Cyanidin-3-glucoside protects the photooxidative damage of retinal pigment epithelium cells by regulating sphingolipid signaling and inhibiting MAPK pathway

Cyanidin-3-glucoside(C3G)is the most common anthocyanin in dark grains and berries and is a food functional factor to improve visual health.However,the mechanisms of C3G on blue light-induced retinal pigment epithelial(RPE)cell photooxidative damage needs further exploration.We investigated the effects of C3G on blue light-irradiated A2E-containing RPE cells and explored whether sphingolipid,mitogen-activated protein kinase(MAPK),and mitochondria-mediated pathways are involved in this mechanism.Blue light irradiation led to mitochondria and lysosome damage in RPE cells,whereas C3G preserved mitochondrial morphology and function and maintained the lysosomal integrity.C3G suppressed the phosphorylation of JNK and p38 MAPK and mitochondria-mediated pathways to inhibit RPE cell apoptosis.Lipidomics data showed that C3G protected RPE cells against blue light-induced lipid peroxidation and apoptosis by maintaining sphingolipids balance.C3G significantly inhibited ceramide(Cer d18:0/15:0,Cer d18:0/16:0 and Cer d18:0/18:0)accumulation and elevated galactosylceramide(GalCer d18:1/15:0 and GalCer d18:1/16:0)levels in the irradiated A2E-containing RPE cells.Furthermore,C3G attenuated cell membrane damage by increasing phosphatidylcholine and phosphatidylserine levels.C3G inhibited apoptosis and preserved the structure of mitochondria and lysosome by regulating sphingolipid signaling and suppression of MAPK activation in RPE cells.Thus,dietary supplementation of C3G prevents retinal photooxidative damage.

Lactobacillus plantarum AR113 attenuates liver injury in D-galactose-induced aging mice via the inhibition of oxidative stress and endoplasmic reticulum stress

Probiotics could effectively eliminate excess reactive oxygen species(ROS)generated during aging or lipid metabolism disorders,but their mechanism is unclear.The major purpose of this study was to investigate the mechanism of Lactiplantibacillus plantarun AR113 alleviating oxidative stress injury in the D-galactose induced aging mice.The result showed that pretreatment with L.plantarun AR113 significantly relieving H_(2)O_(2)induced cytotoxicity in HepG2 cells by maintain cell membrane integrity and increasing antioxidant enzyme activities.In D-galactose induced aging mice,L.plantarun AR113 could significantly attenuate liver damage and inflammatory infiltration by promoting endogenous glutathione(GSH)synthesis and activating the Nrf2/Keap1 signaling pathway in mice,and increasing the expression of regulated phaseⅡdetoxification enzymes and antioxidant enzymes.Further analysis shown that gavage of L.plantarun AR113 could significantly reduce the expression of G protein-coupled receptor 78(GPR78)and C/EBP homologous protein(CHOP)proteins,and promote the restoration of endoplasmic reticulum(ER)homeostasis,thereby activating cell anti-apoptotic pathways.These results were also confirmed in H_(2)O_(2)-treated HepG2 experiments.It indicated that L.plantarun AR113 could inhibit D-galactose-induced liver injury through dual inhibition of ER stress and oxidative stress.L.plantarun AR113 have good application potential in anti-aging and alleviating metabolic disorders.

Lactiplantibacillus plantarum AR113 alleviates microbiota dysbiosis of tongue coating and cerebral ischemia/reperfusion injury in rat

Stroke is one of the leading causes of death and disability worldwide.However,information on stroke-related tongue coating microbiome(TCM)is limited,and whether TCM modulation could benefit for stroke prevention and rehabilitation is unknown.Here,TCM from stroke patients(SP)was characterized using molecular techniques.The occurrence of stroke resulted in TCM dysbiosis with significantly reduced species richness and diversity.The abundance of Prevotella,Leptotrichia,Actinomyces,Alloprevotella,Haemophilus,and TM7_[G-1]were greatly reduced,but common infection Streptococcus and Pseudomonas were remarkably increased.Furthermore,an antioxidative probiotic Lactiplantibacillus plantarum AR113 was used for TCM intervention in stroke rats with cerebral ischemia/reperfusion(I/R).AR113 partly restored I/R induced change of TCM and gut microbiota with significantly improved neurological deficit,relieved histopathologic change,increased activities of antioxidant enzymes,and decreased contents of oxidative stress biomarkers.Moreover,the gene expression of antioxidant-related proteins and apoptosis-related factors heme oxygenase-1(HO-1),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),nuclear factor erythroid 2-related factor 2(Nrf2),NAD(P)H:quinone oxidoreductase-1(NQO-1),and Bcl-2 was significantly increased,but cytochrome C,cleaved caspase-3,and Bax were markedly decreased in the brain by AR113 treatment.The results suggested that AR113 could ameliorate cerebral I/R injury through antioxidation and anti-apoptosis pathways,and AR113 intervention of TCM may have the application potential for stroke prevention and control.

