Kinesins are a superfamily of proteins widely present in eukaryotes,playing crucial roles in plant cell wall assembly,cell elongation regulation,gravity sensing,and fertility control.In this study,bioinformatics analy...Kinesins are a superfamily of proteins widely present in eukaryotes,playing crucial roles in plant cell wall assembly,cell elongation regulation,gravity sensing,and fertility control.In this study,bioinformatics analysis of the OsKMP2 gene(LOC_Os02g28850)was performed using online tools such as ExPASy-ProtParam,ProtScale,CD-search,and DNAMAN software.Additionally,qRT-PCR was employed to analyze the tissue expression pattern of OsKMP2.The results showed that the molecular weight of the OsKMP2 is 118.39728 kDa,and it is a hydrophilic and unstable acidic protein.Secondary structure prediction revealed that it primarily consists ofα-helices(69.45%),random coils(25.19%),and extended strands(5.36%).The gene was expressed in various rice tissues,with the highest expression level observed in leaves.These results indicate that the OsKMP2 gene exhibits high evolutionary conservation and functional diversity in rice.展开更多
Mytilus contain abundant antimicrobial peptides(AMPs)that play a key role in the innate immunity.However,heterologous production of these AMPs remains challenging due to their short sequences,multiple disulfide bonds,...Mytilus contain abundant antimicrobial peptides(AMPs)that play a key role in the innate immunity.However,heterologous production of these AMPs remains challenging due to their short sequences,multiple disulfide bonds,and high content of cationic amino acids,which hinder functional expression in prokaryotic systems such as Escherichia coli.To establish a eukaryotic recombinant expression system for the AMPs of mussel and obtain recombinant mussel AMPs for subsequent studies,we reported the successful recombinant expression of myticofensin B1,a novel defensin-like AMP identified previously in Mytiluscoruscus,using the eukaryotic host Pichia pastoris.The codon-optimized gene encoding the mature myticofensin-B1(composed of 65 amino acid residues,including 6 conserved cysteine residues)was cloned into a pPICZαA vector and expressed in P.pastoris GS115.Structural fidelity of the recombinant peptide was confirmed by liquid chromatography-tandem mass spectrometry(LC-MS/MS),showing a molecular weight of 8849.9 Da,which was consistent with the theoretical prediction.Functional assays demonstrated a broad-spectrum antimicrobial activity of the recombinant myticofensin-B1,with stronger inhibition against Gram-negative bacteria.Scanning electron microscopy revealed different effects of the recombinant myticofensin-B1 against different bacteria.In addition,the recombinant myticofensin-B1 exhibited a very low hemolytic activity against sheep red blood cells and weak cytotoxicity against human A549 lung cancer cells.This study establishes P.pastoris as a powerful platform to produce functional mussel AMP and highlights the potential of the recombinant myticofensin-B1 as a therapeutic agent for aquaculture pathogens and infections.展开更多
Regulatory T cells,a subset of CD4^(+)T cells,play a critical role in maintaining immune tolerance and tissue homeostasis due to their potent immunosuppressive properties.Recent advances in research have highlighted t...Regulatory T cells,a subset of CD4^(+)T cells,play a critical role in maintaining immune tolerance and tissue homeostasis due to their potent immunosuppressive properties.Recent advances in research have highlighted the important therapeutic potential of Tregs in neurological diseases and tissue repair,emphasizing their multifaceted roles in immune regulation.This review aims to summarize and analyze the mechanisms of action and therapeutic potential of Tregs in relation to neurological diseases and neural regeneration.Beyond their classical immune-regulatory functions,emerging evidence points to non-immune mechanisms of regulatory T cells,particularly their interactions with stem cells and other non-immune cells.These interactions contribute to optimizing the repair microenvironment and promoting tissue repair and nerve regeneration,positioning non-immune pathways as a promising direction for future research.By modulating immune and non-immune cells,including neurons and glia within neural tissues,Tregs have demonstrated remarkable efficacy in enhancing regeneration in the central and peripheral nervous systems.Preclinical studies have revealed that Treg cells interact with neurons,glial cells,and other neural components to mitigate inflammatory damage and support functional recovery.Current mechanistic studies show that Tregs can significantly promote neural repair and functional recovery by regulating inflammatory responses and the local immune microenvironment.However,research on the mechanistic roles of regulatory T cells in other diseases remains limited,highlighting substantial gaps and opportunities for exploration in this field.Laboratory and clinical studies have further advanced the application of regulatory T cells.Technical advances have enabled efficient isolation,ex vivo expansion and functionalization,and adoptive transfer of regulatory T cells,with efficacy validated in animal models.Innovative strategies,including gene editing,cell-free technologies,biomaterial-based recruitment,and in situ delivery have expanded the therapeutic potential of regulatory T cells.Gene editing enables precise functional optimization,while biomaterial and in situ delivery technologies enhance their accumulation and efficacy at target sites.These advancements not only improve the immune-regulatory capacity of regulatory T cells but also significantly enhance their role in tissue repair.By leveraging the pivotal and diverse functions of Tregs in immune modulation and tissue repair,regulatory T cells–based therapies may lead to transformative breakthroughs in the treatment of neurological diseases.展开更多
Quantitative real-time PCR(qPCR)is widely used for gene expression analysis,but its accuracy critically depends on stable internal reference genes for normalization.In marine invertebrates,especially non-model taxa su...Quantitative real-time PCR(qPCR)is widely used for gene expression analysis,but its accuracy critically depends on stable internal reference genes for normalization.In marine invertebrates,especially non-model taxa such as cephalopods,systematic evaluation of reference genes is limited,leading to potential bias.The cuttlefish Sepiella japonica is ecologically and economically important in China,yet previous molecular studies have often relied on single unvalidated reference genes,which may compromise data reliability.This study aimed to systematically evaluate the stability of five commonly used reference genes(18S,ef-1α,ef-1γ,gapdh,andβ-actin)across multiple tissues and sexes of S.japonica,and to identify the most suitable reference genes and optimal number for qPCR normalization.Fifteen to sixteen tissue types were collected from ten healthy adults(five males and five females).Total RNA was extracted,reverse-transcribed,and analyzed by qPCR.Gene stability was assessed using four algorithms(geNorm,NormFinder,BestKeeper,andΔCt)integrated with RefFinder,and the optimal gene number was determined using geNorm pairwise variation(V_(n/n+1)<0.15).Four transcriptome-derived genes(creld2,cd109,acy1,and miox)were used for validation.The C_(t)values of the five genes ranged from 15.47 to 20.83.β-actin and gapdh showed pronounced variability in expression stability among tissues and sexes,indicating their limited suitability for normalization.18S exhibited the highest expression(mean C_(t):15.47-16.29)and lowest variability but displayed sex-biased expression,whereas ef-1αand ef-1γremained consistently stable across most tissues in both sexes,with ef-1αbeing the most robust and showing no sex-related bias.Although specific rankings varied among tissues and sexes,the comprehensive results indicated that ef-1αand ef-1γpossessed the highest overall stability,followed by 18S,whileβ-actin and gapdh were the least stable.The final comprehensive rankings were ef-1γ>ef-1α>18S>gapdh>β-actin(male)and ef-1α>ef-1γ>18S>gapdh>β-actin(female).geNorm analysis(V2/3<0.15)indicated that two genes,mainly ef-1αand ef-1γ,were generally sufficient for reliable normalization in most tissues.Validation confirmed that normalization using the stable ef-1αand ef-1γaccurately reflected the expression differences among tissues,whereasβ-actin and gapdh can bias or confound statistical analyses.ef-1αand ef-1γare identified as the most reliable reference gene combination for qPCR analysis in S.japonica,while 18S can serve as an auxiliary gene for within-sex comparisons.The use ofβ-actin or gapdh alone is not recommended.This study establishes a systematic framework for selecting reliable reference genes in S.japonica,thereby facilitating robust qPCR normalization and providing a foundation for future gene expression research in S.japonica and other cephalopods.展开更多
