Recent progress and future directions of the research on nanoplastic-induced neurotoxicity

Many types of plastic products,including polystyrene,have long been used in commercial and industrial applications.Microplastics and nanoplastics,plastic particles derived from these plastic products,are emerging as environmental pollutants that can pose health risks to a wide variety of living organisms,including humans.However,it is not well understood how microplastics and nanoplastics affect cellular functions and induce stress responses.Humans can be exposed to polystyrene-microplastics and polystyrene-nanoplastics through ingestion,inhalation,or skin contact.Most ingested plastics are excreted from the body,but inhaled plastics may accumulate in the lungs and can even reach the brain via the nose-to-brain route.Small-sized polystyrene-nanoplastics can enter cells by endocytosis,accumulate in the cytoplasm,and cause various cellular stresses,such as inflammation with increased pro-inflammatory cytokine production,oxidative stress with generation of reactive oxygen species,and mitochondrial dysfunction.They induce autophagy activation and autophagosome formation,but autophagic flux may be impaired due to lysosomal dysfunction.Unless permanently exposed to polystyrene-nanoplastics,they can be removed from cells by exocytosis and subsequently restore cellular function.However,neurons are very susceptible to this type of stress,thus even acute exposure can lead to neurodegeneration without recovery.This review focuses specifically on recent advances in research on polystyrene-nanoplastic-induced cytotoxicity and neurotoxicity.Furthermore,in this review,based on mechanistic studies of polystyrene-nanoplastics at the cellular level other than neurons,future directions for overcoming the negative effects of polystyrene-nanoplastics on neurons were suggested.

Overstory functional groups indicate the legacy of land use in a secondary tropical forest in southwestern China

Anthropogenic disturbances are widespread in tropical forests and influence the species composition in the overstory.However,the impacts of historical disturbance on tropical forest overstory recovery are unclear due to a lack of disturbance data,and previous studies have focused on understory species.In this study,the purpose was to deter-mine the influence of historical disturbance on the diver-sity,composition and regeneration of overstory species in present forests.In the 20-ha Xishuangbanna tropical sea-sonal rainforest dynamics plot in southwestern China,the historical disturbance boundaries were delineated based on panchromatic photographs from 1965.Factors that drove species clustering in the overstory layer(DBH≥40 cm)were analyzed and the abundance,richness and composition of these species were compared among different tree groups based on multiple regression tree analysis.The coefficient of variation of the brightness value in historical panchro-matic photographs from 1965 was the primary driver of spe-cies clustering in the overstory layer.The abundance and richness of overstory species throughout the regeneration process were similar,but species composition was always different.Although the proportion of large-seeded and vigorous-sprouting species showed no significant differ-ence between disturbed and undisturbed forests in the tree-let layer(DBH<20 cm),the difference became significant when DBH increased.The findings highlight that historical disturbances have strong legacy effects on functional group composition in the overstory and the recovery of overstory species was multidimensional.Functional group composi-tion can better indicate the dynamics of overstory species replacement during secondary succession.

Multidimensional evaluation of salt tolerance in groundnut genotypes through biochemical responses

The manuscript explores the complex interplay between groundnut genotypes,salt tolerance and hormonal influence,shedding light on the dynamic responses of three specific groundnut genotypes,KDG-128,TG-37 A and GG-20,to salt treatments and gibberellic acid(GA3).The study encompasses germination,plant growth,total protein content and oil content as key parameters.Through comprehensive analysis,it identifies TG-37 A and KDG-128 as salt-tolerant genotypes,and GG-20 as salt-susceptible genotypes,which highlighting the potential for targeted breeding efforts to develop more resilient groundnut varieties.Moreover,the quantification of protein and oil content under different treatments provides vital data for optimizing nutritional profiles in groundnut cultivars.Principal Component Analysis(PCA) underscores the significance of the first principal component(PC1)in explaining the majority of variance,capturing primary trends and differences in plant length.Analysis of Variance(ANOVA) and hierarchical analysis confirm the presence of statistically significant differences in protein and oil content among the genotypes.Pearson's correlation coefficient matrix analysis reveals strong positive correlations between plant length and protein content,plant length and oil content,and a moderately positive correlation between protein content and oil content.These findings provide valuable insights into groundnut physiology,salt tolerance,and nutritional composition,with implications for future research in sustainable agriculture and crop improvement.