Microbiomics and metabolomics insights into the microbial regulation on the formation of flavor components in the traditional fermentation process of Chinese Hongqu aged vinegar

This study aimed to investigate microbial succession and metabolic dynamics during the traditional fermentation of Hongqu aged vinegar,and explore the core functional microbes closely related to the formation of flavor components.Microbiome analysis demonstrated that Lactobacillus,Acetobacter,Bacillus,Enterobacter,Lactococcus,Leuconostoc and Weissella were the predominant bacterial genera,while Aspergillus piperis,Aspergillus oryzae,Monascus purpureus,Candida athensensis,C.xylopsoci,Penicillium ochrosalmoneum and Simplicillium aogashimaense were the predominant fungal species.Correlation analysis revealed that Acetobacter was positively correlated with the production of tetramethylpyrazine,acetoin and acetic acid,Lactococcus showed positive correlation with the production of 2-nonanone,2-heptanone,ethyl caprylate,ethyl caprate,1-hexanol,1-octanol and 1-octen-3-ol,C.xylopsoci and C.rugosa were positively associated with the production of diethyl malonate,2,3-butanediyl diacetate,acetoin,benzaldehyde and tetramethylpyrazine.Correspondingly,non-volatile metabolites were also detected through ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry.A variety of amino acids and functional dipeptides were identified during the traditional brewing of Hongqu aged vinegar.Correlation analysis revealed that Lactobacillus was significantly associated with DL-lactate,indolelactic acid,D-(+)-3-phenyllactic acid,pimelic acid,pregabalin and 3-aminobutanoic acid.This study is useful for understanding flavor formation mechanism and developing effective strategies for the suitable strains selection to improve the flavor quality of Hongqu aged vinegar.

Milling degree affects the fermentation properties of rice:perspectives from the composition of nutrients and gut microbiota via in vitro fermentation

Fermentation substrates of rice with different milling degrees(MDs) were prepared and fermented with human feces to compare their fermentation properties and effects on gut microbiota.MD 0s,MD 5s and MD 60s represented brown rice,moderately-milled rice and white rice,respectively.After in vitro fermentation,the MD 5s group showed higher starch utilization,compared with the MD 0s and 60s groups evaluated by Fourier transform infrared spectrometer,and confocal laser scanning microscope.Effects of fermentation substrates of rice with different MDs on gut microbiota were evaluated by 16S rDNA sequencing.All the sample groups reduced the pH and produced short-chain fatty acids(SCFAs) and branched-chain fatty acids.The MD 5s group exhibited higher α-diversity than the MD 0s and 60s groups.Abundances of Phascolarctobacterium,Blautia and norank_f_Ruminococcaceae were higher in the MD 0s and 5s groups,compared with the MD 60s group.These bacteria were also positively correlated with the SCFAs production via Spearman correlation analysis.In vitro culture assay revealed that fermentation substrates of MD 0s and 5s promoted the growth of two probiotics(Akkermansia muciniphila and Bifidobacterium adolescentis).Our results showed that moderate milling might be an appropriate way to produce rice products with richer nutrients and better fermentation properties.

Proliferative potential and angiogenic characteristics of blood outgrowth endothelial cells derived from middle-aged and older adults