OBJECTIVE:To investigate the hypoglycemic mechanism of modified Gegen Qinlian decoction(加味葛根芩连汤,MGQD)by examining its regulation of cholesterol transporter expression and DNA methylation,specifically the low-de...OBJECTIVE:To investigate the hypoglycemic mechanism of modified Gegen Qinlian decoction(加味葛根芩连汤,MGQD)by examining its regulation of cholesterol transporter expression and DNA methylation,specifically the low-density lipoprotein receptor(LDLR)and scavenger receptor class B type 1(SR-B1),in epididymal white adipose tissue(e WAT)and inguinal white adipose tissue(i WAT)of rats with type 2 diabetes mellitus(T2DM).METHODS:The control group(CON)consisted of ten Sprague-Dawley(SD)rats fed a standard chow diet,while 80 SD rats were fed a high-fat diet and administered streptozotocin intraperitoneally to induce diabetes.The diabetic rats were randomly assigned to four groups:T2DM,metformin(MET,200 mg/kg),low-dose MGQD(MGQDL,5 g/kg),and high-dose MGQD(MGQDH,10 g/kg),and received treatment via gavage for 14 weeks.Western blot(WB),quantitative real-time polymerase chain reaction(q PCR),and bisulfite sequencing PCR(BSP)were used to analyze protein levels,m RNA expression,and DNA methylation of Ldlr(gene encoding LDLR)and Srb1(gene encoding SR-B1).RESULTS:MGQD and metformin treatment significantly reduced blood glucose levels,restored LDLR and SR-B1 protein levels in e WAT,and effectively regulated the m RNA expression and non-cytosine-p-guanine(nonCp G)methylation of Srb1 in e WAT.A significant negative correlation was observed between the methylation of Srb1 in e WAT and its m RNA expression.However,MGQD and metformin had no significant effect on the protein levels,m RNA expression,or DNA methylation of Ldlr and Srb1 in i WAT.CONCLUSIONS:MGQD did not significantly affect LDLR and SR-B1 expression or gene methylation in i WAT.However,its hypoglycemic effect may be linked to cholesterol regulation in e WAT.Potential mechanisms include increased LDLR protein levels,which may enhance cholesterol uptake,and increased Srb1 methylation,which may suppress its expression and consequently reduce cholesterol efflux.展开更多
Bamei pigs,an indigenous Chinese breed,yield meat with a delectable flavor and boast higher carcass fat content compared to commercial breeds,making them a rich food source for humans.However,the differences in lipid ...Bamei pigs,an indigenous Chinese breed,yield meat with a delectable flavor and boast higher carcass fat content compared to commercial breeds,making them a rich food source for humans.However,the differences in lipid and nutrient components between the adipose tissue of Bamei pigs and commercial pigs are still unclear.The study employed UPLC-MS/MS to quantify the composition of lipids and metabolites in the backfat of both Bamei and Large White pigs.A total of 428 lipids and 193 metabolites were significantly different between the 2 groups.Specifically,Bamei pig backfat exhibited altered levels of various lipids and metabolites that may potentially contribute to nutritional and flavor differences,including unsaturated triglycerides,free fatty acids,medium-chain triglycerides,essential amino acids,vitamins and antioxidants,while maintaining reduced cholesterol levels.Furthermore,we delved into the molecular mechanisms underlying these nutritional differences by analyzing significantly different 431 m RNAs and 865 proteins and integrating the regulatory network of protein-metabolite-lipid pathway.Importantly,in the pyruvate metabolic pathway of Bamei pigs,the bioprocess of lactate production was inhibited but the acetyl-Co A production was activated,suggesting the possibility that energy allocation favors the biogenesis of lipid precursors.These findings may contribute to guiding industrial food producers in enhancing the quality of lard at the genetic and molecular levels.展开更多
[Objectives]To investigate the structure and function of the lipoxygenase(LOX)gene family in Physcomitrella patens.[Methods]This study employed bioinformatics methods to identify and predict LOX gene family members.Qu...[Objectives]To investigate the structure and function of the lipoxygenase(LOX)gene family in Physcomitrella patens.[Methods]This study employed bioinformatics methods to identify and predict LOX gene family members.Quantitative real-time PCR(qRT-PCR)was utilized to analyze the expression patterns of LOX genes at different stages of Botrytis cinerea infection.[Results]The P.patens LOX gene family comprises eight putative proteins,including two 12-LOX-type members and six 13-LOX-type members.Among the eight LOX proteins,PpLOX7 exhibited the lowest molecular weight and shortest amino acid sequence.PpLOX7 was identified as a basic protein with an isoelectric point(pI)of 8.54,while all other members were acidic.Subcellular localization analysis indicated that PpLOX7 was localized to the chloroplast,whereas the remaining members were distributed in the cytoplasm.Secondary structure prediction showed that all eight proteins were predominantly composed of random coils andα-helixes.Chromosomal mapping revealed that the LOX genes were distributed across 7 of the 27 chromosomes in P.patens,with PpLOX1 and PpLOX2 tandemly arranged on chromosome 15.The qRT-PCR analysis demonstrated distinct expression patterns among the eight PpLOX genes following B.cinerea infection.PpLOX1-3 and PpLOX7 were upregulated to varying degrees,suggesting their potential involvement in the early defense response of P.patens against B.cinerea.Notably,PpLOX2 exhibited highly significant differential expression,making it a key candidate for further investigation.[Conclusions]This study provides foundational insights into the functional roles of the LOX gene family in P.patens during biotic stress responses.展开更多
Silicosis is one of the most serious and prevalent occupational diseases globally,characterized by typical silicotic nodules and fibrosis.Recent studies suggest that the perinodular zone of the lung shares certain cha...Silicosis is one of the most serious and prevalent occupational diseases globally,characterized by typical silicotic nodules and fibrosis.Recent studies suggest that the perinodular zone of the lung shares certain characteristics with the nodules themselves.In this study,a silicotic rat model was established via a single intratracheal in-stillation of a 50 mg/mL silica suspension.Pulmonary anatomical and pathological examinations revealed that silica deposition induced severe alterations in both the nodular and perinodular tissues.Subsequently,pseudo-targeted metabolomics analysis revealed that abnormally elevated ornithine levels were closely associated with the progression of silicosis,from normal to perinodular and finally to nodular tissues.Immunofluorescent stain-ing demonstrated that,in addition to M2 macrophages,silica exposure increased the protein levels of ARG1 in epithelial cells,a finding further confirmed by in vitro experiments using A549 and BEAS-2B cells.Moreover,accumulated ornithine induced epithelial-mesenchymal transition in vitro,increased extracellular matrix expres-sion in NIH 3T3 fibroblasts,and enhanced TGF-β1 levels in RAW264.7 cells.Co-exposure to ornithine and silica significantly induced the aberrant expression of fibrosis-associated proteins compared to silica exposure alone,characterized by increased levels of FN and𝛼-SMA,as well as decreased E-cad expression.These findings sug-gest that silica exposure up-regulates ARG1 in various cells,leading to ornithine accumulation,which in turn accelerates the progression of fibrosis.展开更多
Soft-tissue motion introduces significant challenges in robotic teleoperation,especially in medical scenarios where precise target tracking is critical.Latency across sensing,computation,and actuation chains leads to ...Soft-tissue motion introduces significant challenges in robotic teleoperation,especially in medical scenarios where precise target tracking is critical.Latency across sensing,computation,and actuation chains leads to degraded tracking performance,particularly around high-acceleration segments and trajectory inflection points.This study investigates machine learning-based predictive compensation for latency mitigation in soft-tissue tracking.Three models—autoregressive(AR),long short-term memory(LSTM),and temporal convolutional network(TCN)—were implemented and evaluated on both synthetic and real datasets.By aligning the prediction horizon with the end-to-end system delay,we demonstrate that prediction-based compensation significantly reduces tracking errors.Among the models,TCN achieved superior robustness and accuracy on complex motion patterns,particularly in multi-step prediction tasks,and exhibited better latency–horizon compatibility.The results suggest that TCN is a promising candidate for real-time latency compensation in teleoperated robotic systems involving dynamic soft-tissue interaction.展开更多
In this study,we present the development of a cryobioink designed to fabricate anisotropic scaffolds that support both neural and muscle cell-alignment.Given the critical role of cellular organization in nerve fibers ...In this study,we present the development of a cryobioink designed to fabricate anisotropic scaffolds that support both neural and muscle cell-alignment.Given the critical role of cellular organization in nerve fibers and neuromuscular junctions,we employed a vertical cryobioprinting-enabled ice-templating technique to create scaffolds with aligned microchannels.These channels facilitated cell-alignment,which is important in modeling neural and neuromuscular tissues.By integrating hyaluronic acid-methacrylate(HAMA)with gelatin methacryloyl and the necessary cryoprotective agent melezitose,we showcased that the cryobioink could preserve cell viability during freezing/thawing processes,even at low temperatures employed during cryobioprinting.We optimized HAMA concentration to enhance neural cell viability and alignment,and successfully constructed anisotropic scaffolds featuring distinct sections that contained muscle and neural cells,establishing a model for neuromuscular junctions.The resulting models provide a versatile platform for studying nerve fibers and neuromuscular dysfunctions,offering potential advancements in neural regeneration research.展开更多
