Nucleic acid-based therapies have emerged as promising strategies for the regulation of gene expression and the production of therapeutic antigens or proteins for a series of diseases, including cancers, rare diseases...Nucleic acid-based therapies have emerged as promising strategies for the regulation of gene expression and the production of therapeutic antigens or proteins for a series of diseases, including cancers, rare diseases, and infectious diseases. However, their clinical application faces challenges. These include high molecular weight, limited cellular uptake,and susceptibility to enzymatic degradation by nucleases in vivo. Both viral and non-viral delivery vectors have been developed as a means of addressing these limitations, including lipid nanoparticles(LNPs), exosomes, polymers, and inorganic nanoparticles. Among these,LNPs have garnered significant attention due to their superior biocompatibility, high delivery efficiency and customizable design potential, as demonstrated by the clinical success of the FDA-approved si RNA drug Onpattro®. The critical role of nucleic acid drug carriers is discussed in this review. It also outlines the major types of carriers under development and examines the advancements and applications in LNP-based systems for nucleic acid delivery. By conducting a review of recent advancements in LNP design, delivery mechanisms, and clinical applications, this article aims to clarify the ways in which LNPs overcome delivery barriers, compare LNPs with other carriers, and identify key trends that can inform the development of next-generation LNP platforms for nucleic acid therapeutics.展开更多
The application of DNA hybridization technology,grounded in Watson-Crick base pairing,has facilitated the rational design of framework nucleic acids(FNAs)featuring adaptable shapes and dimensions.These nanostructures ...The application of DNA hybridization technology,grounded in Watson-Crick base pairing,has facilitated the rational design of framework nucleic acids(FNAs)featuring adaptable shapes and dimensions.These nanostructures exhibit remarkable stability and reproducibility,making them promising candidates for biomedical applications.Among various FNAs,tetrahedral FNAs(tFNAs),first introduced by Turberfield,are nanoscale assemblies of oligonucleotides that possess unique physical,chemical,and biological properties.Previous studies have demonstrated that tFNAs exhibit excellent cellular uptake,enhanced tissue permeability,and strong capabilities to promote cell migration,proliferation,and differentiation.Moreover,the intrinsic ability of tFNAs to efficiently penetrate cell membranes allows tFNAs to serve as versatile carriers for small-molecule drugs or functional oligonucleotides,thereby exerting significant anti-inflammatory,antioxidant,antibacterial,and immunomodulatory effects.These features highlight the therapeutic potential of tFNA-based complexes in skin,mucosal,and barrier tissue repair and regeneration.This review provides a comprehensive analysis of recent advances in the application of tFNAs for the prevention and treatment of skin,mucosal,and barrier tissue diseases,with a focus on their mechanisms of action and future prospects in regenerative medicine and targeted therapies.展开更多
Precision medicine has become a cornerstone in modern therapeutic strategies, with nucleic acid aptamers emerging aspivotal tools due to their unique properties. These oligonucleotide fragments, selected through the S...Precision medicine has become a cornerstone in modern therapeutic strategies, with nucleic acid aptamers emerging aspivotal tools due to their unique properties. These oligonucleotide fragments, selected through the Systematic Evolution ofLigands by Exponential Enrichment process, exhibit high affinity and specificity toward their targets, such as DNA, RNA,proteins, and other biomolecules. Nucleic acid aptamers offer significant advantages over traditional therapeutic agents,including superior biological stability, minimal immunogenicity, and the capacity for universal chemical modifications thatenhance their in vivo performance and targeting precision. In the realm of osseous tissue repair and regeneration, a complexphysiological process essential for maintaining skeletal integrity, aptamers have shown remarkable potential in influencingmolecular pathways crucial for bone regeneration, promoting osteogenic differentiation and supporting osteoblast survival. Byengineering aptamers to regulate inflammatory responses and facilitate the proliferation and differentiation of fibroblasts,these oligonucleotides can be integrated into advanced drug delivery systems, significantly improving bone repair efficacywhile minimizing adverse effects. Aptamer-mediated strategies, including the use of siRNA and miRNA mimics or inhibitors,have shown efficacy in enhancing bone mass and microstructure. These approaches hold transformative potential for treatinga range of orthopedic conditions like osteoporosis, osteosarcoma, and osteoarthritis. This review synthesizes the molecularmechanisms and biological roles of aptamers in orthopedic diseases, emphasizing their potential to drive innovative andeffective therapeutic interventions.展开更多