Objectives Autologous blood outgrowth endothelial cells(BOECs)have been proposed to induce therapeutic angiogenesis for treating cardiovascular diseases(CVDs).The aim of the present study was to investigate the proliferative potential and angiogenic characteristics of BOECs among middle-aged and older adults,the population particularly susceptible to CVDs.Methods BOECs were isolated from 48 peripheral blood samples of subjects aged 56±4 years.The cells were then distinguished based on their proliferative abilities,and their phenotype,tube formation capacity,and migratory activity were compared using immunofluorescence staining,flow cytometry,tube formation assay,and wound healing assay,respectively.Correlations between demographic,clinical,and dietary parameters with the number of BOECs were also assessed.Results A total of 132 BOEC colonies with different proliferative potentials were obtained,including colonies lost proliferative ability before passage 3(named LPA),stopped proliferating during passage 3–8(HPA(3–8)),and proliferated after passage 8(HPA(>8)).LPA cells appeared later and displayed abnormal morphology,while HPA(3–8)cells exhibited alterations in von Willebrand factor morphology and lower KDR expression.HPA(>8)cells obtained higher branching intervals and individual cell migration velocity compared with those of HPA(3–8)cells.Correlation analysis showed that the number of both LPA and HPA colonies were positively associated with several CVD risk factors.Additionally,the number of LPA colonies was positively associated with servings of meats and alternatives,fruits,fruits and vegetables,as well as the protein intake.Conclusions Our findings provide evidence that the middle-aged and older populations possess BOECs with different proliferative and angiogenic potentials,exhibiting distinctions in cell morphology,appearance dates,VWF morphology,and KDR expression.Strikingly,a higher number of BOECs is likely associated with an increased risk of CVDs,while the number of BOECs with low proliferative ability may be regulated by diet.

Effect of β-Glucan (Angel Yeast) Compared to a Placebo on Cold and Flu Incidence and Symptoms in an Adult Population—A Double Blind, Randomised Controlled Trial

Background: 1-3, 1-6 β-glucan derived from Baker’s yeast (Saccharomyces cerevisiae) has been widely studied for its immune stimulatory capabilities and safety. Previous studies found β-glucan to have efficacy at reducing incidence of URTIs as well as being a low risk for negative side effects. The current study aimed to examine the effects of yeast β-glucan (Angel Yeast) on cold and flu incidences and symptoms in healthy adults. Methods: Two hundred and thirty-one males and females aged 18 to 65 years old supplemented with either β-glucan or a placebo for 3-months. Participants completed a general health questionnaire every 4 weeks and in addition, if participants experienced any cold or flu symptoms, these were recorded daily (along with severity) until resolved or up to 2 weeks. Results: Supplementation with β-glucan reduced the self-reported severity of sore throats and improved sleep quality compared to the placebo group. Conclusions: Yeast β-glucan supplementation appears to be able to help reduce certain symptoms experienced during a cold or flu episode and is safe and well tolerated.

A Novel Defibrillator-Specific Coprocessor Capable of Running Entropy and CNN Integration Algorithms

It is difficult for the existing Automated External Defibrillator (AED) on-board microprocessors to accurately classify electrocardiographic signals (ECGs) mixed with Cardiopulmonary Resuscitation artifacts in real-time. In order to improve recognition speed and accuracy of electrocardiographic signals containing Cardiopulmonary Resuscitation artifacts, a new special coprocessor system-on-chip (SoC) for defibrillators was designed. In this study, a microprocessor was designed based on the RISC-V architecture to achieve hardware acceleration for ECGs classification;Besides, an Approximate Entropy (ApEn) and Convolutional neural networks (CNNs) integrated algorithm capable of running on it was designed. The algorithm differs from traditional electrocardiographic (ECG) classification algorithms. It can be used to perform ECG classification while chest compressions are applied. The proposed co-processor can be used to accelerate computation rate of ApEn by 34 times compared with pure software computation. It can also be used to accelerate the speed of CNNs ECG recognition by 33 times. The combined algorithm was used to classify ECGs with CPR artifacts. It achieved a precision of 96%, which was significantly superior to that of simple CNNs. The coprocessor can be used to significantly improve the recognition efficiency and accuracy of ECGs containing CPR artifacts. It is suitable for automatic external defibrillator and other medical devices in which one-dimensional physiological signals.

Simulation and Experimental Study of Staple Line Reinforcement Surgery

The aim of this study was to evaluate the effectiveness of BM (basement membrane) and SIS (small intestine submucosa) composite extracellular matrix staple line reinforcement in surgical procedures through finite element modelling simulations and leak-proof performance experiments. The mechanical analyses of soft tissues with and without staple line reinforcement were performed by establishing finite element models of three tissues, namely, stomach, intestine and lungs, under the use scenarios of different anastomosis staple models;and the leak-proof performance of the staple line reinforcement was evaluated by simulating leak-proof experiments of gastric incision margins, intestinal sections, and lung incision margins in vitro. The results showed that the equivalent average stresses of the staple line reinforcement were increased by 20 kPa-68 kPa in gastric and intestinal tissues, and 8 kPa-22 kPa in lung tissues. and that the BM and SIS composite extracellular matrix staple line reinforcement could strengthen the anastomotic structure, and at the same time disperse the high stresses of the anastomosed tissues, which could effectively reduce the postoperative complications such as anastomotic bleeding and anastomotic leakage, and provide a safer and more effective optimized design for surgical mechanical anastomosis. It can effectively reduce postoperative complications such as anastomotic bleeding and anastomotic leakage, and provide a safer and more effective optimized design for surgical mechanical anastomosis.

