair follicle,represents an emerging stem cell model for regenerative medicine.The interaction between HFSC and dermal papilla cell(DPC)gove...Hair follicle stem cell(HFSC),capable of self-renewal and differentiation in hair follicle,represents an emerging stem cell model for regenerative medicine.The interaction between HFSC and dermal papilla cell(DPC)governs hair follicle development.FGF7 functions as a paracrine protein regulating epithelial proliferation,differentiation and migration.The single-cell transcriptome profling and immunofuorescence analysis demonstrated that FGF7 localizes at DPC,while FGF7 receptor(FGFR2)expresses in both DPC and HFSC.Through co-culture experiments of HFSC and DPC,the results indicated that FGF7 secreted from DPC promotes the proliferation of DPC and HFSC via Wnt signaling pathway and induces HFSC differentiation.Furthermore,CUT&Tag assay revealed genomic colocalization between FGF7 and pluripotency-related genes and GSK3β.Electrophoretic mobility shift assay(EMSA)demonstrated that FGF7 interacts with the promoter region of CISH and PRKX.This research provides valuable insights into the molecular mechanisms underlying the hair cycle.Understanding the interaction between HFSC and DPC,as well as the role of FGF7,may advance regenerative medicine and hair loss treatment.展开更多
BACKGROUND Neurodegeneration refers to the progressive loss of neurons,affecting both their structure and function.It is driven by synaptic dysfunction,disruptions in neural networks,and the accumulation of abnormal p...BACKGROUND Neurodegeneration refers to the progressive loss of neurons,affecting both their structure and function.It is driven by synaptic dysfunction,disruptions in neural networks,and the accumulation of abnormal protein variants.Endoplasmic reticulum(ER)stress,caused by the accumulation of misfolded or unfolded protein,is a major contributor to neurodegeneration.Dithiothreitol(DTT)is a widely used redox reagent that disrupts the oxidative protein folding environment,inducing ER stress and leading to the imbalance in protein homeostasis can activate stress response pathway,potentially contributing to neurodegenerative processes.Caenorhabditis elegans(C.elegans)is a widely used model organism for studying neurodegeneration due to its well-mapped nervous system,approximately onethird of neuron cells in their body,complete genome sequenced,and conserved stress response pathway.AIM To study the neurodegeneration in C.elegans caused by DTT-induced ER stress,assessed by behavioral,molecular,and lifespan changes.METHODS C.elegans were cultured on nematode growth medium plates with OP50,and ER stress was induced using DTT.Effects were assessed via behavioral assays such as locomotion,chemotaxis,lifespan assay,and molecular studies.RESULTS DTT exposure led to a significant decline in locomotion and chemotaxis response,indicating neurotoxicity.A reduction in lifespan was observed,suggesting an overall impact on health.Molecular analysis confirmed ER stress activation.DTT-induced ER stress negatively affects C.elegans,leading to behavioral impairments and molecular alterations associated with neurodegeneration.CONCLUSION These findings establish C.elegans as a potential model for studying ER stress-mediated neurotoxicity and its implications in neurodegenerative diseases.展开更多
Histamine(HA)is a biogenic amine formed during the metabolism of microorganisms,excessive intake of which can cause headache,diarrhea,and,in severe cases,food poisoning.The commonly used instrumental detection methods...Histamine(HA)is a biogenic amine formed during the metabolism of microorganisms,excessive intake of which can cause headache,diarrhea,and,in severe cases,food poisoning.The commonly used instrumental detection methods are expensive and time-consuming,making it challenging to realize on-site inspection.In this work,we reported a colorimetric test strip-based detection method for HA by chemical recognition of the amine and imidazole groups of HA.Polydopamine-coated gold nanoparticles that were modified with carboxyl group-terminated polyethylene glycol(Au@PDA-C NPs)were utilized as labels to capture HA through the N-hydroxysuccinimide/1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(NHS/EDC)reaction between HA and the carboxyl group of the Au@PDA-C NPs.The chelate from nitrilotriacetic acid(NTA)and Cu^(2+)at the T-line could then specifically bind the imidazole moiety of HA in the complex of HA and Au@PDA-C NPs,enabling the T-line to present the colorimetric characteristics of Au@PDA-C NPs.The gray value of the T-line in the test strip showed a good linear relationship with the HA concentration in the range of 0–100μg/m L,with the standard curve equation of Y=27.94X+463.0 and the correlation coefficient of R^(2)=0.944.The detection limit and quantification limit of HA were 1.16μg/m L(signal-to-noise ratio(S/N)=3)and 3.88μg/m L(S/N=10),respectively.The two-site chemical recognition method could also achieve specific detection of HA in commercially available samples,showing potential application as an on-site analysis tool for HA.展开更多
Diabetes remains one of the most pressing global metabolic disorders,necessitating regular and precise monitoring of blood glucose levels for effective disease management.In this study,we developed a rapid and reliabl...Diabetes remains one of the most pressing global metabolic disorders,necessitating regular and precise monitoring of blood glucose levels for effective disease management.In this study,we developed a rapid and reliable method for quantifying glucose in whole blood using near-infrared(NIR)spectroscopy.A diabetic rat model was established through a high-fat,high-sugar diet followed by administration of streptozotocin(STZ)over a period of 4 weeks.To construct the NIR calibration model,partial least-squares(PLS)regression was employed,with optimization tailored to spectral range,preprocessing techniques,and the number of latent variables.The optimal model was achieved within the spectral window of 7502.0-5446.2 cm^(-1),using Constant Offset Elimination for spectral pretreatment and a factor number of 10.This optimized model yielded a strong correlation coefficient(R)of 0.9621,with a root mean square error of cross-validation(RMSECV)of 0.612,a residual predictive deviation(RPD)of 3.48,and a root mean square error of prediction(RMSEP)of 0.420.Additionally,biochemical indices were evaluated across all experimental groups to validate the model’s performance.Overall,the proposed NIR-based analytical strategy demonstrated high accuracy,robustness,and reproducibility,offering a promising tool for rapid glucose assessment in whole blood.展开更多