Compared to the single-stranded and double-stranded types of classical nucleic acid structures,atypical nucleic acid structures(such as G4s,i-motif,Triplex,and cyclic nucleic acids)are gradually becoming hotspots in b...Compared to the single-stranded and double-stranded types of classical nucleic acid structures,atypical nucleic acid structures(such as G4s,i-motif,Triplex,and cyclic nucleic acids)are gradually becoming hotspots in biomedical research due to their important biological functions and the close correlation between their abnormal dynamics equilibrium in physiological environments and a variety of hard-tackle diseases.The traditional gel electrophoresis,nuclear magnetic resonance,and circular dichroism detection techniques have shortcomings such as low spatial resolution,high destructiveness,and lack of real-time dynamic monitoring capability.In recent years,fluorescence imaging has gradually become a cutting-edge tool for non-classical nucleic acid structure detection due to their high sensitivity,fast response and dynamic real-time observation performance.In this contribution,we review the fluorescence materials for lighting-up imaging of non-classical nucleic acid structures,including traditional fluorescent small molecules and aggregation-induced emission luminogens(AIEgens).The design principles,detection mechanisms and application scenarios are detailed.Current fluorescence probes have already improved qualities in recognition targetability and signal-to-noise ratio by tuning and optimizing molecular structure-property relationships,but still face challenges such as insufficient selectivity and poor penetration capability in vivo.In the future,it is necessary to integrate multimodal imaging,artificial intelligence-assisted design and targeted delivery system to build a highly sensitive and multi-channel responsive platform to thoroughly disclose the association between the dynamic conformation of nucleic acid and disease,and to promote the development of precise and novel therapeutic strategies.展开更多
Metabolism is a general term for a series of ordered chemical reactions in an organism used to maintain life,mainly divided into anabolic and catabolic metabolism.Nucleic acid therapy can not only precisely up-regulat...Metabolism is a general term for a series of ordered chemical reactions in an organism used to maintain life,mainly divided into anabolic and catabolic metabolism.Nucleic acid therapy can not only precisely up-regulate and down-regulate the expression of target genes but also correct mutated disease-causing genes,which demonstrates irreplaceable and outstanding advantages in the treatment of metabolismrelated diseases and has been applied to the clinical treatment of metabolism-related diseases.In this review,we introduce the structures of several major nucleic acid drugs and the mechanism of nucleic acid therapy.Subsequently,we describe the mechanisms of various biomolecular and tissue metabolisms and the etiology of metabolic disorders,classified according to metabolic substrates.We analyze the signal pathways and potential targets affecting the metabolism of each substrate and describe the nucleic acid drugs applied to these targets and their delivery technologies.This review aims to provide new ideas and targets for treating these diseases by investigating the role played by metabolism in developing diseases and providing guidance for the selection and design of nucleic acid drugs.展开更多
Ultrasensitive detection of nucleic acids is of great significance for precision medicine.Digital polymerase chain reaction(dPCR)is the most sensitive method but requires sophisticated and expensive instruments and a ...Ultrasensitive detection of nucleic acids is of great significance for precision medicine.Digital polymerase chain reaction(dPCR)is the most sensitive method but requires sophisticated and expensive instruments and a long reaction time.Digital PCR-free technologies,which mean the digital assay not relying on thermal cycling to amplify the signal for quantitative detection of nucleic acids at the singlemolecule level,include the digital isothermal amplification techniques(d IATs)and the digital clustered regularly interspaced short palindromic repeats(CRISPR)technologies.They combine the advantages of d PCR and IATs,which could be fast and simple,enabling absolute quantification of nucleic acids at a single-molecule level with minimum instrument,representing the next-generation molecular diagnostic technology.Herein,we systematically summarized the strategies and applications of various dIATs,including the digital loop-mediated isothermal amplification(dLAMP),the digital recombinase polymerase amplification(dRPA),the digital rolling circle amplification(dRCA),the digital nucleic acid sequencebased amplification(d NASBA)and the digital multiple displacement amplification(d MDA),and evaluated the pros and cons of each method.The emerging digital CRISPR technologies,including the detection mechanism of CRISPR and the various strategies for signal amplification,are also introduced comprehensively in this review.The current challenges as well as the future perspectives of the digital PCR-free technology were discussed.展开更多
Neuropathic pain(NP)is one of the most common pathological pain types and is associated with limited treatment options;moreover,it affects patients’quality of life and causes a heavy social burden.Despite the emphasi...Neuropathic pain(NP)is one of the most common pathological pain types and is associated with limited treatment options;moreover,it affects patients’quality of life and causes a heavy social burden.Despite the emphasis on inhibiting neuronal apoptosis to relieve NP,the crucial role of a neuroinflammation is often overlooked.Therefore,refocusing on the regulation of microglia polarization to create a more conducive environment for neuron holds great potential in NP treatment.In recent years,small interfering RNAs(siRNAs)had become an attractive therapeutic option.However,an efficient loading and delivery system for siRNA is still in lack.In our study,a nanostructured tetrahedral framework nucleic acid loaded with the small interfering RNA C–C chemokine receptor 2(T-siCCR2)was successfully designed and synthesized for use in NP rat model in vivo and in a lipopolysaccharide(LPS)-induced inflammatory environment in vitro.This nanoscale complex is endowed with structural stability and satisfactory delivery efficiency while assuring the silencing effect of siRNA-CCR2.In vivo,T-siCCR2 treatment exhibited favorable effects on pain relief and functional improvement in the NP animal model by directly targeting microglia.In vitro,T-siCCR2 counteracts LPS-induced inflammation by inhibiting the differentiation of microglia toward the M1 phenotype,thus playing a neuroprotective role.RNA sequencing was subsequently performed to elucidate the underlying mechanism involved.These results indicate that T-siCCR2 may serve as a potential treatment option for NP in the future.展开更多