Development and Innovation of Modern Microvascular Anastomoses

High-quality microvascular anastomosis is the foundation of successful microsurgery and one of the most important basic skills for microsurgeons. The traditional manual suture is recognized as the “gold standard” for microvascular anastomosis, but it still has problems such as long operation time and easy to cause vascular damage. In order to improve the success rate of microvascular anastomosis, reduce surgical complications and make the prognosis of patients better. In order to improve the success rate of microvascular anastomosis and reduce the surgical complications, scholars at home and abroad have developed some new vascular anastomosis techniques that are simple, fast and minimally invasive while improving the traditional surgical suturing methods. In this paper, we review the microvascular anastomosis, and its application research in two methods of traditional hand suture and non-suture anastomosis, in order to promote the application development of microvascular anastomosis.

Nanomaterials-mediated lysosomal regulation:a robust protein-clearance approach for the treatment of Alzheimer’s disease

Alzheimer’s disease is a debilitating,progressive neurodegenerative disorder characterized by the progressive accumulation of abnormal proteins,including amyloid plaques and intracellular tau tangles,primarily within the brain.Lysosomes,crucial intracellular organelles responsible for protein degradation,play a key role in maintaining cellular homeostasis.Some studies have suggested a link between the dysregulation of the lysosomal system and pathogenesis of neurodegenerative diseases,including Alzheimer’s disease.Restoring the normal physiological function of lysosomes hold the potential to reduce the pathological burden and improve the symptoms of Alzheimer’s disease.Currently,the efficacy of drugs in treating Alzheimer’s disease is limited,with major challenges in drug delivery efficiency and targeting.Recently,nanomaterials have gained widespread use in Alzheimer’s disease drug research owing to their favorable physical and chemical properties.This review aims to provide a comprehensive overview of recent advances in using nanomaterials(polymeric nanomaterials,nanoemulsions,and carbon-based nanomaterials)to enhance lysosomal function in treating Alzheimer’s disease.This review also explores new concepts and potential therapeutic strategies for Alzheimer’s disease through the integration of nanomaterials and modulation of lysosomal function.In conclusion,this review emphasizes the potential of nanomaterials in modulating lysosomal function to improve the pathological features of Alzheimer’s disease.The application of nanotechnology to the development of Alzheimer’s disease drugs brings new ideas and approaches for future treatment of this disease.

Design and performance of a high-speed and low-noise preamplifier for SiPM

Considering the R&D for upgrading the K^(0)_(L) andμdetectors in the Belle II experiment using a scintillator and silicon pho-tomultiplier(SiPM),we designed a compact high-speed and low-noise preamplifier.The preamplifier demonstrated a good gain stability,bandwidth of 426 MHz,baseline noise level ofσ≈0.6 mV,dynamic range of up to170 mV of the input signal amplitude,good time resolution of 20 ps,and it can be comprehensively applied to SiPMs.Adopting pole-zero-cancelation in the preamplifier reduces both the rise and fall times of the SiPM signal,which can significantly improve the time resolution and reduce the pile-up when using a large SiPM or an array of SiPMs.Various combinations of the preamplifier and several types of SiPMs demonstrated time resolutions better than 50 ps for most cases;when the number of detected photons was larger than 60,a time resolution of approximately 25 ps was achieved.

Role of CD36 in central nervous system diseases

CD36 is a highly glycosylated integral membrane protein that belongs to the scavenger receptor class B family and regulates the pathological progress of metabolic diseases.CD36 was recently found to be widely expressed in various cell types in the nervous system,including endothelial cells,pericytes,astrocytes,and microglia.CD36 mediates a number of regulatory processes,such as endothelial dysfunction,oxidative stress,mitochondrial dysfunction,and inflammatory responses,which are involved in many central nervous system diseases,such as stroke,Alzheimer’s disease,Parkinson’s disease,and spinal cord injury.CD36 antagonists can suppress CD36 expression or prevent CD36 binding to its ligand,thereby achieving inhibition of CD36-mediated pathways or functions.Here,we reviewed the mechanisms of action of CD36 antagonists,such as Salvianolic acid B,tanshinone IIA,curcumin,sulfosuccinimidyl oleate,antioxidants,and small-molecule compounds.Moreover,we predicted the structures of binding sites between CD36 and antagonists.These sites can provide targets for more efficient and safer CD36 antagonists for the treatment of central nervous system diseases.