Objectives:Keratin 6A(KRT6A)has been implicated in the progression of multiple malignancies;however,its expression pattern and biological role in cervical cancer(CC)have not been elucidated.This study aims to investig...Objectives:Keratin 6A(KRT6A)has been implicated in the progression of multiple malignancies;however,its expression pattern and biological role in cervical cancer(CC)have not been elucidated.This study aims to investigate KRT6A expression in CC tissues and evaluate its effects on cellular proliferation,migration,and invasion,thereby assessing its potential as a biomarker and therapeutic target.Methods:Differentially expressed genes were screened from the Gene Expression Omnibus(GEO)dataset(GSE9750)using the thresholds∣log2FC∣>2 and false discovery rate(FDR)<0.05.Immunohistochemistry was performed to evaluate KRT6A protein expression in tumor tissues.Stable KRT6A knockdown was established in CC cell lines to assess proliferation,colony formation,migration,and invasion in vitro.A nude-mouse xenograft model was used to evaluate tumor growth in vivo.Results:KRT6A expression was significantly increased in CC tissues relative to adjacent non-tumor tissues(p=0.019).Patients with KRT6A-positive tumors had a significantly lower 3-year progression-free survival(PFS)rate than those with KRT6Anegative tumors(45.16%vs.78.57%;p=0.009).KRT6A was identified as an independent prognostic indicator for CC(p=0.044).In vitro,KRT6A silencing significantly reduced colony formation,proliferation,migration,and invasion in SiHa and HeLa cells in comparison to controls(p<0.05).In vivo,tumors in the KRT6A knockdown group were significantly smaller,with reduced expression of both KRT6A and Ki-67 in tumor tissues(p<0.05).Conclusion:KRT6A is overexpressed in CC and associates with adverse clinicopathological features and poor PFS.Knockdown of KRT6A suppresses cell proliferation,migration,and invasion,indicating that KRT6A may represent a promising prognostic biomarker and potential target for treating CC.展开更多
BACKGROUND The diagnosis of primary biliary cholangitis(PBC)remains challenging,particularly in cases where anti-mitochondrial antibody(AMA),anti-mitochondrial E2 subunit antibody(AMA-M2),anti-glycoprotein 210(anti-gp...BACKGROUND The diagnosis of primary biliary cholangitis(PBC)remains challenging,particularly in cases where anti-mitochondrial antibody(AMA),anti-mitochondrial E2 subunit antibody(AMA-M2),anti-glycoprotein 210(anti-gp210),and anti-speckled protein 100(anti-Sp100)are all negative.In such instances,the condition is often confirmed through a liver needle biopsy.AIM To identify additional plasma biomarkers for non-invasive diagnostic methods of PBC.METHODS We utilized the Sengenics KREX^(TM)immunome protein array to identify potential biomarkers for the diagnosis of PBC.Subsequently,we validated the predictive capability of the RPL30 antibody through an ELISA and retrospectively analyzed its association with the clinical features of 17 autoantibody-negative PBC cases and 45 autoantibody-positive PBC cases.RESULTS In our study we observed that RPL30 demonstrated the highest fold-change difference in PBC,with a penetrance frequency of 40%and a penetrance fold change of 38.30147.The analysis of anti-RPL30 optical density values between patients with AMA/AMA-M2/anti-gp210/anti-Sp100-negative PBC(autoantibody-negative PBC)and healthy controls using a receiver operating characteristic curve yielded an area under the curve of 0.853.This analysis established an optimal cutoff value of 0.0708,achieving 100%specificity and 75%sensitivity.The combination of anti-RPL30 and other autoantibodies elevated the diagnosis rate of PBC from 61.29%to 79.00%(P=0.0489).Anti-RPL30 demonstrated a high positive rate in antibody-negative PBC cases,including AMA/AMAM2/anti-gp210/anti-Sp100-negative cases.Correlation analysis of anti-RPL30 optical density values with clinical data from patients with PBC revealed a positive association with both the international normalized ratio(P=0.008)and the Model for End-Stage Liver Disease score(P=0.003).CONCLUSION Our study highlighted the potential of anti-RPL30 as a promising biomarker for diagnosing PBC,particularly in autoantibody-negative cases.展开更多
Soft rot is a destructive disease that inflicts significant losses on agricultural production and the economy post-harvest.Biocontrol strategies based on antagonistic microorganisms have a broad application prospect t...Soft rot is a destructive disease that inflicts significant losses on agricultural production and the economy post-harvest.Biocontrol strategies based on antagonistic microorganisms have a broad application prospect to fight against plant pathogens.This study utilized fluorescence-activated droplet sorting(FADS)technology as an alternative to traditional plate culture methods to isolate microorganisms with antagonistic activity against the soft rot pathogen Erwinia carotovora Ecc15.Initially,the culture performance of the FADS platform was evaluated by analyzing bacterial diversity in droplet culture samples and agar plate culture samples,our data showed that droplet culture exhibited higher species richness and diversity than plate culture,and more than 95%of the operational taxonomic units(OTUs)in the droplet samples belonged to the rare biosphere.Additionally,we developed a green fluorescent protein(GFP)-Ecc15-based FADS screening system,which achieved an enrichment ratio of up to 148.Using this system,we successfully screened 32 antagonistic bacteria from rhizosphere soil sample of healthy konjac plants,and some may be novel microbial resources,including the genera Lelliottia,Buttiauxella and Leclercia.Notably,strain D-62 exhibited the strongest antibacterial ability against Ecc15,with an inhibition zone diameter of(20.86±1.56)mm.In vivo experiments conducted on the corms of Amorphophallus konjac demonstrated that strain D-62 could effectively reduce the infection ability of Ecc15 to the corms,indicating that strain D-62 has the potential to be developed as a biocontrol agent.Our findings suggested that the FADS screening system showed a screening efficiency approximately 3×10^(3)times higher than plate screening system,while significantly reducing costs of infrastructure,labor and consumables,it provides theoretical guidance for the screening of other plant pathogen biocontrol bacteria.展开更多