Diabetic kidney disease(DKD)is recognized as a severe complication in the development of diabetes mellitus(DM),posing a significant burden for global health.Major characteristics of DKD kidneys include tubulointerstit...Diabetic kidney disease(DKD)is recognized as a severe complication in the development of diabetes mellitus(DM),posing a significant burden for global health.Major characteristics of DKD kidneys include tubulointerstitial oxidative stress,inflammation,excessive extracellular matrix deposition,and progressing renal fibrosis.However,current treatment options are limited and cannot offer enough efficacy,thus urgently requiring novel therapeutic approaches.Tetrahedral framework nucleic acids(tFNAs)are a novel type of self-assembled DNA nanomaterial with excellent structural stability,biocompatibility,tailorable functionality,and regulatory effects on cellular behaviors.In this study,we established an in vitro high glucose(HG)-induced human renal tubular epithelial cells(HK-2 cells)pro-fibrogenic model and explored the antioxidative,anti-inflammatory,and antifibrotic capacity of tFNAs and the potential molecular mechanisms.tFNAs not only effectively alleviated oxidative stress through reactive oxygen species(ROS)-scavenging and activating the serine and threonine kinase(Akt)/nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway but also inhibited the production of proinflammatory factors such as tumor necrosis factor(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)in diabetic HK-2 cells.Additionally,tFNAs significantly downregulated the expression of Collagen I andα-smooth muscle actin(α-SMA),two representative biomarkers of pro-fibrogenic myofibroblasts in the renal tubular epithelial-mesenchymal transition(EMT).Furthermore,we found that tFNAs exerted this function by inhibiting the Wnt/β-catenin signaling pathway,preventing the occurrence of EMT and fibrosis.The findings of this study demonstrated that tFNAs are naturally endowed with great potential to prevent fibrosis progress in DKD kidneys and can be further combined with emerging pharmacotherapies,providing a secure and efficient drug delivery strategy for future DKD therapy.展开更多
The potential of messenger RNA(m RNA)as a therapeutic tool for treating diseases has garnered considerable interest,especially in the wake of the successful creation of m RNA vaccines to counter corona virus disease 2...The potential of messenger RNA(m RNA)as a therapeutic tool for treating diseases has garnered considerable interest,especially in the wake of the successful creation of m RNA vaccines to counter corona virus disease 2019(COVID-19).Nucleic acid-based drug gene therapies have emerged as exceptionally promising avenues for combating disease.Furthermore,lipid nanoparticles(LNPs)are ideal carriers for nucleic acid delivery owing to their ionic nature,which enables nucleic acids to electrostatically interact with intracellular membranes,thereby promoting efficient intracellular nucleic acid release.Unfortunately,the effectiveness of LNPs in targeting organs beyond the liver is relatively poor.Thus,enhanced extrahepatic targeting is another important property that would lead to improved in vivo delivery by LNPs.This review focuses on the fundamental characteristics and functions of LNPs developed to facilitate cellular uptake and ensure effective intracellular release of m RNAs.Promising applications,possible advantages and potential challenges associated with use of LNPs in organ specific delivery and release of m RNAs are summarized.Furthermore,the need for future research to address limitations of currently developed LNPs for clinical applications of the m RNA technology is emphasized.展开更多
Hyperglycemia resulting from diabetes mellitus(DM)exacerbates osteoporosis and fractures,damaging bone regeneration due to impaired healing capacity.Stem cell therapy offers the potential for bone repair,accelerating ...Hyperglycemia resulting from diabetes mellitus(DM)exacerbates osteoporosis and fractures,damaging bone regeneration due to impaired healing capacity.Stem cell therapy offers the potential for bone repair,accelerating the healing of bone defects by introducing stem cells with osteogenic differentiation ability.Dental follicle stem cells(DFSCs)are a newly emerging type of dental stem cells that not only have the potential for multipotent differentiation but also hold easy accessibility and can stand longterm storage.However,DM-associated oxidative stress and inflammation elevate the risk of DFSCs dysfunction and apoptosis,diminishing stem cell therapy efficacy.Recent nanomaterial advances,particularly in DNA nanostructures like tetrahedral framework nucleic acids(tFNAs),have been promising candidates for modulating cellular behaviors.Accumulating experiments have shown that tFNAs’cell proliferation and migration-promoting ability and induce osteogenic differentiation of stem cells.Meanwhile,tFNAs can scavenge reactive oxygen species(ROS)and downregulate the secretion of inflammatory factors by inhibiting various inflammation-related signaling pathways.Here,we applied tFNAs to modify DFSCs and observed enhanced osteogenic differentiation alongside ROS scavenging and anti-inflammatory effects mediated by suppressing the ROS/mitogen-activated protein kinases(MAPKs)/nuclear factor kappa-B(NF-κB)signaling pathway.This intervention reduced stem cell apoptosis,bolstering stem cell therapy efficacy in DM.Our study establishes a simple yet potent tFNAs-DFSCs system,offering potential as a bone repair agent for future DM treatment.展开更多