Rolling velocity and relative motion of particle detector in local granular flow

The velocity of a particle detector in granular flow can be regarded as the combination of rolling and sliding velocities.The study of the contribution of rolling velocity and sliding velocity provides a new explanation to the relative motion between the detector and the local granular flow.In this study,a spherical detector using embedded inertial navigation technology is placed in the chute granular flow to study the movement of the detector relative to the granular flow.It is shown by particle image velocimetry(PIV)that the velocity of chute granular flow conforms to Silbert’s formula.And the velocity of the detector is greater than that of the granular flow around it.By decomposing the velocity into sliding and rolling velocity,it is indicated that the movement of the detector relative to the granular flow is mainly caused by rolling.The rolling detail shown by DEM simulation leads to two potential mechanisms based on the position and drive of the detector.

Computational Simulation of Aptamer-target Binding Mechanisms

Aptamers are a type of single-chain oligonucleotide that can combine with a specific target.Due to their simple preparation,easy modification,stable structure and reusability,aptamers have been widely applied as biochemical sensors for medicine,food safety and environmental monitoring.However,there is little research on aptamer-target binding mechanisms,which limits their application and development.Computational simulation has gained much attention for revealing aptamer-target binding mechanisms at the atomic level.This work summarizes the main simulation methods used in the mechanistic analysis of aptamer-target complexes,the characteristics of binding between aptamers and different targets(metal ions,small organic molecules,biomacromolecules,cells,bacteria and viruses),the types of aptamer-target interactions and the factors influencing their strength.It provides a reference for further use of simulations in understanding aptamer-target binding mechanisms.

Lactobacillus plantarum AR495 improves stress-induced irritable bowel syndrome in rats by targeting gut microbiota and Mast cell-PAR2-TRPV1 signaling pathway

Probiotics have great potential in regulating intestinal pain.In this study,the effects of Lactobacillus plantarum AR495 on the visceral sensitivity and gut microbiota of irritable bowel syndrome(IBS)rats were studied.The results showed that tryptase released after mast cell activation and degranulation plays a key role in visceral pain,and L.plantarum AR495 reduced the stimulation of colonic mast cells and the expression of protease-activated receptor 2(PAR2)and TRPV1 in dorsal root ganglia.Research further showed that supplementation with L.plantarum AR495 increased the level of short-chain fatty acids(SCFAs)and enhanced the barrier function of the colon.In addition,the microbiota analysis of the colon indicated that L.plantarum AR495 promoted the proliferation of Bifidobacterium and inhibited the proliferation of Lachnospiraceae,which alleviated the imbalance of the intestinal microbiota caused by IBS to a certain extent.In total,L.plantarum AR495 might reduce visceral sensitivity through the Mast cell-PAR2-TRPV1 signaling pathway by maintaining the homeostasis of the intestinal barrier.

Nomogram prediction of vessels encapsulating tumor clusters in small hepatocellular carcinoma≤3 cm based on enhanced magnetic resonance imaging

BACKGROUND Vessels encapsulating tumor clusters(VETC)represent a recently discovered vascular pattern associated with novel metastasis mechanisms in hepatocellular carcinoma(HCC).However,it seems that no one have focused on predicting VETC status in small HCC(sHCC).This study aimed to develop a new nomogram for predicting VETC positivity using preoperative clinical data and image features in sHCC(≤3 cm)patients.AIM To construct a nomogram that combines preoperative clinical parameters and image features to predict patterns of VETC and evaluate the prognosis of sHCC patients.METHODS A total of 309 patients with sHCC,who underwent segmental resection and had their VETC status confirmed,were included in the study.These patients were recruited from three different hospitals:Hospital 1 contributed 177 patients for the training set,Hospital 2 provided 78 patients for the test set,and Hospital 3 provided 54 patients for the validation set.Independent predictors of VETC were identified through univariate and multivariate logistic analyses.These independent predictors were then used to construct a VETC prediction model for sHCC.The model’s performance was evaluated using the area under the curve(AUC),calibration curve,and clinical decision curve.Additionally,Kaplan-Meier survival analysis was performed to confirm whether the predicted VETC status by the model is associated with early recurrence,just as it is with the actual VETC status and early recurrence.RESULTS Alpha-fetoprotein_lg10,carbohydrate antigen 199,irregular shape,non-smooth margin,and arterial peritumoral enhancement were identified as independent predictors of VETC.The model incorporating these predictors demonstrated strong predictive performance.The AUC was 0.811 for the training set,0.800 for the test set,and 0.791 for the validation set.The calibration curve indicated that the predicted probability was consistent with the actual VETC status in all three sets.Furthermore,the decision curve analysis demonstrated the clinical benefits of our model for patients with sHCC.Finally,early recurrence was more likely to occur in the VETC-positive group compared to the VETC-negative group,regardless of whether considering the actual or predicted VETC status.CONCLUSION Our novel prediction model demonstrates strong performance in predicting VETC positivity in sHCC(≤3 cm)patients,and it holds potential for predicting early recurrence.This model equips clinicians with valuable information to make informed clinical treatment decisions.