BACKGROUND The use of apixaban in chronic hemodialysis(HD)patients for non-valvular atrial fibrillation(NVAF)is still controversial regarding benefit of stroke protection vs risk of bleeding.Rotational thromboelastome...BACKGROUND The use of apixaban in chronic hemodialysis(HD)patients for non-valvular atrial fibrillation(NVAF)is still controversial regarding benefit of stroke protection vs risk of bleeding.Rotational thromboelastometry(ROTEM)is a point of care method that evaluates clot formation in whole blood and has been used as an evaluation tool for bleeding risk assessment in non-HD apixaban users.AIM To determine whether bleeding risk can be predicted using ROTEM activated with tissue factor(EXTEM)in HD patients receiving apixaban for NVAF.METHODS Nineteen HD patients with NVAF treated with apixaban for at least 8 days were enrolled.Four dosing regimens were recorded as prescribed by their physician,from 2.5 mg once daily on a non-dialysis day to 5 mg twice daily.Standard coagulation tests,along with ROTEM and apixaban drug levels(using liquid anti-Xa assay)were performed once on a non-dialysis day before and two hours after apixaban morning pill administration.Patients were subsequently monitored for thrombotic/bleeding events and all-cause mortality.RESULTS Over a median follow-up period of 36 months,six patients experienced a bleeding event(group A)and 13 remained free of bleeding(group B).Six deaths were recorded:Three due to major bleeding,one from thrombotic stroke,and two unrelated to coagulopathy.EXTEM clotting time(CT)-post was the only parameter that significantly differed between group A and group B(P=0.013).Each 1-second increase in CT-post was linked to an 8%higher likelihood of a bleeding event(odds ratio=1.08,95%confidence interval:1.0-1.17;P=0.048).A significant progressive increase was observed with the drug’s trough and peak levels(P<0.05)across the four dosing regimens but no significant relationship was found between CT and apixaban dose groups.CONCLUSION Early detection of bleeding risk in HD patients with NVAF on Apixaban maybe be effectively achieved through frequent monitoring using ROTEM EXTEM post CT,thereby helping to reduce associated morbidity.展开更多
Functional recovery in penetrating neurological injury is hampered by a lack of clinical regenerative therapies.Biomaterial therapies show promise as medical materials for neural repair through immunomodulation,struct...Functional recovery in penetrating neurological injury is hampered by a lack of clinical regenerative therapies.Biomaterial therapies show promise as medical materials for neural repair through immunomodulation,structural support,and delivery of therapeutic biomolecules.However,a lack of facile and pathology-mimetic models for therapeutic testing is a bottleneck in neural tissue engineering research.We have deployed a two-dimensional,high-density multicellular cortical brain sheet to develop a facile model of injury(macrotransection/scratch wound)in vitro.The model encompasses the major neural cell types involved in pathological responses post-injury.Critically,we observed hallmark pathological responses in injury foci including cell scarring,immune cell infiltration,precursor cell migration,and shortrange axonal sprouting.Delivering test magnetic particles to evaluate the potential of the model for biomaterial screening shows a high uptake of introduced magnetic particles by injury-activated immune cells,mimicking in vivo findings.Finally,we proved it is feasible to create reproducible traumatic injuries in the brain sheet(in multielectrode array devices in situ)characterized by focal loss of electrical spiking in injury sites,offering the potential for longer term,electrophysiology plus histology assays.To our knowledge,this is the first in vitro simulation of transecting injury in a two-dimensional multicellular cortical brain cell sheet,that allows for combined histological and electrophysiological readouts of damage/repair.The patho-mimicry and adaptability of this simplified model of brain injury could benefit the testing of biomaterial therapeutics in regenerative neurology,with the option for functional electrophysiological readouts.展开更多
The interest in using the Datura stramonium plant is due to its natural products,which are used in many pharmaceutical industries.The objective of the current study was to assess the therapeutic and cytotoxic effects ...The interest in using the Datura stramonium plant is due to its natural products,which are used in many pharmaceutical industries.The objective of the current study was to assess the therapeutic and cytotoxic effects of the D.stramonium plant on two types of human cancer cell models(MCF7 and HT29)in vitro.A soxhlet apparatus was used to obtain methanolic extract from dried plant leaves.The recovered crude,after the solvent had evaporated,was then dispersed at varied concentrations of extract 100,50,20,and 0.0µg/mL and tested to see how the cells responded.Also,the cancer-testis antigen(CTA)gene transcription in the two cell types exposed to the plant extract was examined using a semi-quantitative real-time polymerase chain reaction.Gas chromatography–mass spectrometry(GC-MS)results produced the significant main metabolites Nonanoic acid,Tropine N-Oxide,3,6-Ditigloyloxy-7-hydroxytropane,Hexadecanoic acid,2-Pentadecanone,6,10,14-trimethyl-,Carvenone,methyl ester,Phytol,Aposcopolamine,Hyoscyamine,4,8,12,16-Tetramethylheptadecan-4-olide,Scopolamine,Alpha.-Tocospiro A,1,2-Cycloheptanedione,3,3,7,7-tetramethyl-,dihydrazone,Campesterol,Stigmasterol,Gamma-Sitosterol and dl-.alpha.-Tocopherol.The results showed that the two types of cell lines impacted by D.stramonium extract,through untreated type 1 cells(MCF7)gave a highly significant transcription according to all applicable genes.All implemented analyses cleared the strong genetic impacts of Datura extract on cancer cells’genomes.TGIF2LY and C2orf63 transcript accumulation were also significantly elevated when exposed to plant extract at a level of 50µg/mL in cell line type 2(HT29),but TGIF2LY and P53 had the lowest relative expression at a level of 100µg/mL when treated the same cell line type.展开更多
基金National Institute of Arthritis and Musculoskeletal and Skin Diseases of the National Institutes of Health,Grant/Award Number:R01AR069044Rutgers-New Jersey Medical School Department of Orthopaedics。
文摘Osteoclasts are essential for maintaining healthy bone.Pathological elevation of os-teoclastogenesis or osteoclast activity can cause osteoporosis and increase the risk of bone fracture.However,a few options are available for directly measuring osteoclast activity in vivo to test interventions that may affect osteoclasts.Here,we describe an in vivo method to measure osteoclast-mediated bone loss targeted at normal mouse calvaria.The method employs a novel procedure for measuring osteoclast resorption pits using micro-computed tomography.The potential utility of this mouse calvaria model to assess therapies targeting osteoclasts was validated using zoledronic acid,which is a nitrogen-containing bisphosphonate drug used to treat osteoporosis.