We report the development of a triplex nucleic acid lateral flow immunoassay(NALFIA)for the detection of the genomes of Nipah virus(NiV),Middle East respiratory syndrome coronavirus(MERS-CoV)and Reston ebolavirus(REBO...We report the development of a triplex nucleic acid lateral flow immunoassay(NALFIA)for the detection of the genomes of Nipah virus(NiV),Middle East respiratory syndrome coronavirus(MERS-CoV)and Reston ebolavirus(REBOV),which are intended for screening bats as well as other hosts and reservoirs of these three viruses.Our triplex NALFIA is a two-step assay format:the target nucleic acid in the sample is first amplified using tagged primers,and the tagged dsDNA amplicons are captured by antibodies immobilized on the NALFIA device,resulting in signal development from the binding of a streptavidin-colloidal gold conjugate to a biotin tag on the captured amplicons.Triplex amplification of the N gene of NiV,the UpE gene of MERS-CoV,and the Vp40 gene of REBOV was optimized,and three compatible combinations of hapten labels and antibodies were identified for end point detection.The lowest RNA copy numbers detected by the triplex NALFIA were 8.21e4 for the NiV N target,7.09e1 for the MERS-CoV UpE target,and 1.83e4 for the REBOV Vp40 target.Using simulated samples,the sensitivity and specificity for MERS-CoV and REBOV targets were estimated to be 100%,while the sensitivity and specificity for the NiV target were 91%and 93.3%,respectively.The compliance rate between triplex NALFIA and real-time RT‒PCR was 92%for the NiV N target and 100%for the MERS-CoV UpE and REBOV Vp40 targets.展开更多
[Objective] The aim of this study was to provide basis for deeply understanding the diapause mechanism of Papilio memnon L. [Method] RNA and DNA content of non-diapause pupae, diapause pupae and eclosion-adult from di...[Objective] The aim of this study was to provide basis for deeply understanding the diapause mechanism of Papilio memnon L. [Method] RNA and DNA content of non-diapause pupae, diapause pupae and eclosion-adult from diapause pupae at different development stages were detected by the colorimetry. [Result] RNA content of non-diapause pupae was 4.614 0-7.946 3 μg/mg, while diapause pupae was 4.326 0-5.885 3 μg/mg and eclosion-adult from diapause pupae was 20.779 3 μg/mg at initial stage. DNA content of non-diapause pupae was 0.448 7-0.535 0 μg/mg, while diapause pupae was 0.452 0-0.828 3 μg/mg and eclosion-adult from diapause pupae was 1.727 0 μg/mg at initial stage. [Conclusion] The nucleic acid content and change is related to the development stage.展开更多
Real-time PCR is a closed DNA amplification system that skillfully integrates biochemical, photoelectric and computer techniques. Fluorescence data acquired once per cycle provides rapid absolute quantification of ini...Real-time PCR is a closed DNA amplification system that skillfully integrates biochemical, photoelectric and computer techniques. Fluorescence data acquired once per cycle provides rapid absolute quantification of initial template copy numbers as PCR products are generated. This technique significantly simplifies and accelerates the process of producing reproducible quantification of nucleic acid molecules. It not only is a sensitive, accurate and rapid quantitative method, but it also provides an easier way to calculate the absolute starting copy number of nucleic acid molecules to be tested. Together with molecular bio-techniques, like microarray, real-time PCR will play a very important role in many aspects of molecular life science such as functional gene analysis and disease molecular diagnostics. This review introduces the detailed principles and application of the real-time PCR technique, describes a recently developed system for exact quantification of AUX/IAA genes In Arabidopsis, and discusses the problems with the real-time PCR process.展开更多
基金supported by the Regional University-Industry Technology Transfer Center for Biopharmaceuticals (Nanjing,Jiangsu) Early-Stage Translational Grant (JB2025211)。
文摘Nucleic acid-based therapies have emerged as promising strategies for the regulation of gene expression and the production of therapeutic antigens or proteins for a series of diseases, including cancers, rare diseases, and infectious diseases. However, their clinical application faces challenges. These include high molecular weight, limited cellular uptake,and susceptibility to enzymatic degradation by nucleases in vivo. Both viral and non-viral delivery vectors have been developed as a means of addressing these limitations, including lipid nanoparticles(LNPs), exosomes, polymers, and inorganic nanoparticles. Among these,LNPs have garnered significant attention due to their superior biocompatibility, high delivery efficiency and customizable design potential, as demonstrated by the clinical success of the FDA-approved si RNA drug Onpattro®. The critical role of nucleic acid drug carriers is discussed in this review. It also outlines the major types of carriers under development and examines the advancements and applications in LNP-based systems for nucleic acid delivery. By conducting a review of recent advancements in LNP design, delivery mechanisms, and clinical applications, this article aims to clarify the ways in which LNPs overcome delivery barriers, compare LNPs with other carriers, and identify key trends that can inform the development of next-generation LNP platforms for nucleic acid therapeutics.