Tuning beneficial calcineurin phosphatase activation to counterα-synuclein toxicity in a yeast model of Parkinson’s disease

Calcineurin(CN)is a calcium-and calmodulindependent serine/threonine that has been studied in many model organisms including yeast,filamentous fungi,plants,and mammals.Its biological functions range from ion homeostasis and virulence in lower eukaryotes to T-cell activation in humans by human nuclear factors of activated T-cells.CN is a heterodimeric protein consisting of a catalytic subunit,calcineurin A(Cna1p),which contains an active site with a dinuclear metal center,and a regulatory Ca^(2+) binding subunit called calcineurin B(Cnb1p)required to activate Cna1p.The calcineurin B subunit has been highly conserved through evolution:For example,the mammalian calcineurin B shows 54%identity with calcineurin B from Saccharomyces cerevisiae.

Isolation of a feline-derived feline panleukopenia virus with an A300P substitution in the VP2 protein and confrmation of its pathogenicity in dogs

Feline panleukopenia virus(FPV)is a single-stranded DNA virus that can infect cats and cause feline panleukopenia,which is a highly contagious and fatal disease in felines.The sequence of FPV is highly variable,and mutations in the amino acids of its capsid protein play crucial roles in altering viral virulence,immunogenicity,host selection,and other abilities.In this study,the epidemiology of FPV was studied using 746 gastrointestinal swab samples derived from cats that presented gastrointestinal symptoms specifcally,diarrhea or vomiting during the period spanning from 2018 to 2022.The overall prevalence of FPV-positive patients among these samples was determined to be 45.4%.Capsid(virion)protein 2(VP2)gene of each FPV-positive sample was sequenced and amplifed,yielding 65 VP2 sequences.Among them,six VP2 gene sequences were detected in the majority of the samples test positive for FPV,and these positive samples originated from a diverse range of geographical locations.These isolates were named FPV-6,FPV-10,FPV-15,FPV-251,FPV-271 and FPV-S2.Additionally,the substitution of Ala300Pro(A300P)in VP2 was detected for the frst time in feline-derived FPV(FPV-251).FPV-251 isolate,with this substitution in VP2 protein,exhibited stable proliferative capacity in Madin-Darby canine kidney(MDCK)cells and A72 cells.FPV-271 was selected as the FPV control isolate due to its single amino acid diference from VP2 protein of FPV-251 at position 300(FPV-271 has alanine,while FPV-251 has proline).After oral infection,both FPV-251 and FPV-271 isolates caused feline panleukopenia,which is characterized by clinical signs of enterocolitis.However,FPV-251 can infect dogs through the oral route and cause gastrointestinal(GI)symptoms with lesions in the intestine and mesenteric lymph nodes(MLNs)of infected dogs.This is the frst report on the presence of an A300P substitution in VP2 protein of feline-derived FPV.Additionally,FPV isolate with a substitution of A300P at VP2 protein demonstrated efcient replication capabilities in canine cell lines and the ability to infect dogs.

Asexual Reproduction of Aurelia coerulea Polyps in situ on Settling Plates in Jiaozhou Bay,China

Jellyfish blooms are increasingly prevalent coastal phenomena,with Aurelia coerulea being a significant contributor.The complex life history of jellyfish,featuring a sessile polyp stage,is a key factor in bloom formation,yet it is challenging to study in the field.This study aimed to describe the asexual reproduction of A.coerulea polyps in Jiaozhou Bay,China,and to assess their ability to form sedentary colonies and their relationship with environmental factors.Using settling plates,we observed polyps in situ and documented three asexual reproduction modes:budding,strobilation,and podocyst formation.Environmental parameters including temperature,salinity,dissolved oxygen(DO),chlorophyll a(Chl a),and the biomass of phytoplankton and zooplankton were monitored.Budding was the predominant reproductive method,featuring stolon budding in autumn and direct budding during the winter and spring seasons.Strobilation took place in the colder months,releasing ephyrae between April and May.Polyp abundance was positively correlated with seawater temperature and negatively correlated with DO concentration.Polyps at 1.0 m depth showed higher survivorship and proliferation over 8 months,whereas those at 2.0 m depth experienced a steady decline and died within 4 months.The study enhances our understanding of the life cycle and asexual reproduction strategies of A.coerulea polyps in the field,with implications for predicting and managing jellyfish blooms.The findings underscore the importance of temperature and DO in polyp survival and suggest that initial polyp population proliferation is crucial for long-term survival.