基金financially supported by ARC Linkage project(LP210200642)ARC Center of Excellence for Quantum Biotechnology(grant no.CE230100021)+1 种基金National Health and Medical Research Council Investigator Fellowship—(grant no.APP2017499)Chan Zuckerberg Initiative Deep Tissue Imaging Phase 2(grant no.DT12-0000000182).
文摘Lateral flow immunoassays(LFIAs)are low-cost,rapid,and easy to use for pointof-care testing(POCT),but the majority of the available LFIA tests are indicative,rather than quantitative,and their sensitivity in antigen tests are usually limited at the nanogram range,which is primarily due to the passive capillary fluidics through nitrocellulose membranes,often associated with non-specific bindings and high background noise.To overcome this challenge,we report a Beads-on-a-Tip design by replacing nitrocellulose membranes with a pipette tip loaded with magnetic beads.The beads are pre-conjugated with capture antibodies that support a typical sandwich immunoassay.This design enriches the low-abundant antigen proteins and allows an active washing process to significantly reduce non-specific bindings.To further improve the detection sensitivity,we employed upconversion nanoparticles(UCNPs)as luminescent reporters and SARS-CoV-2 spike(S)antigen as a model analyte to benchmark the performance of this design against our previously reported methods.We found that the key to enhance the immunocomplex formation and signal-to-noise ratio lay in optimizing incubation time and the UCNP-to-bead ratio.We therefore successfully demonstrated that the new method can achieve a very large dynamic range from 500 fg/mL to 10μg/mL,across over 7 digits,and a limit of detection of 706 fg/mL,nearly another order of magnitude lower than the best reported LFIA using UCNPs in COVID-19 spike antigen detection.Our system offers a promising solution for ultra-sensitive and quantitative POCT diagnostics.
基金supported by grants from the National Science and Technology Council(NSTC),Taiwan(NSTC 114-2320-B-037-020-MY3,113-2320-B-037-023,and 112-2320-B-037-012 granted to M.K.,113-2321-B-255-001,113-2321-B-182-003,112-2321-B-182-003,112-2321-B-255-001,111-2320-B-255-006-MY3,and 111-2321-B-255-001 granted to T.L.H.)Kaohsiung Medical University Research Foundation[KMU-Q113011 awarded to M.K.and KMU-M114020 awarded to B.H.C.]NSYSU-KMU joint research project(NSYSU-KMU-114-P16)awarded to M.K.
文摘The sustainable sourcing of novel bioactive compounds from natural sources is crucial to the success of the pharmaceutical,food,and cosmetics industries.Iris bucharica Foster(syn.Juno bucharica(Foster)Vved.)is a promising source of novel bioactive molecules,particularly phenolic compounds,which are renowned for their antioxidant properties.In this study,we developed a reliable HPLC-UV-DAD method to identify and quantify phenolic compounds in the leaves and bulbs of I.bucharica,establishing the first set of quality control markers for this species.A total of 21 phenolic compounds were identified in the leaves,with flavonoids isoorientin,guaijaverin,hyperoside,and cosmosiin,the isoflavonoid biochanin A,and the simple phenolic ferulic acid being the most prominent.In comparison,14 compounds were identified in the bulbs,primarily isoflavonoids,including tectoridin and germanaism B,and flavonoid apigenin.The leaves extracts exhibited significant antioxidant activity,whereas the anti-allergic and anti-inflammatory effects were mild.These findings highlight I.bucharica as a sustainable source of bioactive compounds with potential industrial applications.However,further studies are needed to evaluate bioavailability and in vivo efficacy,as well as to optimise extraction methods to realize its industrial potential fully.
基金supported by the National Natural Science Foundation of China (Nos.22234005,22494632,22404081)the Natural Science Foundation of Jiangsu Province (Nos.BK20222015,BK20240534)。
文摘Detecting biomarkers in body fluids by optical lateral flow immune assay(LFIA) technology provides rapid access to disease information for early diagnosis.LFIA is based on an antigen-antibody reaction and is rapidly becoming the preferred choice of physicians and patients for point-of-care testing due to its simplicity,cost-effectiveness,and rapid detection.Observing the optical signal change from the colloidal gold of the traditional LFIA strip has been widely applied for various biomarkers detection in body fluids.Despite the significant progress,rapid real-time detection of color changes in the colloidal gold by the naked eye still faces many limitations,such as large errors and the inability to quantify and accurately detect.New optical LFIA strip technology has emerged in recent years to extend its application scenarios for achieving quantitative detection such as fluorescence,afterglow,and chemiluminescence.Herein,we summarized the development of optical LFIA technology from single to hyphenated optical signals for biomarkers detection in body fluids from invasive and non-invasive sources.Moreover,the challenge and outlook of optical LFIA strip technology are highlighted to inspire the designing of next-generation diagnostic platforms.
文摘Background:Pistacia chinensis Bunge has been traditionally used to manage various conditions,including asthma,pain,inflammation,hepatoprotection,and diabetes.The study was conducted to investigate the antioxidant and anti-lipoxygenase(LOX)properties of the isolated compound 2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-4H-chromen-4-one from Pistacia chinensis.Methods:LOX assay and antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl(DPPH)assay were performed.Molecular docking studies were conducted using a molecular operating environment.Results:The LOX assay revealed significant inhibitory effects at 0.2µM concentration,with an IC50 value of 37.80µM.The antioxidant effect demonstrated dose-dependency across 5 to 100µg/mL concentrations,reaching 93.09%at 100µg/mL,comparable to ascorbic acid’s 95.43%effect.Molecular docking studies highlighted strong interactions with the lipoxygenase enzyme,presenting an excellent docking score of-10.98 kcal/mol.Conclusion:These findings provide valuable insights into Pistacia chinensis’chemical components and biological effects,reinforcing its traditional medicinal applications.