BACKGROUND Mesenchymal stem cells(MSCs)are considered a promising therapy for various diseases due to their strong potential in regenerative medicine and immunomodulation.The tissue source of MSCs has gained attention...BACKGROUND Mesenchymal stem cells(MSCs)are considered a promising therapy for various diseases due to their strong potential in regenerative medicine and immunomodulation.The tissue source of MSCs has gained attention for its role in influencing their function,accessibility,and readiness for clinical use.AIM To identify the most suitable adipose source for MSC isolation and expansion for further applications.METHODS We isolated MSCs from solid adipose tissue and liposuction aspirates using the enzyme method.The MSCs were examined for their expansion using population doubling time,differentiation capacity using multilineage differentiation induction,surface markers using flow cytometry,and stability of chromosomes using the karyotyping method.Growth factors and cytokines in MSC-conditioned media were analyzed using the Luminex assay.RESULTS MSCs were isolated from solid adipose tissue and lipoaspirates and expanded from passage 0 to passage 2.All adipose-derived MSCs(AD-MSCs)exhibited the typical elongated,spindle-shaped morphology and comparable proliferation rate.They expressed positive surface markers(cluster of differentiation 73[CD73]:>97%,CD90:>98%,and CD105:>95%),and negative markers(<1%).All MSCs expressed similar levels of stemness genes(octamer-binding transcription factor 4,SRY-box 2,Krüppel-like factor,and MYC),colonyforming,and trilineage differentiation potential.Karyotyping analysis revealed normal chromosomal patterns in all samples,except one sample exhibiting a polymorphism(1qh+).Furthermore,the growth factors and cytokines of hepatocyte growth factor,vascular endothelial growth factor A,interleukin 6(IL-6),and IL-8 were detected in all AD-MSC conditioned media;but fibroblast growth factor-2 and keratinocyte growth factor were selectively expressed in conditioned media from solid or lipoaspirate AD-MSCs,respectively.CONCLUSION These findings indicate that AD-MSCs from both adipose sources possess all of the characteristic features of MSCs with source-specific secretome differences,which are suitable for further expansion and various clinical applications.展开更多
Objective To develop a depression recognition model by integrating the spirit-expression diagnostic framework of traditional Chinese medicine(TCM)with machine learning algorithms.The proposed model seeks to establish ...Objective To develop a depression recognition model by integrating the spirit-expression diagnostic framework of traditional Chinese medicine(TCM)with machine learning algorithms.The proposed model seeks to establish a TCM-informed tool for early depression screening,thereby bridging traditional diagnostic principles with modern computational approaches.Methods The study included patients with depression who visited the Shanghai Pudong New Area Mental Health Center from October 1,2022 to October 1,2023,as well as students and teachers from Shanghai University of Traditional Chinese Medicine during the same period as the healthy control group.Videos of 3–10 s were captured using a Xiaomi Pad 5,and the TCM spirit and expressions were determined by TCM experts(at least 3 out of 5 experts agreed to determine the category of TCM spirit and expressions).Basic information,facial images,and interview information were collected through a portable TCM intelligent analysis and diagnosis device,and facial diagnosis features were extracted using the Open CV computer vision library technology.Statistical analysis methods such as parametric and non-parametric tests were used to analyze the baseline data,TCM spirit and expression features,and facial diagnosis feature parameters of the two groups,to compare the differences in TCM spirit and expression and facial features.Five machine learning algorithms,including extreme gradient boosting(XGBoost),decision tree(DT),Bernoulli naive Bayes(BernoulliNB),support vector machine(SVM),and k-nearest neighbor(KNN)classification,were used to construct a depression recognition model based on the fusion of TCM spirit and expression features.The performance of the model was evaluated using metrics such as accuracy,precision,and the area under the receiver operating characteristic(ROC)curve(AUC).The model results were explained using the Shapley Additive exPlanations(SHAP).Results A total of 93 depression patients and 87 healthy individuals were ultimately included in this study.There was no statistically significant difference in the baseline characteristics between the two groups(P>0.05).The differences in the characteristics of the spirit and expressions in TCM and facial features between the two groups were shown as follows.(i)Quantispirit facial analysis revealed that depression patients exhibited significantly reduced facial spirit and luminance compared with healthy controls(P<0.05),with characteristic features such as sad expressions,facial erythema,and changes in the lip color ranging from erythematous to cyanotic.(ii)Depressed patients exhibited significantly lower values in facial complexion L,lip L,and a values,and gloss index,but higher values in facial complexion a and b,lip b,low gloss index,and matte index(all P<0.05).(iii)The results of multiple models show that the XGBoost-based depression recognition model,integrating the TCM“spirit-expression”diagnostic framework,achieved an accuracy of 98.61%and significantly outperformed four benchmark algorithms—DT,BernoulliNB,SVM,and KNN(P<0.01).(iv)The SHAP visualization results show that in the recognition model constructed by the XGBoost algorithm,the complexion b value,categories of facial spirit,high gloss index,low gloss index,categories of facial expression and texture features have significant contribution to the model.Conclusion This study demonstrates that integrating TCM spirit-expression diagnostic features with machine learning enables the construction of a high-precision depression detection model,offering a novel paradigm for objective depression diagnosis.展开更多
The esophagus is a tubular organ essential for maintaining normal eating function in humans.However,the replacement of the esophagus remains challenging in clinical settings.Although tissue engineering scaffolds are a...The esophagus is a tubular organ essential for maintaining normal eating function in humans.However,the replacement of the esophagus remains challenging in clinical settings.Although tissue engineering scaffolds are a promising alternative solution,their fabrication is difficult due to the complex structure and function of the esophagus.This review describes the existing fabrication methods for esophageal tubular scaffolds,including decellularization,casting,electrospinning,three dimensional(3 D)bioprinting,and pin-frogging.Also discussed are the stimulation cues of the fabricated esophageal tubular scaffold that induce esophageal muscle and epithelial cells.Finally,this review emphasizes three important concerns for esophageal tubular scaffolds:leakage and porosity,elasticity and proliferation of smooth muscle cells,and biocompatibility and structural fidelity of biomaterials.展开更多
[Objectives]The present study was conducted to investigate the change rule ofβ-fructofuranosidase gene expression and its enzyme activity in the midgut of 5 th instar silkworm(Bombyx mori),in order to provide a refer...[Objectives]The present study was conducted to investigate the change rule ofβ-fructofuranosidase gene expression and its enzyme activity in the midgut of 5 th instar silkworm(Bombyx mori),in order to provide a reference for illustrating the enzymatic mechanism of usingβ-fructofuranosidase to absorb sucrose nutrition from mulberry leaves.[Methods]Real-time fluorescent quantitative PCR was applied to analyze the expression of BmSuc1 and BmSuc2 in midgut of 5 th-instar silkworm larvae,meanwhile the activities ofβ-fructofuranosidase was determined.[Results]BmSuc1 was expressed in the midgut of 5 th-instar silkworm larvae at different developmental stages.Its expression was upregulated at the beginning of the 5 th instar and during the peak feeding period,whereas BmSuc2 expression remained very low throughout the entire 5 th instar.The activity ofβ-fructofuranosidase was relatively high during the peak feeding period of 5 th-instar larvae,showing a trend of increasing first and then decreasing.[Conclusions]The expression pattern of the BmSuc1 gene and the changes inβ-fructofuranosidase activity were generally consistent with the physiological process of sugar nutrient absorption and utilization from mulberry leaves in 5 th-instar silkworms.It suggests that BmSuc1,as a sucrose hydrolase gene,plays a major role in the digestion and absorption of sucrose nutrients from mulberry leaves in the midgut tissue.展开更多