Overview on signal transduction cascades regulation roles of garlic and its bioactive constituents

High-throughput technologies in combination with modern exciting advancements in mass spectrometry-based proteomics and data analysis pipelines have empowered comprehensive characterization of disease phenotypes and their mechanistic regulation by dietary agents and bioactive molecules at unprecedented dimensionality and resolution.Extra-ordinary breakthroughs in the field of nutrigenomics have leveraged our understanding altogether to a new level of maturity.Interdisciplinary researchers have extensively analyzed health promoting and pharmacologically significant properties of garlic(Allium sativum).Importantly,garlic and its biologically active chemicals targeted oncogenic signaling cascades.In this mini-review we have attempted to summarize how garlic and its bioactive constituents regulated signal transduction cascades in cell culture studies and tumor-bearing mice.

When Fleeing Matters:Differences in Escape Behaviours of Three Northeast Asian Anurans

Prey species may have their own optimal escape strategy to balance predation risks and the energetic cost of fleeing.Some species have an advantage when maintaining a short fleeing distance,while others may favour an earlier escape based on microhabitat,size,or body condition.Here,we examined the escape behaviour of the three syntopic Northeast Asian anuran species:Mongolian toads(Strauchbufo raddei),Amur brown frogs(Rana amurensis),and Japanese treefrogs(Dryophytes japonicus)in Mongolia,Russia,China and DPR Korea.We examined flight initiation distance(FID;the distance from a potential predator to the point when the individual starts to flee)and distance fled(DF;distance between flight initiation and flight termination points)of each species and the effects of microhabitat,sex,and body size.Strauchbufo raddei and R.amurensis had a longer FID than D.japonicus,and S.raddei also had a longer DF than D.japonicus.These trends remained similar when dividing FID and DF by a size proxy(snout-vent length)for all individuals.This suggests that the treefrog D.japonicus used a strategy to stay immobile even when they were detected,and the toad S.raddei reacted quicker and more sensitively to predators despite the presence of toxin.Female S.raddei had a significantly longer FID than males suggesting that females are more sensitive to predation risk in this species,but body size was not significant for any of the three species.Our results indicate that the three sympatric species have different escaping strategies,likely related to differences in physiology and crypticity.

Development of biofertilizers for sustainable agriculture over four decades(1980-2022)

The application of biofertilizers is becoming an inevitable trend to substitute chemical fertilizers for sustainable agriculture.To better understand the development of biofertilizers from 1980 to 2022,we used bibliometric mining to analyze 12,880 journal articles related to biofertilizer.The network cooccurrence analysis suggested that the biofertilizers research can be separated into three stages.The first stage(1980-2005)focused on nitrogen fixation.The second stage(2006-2015)concentrated on the mechanisms for increasing plant yield.The third stage(2016-2022)was the application of biofertilizers to improve the soil environment.The keyword analysis revealed the mechanisms of biofertilizers to improve plant-growth:biofertilizers can impact the nutritional status of plants,regulate plant hormones,and improve soil environments and the microbiome.The bacteria use as biofertilizers,included Pseudomonas,Azospirillum,and Bacillus,were also identified through bibliometric mining.These findings provide critical discernment to aid further study of biofertilizers for sustainable agriculture.