基金supported by the National Natural Science Foundation of China(No.81960199)Clinical Translational Innovation Cultivating Fund 550 Project of Hainan General Hospital,Joint Program on Health Science&Technology Innovation of Hainan Province(No.WSJK2024MS127)Academic Enhancement Support Program of Hainan Medical University(No.XSTS2025093).
文摘The application of DNA hybridization technology,grounded in Watson-Crick base pairing,has facilitated the rational design of framework nucleic acids(FNAs)featuring adaptable shapes and dimensions.These nanostructures exhibit remarkable stability and reproducibility,making them promising candidates for biomedical applications.Among various FNAs,tetrahedral FNAs(tFNAs),first introduced by Turberfield,are nanoscale assemblies of oligonucleotides that possess unique physical,chemical,and biological properties.Previous studies have demonstrated that tFNAs exhibit excellent cellular uptake,enhanced tissue permeability,and strong capabilities to promote cell migration,proliferation,and differentiation.Moreover,the intrinsic ability of tFNAs to efficiently penetrate cell membranes allows tFNAs to serve as versatile carriers for small-molecule drugs or functional oligonucleotides,thereby exerting significant anti-inflammatory,antioxidant,antibacterial,and immunomodulatory effects.These features highlight the therapeutic potential of tFNA-based complexes in skin,mucosal,and barrier tissue repair and regeneration.This review provides a comprehensive analysis of recent advances in the application of tFNAs for the prevention and treatment of skin,mucosal,and barrier tissue diseases,with a focus on their mechanisms of action and future prospects in regenerative medicine and targeted therapies.
基金Key research and development projects of Sichuan Science and Technology Plan Project(2024YFFK0135)Fujian Provincial Natural Science Foundation of China(2024J011450).
文摘Precision medicine has become a cornerstone in modern therapeutic strategies, with nucleic acid aptamers emerging aspivotal tools due to their unique properties. These oligonucleotide fragments, selected through the Systematic Evolution ofLigands by Exponential Enrichment process, exhibit high affinity and specificity toward their targets, such as DNA, RNA,proteins, and other biomolecules. Nucleic acid aptamers offer significant advantages over traditional therapeutic agents,including superior biological stability, minimal immunogenicity, and the capacity for universal chemical modifications thatenhance their in vivo performance and targeting precision. In the realm of osseous tissue repair and regeneration, a complexphysiological process essential for maintaining skeletal integrity, aptamers have shown remarkable potential in influencingmolecular pathways crucial for bone regeneration, promoting osteogenic differentiation and supporting osteoblast survival. Byengineering aptamers to regulate inflammatory responses and facilitate the proliferation and differentiation of fibroblasts,these oligonucleotides can be integrated into advanced drug delivery systems, significantly improving bone repair efficacywhile minimizing adverse effects. Aptamer-mediated strategies, including the use of siRNA and miRNA mimics or inhibitors,have shown efficacy in enhancing bone mass and microstructure. These approaches hold transformative potential for treatinga range of orthopedic conditions like osteoporosis, osteosarcoma, and osteoarthritis. This review synthesizes the molecularmechanisms and biological roles of aptamers in orthopedic diseases, emphasizing their potential to drive innovative andeffective therapeutic interventions.