The pathogenesis-related protein PR10 plays a vital role in plant growth,development,and stress responses.This study systematically identified and analyzed PR10 genes in cultivated peanut(Arachis hypogaea L.),examinin...The pathogenesis-related protein PR10 plays a vital role in plant growth,development,and stress responses.This study systematically identified and analyzed PR10 genes in cultivated peanut(Arachis hypogaea L.),examining their phylogenetic relationships,conserved motifs,gene structures,and syntenic relationships.The analysis identified 54 Ah PR10 genes,which were classified into eight groups based on phylogenetic relationships,supported by gene structure and conserved motif characterization.Analysis of chromosomal distribution and synteny demonstrated that segmental duplications played a crucial role in the expansion of the Ah PR10 gene family.The identified Ah PR10 genes exhibited both constitutive and inducible expression patterns.Significantly,Ah PR10-7,Ah PR10-33,and Ah PR10-41 demonstrated potential importance in peanut resistance to Aspergillus flavus.In vitro fungistatic experiments demonstrated that recombinant Ah PR10-33 effectively inhibited A.flavus mycelial growth.These findings provide valuable insights for future investigations into Ah PR10 functions in protecting peanut from A.flavus infection.展开更多
Microwave ablation(MWA)is a minimally invasive technique for treating hepatic tumors,necessitating precise monitoring to ensure treatment efficacy and minimize damage to surrounding tissues.This study explores the pot...Microwave ablation(MWA)is a minimally invasive technique for treating hepatic tumors,necessitating precise monitoring to ensure treatment efficacy and minimize damage to surrounding tissues.This study explores the potential of photoacoustic imaging(PAI)in monitoring MWA by examining ex vivo porcine liver tissues.In this study,a comprehensive analysis of photoacoustic signals was performed to compare the main lobe width(MLW)between ablated and normal regions in porcine liver tissue.Histological staining with succinate dehydrogenase(SDH)and shear wave elastography(SWE)were employed to validate the changes in tissue elasticity after ablation.The analysis demonstrated a notable reduction in the MLW of the average A-lines in ablated tissues compared to nonablated regions(p<0.01).This reduction,attributed to increased tissue density and enhanced elasticity,indicates accelerated sound propagation in thermally ablated areas,which then serves as a critical parameter for mapping tissue characteristics.The reconstruction of the MLW distribution successfully delineated the ablated regions,and was consistent with the results of SDH staining and SWE.In addition,MLW-based imaging exhibited higher spatial resolution compared to SWE.Incorporating MLW analysis into PAI may be a promising strategy to improve the accuracy and effectiveness of MWA monitoring in clinical settings.展开更多
Due to the unique microstructure and diverse opsin genes of the trinocular compound eye,stomatopoda possess an extraordinary ability to perceive multiple properties of light.They not only can detect natural light(NL)a...Due to the unique microstructure and diverse opsin genes of the trinocular compound eye,stomatopoda possess an extraordinary ability to perceive multiple properties of light.They not only can detect natural light(NL)and linearly polarized light(LPL),but also are the only animals capable of recognizing circularly polarized light(CPL).Here,we integrated single-cell RNA sequencing,previously published Illumina data,and in-situ hybridization(ISH)to quantify and localize functional opsin genes in Oratosquilla oratoria,a common stomatopoda species in the China Sea.A total of high-quality 31777 cells were captured for the first time in the O.oratoria compound eye,which were classified into 25 cell subpopulations,and hypothesized that cluster 22 is a critical cell subpopulation responsible for light(whether NL,LPL,or CPL)response in O.oratoria.Furthermore,we propose that the long-wavelengthsensitive opsin gene(lws)gene family,retinol dehydrogenase(rdh),voltage-gated ion channel(vgic),arrestin(arr),and myosin(myo)collectively mediate the light response in O.oratoria.Considering that very few vision-related opsin genes show differential expression in right-handed CPL(RCPL)-vs.-dark(DL),which provides additional evidence that stomatopoda cannot recognize RCPL.Meanwhile,we believe that UV-stimulated scaffold protein A(uvssa)and red pigment concentrating hormone(rpch)play special contributions in the left-handed CPL(LCPL)environment response.ISH revealing that 16 lws,6 middle-wavelength-sensitive(mws),and 2 ultraviolet(uv)opsin genes were expressed in the photoreceptors of the O.oratoria compound eye.Although the inability to determine the functional types of cell subpopulations limits the resolution of opsin genes,these findings systematically elucidate the specific expression patterns of opsin genes in O.oratoria and represent a significant step toward refining the visual ecological theory of O.oratoria and other stomatopod species.展开更多
While methodology for determining the mode of evolution in coding sequences has been well established,evaluation of adaptation events in emerging types of phenotype data needs further development.Here,we propose an an...While methodology for determining the mode of evolution in coding sequences has been well established,evaluation of adaptation events in emerging types of phenotype data needs further development.Here,we propose an analysis framework(expression variance decomposition,EVaDe)for comparative single-cell expression data based on phenotypic evolution theory.After decomposing the gene expression variance into separate components,we use two strategies to identify genes exhibiting large between-taxon expression divergence and small within-cell-type expression noise in certain cell types,attributing this pattern to putative adaptive evolution.In a dataset of primate prefrontal cortex,we find that such humanspecific key genes enrich with neurodevelopment-related functions,while most other genes exhibit neutral evolution patterns.Specific neuron types are found to harbor more of these key genes than other cell types,thus likely to have experienced more extensive adaptation.Reassuringly,at the molecular sequence level,the key genes are significantly associated with the rapidly evolving conserved non-coding elements.An additional case analysis comparing the naked mole-rat(NMR)with the mouse suggests that innateimmunity-related genes and cell types have undergone putative expression adaptation in NMR.Overall,the EVaDe framework may effectively probe adaptive evolution mode in single-cell expression data.展开更多
Contrary to the adult central nervous system,the peripheral nervous system has an intrinsic ability to regenerate that relies on the expression of regenerationassociated genes,such as some kinesin family members.Kines...Contrary to the adult central nervous system,the peripheral nervous system has an intrinsic ability to regenerate that relies on the expression of regenerationassociated genes,such as some kinesin family members.Kinesins contribute to nerve regeneration through the transport of specific cargo,such as proteins and membrane components,from the cell body towards the axon periphery.We show here that KIF4A,associated with neurodevelopmental disorders and previously believed to be only expressed during development,is also expressed in the adult vertebrate nervous system and up-regulated in injured peripheral nervous system cells.KIF4A is detected both in the cell bodies and regrowing axons of injured neurons,consistent with its function as an axonal transporter of cargoes such asβ1-integrin and L1CAM.Our study further demonstrates that KIF4A levels are greatly increased in Schwann cells from injured distal nerve stumps,particularly at a time when they are reprogrammed into an essential proliferative repair phenotype.Moreover,Kif4a m RNA levels were approximately~6-fold higher in proliferative cultured Schwann cells compared with non-proliferative ones.A hypothesized function for Kif4a in Schwann cell proliferation was further confirmed by Kif4a knockdown,as this significantly reduced Schwann cell proliferation in vitro.Our findings show that KIF4A is expressed in adult vertebrate nervous systems and is up-regulated following peripheral injury.The timing of KIF4A up-regulation,its location during regeneration,and its proliferative role,all suggest a dual role for this protein in neuroregeneration that is worth exploring in the future.展开更多
Neuroinflammation is a key process in the pathogenesis of various neurodegenerative diseases,such as multiple sclerosis(MS),Alzheimer's disease,and traumatic brain injury.Even for disorders historically unrelated ...Neuroinflammation is a key process in the pathogenesis of various neurodegenerative diseases,such as multiple sclerosis(MS),Alzheimer's disease,and traumatic brain injury.Even for disorders historically unrelated to neuroinflammation,such as Alzheimer's disease,it is now shown to precede pathological protein aggregations.展开更多
基金Supported by College Student Innovation and Entrepreneurship Training Program(S202210553003)Hunan Provincial Education Department Outstanding Youth Research Project(23B0820).
文摘Kinesins are a superfamily of proteins widely present in eukaryotes,playing crucial roles in plant cell wall assembly,cell elongation regulation,gravity sensing,and fertility control.In this study,bioinformatics analysis of the OsKMP2 gene(LOC_Os02g28850)was performed using online tools such as ExPASy-ProtParam,ProtScale,CD-search,and DNAMAN software.Additionally,qRT-PCR was employed to analyze the tissue expression pattern of OsKMP2.The results showed that the molecular weight of the OsKMP2 is 118.39728 kDa,and it is a hydrophilic and unstable acidic protein.Secondary structure prediction revealed that it primarily consists ofα-helices(69.45%),random coils(25.19%),and extended strands(5.36%).The gene was expressed in various rice tissues,with the highest expression level observed in leaves.These results indicate that the OsKMP2 gene exhibits high evolutionary conservation and functional diversity in rice.