RUFY4 deletion prevents pathological bone loss by blocking endo-lysosomal trafficking of osteoclasts

Mature osteoclasts degrade bone matrix by exocytosis of active proteases from secretory lysosomes through a ruffled border.However,the molecular mechanisms underlying lysosomal trafficking and secretion in osteoclasts remain largely unknown.Here,we show with GeneChip analysis that RUN and FYVE domain-containing protein 4(RUFY4)is strongly upregulated during osteoclastogenesis.Mice lacking Rufy4 exhibited a high trabecular bone mass phenotype with abnormalities in osteoclast function in vivo.Furthermore,deleting Rufy4 did not affect osteoclast differentiation,but inhibited bone-resorbing activity due to disruption in the acidic maturation of secondary lysosomes,their trafficking to the membrane,and their secretion of cathepsin K into the extracellular space.Mechanistically,RUFY4 promotes late endosome-lysosome fusion by acting as an adaptor protein between Rab7 on late endosomes and LAMP2 on primary lysosomes.Consequently,Rufy4-deficient mice were highly protected from lipopolysaccharide-and ovariectomy-induced bone loss.Thus,RUFY4 plays as a new regulator in osteoclast activity by mediating endo-lysosomal trafficking and have a potential to be specific target for therapies against bone-loss diseases such as osteoporosis.

The role of tazarotene-induced gene 1 in carcinogenesis:is it a tumor suppressor gene or an oncogene?

Tazarotene-induced gene 1(TIG1)is induced by a derivative of vitamin A and is known to regulate many important biological processes and control the development of cancer.TIG1 is widely expressed in various tissues;yet in many cancer tissues,it is not expressed because of the methylation of its promoter.Additionally,the expression of TIG1 in cancer cells inhibits their growth and invasion,suggesting that TIG1 acts as a tumor suppressor gene.However,in some cancers,poor prognosis is associated with TIG1 expression,indicating its protumor growth characteristics,especially in promoting the invasion of inflammatory breast cancer cells.This review comprehensively summarizes the roles of the TIG1 gene in cancer development and details the mechanisms through which TIG1 regulates cancer development,with the aim of understanding its various roles in cancer development.

Research on Total Electric Field Prediction Method of Ultra-High Voltage Direct Current Transmission Line Based on Stacking Algorithm

Ultra-high voltage(UHV)transmission lines are an important part of China’s power grid and are often surrounded by a complex electromagnetic environment.The ground total electric field is considered a main electromagnetic environment indicator of UHV transmission lines and is currently employed for reliable long-term operation of the power grid.Yet,the accurate prediction of the ground total electric field remains a technical challenge.In this work,we collected the total electric field data from the Ningdong-Zhejiang±800 kV UHVDC transmission project,as of the Ling Shao line,and perform an outlier analysis of the total electric field data.We show that the Local Outlier Factor(LOF)elimination algorithm has a small average difference and overcomes the performance of Density-Based Spatial Clustering of Applications with Noise(DBSCAN)and Isolated Forest elimination algorithms.Moreover,the Stacking algorithm has been found to have superior prediction accuracy than a variety of similar prediction algorithms,including the traditional finite element.The low prediction error of the Stacking algorithm highlights the superior ability to accurately forecast the ground total electric field of UHVDC transmission lines.

Emergence of taurine as a therapeutic agent for neurological disorders

Taurine is a sulfur-containing,semi-essential amino acid that occurs naturally in the body.It alternates between inflammation and oxidative stress-mediated injury in various disease models.As part of its limiting functions,taurine also modulates endoplasmic reticulum stress,Ca^(2+)homeostasis,and neuronal activity at the molecular level.Taurine effectively protects against a number of neurological disorders,including stro ke,epilepsy,cerebral ischemia,memory dysfunction,and spinal cord injury.Although various therapies are available,effective management of these disorders remains a global challenge.Approximately 30 million people are affected worldwide.The design of taurine fo rmation co uld lead to potential drugs/supplements for the health maintenance and treatment of central nervous system disorders.The general neuroprotective effects of taurine and the various possible underlying mechanisms are discussed in this review.This article is a good resource for understanding the general effects of taurine on various diseases.Given the strong evidence for the neuropharmacological efficacy of taurine in various experimental paradigms,it is concluded that this molecule should be considered and further investigated as a potential candidate for neurotherapeutics,with emphasis on mechanism and clinical studies to determine efficacy.