文摘Compared to the single-stranded and double-stranded types of classical nucleic acid structures,atypical nucleic acid structures(such as G4s,i-motif,Triplex,and cyclic nucleic acids)are gradually becoming hotspots in biomedical research due to their important biological functions and the close correlation between their abnormal dynamics equilibrium in physiological environments and a variety of hard-tackle diseases.The traditional gel electrophoresis,nuclear magnetic resonance,and circular dichroism detection techniques have shortcomings such as low spatial resolution,high destructiveness,and lack of real-time dynamic monitoring capability.In recent years,fluorescence imaging has gradually become a cutting-edge tool for non-classical nucleic acid structure detection due to their high sensitivity,fast response and dynamic real-time observation performance.In this contribution,we review the fluorescence materials for lighting-up imaging of non-classical nucleic acid structures,including traditional fluorescent small molecules and aggregation-induced emission luminogens(AIEgens).The design principles,detection mechanisms and application scenarios are detailed.Current fluorescence probes have already improved qualities in recognition targetability and signal-to-noise ratio by tuning and optimizing molecular structure-property relationships,but still face challenges such as insufficient selectivity and poor penetration capability in vivo.In the future,it is necessary to integrate multimodal imaging,artificial intelligence-assisted design and targeted delivery system to build a highly sensitive and multi-channel responsive platform to thoroughly disclose the association between the dynamic conformation of nucleic acid and disease,and to promote the development of precise and novel therapeutic strategies.
基金financially supported by the National Natural Science Foundation of China(Nos.32225029,22205240,52073287,22075289,82071552 and 22376006)National Key R&D Program of China(No.2023YFC2605003)。
文摘Metabolism is a general term for a series of ordered chemical reactions in an organism used to maintain life,mainly divided into anabolic and catabolic metabolism.Nucleic acid therapy can not only precisely up-regulate and down-regulate the expression of target genes but also correct mutated disease-causing genes,which demonstrates irreplaceable and outstanding advantages in the treatment of metabolismrelated diseases and has been applied to the clinical treatment of metabolism-related diseases.In this review,we introduce the structures of several major nucleic acid drugs and the mechanism of nucleic acid therapy.Subsequently,we describe the mechanisms of various biomolecular and tissue metabolisms and the etiology of metabolic disorders,classified according to metabolic substrates.We analyze the signal pathways and potential targets affecting the metabolism of each substrate and describe the nucleic acid drugs applied to these targets and their delivery technologies.This review aims to provide new ideas and targets for treating these diseases by investigating the role played by metabolism in developing diseases and providing guidance for the selection and design of nucleic acid drugs.
基金supported by the National Key Research and Development Program of China(Nos.2023YFC2307305,2021YFF0703300)the Shenzhen Medical Research Fund(No.B2303003)+3 种基金Shenzhen Research Funding Program(Nos.JCYJ20220818102014028,RCBS20210609104339043)National Natural Science Foundation of China(No.22174167)Guangdong Basic and Applied Basic Research(No.2024A1515011281)Fundamental Research Funds for the Central Universities(No.24qnpy087)from Sun Yat-sen University。
文摘Ultrasensitive detection of nucleic acids is of great significance for precision medicine.Digital polymerase chain reaction(dPCR)is the most sensitive method but requires sophisticated and expensive instruments and a long reaction time.Digital PCR-free technologies,which mean the digital assay not relying on thermal cycling to amplify the signal for quantitative detection of nucleic acids at the singlemolecule level,include the digital isothermal amplification techniques(d IATs)and the digital clustered regularly interspaced short palindromic repeats(CRISPR)technologies.They combine the advantages of d PCR and IATs,which could be fast and simple,enabling absolute quantification of nucleic acids at a single-molecule level with minimum instrument,representing the next-generation molecular diagnostic technology.Herein,we systematically summarized the strategies and applications of various dIATs,including the digital loop-mediated isothermal amplification(dLAMP),the digital recombinase polymerase amplification(dRPA),the digital rolling circle amplification(dRCA),the digital nucleic acid sequencebased amplification(d NASBA)and the digital multiple displacement amplification(d MDA),and evaluated the pros and cons of each method.The emerging digital CRISPR technologies,including the detection mechanism of CRISPR and the various strategies for signal amplification,are also introduced comprehensively in this review.The current challenges as well as the future perspectives of the digital PCR-free technology were discussed.
基金supported by National Natural Science Foundation of China(Nos.81874027,82370929,81970916)Sichuan Science and Technology Program(Nos.2019YFQ0003,2022YFS0051,2022NSFSC0002)+3 种基金Sichuan Province Youth Science and Technology Innovation Team(No.2022JDTD0021)Research and Develop Program,West China Hospital of Stomatology Sichuan University(Nos.RD03202302,RCDWJS2024–1)135-project for disciplines of excellenceClinical Research Incubation project of West China Hospital of Sichuan University(No.2021HXFH036)。
文摘Neuropathic pain(NP)is one of the most common pathological pain types and is associated with limited treatment options;moreover,it affects patients’quality of life and causes a heavy social burden.Despite the emphasis on inhibiting neuronal apoptosis to relieve NP,the crucial role of a neuroinflammation is often overlooked.Therefore,refocusing on the regulation of microglia polarization to create a more conducive environment for neuron holds great potential in NP treatment.In recent years,small interfering RNAs(siRNAs)had become an attractive therapeutic option.However,an efficient loading and delivery system for siRNA is still in lack.In our study,a nanostructured tetrahedral framework nucleic acid loaded with the small interfering RNA C–C chemokine receptor 2(T-siCCR2)was successfully designed and synthesized for use in NP rat model in vivo and in a lipopolysaccharide(LPS)-induced inflammatory environment in vitro.This nanoscale complex is endowed with structural stability and satisfactory delivery efficiency while assuring the silencing effect of siRNA-CCR2.In vivo,T-siCCR2 treatment exhibited favorable effects on pain relief and functional improvement in the NP animal model by directly targeting microglia.In vitro,T-siCCR2 counteracts LPS-induced inflammation by inhibiting the differentiation of microglia toward the M1 phenotype,thus playing a neuroprotective role.RNA sequencing was subsequently performed to elucidate the underlying mechanism involved.These results indicate that T-siCCR2 may serve as a potential treatment option for NP in the future.