文摘Succinylcholine(SC)is a widely used depolarizing muscle relaxant,but improper use can lead to arrhythmias and,in severe cases,pose a life-threatening risk.Additionally,some criminals exploit SC for illicit activities.Therefore,rapid SC detection is paramount for clinical practice and public safety.Currently,however,limited methods are available for the rapid detection of SC.A fluorescent indicator displacement assay sensor based on molecular recognition of an amide naphthotube was developed.This sensor enabled the rapid fluorescent detection of SC through competitive binding between SC and methylene blue with the amide naphthotube.The sensor exhibited exceptional sensitivity with a detection limit as low as 1.1μmol/L and a detection range of 1.1~60μmol/L,coupled with outstanding selectivity and robust stability.Furthermore,this sensor accurately determined SC levels in biological samples such as serum.In summary,this research provides a new solution for the rapid and accurate sensing of SC in complex matrices and offers new insights for the swift identification and detection of toxins.
基金financially supported by National Natural Science Foundation of China (Nos. 22074033, 22374035)。
文摘Micro RNA-133a(mi RNA-133a) and cardiac troponin I(c Tn I) are different-type crucial biomarkers of acute myocardial infarction(AMI), whose levels are great significance for AMI diagnosis and treatment. Herein,a novel photoelectrochemical-electrochemical(PEC-EC) dual-mode biosensing platform for dual-target assays of mi RNA-133a and c Tn I was developed. In which, a PEC-EC dual-mode sensing platform for mi RNA-133a was constructed based on the changes of the photocurrent inhibition effect and the electrochemical signal of Fc on the Fc-hairpin DNA probe(Fc-HP)/Zn Cd S-quantum dots(QDs)/ITO electrode. Furthermore, under magnetic separation and the specific interaction between c Tn I and its aptamer, the N-doped porous carbon-Zn O polyhedra(NPC-Zn O)-hemin-capture DNA probe hybrid(NH-CP) was obtained and introduced to the Fc-HP/Zn Cd S-QDs/ITO electrode via hybridization between NH-CP and Fc-HP. The hemin molecules encapsulated in NH-CP could effectively induce the photocurrent-polarity-switching of the FcHP/Zn Cd S-QDs/ITO electrode and generate a new electrochemical signal originating from hemin. Thus,c Tn I was assayed sensitively and selectively by the PEC-EC dual-mode biosensing platform. Here, Fc and hemin not only serve as the electrochemical indicators, but also respectively inhibit the photocurrent and switch the photocurrent polarity of Zn Cd S-QDs. Furthermore, the proposed biosensing platform could be easily expanded to the detection of other multiplex-type biomarkers via the change of the sequences of the related DNA probes, implying its significant potential in clinical diagnosis and biological analysis.
基金supported by the National Natural Science Foundation of China(No.81600459)National Natural Science Foundation of Hubei Province(No.2016CFB312)+3 种基金Talent Introduction Project of Hubei Polytechnic University(No.15xjz03R)Industry-University Collaboration(No.KY2023-269)Hubei Key Laboratory of Kidney Disease Pathogenesis and Intervention Foundation(No.SB202103)Provincial College Student Innovation and Entrepreneurship Training Program(No.S202210920011).
文摘Background:Ethylhexyl triazone(EHT)and diethylhexyl butamido triazone(HEB)both belong to the recently developed class of triazine ultraviolet filters.However,their toxicity profiles remain unclear.Objective:To assess the genotoxic and phototoxic effects of EHT and HEB.Methods:The genotoxicity of EHT and HEB was assessed using in vitro bacterial reverse mutation assays,chromosomal aberration assays,and micronucleus assays.Meanwhile,their phototoxicity was evaluated using in vitro 3T3 neutral red uptake(NRU)phototoxicity assays and in vivo skin phototoxicity tests.Results:In the bacterial reverse mutation assay,the number of bacterial colonies was not significantly higher in the EHT and HEB groups than in the solvent control group.Similarly,the chromosomal aberration assay revealed no increase in aberration rates after either EHT or HEB treatment.In the micronucleus assay,the frequency of micronuclei was comparable between the treatment and control groups.Finally,based on the 3T3 NRU phototoxicity assay,both EHT and HEB(photo-irritation factor<2 and mean photo effect value<0.1)were classified as non-phototoxic.The skin phototoxicity test in vivo showed the same results as in vitro.Conclusion:Results from a series of genotoxicity and phototoxicity assays indicate that EHT and HEB possess neither genotoxic nor phototoxic potential.These findings provide experimental evidence supporting the safety of EHT and HEB for topical applications.
基金supported by the National Natural Science Foundation of China(22067016)the Project of Qinghai Outstanding Youth Fund Project(2023-ZJ-964J).
文摘Objective:To explore the mechanism of action of Zangjiangzhi capsule(ZJZC)in treating hyperlipidemia(HLP).Methods:The components of ZJZC were analyzed and identified using ultra-high performance liquid chromatography with Q-Exactive Orbitrap tandem mass spectrometry(UHPLC-Q-Exactive-Orbitrap-MS/MS).Network pharmacology analysis was used to explore the mechanism of action of ZJZC in HLP treatment.The SwissTargetPrediction database was used to predict compound targets,and GeneCards,DisGeNet,OMIM,and DRUGBANK databases were used to identify HLP-related targets.Proteineprotein interaction diagrams were constructed using the STRING database.The targets were subjected to gene ontology and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.The“herbingredient-target”network was visualized using Cytoscape.Preliminary validation was performed using molecular docking and enzyme-linked immunosorbent assay.Results:Ninety compounds were identified in ZJZC,including 34 flavonoids,12 phenols,10 terpenoids,10 alkaloids,8 organic acids,8 anthraquinones,and 9 other compounds.In total,904 targets were identified for these compounds.Among them,158 targets intersected with the HLP target network.Network pharmacology analysis showed that MAPK1,PPAR-a,RXRA,HSP90AA1,PIK3R1,AKT1,PIK3CA,IL6,TNF,and ESR1 are the key targets of action.KEGG enrichment analysis identified 164 pathways.Among these,the AGE-RAGE signaling pathway in diabetic complications,lipid and atherosclerosis pathways,regulation of lipids in adipocytes,and insulin resistance are related to HLP.Molecular docking showed good affinity between the key targets and ingredients.Further,ZJZC treatment in mice resulted in lower expression of MAPK1 protein and increased expression of PPAR-a protein,which have been shown to be strongly associated with HLP.Conclusions:This study showed that ZJZC contains various active ingredients and can modulate multiple targets and pathways associated with HLP,providing evidence at the molecular level for its clinical application in the treatment of HLP.