基金supported by the National Natural Science Foundation of China(32271580,42020104009)the Fundamental Research Funds for Zhejiang Provincial Universities and Research Institutes(JX6311101923)。
文摘Mytilus contain abundant antimicrobial peptides(AMPs)that play a key role in the innate immunity.However,heterologous production of these AMPs remains challenging due to their short sequences,multiple disulfide bonds,and high content of cationic amino acids,which hinder functional expression in prokaryotic systems such as Escherichia coli.To establish a eukaryotic recombinant expression system for the AMPs of mussel and obtain recombinant mussel AMPs for subsequent studies,we reported the successful recombinant expression of myticofensin B1,a novel defensin-like AMP identified previously in Mytiluscoruscus,using the eukaryotic host Pichia pastoris.The codon-optimized gene encoding the mature myticofensin-B1(composed of 65 amino acid residues,including 6 conserved cysteine residues)was cloned into a pPICZαA vector and expressed in P.pastoris GS115.Structural fidelity of the recombinant peptide was confirmed by liquid chromatography-tandem mass spectrometry(LC-MS/MS),showing a molecular weight of 8849.9 Da,which was consistent with the theoretical prediction.Functional assays demonstrated a broad-spectrum antimicrobial activity of the recombinant myticofensin-B1,with stronger inhibition against Gram-negative bacteria.Scanning electron microscopy revealed different effects of the recombinant myticofensin-B1 against different bacteria.In addition,the recombinant myticofensin-B1 exhibited a very low hemolytic activity against sheep red blood cells and weak cytotoxicity against human A549 lung cancer cells.This study establishes P.pastoris as a powerful platform to produce functional mussel AMP and highlights the potential of the recombinant myticofensin-B1 as a therapeutic agent for aquaculture pathogens and infections.
基金supported by the National Natural Science Foundation of China,Nos.32271389,31900987(both to PY)the Natural Science Foundation of Jiangsu Province,No.BK20230608(to JJ)。
文摘Regulatory T cells,a subset of CD4^(+)T cells,play a critical role in maintaining immune tolerance and tissue homeostasis due to their potent immunosuppressive properties.Recent advances in research have highlighted the important therapeutic potential of Tregs in neurological diseases and tissue repair,emphasizing their multifaceted roles in immune regulation.This review aims to summarize and analyze the mechanisms of action and therapeutic potential of Tregs in relation to neurological diseases and neural regeneration.Beyond their classical immune-regulatory functions,emerging evidence points to non-immune mechanisms of regulatory T cells,particularly their interactions with stem cells and other non-immune cells.These interactions contribute to optimizing the repair microenvironment and promoting tissue repair and nerve regeneration,positioning non-immune pathways as a promising direction for future research.By modulating immune and non-immune cells,including neurons and glia within neural tissues,Tregs have demonstrated remarkable efficacy in enhancing regeneration in the central and peripheral nervous systems.Preclinical studies have revealed that Treg cells interact with neurons,glial cells,and other neural components to mitigate inflammatory damage and support functional recovery.Current mechanistic studies show that Tregs can significantly promote neural repair and functional recovery by regulating inflammatory responses and the local immune microenvironment.However,research on the mechanistic roles of regulatory T cells in other diseases remains limited,highlighting substantial gaps and opportunities for exploration in this field.Laboratory and clinical studies have further advanced the application of regulatory T cells.Technical advances have enabled efficient isolation,ex vivo expansion and functionalization,and adoptive transfer of regulatory T cells,with efficacy validated in animal models.Innovative strategies,including gene editing,cell-free technologies,biomaterial-based recruitment,and in situ delivery have expanded the therapeutic potential of regulatory T cells.Gene editing enables precise functional optimization,while biomaterial and in situ delivery technologies enhance their accumulation and efficacy at target sites.These advancements not only improve the immune-regulatory capacity of regulatory T cells but also significantly enhance their role in tissue repair.By leveraging the pivotal and diverse functions of Tregs in immune modulation and tissue repair,regulatory T cells–based therapies may lead to transformative breakthroughs in the treatment of neurological diseases.
文摘Quantitative real-time PCR(qPCR)is widely used for gene expression analysis,but its accuracy critically depends on stable internal reference genes for normalization.In marine invertebrates,especially non-model taxa such as cephalopods,systematic evaluation of reference genes is limited,leading to potential bias.The cuttlefish Sepiella japonica is ecologically and economically important in China,yet previous molecular studies have often relied on single unvalidated reference genes,which may compromise data reliability.This study aimed to systematically evaluate the stability of five commonly used reference genes(18S,ef-1α,ef-1γ,gapdh,andβ-actin)across multiple tissues and sexes of S.japonica,and to identify the most suitable reference genes and optimal number for qPCR normalization.Fifteen to sixteen tissue types were collected from ten healthy adults(five males and five females).Total RNA was extracted,reverse-transcribed,and analyzed by qPCR.Gene stability was assessed using four algorithms(geNorm,NormFinder,BestKeeper,andΔCt)integrated with RefFinder,and the optimal gene number was determined using geNorm pairwise variation(V_(n/n+1)<0.15).Four transcriptome-derived genes(creld2,cd109,acy1,and miox)were used for validation.The C_(t)values of the five genes ranged from 15.47 to 20.83.β-actin and gapdh showed pronounced variability in expression stability among tissues and sexes,indicating their limited suitability for normalization.18S exhibited the highest expression(mean C_(t):15.47-16.29)and lowest variability but displayed sex-biased expression,whereas ef-1αand ef-1γremained consistently stable across most tissues in both sexes,with ef-1αbeing the most robust and showing no sex-related bias.Although specific rankings varied among tissues and sexes,the comprehensive results indicated that ef-1αand ef-1γpossessed the highest overall stability,followed by 18S,whileβ-actin and gapdh were the least stable.The final comprehensive rankings were ef-1γ>ef-1α>18S>gapdh>β-actin(male)and ef-1α>ef-1γ>18S>gapdh>β-actin(female).geNorm analysis(V2/3<0.15)indicated that two genes,mainly ef-1αand ef-1γ,were generally sufficient for reliable normalization in most tissues.Validation confirmed that normalization using the stable ef-1αand ef-1γaccurately reflected the expression differences among tissues,whereasβ-actin and gapdh can bias or confound statistical analyses.ef-1αand ef-1γare identified as the most reliable reference gene combination for qPCR analysis in S.japonica,while 18S can serve as an auxiliary gene for within-sex comparisons.The use ofβ-actin or gapdh alone is not recommended.This study establishes a systematic framework for selecting reliable reference genes in S.japonica,thereby facilitating robust qPCR normalization and providing a foundation for future gene expression research in S.japonica and other cephalopods.
基金National Natural Science Foundation of China:Investigation on the Mechanism of Gegen Qinlian Decoction in Lowering Blood Glucose Based on Improving Cholesterol Homeostasis of Lipid Rafts in Adipocytes(No.82060741)Study on the Cause of“Dampness Stagnating in Pi”in Type 2 Diabetes Mellitus by Overeating High Fat and Sugar Diet Based on Glycerol Transport Which Is Affected by Methylation/Hormones Signal(No.82160830)+4 种基金Investigation on the Mechanism of Gegen Qinlian Decoction in Lowering Blood Glucose Based on Improving Phosphatidylcholine Biosynthesis in Adipose Tissue(No.82360799)Jiangxi Provincial Administration of Traditional Chinese Medicine Science and Technology Program:the Relationship between Adipose Tissue and Spleen Image Based on Multivariate Analysis of Modern Literature(No.2020B0328)Ganpo Juncai-Training Program for Academic and Technical Leaders in Major Disciplines of the Province(Leading Talents-Academic Category)(20232BCJ22022)Jiangxi University of Chinese Medicine,First-level Discipline of Integrative Medicine(Jiangxi Province Double First-class Discipline)(zxyylxk20220103)Jiangxi University of Chinese Medicine Science and Technology Innovation Team Development Program(CXTD22007)。
文摘OBJECTIVE:To investigate the hypoglycemic mechanism of modified Gegen Qinlian decoction(加味葛根芩连汤,MGQD)by examining its regulation of cholesterol transporter expression and DNA methylation,specifically the low-density lipoprotein receptor(LDLR)and scavenger receptor class B type 1(SR-B1),in epididymal white adipose tissue(e WAT)and inguinal white adipose tissue(i WAT)of rats with type 2 diabetes mellitus(T2DM).METHODS:The control group(CON)consisted of ten Sprague-Dawley(SD)rats fed a standard chow diet,while 80 SD rats were fed a high-fat diet and administered streptozotocin intraperitoneally to induce diabetes.The diabetic rats were randomly assigned to four groups:T2DM,metformin(MET,200 mg/kg),low-dose MGQD(MGQDL,5 g/kg),and high-dose MGQD(MGQDH,10 g/kg),and received treatment via gavage for 14 weeks.Western blot(WB),quantitative real-time polymerase chain reaction(q PCR),and bisulfite sequencing PCR(BSP)were used to analyze protein levels,m RNA expression,and DNA methylation of Ldlr(gene encoding LDLR)and Srb1(gene encoding SR-B1).RESULTS:MGQD and metformin treatment significantly reduced blood glucose levels,restored LDLR and SR-B1 protein levels in e WAT,and effectively regulated the m RNA expression and non-cytosine-p-guanine(nonCp G)methylation of Srb1 in e WAT.A significant negative correlation was observed between the methylation of Srb1 in e WAT and its m RNA expression.However,MGQD and metformin had no significant effect on the protein levels,m RNA expression,or DNA methylation of Ldlr and Srb1 in i WAT.CONCLUSIONS:MGQD did not significantly affect LDLR and SR-B1 expression or gene methylation in i WAT.However,its hypoglycemic effect may be linked to cholesterol regulation in e WAT.Potential mechanisms include increased LDLR protein levels,which may enhance cholesterol uptake,and increased Srb1 methylation,which may suppress its expression and consequently reduce cholesterol efflux.