Intelligent diagnosis of atrial septal defect in children using echocardiography with deep learning

Background Atrial septal defect(ASD)is one of the most common congenital heart diseases.The diagnosis of ASD via transthoracic echocardiography is subjective and time-consuming.Methods The objective of this study was to evaluate the feasibility and accuracy of automatic detection of ASD in children based on color Doppler echocardiographic static images using end-to-end convolutional neural networks.The proposed depthwise separable convolution model identifies ASDs with static color Doppler images in a standard view.Among the standard views,we selected two echocardiographic views,i.e.,the subcostal sagittal view of the atrium septum and the low parasternal four-chamber view.The developed ASD detection system was validated using a training set consisting of 396 echocardiographic images corresponding to 198 cases.Additionally,an independent test dataset of 112 images corresponding to 56 cases was used,including 101 cases with ASDs and 153 cases with normal hearts.Results The average area under the receiver operating characteristic curve,recall,precision,specificity,F1-score,and accuracy of the proposed ASD detection model were 91.99,80.00,82.22,87.50,79.57,and 83.04,respectively.Conclusions The proposed model can accurately and automatically identify ASD,providing a strong foundation for the intelligent diagnosis of congenital heart diseases.

Trend analysis of stroke risk research based on three-dimensional metrics

This study comprehensively analyzes the status,characteristics,focal points,and evolving trends of global research on“stroke risk analysis”over the past four years(2020–2023),aiming to provide insights for directing future research endeavors.By utilizing the Newcastle-Ottawa Scale,63 high-quality research papers were selected and subjected to a systematic literature review.In terms of research methods,stroke risk analysis research has evolved from clinical trials(e.g.,establishing control groups,using authoritative scales)towards statistical and data analysis methods(e.g.,decision tree analysis).Regarding research factors,early studies primarily focused on pathological factors associated with hemorrhagic and ischemic stroke,such as hypertension,hyperlipidemia,and diabetes.Recent research from the past two years indicates a shift towards emerging factors,including temperature conditions,air quality,and Corona Virus Disease 2019(COVID-19).In terms of application domains,stroke research covers a broad range of fields but mainly focuses on exploring risk factors,interventions during diagnosis and treatment stages,and rehabilitation,with clinical diagnosis,treatment,and drug intervention studies being predominant.While the research landscape is becoming increasingly diversified and comprehensive,there remains a need for more comprehensive and in-depth studies on novel topics,as well as integrated applications of research methods,presenting ample opportunities for exploring dependent variables in future stroke.

The protective effect of oleic acid on Lactiplantibacillus plantarum during freeze-drying

Membrane characteristics are determined by the fatty acids composition,which affects survival rates after freeze-drying.However,it is unknown which composition provides the greatest effect.In this study,we found that the addition of Tween 80 and Tween 20 significantly increased survival rates of Lactiplantibacillus plantarum,which reached a maximum of 93.1%.Conversely,Tween 60 caused a significant decrease.We further found that the difference between the effects of adding Tween 80 and Tween 60 was the change in oleic acid contents.To verify the role of oleic acid,we used CRISPR-Cas9 to knock-out the key synthesis gene cla-er.The survival rates of L.plantarum AR113Δcla-er declined to 5.48%.The addition of oleic acid restored the rates to those of wild type strains.Moreover,the membrane integrity and fl uidity of knockout strains markedly decreased.This is the fi rst confi rmation that Tween 80 or Tween 20 increases the survival rate by increasing the content of oleic acid in the cell membrane.These fi ndings also indicated that oleic acid in cell membranes has a substantial protective effect on L.plantarum during freeze-drying.