基金Supported by the National Natural Science Foundation of China(21775114,21874102)。
文摘Silver ion(Ag^(+))is a highly toxic metal ion,and its monitoring in water or food resources has become extraordinarily necessary within the scope of human health.In the light of the fact of Ag^(+)-induced folding structure of specific peptides,an unlabeled and highselectivity Ag^(+)assay is presented by means of intrinsic fluorescence of peptides.Under the quenching effect of gold nanoparticles(AuNPs),characteristic fluorescence of peptides could be considerably reduced by rapid modification.Along with the Ag adding,the fluorescence signals of peptide-AuNPs are largely enhanced by the behavior between peptides and Agt.This is basically involving the formation of 4-coordinated complexes,generating the changes of peptides in structure and fluorescence properties.Under this circumstance,the adverse influence of plenty of interfering ions is suppressed,including the toxic Hg^(2+),Pb^(2+).The results highlight that Ag ions could be selectively recognized as low as 2.4 nmol/L with a linear range of 5 to 800 nmol/L.In comparison with other programs,the given approach declares simplicity,sensitivity,and superior selectivity.Furthermore,the biosensor excels in the practical application in water samples(e.g.,lake,tap and drinking water)owing to its non-interference and on-site rapid determination.
基金supported by the National Key Research And Development Program of China(2022YFD1300204)。
文摘Hair follicle stem cell(HFSC),capable of self-renewal and differentiation in hair follicle,represents an emerging stem cell model for regenerative medicine.The interaction between HFSC and dermal papilla cell(DPC)governs hair follicle development.FGF7 functions as a paracrine protein regulating epithelial proliferation,differentiation and migration.The single-cell transcriptome profling and immunofuorescence analysis demonstrated that FGF7 localizes at DPC,while FGF7 receptor(FGFR2)expresses in both DPC and HFSC.Through co-culture experiments of HFSC and DPC,the results indicated that FGF7 secreted from DPC promotes the proliferation of DPC and HFSC via Wnt signaling pathway and induces HFSC differentiation.Furthermore,CUT&Tag assay revealed genomic colocalization between FGF7 and pluripotency-related genes and GSK3β.Electrophoretic mobility shift assay(EMSA)demonstrated that FGF7 interacts with the promoter region of CISH and PRKX.This research provides valuable insights into the molecular mechanisms underlying the hair cycle.Understanding the interaction between HFSC and DPC,as well as the role of FGF7,may advance regenerative medicine and hair loss treatment.
文摘BACKGROUND Neurodegeneration refers to the progressive loss of neurons,affecting both their structure and function.It is driven by synaptic dysfunction,disruptions in neural networks,and the accumulation of abnormal protein variants.Endoplasmic reticulum(ER)stress,caused by the accumulation of misfolded or unfolded protein,is a major contributor to neurodegeneration.Dithiothreitol(DTT)is a widely used redox reagent that disrupts the oxidative protein folding environment,inducing ER stress and leading to the imbalance in protein homeostasis can activate stress response pathway,potentially contributing to neurodegenerative processes.Caenorhabditis elegans(C.elegans)is a widely used model organism for studying neurodegeneration due to its well-mapped nervous system,approximately onethird of neuron cells in their body,complete genome sequenced,and conserved stress response pathway.AIM To study the neurodegeneration in C.elegans caused by DTT-induced ER stress,assessed by behavioral,molecular,and lifespan changes.METHODS C.elegans were cultured on nematode growth medium plates with OP50,and ER stress was induced using DTT.Effects were assessed via behavioral assays such as locomotion,chemotaxis,lifespan assay,and molecular studies.RESULTS DTT exposure led to a significant decline in locomotion and chemotaxis response,indicating neurotoxicity.A reduction in lifespan was observed,suggesting an overall impact on health.Molecular analysis confirmed ER stress activation.DTT-induced ER stress negatively affects C.elegans,leading to behavioral impairments and molecular alterations associated with neurodegeneration.CONCLUSION These findings establish C.elegans as a potential model for studying ER stress-mediated neurotoxicity and its implications in neurodegenerative diseases.
基金supported by the Zhejiang Gongshang University(QRK22006)the Eagle Plan Cultivation Project of Zhejiang Provincial Administration for Market Regulation(CY2023005)。
文摘Histamine(HA)is a biogenic amine formed during the metabolism of microorganisms,excessive intake of which can cause headache,diarrhea,and,in severe cases,food poisoning.The commonly used instrumental detection methods are expensive and time-consuming,making it challenging to realize on-site inspection.In this work,we reported a colorimetric test strip-based detection method for HA by chemical recognition of the amine and imidazole groups of HA.Polydopamine-coated gold nanoparticles that were modified with carboxyl group-terminated polyethylene glycol(Au@PDA-C NPs)were utilized as labels to capture HA through the N-hydroxysuccinimide/1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(NHS/EDC)reaction between HA and the carboxyl group of the Au@PDA-C NPs.The chelate from nitrilotriacetic acid(NTA)and Cu^(2+)at the T-line could then specifically bind the imidazole moiety of HA in the complex of HA and Au@PDA-C NPs,enabling the T-line to present the colorimetric characteristics of Au@PDA-C NPs.The gray value of the T-line in the test strip showed a good linear relationship with the HA concentration in the range of 0–100μg/m L,with the standard curve equation of Y=27.94X+463.0 and the correlation coefficient of R^(2)=0.944.The detection limit and quantification limit of HA were 1.16μg/m L(signal-to-noise ratio(S/N)=3)and 3.88μg/m L(S/N=10),respectively.The two-site chemical recognition method could also achieve specific detection of HA in commercially available samples,showing potential application as an on-site analysis tool for HA.
基金The University Key Research Projects of Henan Province(Grant No.25B360004)the Backbone Teachers Program of North Henan Medical University(Sanquan College of Xinxiang Medical University)(Grant No.SQ2025GGJS08).