基金supported by the National Key Research and Development Program of China(2021YFF1000602)the National Natural Science Foundations(32202642)the earmarked fund for CARS-35-PIG.
文摘Bamei pigs,an indigenous Chinese breed,yield meat with a delectable flavor and boast higher carcass fat content compared to commercial breeds,making them a rich food source for humans.However,the differences in lipid and nutrient components between the adipose tissue of Bamei pigs and commercial pigs are still unclear.The study employed UPLC-MS/MS to quantify the composition of lipids and metabolites in the backfat of both Bamei and Large White pigs.A total of 428 lipids and 193 metabolites were significantly different between the 2 groups.Specifically,Bamei pig backfat exhibited altered levels of various lipids and metabolites that may potentially contribute to nutritional and flavor differences,including unsaturated triglycerides,free fatty acids,medium-chain triglycerides,essential amino acids,vitamins and antioxidants,while maintaining reduced cholesterol levels.Furthermore,we delved into the molecular mechanisms underlying these nutritional differences by analyzing significantly different 431 m RNAs and 865 proteins and integrating the regulatory network of protein-metabolite-lipid pathway.Importantly,in the pyruvate metabolic pathway of Bamei pigs,the bioprocess of lactate production was inhibited but the acetyl-Co A production was activated,suggesting the possibility that energy allocation favors the biogenesis of lipid precursors.These findings may contribute to guiding industrial food producers in enhancing the quality of lard at the genetic and molecular levels.
基金Supported by the Science and Technology Program of Guizhou Provence(Qiankehejichu-ZK[2023]Yiban 271).
文摘[Objectives]To investigate the structure and function of the lipoxygenase(LOX)gene family in Physcomitrella patens.[Methods]This study employed bioinformatics methods to identify and predict LOX gene family members.Quantitative real-time PCR(qRT-PCR)was utilized to analyze the expression patterns of LOX genes at different stages of Botrytis cinerea infection.[Results]The P.patens LOX gene family comprises eight putative proteins,including two 12-LOX-type members and six 13-LOX-type members.Among the eight LOX proteins,PpLOX7 exhibited the lowest molecular weight and shortest amino acid sequence.PpLOX7 was identified as a basic protein with an isoelectric point(pI)of 8.54,while all other members were acidic.Subcellular localization analysis indicated that PpLOX7 was localized to the chloroplast,whereas the remaining members were distributed in the cytoplasm.Secondary structure prediction showed that all eight proteins were predominantly composed of random coils andα-helixes.Chromosomal mapping revealed that the LOX genes were distributed across 7 of the 27 chromosomes in P.patens,with PpLOX1 and PpLOX2 tandemly arranged on chromosome 15.The qRT-PCR analysis demonstrated distinct expression patterns among the eight PpLOX genes following B.cinerea infection.PpLOX1-3 and PpLOX7 were upregulated to varying degrees,suggesting their potential involvement in the early defense response of P.patens against B.cinerea.Notably,PpLOX2 exhibited highly significant differential expression,making it a key candidate for further investigation.[Conclusions]This study provides foundational insights into the functional roles of the LOX gene family in P.patens during biotic stress responses.
基金supported by the National Natural Science Foundation of China(No.81973466)the National Administration of Traditional Chinese Medicine Youth Qihuang Scholars Support Project,and the Program of Graduate Innovation Research in Shanxi Province(No.2023KY019).
文摘Silicosis is one of the most serious and prevalent occupational diseases globally,characterized by typical silicotic nodules and fibrosis.Recent studies suggest that the perinodular zone of the lung shares certain characteristics with the nodules themselves.In this study,a silicotic rat model was established via a single intratracheal in-stillation of a 50 mg/mL silica suspension.Pulmonary anatomical and pathological examinations revealed that silica deposition induced severe alterations in both the nodular and perinodular tissues.Subsequently,pseudo-targeted metabolomics analysis revealed that abnormally elevated ornithine levels were closely associated with the progression of silicosis,from normal to perinodular and finally to nodular tissues.Immunofluorescent stain-ing demonstrated that,in addition to M2 macrophages,silica exposure increased the protein levels of ARG1 in epithelial cells,a finding further confirmed by in vitro experiments using A549 and BEAS-2B cells.Moreover,accumulated ornithine induced epithelial-mesenchymal transition in vitro,increased extracellular matrix expres-sion in NIH 3T3 fibroblasts,and enhanced TGF-β1 levels in RAW264.7 cells.Co-exposure to ornithine and silica significantly induced the aberrant expression of fibrosis-associated proteins compared to silica exposure alone,characterized by increased levels of FN and𝛼-SMA,as well as decreased E-cad expression.These findings sug-gest that silica exposure up-regulates ARG1 in various cells,leading to ornithine accumulation,which in turn accelerates the progression of fibrosis.
基金Support by Sichuan Science and Technology Program[2023YFSY0026,2023YFH0004]Guangzhou Huashang University[2024HSZD01,HS2023JYSZH01].
文摘Soft-tissue motion introduces significant challenges in robotic teleoperation,especially in medical scenarios where precise target tracking is critical.Latency across sensing,computation,and actuation chains leads to degraded tracking performance,particularly around high-acceleration segments and trajectory inflection points.This study investigates machine learning-based predictive compensation for latency mitigation in soft-tissue tracking.Three models—autoregressive(AR),long short-term memory(LSTM),and temporal convolutional network(TCN)—were implemented and evaluated on both synthetic and real datasets.By aligning the prediction horizon with the end-to-end system delay,we demonstrate that prediction-based compensation significantly reduces tracking errors.Among the models,TCN achieved superior robustness and accuracy on complex motion patterns,particularly in multi-step prediction tasks,and exhibited better latency–horizon compatibility.The results suggest that TCN is a promising candidate for real-time latency compensation in teleoperated robotic systems involving dynamic soft-tissue interaction.
基金support from the National Institutes of Technology(R21EB030257,R01EB028143,R01HL153857,R01HL166522,R01CA282451,R56EB034702)National Science Foundation(CBET-EBMS-1936105,CISE-IIS-2225698)+1 种基金Chan Zuckerberg Initiative(2022316712,2024-347836)the Brigham Research Institute.
文摘In this study,we present the development of a cryobioink designed to fabricate anisotropic scaffolds that support both neural and muscle cell-alignment.Given the critical role of cellular organization in nerve fibers and neuromuscular junctions,we employed a vertical cryobioprinting-enabled ice-templating technique to create scaffolds with aligned microchannels.These channels facilitated cell-alignment,which is important in modeling neural and neuromuscular tissues.By integrating hyaluronic acid-methacrylate(HAMA)with gelatin methacryloyl and the necessary cryoprotective agent melezitose,we showcased that the cryobioink could preserve cell viability during freezing/thawing processes,even at low temperatures employed during cryobioprinting.We optimized HAMA concentration to enhance neural cell viability and alignment,and successfully constructed anisotropic scaffolds featuring distinct sections that contained muscle and neural cells,establishing a model for neuromuscular junctions.The resulting models provide a versatile platform for studying nerve fibers and neuromuscular dysfunctions,offering potential advancements in neural regeneration research.