基金supported by the National Natural Science Foundation of China(No.82101077)Sichuan Science and Technology Program(No.2023NSFSC1516)+2 种基金Postdoctoral Science Foundation of China(Nos.2021M692271,2023T160455)West China School/Hospital of Stomatology Sichuan University,No.RCDWJS2023-5,Fundamental Research Funds for the Central UniversitiesResearch and Develop Program,West China Hospital of Stomatology Sichuan University.
文摘Diabetic kidney disease(DKD)is recognized as a severe complication in the development of diabetes mellitus(DM),posing a significant burden for global health.Major characteristics of DKD kidneys include tubulointerstitial oxidative stress,inflammation,excessive extracellular matrix deposition,and progressing renal fibrosis.However,current treatment options are limited and cannot offer enough efficacy,thus urgently requiring novel therapeutic approaches.Tetrahedral framework nucleic acids(tFNAs)are a novel type of self-assembled DNA nanomaterial with excellent structural stability,biocompatibility,tailorable functionality,and regulatory effects on cellular behaviors.In this study,we established an in vitro high glucose(HG)-induced human renal tubular epithelial cells(HK-2 cells)pro-fibrogenic model and explored the antioxidative,anti-inflammatory,and antifibrotic capacity of tFNAs and the potential molecular mechanisms.tFNAs not only effectively alleviated oxidative stress through reactive oxygen species(ROS)-scavenging and activating the serine and threonine kinase(Akt)/nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway but also inhibited the production of proinflammatory factors such as tumor necrosis factor(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)in diabetic HK-2 cells.Additionally,tFNAs significantly downregulated the expression of Collagen I andα-smooth muscle actin(α-SMA),two representative biomarkers of pro-fibrogenic myofibroblasts in the renal tubular epithelial-mesenchymal transition(EMT).Furthermore,we found that tFNAs exerted this function by inhibiting the Wnt/β-catenin signaling pathway,preventing the occurrence of EMT and fibrosis.The findings of this study demonstrated that tFNAs are naturally endowed with great potential to prevent fibrosis progress in DKD kidneys and can be further combined with emerging pharmacotherapies,providing a secure and efficient drug delivery strategy for future DKD therapy.
基金supported by Guang Dong Basic and Applied Basic Research Foundation(No.2023B1515120001)Shenzhen University 2035 Program for Excellent Research(Nos.00000208 and 00000225)。
文摘The potential of messenger RNA(m RNA)as a therapeutic tool for treating diseases has garnered considerable interest,especially in the wake of the successful creation of m RNA vaccines to counter corona virus disease 2019(COVID-19).Nucleic acid-based drug gene therapies have emerged as exceptionally promising avenues for combating disease.Furthermore,lipid nanoparticles(LNPs)are ideal carriers for nucleic acid delivery owing to their ionic nature,which enables nucleic acids to electrostatically interact with intracellular membranes,thereby promoting efficient intracellular nucleic acid release.Unfortunately,the effectiveness of LNPs in targeting organs beyond the liver is relatively poor.Thus,enhanced extrahepatic targeting is another important property that would lead to improved in vivo delivery by LNPs.This review focuses on the fundamental characteristics and functions of LNPs developed to facilitate cellular uptake and ensure effective intracellular release of m RNAs.Promising applications,possible advantages and potential challenges associated with use of LNPs in organ specific delivery and release of m RNAs are summarized.Furthermore,the need for future research to address limitations of currently developed LNPs for clinical applications of the m RNA technology is emphasized.