文摘Diabetes remains one of the most pressing global metabolic disorders,necessitating regular and precise monitoring of blood glucose levels for effective disease management.In this study,we developed a rapid and reliable method for quantifying glucose in whole blood using near-infrared(NIR)spectroscopy.A diabetic rat model was established through a high-fat,high-sugar diet followed by administration of streptozotocin(STZ)over a period of 4 weeks.To construct the NIR calibration model,partial least-squares(PLS)regression was employed,with optimization tailored to spectral range,preprocessing techniques,and the number of latent variables.The optimal model was achieved within the spectral window of 7502.0-5446.2 cm^(-1),using Constant Offset Elimination for spectral pretreatment and a factor number of 10.This optimized model yielded a strong correlation coefficient(R)of 0.9621,with a root mean square error of cross-validation(RMSECV)of 0.612,a residual predictive deviation(RPD)of 3.48,and a root mean square error of prediction(RMSEP)of 0.420.Additionally,biochemical indices were evaluated across all experimental groups to validate the model’s performance.Overall,the proposed NIR-based analytical strategy demonstrated high accuracy,robustness,and reproducibility,offering a promising tool for rapid glucose assessment in whole blood.
基金supported by Yixing Taodu Light Science and Technology Research Plan,Grant Number:2024SF14.
文摘Objectives:Keratin 6A(KRT6A)has been implicated in the progression of multiple malignancies;however,its expression pattern and biological role in cervical cancer(CC)have not been elucidated.This study aims to investigate KRT6A expression in CC tissues and evaluate its effects on cellular proliferation,migration,and invasion,thereby assessing its potential as a biomarker and therapeutic target.Methods:Differentially expressed genes were screened from the Gene Expression Omnibus(GEO)dataset(GSE9750)using the thresholds∣log2FC∣>2 and false discovery rate(FDR)<0.05.Immunohistochemistry was performed to evaluate KRT6A protein expression in tumor tissues.Stable KRT6A knockdown was established in CC cell lines to assess proliferation,colony formation,migration,and invasion in vitro.A nude-mouse xenograft model was used to evaluate tumor growth in vivo.Results:KRT6A expression was significantly increased in CC tissues relative to adjacent non-tumor tissues(p=0.019).Patients with KRT6A-positive tumors had a significantly lower 3-year progression-free survival(PFS)rate than those with KRT6Anegative tumors(45.16%vs.78.57%;p=0.009).KRT6A was identified as an independent prognostic indicator for CC(p=0.044).In vitro,KRT6A silencing significantly reduced colony formation,proliferation,migration,and invasion in SiHa and HeLa cells in comparison to controls(p<0.05).In vivo,tumors in the KRT6A knockdown group were significantly smaller,with reduced expression of both KRT6A and Ki-67 in tumor tissues(p<0.05).Conclusion:KRT6A is overexpressed in CC and associates with adverse clinicopathological features and poor PFS.Knockdown of KRT6A suppresses cell proliferation,migration,and invasion,indicating that KRT6A may represent a promising prognostic biomarker and potential target for treating CC.
基金Supported by National Natural Science Foundation of China,No.82172257Health Care Major Project of Guangzhou,No.202206080001Science and Technology Cooperation Program of Fujian Province,No.2021I0036。
文摘BACKGROUND The diagnosis of primary biliary cholangitis(PBC)remains challenging,particularly in cases where anti-mitochondrial antibody(AMA),anti-mitochondrial E2 subunit antibody(AMA-M2),anti-glycoprotein 210(anti-gp210),and anti-speckled protein 100(anti-Sp100)are all negative.In such instances,the condition is often confirmed through a liver needle biopsy.AIM To identify additional plasma biomarkers for non-invasive diagnostic methods of PBC.METHODS We utilized the Sengenics KREX^(TM)immunome protein array to identify potential biomarkers for the diagnosis of PBC.Subsequently,we validated the predictive capability of the RPL30 antibody through an ELISA and retrospectively analyzed its association with the clinical features of 17 autoantibody-negative PBC cases and 45 autoantibody-positive PBC cases.RESULTS In our study we observed that RPL30 demonstrated the highest fold-change difference in PBC,with a penetrance frequency of 40%and a penetrance fold change of 38.30147.The analysis of anti-RPL30 optical density values between patients with AMA/AMA-M2/anti-gp210/anti-Sp100-negative PBC(autoantibody-negative PBC)and healthy controls using a receiver operating characteristic curve yielded an area under the curve of 0.853.This analysis established an optimal cutoff value of 0.0708,achieving 100%specificity and 75%sensitivity.The combination of anti-RPL30 and other autoantibodies elevated the diagnosis rate of PBC from 61.29%to 79.00%(P=0.0489).Anti-RPL30 demonstrated a high positive rate in antibody-negative PBC cases,including AMA/AMAM2/anti-gp210/anti-Sp100-negative cases.Correlation analysis of anti-RPL30 optical density values with clinical data from patients with PBC revealed a positive association with both the international normalized ratio(P=0.008)and the Model for End-Stage Liver Disease score(P=0.003).CONCLUSION Our study highlighted the potential of anti-RPL30 as a promising biomarker for diagnosing PBC,particularly in autoantibody-negative cases.
基金supported by the Guizhou Province High-level Innovative Talent Project(Qiankehe Platform Talent-GCC[2022]027-1)the National Key Research and Development Program of China(2019YFA0904800).
文摘Soft rot is a destructive disease that inflicts significant losses on agricultural production and the economy post-harvest.Biocontrol strategies based on antagonistic microorganisms have a broad application prospect to fight against plant pathogens.This study utilized fluorescence-activated droplet sorting(FADS)technology as an alternative to traditional plate culture methods to isolate microorganisms with antagonistic activity against the soft rot pathogen Erwinia carotovora Ecc15.Initially,the culture performance of the FADS platform was evaluated by analyzing bacterial diversity in droplet culture samples and agar plate culture samples,our data showed that droplet culture exhibited higher species richness and diversity than plate culture,and more than 95%of the operational taxonomic units(OTUs)in the droplet samples belonged to the rare biosphere.Additionally,we developed a green fluorescent protein(GFP)-Ecc15-based FADS screening system,which achieved an enrichment ratio of up to 148.Using this system,we successfully screened 32 antagonistic bacteria from rhizosphere soil sample of healthy konjac plants,and some may be novel microbial resources,including the genera Lelliottia,Buttiauxella and Leclercia.Notably,strain D-62 exhibited the strongest antibacterial ability against Ecc15,with an inhibition zone diameter of(20.86±1.56)mm.In vivo experiments conducted on the corms of Amorphophallus konjac demonstrated that strain D-62 could effectively reduce the infection ability of Ecc15 to the corms,indicating that strain D-62 has the potential to be developed as a biocontrol agent.Our findings suggested that the FADS screening system showed a screening efficiency approximately 3×10^(3)times higher than plate screening system,while significantly reducing costs of infrastructure,labor and consumables,it provides theoretical guidance for the screening of other plant pathogen biocontrol bacteria.