文摘BACKGROUND Mesenchymal stem cells(MSCs)are considered a promising therapy for various diseases due to their strong potential in regenerative medicine and immunomodulation.The tissue source of MSCs has gained attention for its role in influencing their function,accessibility,and readiness for clinical use.AIM To identify the most suitable adipose source for MSC isolation and expansion for further applications.METHODS We isolated MSCs from solid adipose tissue and liposuction aspirates using the enzyme method.The MSCs were examined for their expansion using population doubling time,differentiation capacity using multilineage differentiation induction,surface markers using flow cytometry,and stability of chromosomes using the karyotyping method.Growth factors and cytokines in MSC-conditioned media were analyzed using the Luminex assay.RESULTS MSCs were isolated from solid adipose tissue and lipoaspirates and expanded from passage 0 to passage 2.All adipose-derived MSCs(AD-MSCs)exhibited the typical elongated,spindle-shaped morphology and comparable proliferation rate.They expressed positive surface markers(cluster of differentiation 73[CD73]:>97%,CD90:>98%,and CD105:>95%),and negative markers(<1%).All MSCs expressed similar levels of stemness genes(octamer-binding transcription factor 4,SRY-box 2,Krüppel-like factor,and MYC),colonyforming,and trilineage differentiation potential.Karyotyping analysis revealed normal chromosomal patterns in all samples,except one sample exhibiting a polymorphism(1qh+).Furthermore,the growth factors and cytokines of hepatocyte growth factor,vascular endothelial growth factor A,interleukin 6(IL-6),and IL-8 were detected in all AD-MSC conditioned media;but fibroblast growth factor-2 and keratinocyte growth factor were selectively expressed in conditioned media from solid or lipoaspirate AD-MSCs,respectively.CONCLUSION These findings indicate that AD-MSCs from both adipose sources possess all of the characteristic features of MSCs with source-specific secretome differences,which are suitable for further expansion and various clinical applications.
基金General Program of National Natural Science Foundation of China(82474390)Construction Project of Pudong New Area Famous TCM Studios(National Pilot Zone for TCM Development,Shanghai)(PDZY-2025-0716)Shanghai Municipal Science and Technology Program Project Shanghai Key Laboratory of Health Identification and Assessment(21DZ2271000).
文摘Objective To develop a depression recognition model by integrating the spirit-expression diagnostic framework of traditional Chinese medicine(TCM)with machine learning algorithms.The proposed model seeks to establish a TCM-informed tool for early depression screening,thereby bridging traditional diagnostic principles with modern computational approaches.Methods The study included patients with depression who visited the Shanghai Pudong New Area Mental Health Center from October 1,2022 to October 1,2023,as well as students and teachers from Shanghai University of Traditional Chinese Medicine during the same period as the healthy control group.Videos of 3–10 s were captured using a Xiaomi Pad 5,and the TCM spirit and expressions were determined by TCM experts(at least 3 out of 5 experts agreed to determine the category of TCM spirit and expressions).Basic information,facial images,and interview information were collected through a portable TCM intelligent analysis and diagnosis device,and facial diagnosis features were extracted using the Open CV computer vision library technology.Statistical analysis methods such as parametric and non-parametric tests were used to analyze the baseline data,TCM spirit and expression features,and facial diagnosis feature parameters of the two groups,to compare the differences in TCM spirit and expression and facial features.Five machine learning algorithms,including extreme gradient boosting(XGBoost),decision tree(DT),Bernoulli naive Bayes(BernoulliNB),support vector machine(SVM),and k-nearest neighbor(KNN)classification,were used to construct a depression recognition model based on the fusion of TCM spirit and expression features.The performance of the model was evaluated using metrics such as accuracy,precision,and the area under the receiver operating characteristic(ROC)curve(AUC).The model results were explained using the Shapley Additive exPlanations(SHAP).Results A total of 93 depression patients and 87 healthy individuals were ultimately included in this study.There was no statistically significant difference in the baseline characteristics between the two groups(P>0.05).The differences in the characteristics of the spirit and expressions in TCM and facial features between the two groups were shown as follows.(i)Quantispirit facial analysis revealed that depression patients exhibited significantly reduced facial spirit and luminance compared with healthy controls(P<0.05),with characteristic features such as sad expressions,facial erythema,and changes in the lip color ranging from erythematous to cyanotic.(ii)Depressed patients exhibited significantly lower values in facial complexion L,lip L,and a values,and gloss index,but higher values in facial complexion a and b,lip b,low gloss index,and matte index(all P<0.05).(iii)The results of multiple models show that the XGBoost-based depression recognition model,integrating the TCM“spirit-expression”diagnostic framework,achieved an accuracy of 98.61%and significantly outperformed four benchmark algorithms—DT,BernoulliNB,SVM,and KNN(P<0.01).(iv)The SHAP visualization results show that in the recognition model constructed by the XGBoost algorithm,the complexion b value,categories of facial spirit,high gloss index,low gloss index,categories of facial expression and texture features have significant contribution to the model.Conclusion This study demonstrates that integrating TCM spirit-expression diagnostic features with machine learning enables the construction of a high-precision depression detection model,offering a novel paradigm for objective depression diagnosis.
基金support from the National Natural Science Foundation of China(No.82472440)Hubei Provincial Natural Science Foundation of China(No.2023AFB141)+1 种基金the National Medical Products Administration Key Laboratory for Dental Materials(No.PKUSS20240401)the Cross-Research Support Program from Huazhong University of Science and Technology。
文摘The esophagus is a tubular organ essential for maintaining normal eating function in humans.However,the replacement of the esophagus remains challenging in clinical settings.Although tissue engineering scaffolds are a promising alternative solution,their fabrication is difficult due to the complex structure and function of the esophagus.This review describes the existing fabrication methods for esophageal tubular scaffolds,including decellularization,casting,electrospinning,three dimensional(3 D)bioprinting,and pin-frogging.Also discussed are the stimulation cues of the fabricated esophageal tubular scaffold that induce esophageal muscle and epithelial cells.Finally,this review emphasizes three important concerns for esophageal tubular scaffolds:leakage and porosity,elasticity and proliferation of smooth muscle cells,and biocompatibility and structural fidelity of biomaterials.
基金Supported by General Project of Yunnan Provincial Agricultural Basic Research Joint Special Project(202301BD070001-229)Yunnan Provincial Key R&D Program(202403AK140075)+1 种基金Modern Sericulture Industry Technology System of Yunan Province(KJTX-07)Honghe Comprehensive Test Station of National Sericulture Industry Technology System(CARS-18).
文摘[Objectives]The present study was conducted to investigate the change rule ofβ-fructofuranosidase gene expression and its enzyme activity in the midgut of 5 th instar silkworm(Bombyx mori),in order to provide a reference for illustrating the enzymatic mechanism of usingβ-fructofuranosidase to absorb sucrose nutrition from mulberry leaves.[Methods]Real-time fluorescent quantitative PCR was applied to analyze the expression of BmSuc1 and BmSuc2 in midgut of 5 th-instar silkworm larvae,meanwhile the activities ofβ-fructofuranosidase was determined.[Results]BmSuc1 was expressed in the midgut of 5 th-instar silkworm larvae at different developmental stages.Its expression was upregulated at the beginning of the 5 th instar and during the peak feeding period,whereas BmSuc2 expression remained very low throughout the entire 5 th instar.The activity ofβ-fructofuranosidase was relatively high during the peak feeding period of 5 th-instar larvae,showing a trend of increasing first and then decreasing.[Conclusions]The expression pattern of the BmSuc1 gene and the changes inβ-fructofuranosidase activity were generally consistent with the physiological process of sugar nutrient absorption and utilization from mulberry leaves in 5 th-instar silkworms.It suggests that BmSuc1,as a sucrose hydrolase gene,plays a major role in the digestion and absorption of sucrose nutrients from mulberry leaves in the midgut tissue.
基金supported by the National Key R&D Program of China(2022YFD1200400)the National Natural Science Foundation of China(32301851)。
文摘The pathogenesis-related protein PR10 plays a vital role in plant growth,development,and stress responses.This study systematically identified and analyzed PR10 genes in cultivated peanut(Arachis hypogaea L.),examining their phylogenetic relationships,conserved motifs,gene structures,and syntenic relationships.The analysis identified 54 Ah PR10 genes,which were classified into eight groups based on phylogenetic relationships,supported by gene structure and conserved motif characterization.Analysis of chromosomal distribution and synteny demonstrated that segmental duplications played a crucial role in the expansion of the Ah PR10 gene family.The identified Ah PR10 genes exhibited both constitutive and inducible expression patterns.Significantly,Ah PR10-7,Ah PR10-33,and Ah PR10-41 demonstrated potential importance in peanut resistance to Aspergillus flavus.In vitro fungistatic experiments demonstrated that recombinant Ah PR10-33 effectively inhibited A.flavus mycelial growth.These findings provide valuable insights for future investigations into Ah PR10 functions in protecting peanut from A.flavus infection.