基金supported by the National Natural Science Foundation of China (Nos. 82101077, 82370929)Sichuan Science and Technology Program (Nos. 2023NSFSC1516, 2023NSFSC1706)+3 种基金Postdoctoral Science Foundation of China (Nos. 2021M692271, 2023T160455, BX20220220, 2022M722251)West China School/Hospital of Stomatology Sichuan University (No. RCDWJS2023–5)Fundamental Research Funds for the Central UniversitiesResearch and Develop Program, West China Hospital of Stomatology Sichuan University
文摘Hyperglycemia resulting from diabetes mellitus(DM)exacerbates osteoporosis and fractures,damaging bone regeneration due to impaired healing capacity.Stem cell therapy offers the potential for bone repair,accelerating the healing of bone defects by introducing stem cells with osteogenic differentiation ability.Dental follicle stem cells(DFSCs)are a newly emerging type of dental stem cells that not only have the potential for multipotent differentiation but also hold easy accessibility and can stand longterm storage.However,DM-associated oxidative stress and inflammation elevate the risk of DFSCs dysfunction and apoptosis,diminishing stem cell therapy efficacy.Recent nanomaterial advances,particularly in DNA nanostructures like tetrahedral framework nucleic acids(tFNAs),have been promising candidates for modulating cellular behaviors.Accumulating experiments have shown that tFNAs’cell proliferation and migration-promoting ability and induce osteogenic differentiation of stem cells.Meanwhile,tFNAs can scavenge reactive oxygen species(ROS)and downregulate the secretion of inflammatory factors by inhibiting various inflammation-related signaling pathways.Here,we applied tFNAs to modify DFSCs and observed enhanced osteogenic differentiation alongside ROS scavenging and anti-inflammatory effects mediated by suppressing the ROS/mitogen-activated protein kinases(MAPKs)/nuclear factor kappa-B(NF-κB)signaling pathway.This intervention reduced stem cell apoptosis,bolstering stem cell therapy efficacy in DM.Our study establishes a simple yet potent tFNAs-DFSCs system,offering potential as a bone repair agent for future DM treatment.
基金funded by the Department of Biotechnology,Ministry of Science and Technology,Government of India(DBT)under grant number ADMaC DBT-NER/LIVS/11/2012.
文摘We report the development of a triplex nucleic acid lateral flow immunoassay(NALFIA)for the detection of the genomes of Nipah virus(NiV),Middle East respiratory syndrome coronavirus(MERS-CoV)and Reston ebolavirus(REBOV),which are intended for screening bats as well as other hosts and reservoirs of these three viruses.Our triplex NALFIA is a two-step assay format:the target nucleic acid in the sample is first amplified using tagged primers,and the tagged dsDNA amplicons are captured by antibodies immobilized on the NALFIA device,resulting in signal development from the binding of a streptavidin-colloidal gold conjugate to a biotin tag on the captured amplicons.Triplex amplification of the N gene of NiV,the UpE gene of MERS-CoV,and the Vp40 gene of REBOV was optimized,and three compatible combinations of hapten labels and antibodies were identified for end point detection.The lowest RNA copy numbers detected by the triplex NALFIA were 8.21e4 for the NiV N target,7.09e1 for the MERS-CoV UpE target,and 1.83e4 for the REBOV Vp40 target.Using simulated samples,the sensitivity and specificity for MERS-CoV and REBOV targets were estimated to be 100%,while the sensitivity and specificity for the NiV target were 91%and 93.3%,respectively.The compliance rate between triplex NALFIA and real-time RT‒PCR was 92%for the NiV N target and 100%for the MERS-CoV UpE and REBOV Vp40 targets.
基金Supported by the International Advanced Forestry Science and Technology Project Imported by State Forestry Administration (2005-4-59 and 2008-4-68)~~
文摘[Objective] The aim of this study was to provide basis for deeply understanding the diapause mechanism of Papilio memnon L. [Method] RNA and DNA content of non-diapause pupae, diapause pupae and eclosion-adult from diapause pupae at different development stages were detected by the colorimetry. [Result] RNA content of non-diapause pupae was 4.614 0-7.946 3 μg/mg, while diapause pupae was 4.326 0-5.885 3 μg/mg and eclosion-adult from diapause pupae was 20.779 3 μg/mg at initial stage. DNA content of non-diapause pupae was 0.448 7-0.535 0 μg/mg, while diapause pupae was 0.452 0-0.828 3 μg/mg and eclosion-adult from diapause pupae was 1.727 0 μg/mg at initial stage. [Conclusion] The nucleic acid content and change is related to the development stage.
文摘Real-time PCR is a closed DNA amplification system that skillfully integrates biochemical, photoelectric and computer techniques. Fluorescence data acquired once per cycle provides rapid absolute quantification of initial template copy numbers as PCR products are generated. This technique significantly simplifies and accelerates the process of producing reproducible quantification of nucleic acid molecules. It not only is a sensitive, accurate and rapid quantitative method, but it also provides an easier way to calculate the absolute starting copy number of nucleic acid molecules to be tested. Together with molecular bio-techniques, like microarray, real-time PCR will play a very important role in many aspects of molecular life science such as functional gene analysis and disease molecular diagnostics. This review introduces the detailed principles and application of the real-time PCR technique, describes a recently developed system for exact quantification of AUX/IAA genes In Arabidopsis, and discusses the problems with the real-time PCR process.