文摘BACKGROUND The use of apixaban in chronic hemodialysis(HD)patients for non-valvular atrial fibrillation(NVAF)is still controversial regarding benefit of stroke protection vs risk of bleeding.Rotational thromboelastometry(ROTEM)is a point of care method that evaluates clot formation in whole blood and has been used as an evaluation tool for bleeding risk assessment in non-HD apixaban users.AIM To determine whether bleeding risk can be predicted using ROTEM activated with tissue factor(EXTEM)in HD patients receiving apixaban for NVAF.METHODS Nineteen HD patients with NVAF treated with apixaban for at least 8 days were enrolled.Four dosing regimens were recorded as prescribed by their physician,from 2.5 mg once daily on a non-dialysis day to 5 mg twice daily.Standard coagulation tests,along with ROTEM and apixaban drug levels(using liquid anti-Xa assay)were performed once on a non-dialysis day before and two hours after apixaban morning pill administration.Patients were subsequently monitored for thrombotic/bleeding events and all-cause mortality.RESULTS Over a median follow-up period of 36 months,six patients experienced a bleeding event(group A)and 13 remained free of bleeding(group B).Six deaths were recorded:Three due to major bleeding,one from thrombotic stroke,and two unrelated to coagulopathy.EXTEM clotting time(CT)-post was the only parameter that significantly differed between group A and group B(P=0.013).Each 1-second increase in CT-post was linked to an 8%higher likelihood of a bleeding event(odds ratio=1.08,95%confidence interval:1.0-1.17;P=0.048).A significant progressive increase was observed with the drug’s trough and peak levels(P<0.05)across the four dosing regimens but no significant relationship was found between CT and apixaban dose groups.CONCLUSION Early detection of bleeding risk in HD patients with NVAF on Apixaban maybe be effectively achieved through frequent monitoring using ROTEM EXTEM post CT,thereby helping to reduce associated morbidity.
基金supported by awards from the EPSRC Centre for Doctoral Training in Regenerative Medicine(EP/L014904/1,to JW)an NHS bursary(to RHB)an EPSRC Healthcare Technologies award(EP/T013885/1,to DMC)。
文摘Functional recovery in penetrating neurological injury is hampered by a lack of clinical regenerative therapies.Biomaterial therapies show promise as medical materials for neural repair through immunomodulation,structural support,and delivery of therapeutic biomolecules.However,a lack of facile and pathology-mimetic models for therapeutic testing is a bottleneck in neural tissue engineering research.We have deployed a two-dimensional,high-density multicellular cortical brain sheet to develop a facile model of injury(macrotransection/scratch wound)in vitro.The model encompasses the major neural cell types involved in pathological responses post-injury.Critically,we observed hallmark pathological responses in injury foci including cell scarring,immune cell infiltration,precursor cell migration,and shortrange axonal sprouting.Delivering test magnetic particles to evaluate the potential of the model for biomaterial screening shows a high uptake of introduced magnetic particles by injury-activated immune cells,mimicking in vivo findings.Finally,we proved it is feasible to create reproducible traumatic injuries in the brain sheet(in multielectrode array devices in situ)characterized by focal loss of electrical spiking in injury sites,offering the potential for longer term,electrophysiology plus histology assays.To our knowledge,this is the first in vitro simulation of transecting injury in a two-dimensional multicellular cortical brain cell sheet,that allows for combined histological and electrophysiological readouts of damage/repair.The patho-mimicry and adaptability of this simplified model of brain injury could benefit the testing of biomaterial therapeutics in regenerative neurology,with the option for functional electrophysiological readouts.
文摘The interest in using the Datura stramonium plant is due to its natural products,which are used in many pharmaceutical industries.The objective of the current study was to assess the therapeutic and cytotoxic effects of the D.stramonium plant on two types of human cancer cell models(MCF7 and HT29)in vitro.A soxhlet apparatus was used to obtain methanolic extract from dried plant leaves.The recovered crude,after the solvent had evaporated,was then dispersed at varied concentrations of extract 100,50,20,and 0.0µg/mL and tested to see how the cells responded.Also,the cancer-testis antigen(CTA)gene transcription in the two cell types exposed to the plant extract was examined using a semi-quantitative real-time polymerase chain reaction.Gas chromatography–mass spectrometry(GC-MS)results produced the significant main metabolites Nonanoic acid,Tropine N-Oxide,3,6-Ditigloyloxy-7-hydroxytropane,Hexadecanoic acid,2-Pentadecanone,6,10,14-trimethyl-,Carvenone,methyl ester,Phytol,Aposcopolamine,Hyoscyamine,4,8,12,16-Tetramethylheptadecan-4-olide,Scopolamine,Alpha.-Tocospiro A,1,2-Cycloheptanedione,3,3,7,7-tetramethyl-,dihydrazone,Campesterol,Stigmasterol,Gamma-Sitosterol and dl-.alpha.-Tocopherol.The results showed that the two types of cell lines impacted by D.stramonium extract,through untreated type 1 cells(MCF7)gave a highly significant transcription according to all applicable genes.All implemented analyses cleared the strong genetic impacts of Datura extract on cancer cells’genomes.TGIF2LY and C2orf63 transcript accumulation were also significantly elevated when exposed to plant extract at a level of 50µg/mL in cell line type 2(HT29),but TGIF2LY and P53 had the lowest relative expression at a level of 100µg/mL when treated the same cell line type.