基金supported by the National Natural Science Foundation of China(82427808,61875085)the Jiangsu Provincial University Natural Science Foundation(25KJB413004)+1 种基金the Nanjing Health Science and Technology Development Foundation(ZKX24043)the Fundamental Research Funds for the Central Universities(NJ2024029).
文摘Microwave ablation(MWA)is a minimally invasive technique for treating hepatic tumors,necessitating precise monitoring to ensure treatment efficacy and minimize damage to surrounding tissues.This study explores the potential of photoacoustic imaging(PAI)in monitoring MWA by examining ex vivo porcine liver tissues.In this study,a comprehensive analysis of photoacoustic signals was performed to compare the main lobe width(MLW)between ablated and normal regions in porcine liver tissue.Histological staining with succinate dehydrogenase(SDH)and shear wave elastography(SWE)were employed to validate the changes in tissue elasticity after ablation.The analysis demonstrated a notable reduction in the MLW of the average A-lines in ablated tissues compared to nonablated regions(p<0.01).This reduction,attributed to increased tissue density and enhanced elasticity,indicates accelerated sound propagation in thermally ablated areas,which then serves as a critical parameter for mapping tissue characteristics.The reconstruction of the MLW distribution successfully delineated the ablated regions,and was consistent with the results of SDH staining and SWE.In addition,MLW-based imaging exhibited higher spatial resolution compared to SWE.Incorporating MLW analysis into PAI may be a promising strategy to improve the accuracy and effectiveness of MWA monitoring in clinical settings.
基金Supported by the Natural Science Foundation of Shandong Province(No.ZR2021QD110)the National Natural Science Foundation of China(No.42106128)。
文摘Due to the unique microstructure and diverse opsin genes of the trinocular compound eye,stomatopoda possess an extraordinary ability to perceive multiple properties of light.They not only can detect natural light(NL)and linearly polarized light(LPL),but also are the only animals capable of recognizing circularly polarized light(CPL).Here,we integrated single-cell RNA sequencing,previously published Illumina data,and in-situ hybridization(ISH)to quantify and localize functional opsin genes in Oratosquilla oratoria,a common stomatopoda species in the China Sea.A total of high-quality 31777 cells were captured for the first time in the O.oratoria compound eye,which were classified into 25 cell subpopulations,and hypothesized that cluster 22 is a critical cell subpopulation responsible for light(whether NL,LPL,or CPL)response in O.oratoria.Furthermore,we propose that the long-wavelengthsensitive opsin gene(lws)gene family,retinol dehydrogenase(rdh),voltage-gated ion channel(vgic),arrestin(arr),and myosin(myo)collectively mediate the light response in O.oratoria.Considering that very few vision-related opsin genes show differential expression in right-handed CPL(RCPL)-vs.-dark(DL),which provides additional evidence that stomatopoda cannot recognize RCPL.Meanwhile,we believe that UV-stimulated scaffold protein A(uvssa)and red pigment concentrating hormone(rpch)play special contributions in the left-handed CPL(LCPL)environment response.ISH revealing that 16 lws,6 middle-wavelength-sensitive(mws),and 2 ultraviolet(uv)opsin genes were expressed in the photoreceptors of the O.oratoria compound eye.Although the inability to determine the functional types of cell subpopulations limits the resolution of opsin genes,these findings systematically elucidate the specific expression patterns of opsin genes in O.oratoria and represent a significant step toward refining the visual ecological theory of O.oratoria and other stomatopod species.
文摘While methodology for determining the mode of evolution in coding sequences has been well established,evaluation of adaptation events in emerging types of phenotype data needs further development.Here,we propose an analysis framework(expression variance decomposition,EVaDe)for comparative single-cell expression data based on phenotypic evolution theory.After decomposing the gene expression variance into separate components,we use two strategies to identify genes exhibiting large between-taxon expression divergence and small within-cell-type expression noise in certain cell types,attributing this pattern to putative adaptive evolution.In a dataset of primate prefrontal cortex,we find that such humanspecific key genes enrich with neurodevelopment-related functions,while most other genes exhibit neutral evolution patterns.Specific neuron types are found to harbor more of these key genes than other cell types,thus likely to have experienced more extensive adaptation.Reassuringly,at the molecular sequence level,the key genes are significantly associated with the rapidly evolving conserved non-coding elements.An additional case analysis comparing the naked mole-rat(NMR)with the mouse suggests that innateimmunity-related genes and cell types have undergone putative expression adaptation in NMR.Overall,the EVaDe framework may effectively probe adaptive evolution mode in single-cell expression data.
基金supported by the Portuguese Foundation for Science and Technology(FCT),Centro 2020 and Portugol2020 and the EU FEDER program,via the project GoBack to SIV(PTDC/CVT-CVT/32261/2017,CENTRO-01-0145-FEDER-032261)the doctoral grants of PDC(SFRH/BD/139974/2018)and BMS(2020.06525.BD and DOI 10.54499/2020.06525.BD)+5 种基金the post-doctoral grant to JPF(SFRH/BPD/113359/2015-program-contract described in paragraphs 4,5,6 of art.23 of Law no.100157/2016,of August 29,as amended by Law no.57/2017 of July 2019),the project PTDC/MED-NEU/1677/2021 to JBRthe Institute of Biomedicine iBiMED(UIDB/04501/2020 and DOI 10.54499/UIDB/04501/2020,UIDP/04501/2020 and DOI 10.54499/UIDP/04501/2020)its LiM Bioimaging Facility-a PPBI node(POCI-01-0145-FEDER-022122)supported by the Research Commission of the Medical Faculty of the Heinrich-Heine-University(HHU)Düsseldorf,of the Biologisch-Medizinisches Forschungszentrum(BMFZ)of HHUfinanced by the Spanish"Plan Nacional de Investigacion Cientifica,Desarrollo e Innovacion Tecnologica,Ministerio de Economia y Competitividad(Instituto de Salud CarlosⅢ)",co-financed by the European Union(FEDER program),(grant FIS P/20/00318 and FIS P23/00337 to VC)grant CPP2021-009070 to VC by the"Proyectos de colaboracion publico-privada,Plan de Investigacion Cientifica,Tecnica y de inovacion 2021-2023,Ministerio de Ciencia e Innovacion,Union Europea,Agencia Estatal de Investigacion,Espana"。
文摘Contrary to the adult central nervous system,the peripheral nervous system has an intrinsic ability to regenerate that relies on the expression of regenerationassociated genes,such as some kinesin family members.Kinesins contribute to nerve regeneration through the transport of specific cargo,such as proteins and membrane components,from the cell body towards the axon periphery.We show here that KIF4A,associated with neurodevelopmental disorders and previously believed to be only expressed during development,is also expressed in the adult vertebrate nervous system and up-regulated in injured peripheral nervous system cells.KIF4A is detected both in the cell bodies and regrowing axons of injured neurons,consistent with its function as an axonal transporter of cargoes such asβ1-integrin and L1CAM.Our study further demonstrates that KIF4A levels are greatly increased in Schwann cells from injured distal nerve stumps,particularly at a time when they are reprogrammed into an essential proliferative repair phenotype.Moreover,Kif4a m RNA levels were approximately~6-fold higher in proliferative cultured Schwann cells compared with non-proliferative ones.A hypothesized function for Kif4a in Schwann cell proliferation was further confirmed by Kif4a knockdown,as this significantly reduced Schwann cell proliferation in vitro.Our findings show that KIF4A is expressed in adult vertebrate nervous systems and is up-regulated following peripheral injury.The timing of KIF4A up-regulation,its location during regeneration,and its proliferative role,all suggest a dual role for this protein in neuroregeneration that is worth exploring in the future.
基金supported by FWO(Fonds voor Wetenschappelijk Onderzoek),grant number G07562NFWO(to BB)。
文摘Neuroinflammation is a key process in the pathogenesis of various neurodegenerative diseases,such as multiple sclerosis(MS),Alzheimer's disease,and traumatic brain injury.Even for disorders historically unrelated to neuroinflammation,such as Alzheimer's disease,it is now shown to precede pathological protein aggregations.