Genetic analyses of ancient tea trees provide insights into the breeding history and dissemination of Chinese Assam tea(Camellia sinensis var.assamica)

Chinese Assam tea(Camellia sinensis var.assamica)is an important tea crop with a long history of cultivation in Yunnan,China.Despite its potential value as a genetic resource,its genetic diversity and domestication/breeding history remain unclear.To address this issue,we genotyped 469 ancient tea plant trees representing 26 C.sinensis var.assamica populations,plus two of its wild relatives(six and three populations of C.taliensis and C.crassicolumna,respectively)using 16 nuclear microsatellite loci.Results showed that Chinese Assam tea has a relatively high,but comparatively lower gene diversity(H_(S)=0.638)than the wild relative C.crassicolumna(H_S=0.658).Clustering in STRUCTURE indicated that Chinese Assam tea and its two wild relatives formed distinct genetic groups,with considerable interspecific introgression.The Chinese Assam tea accessions clustered into three gene pools,corresponding well with their geographic distribution.However,New Hybrids analysis indicated that 68.48%of ancient Chinese Assam tea plants from Xishuangbanna were genetic intermediates between the Puer and Lincang gene pools.In addition,10%of the ancient Chinese Assam tea individuals were found to be hybrids between Chinese Assam tea and C.taliensis.Our results suggest that Chinese Assam tea was domesticated separately in three gene pools(Puer,Lincang and Xishuangbanna)in the Mekong River valley and that the hybrids were subsequently selected during the domestication process.Although the domestication history of Chinese Assam tea in southwestern Yunnan remains complex,our results will help to identify valuable genetic resources that may be useful in future tea breeding programs.

Comparative evaluation of artificial intelligence systems'accuracy in providing medical drug dosages:A methodological study

BACKGROUND Medication errors,especially in dosage calculation,pose risks in healthcare.Artificial intelligence(AI)systems like ChatGPT and Google Bard may help reduce errors,but their accuracy in providing medication information remains to be evaluated.AIM To evaluate the accuracy of AI systems(ChatGPT 3.5,ChatGPT 4,Google Bard)in providing drug dosage information per Harrison's Principles of Internal Medicine.METHODS A set of natural language queries mimicking real-world medical dosage inquiries was presented to the AI systems.Responses were analyzed using a 3-point Likert scale.The analysis,conducted with Python and its libraries,focused on basic statistics,overall system accuracy,and disease-specific and organ system accuracies.RESULTS ChatGPT 4 outperformed the other systems,showing the highest rate of correct responses(83.77%)and the best overall weighted accuracy(0.6775).Disease-specific accuracy varied notably across systems,with some diseases being accurately recognized,while others demonstrated significant discrepancies.Organ system accuracy also showed variable results,underscoring system-specific strengths and weaknesses.CONCLUSION ChatGPT 4 demonstrates superior reliability in medical dosage information,yet variations across diseases emphasize the need for ongoing improvements.These results highlight AI's potential in aiding healthcare professionals,urging continuous development for dependable accuracy in critical medical situations.

Human immunodeficiency virus cascade–continuum of care stages and outcomes in a hospital in southern Brazil

BACKGROUND The human immunodeficiency virus(HIV)continuum of care cascade illustrates the 90-90-90 goals defined by the Joint United Nations Program on HIV/acquired immunodeficiency syndrome(UNAIDS).The care cascade includes the following five steps:Diagnosis,linkage to care,retention in care,adherence to antiretroviral therapy(ART),and viral suppression.AIM To elaborate the HIV cascade of patients diagnosed with HIV at the Nossa Senhora da Conceição Hospital(HNSC)and to determine possible local causes for the loss of patients between each step of the cascade.METHODS This retrospective cohort study included patients diagnosed with HIV infection from January 1,2015 to December 31,2016 and followed up until July 31,2019.The data were analyzed by IBM SPSS software version 25,and Poisson regression with simple robust variance was used to analyze variables in relation to each step of the cascade.Variables with P<0.20 were included in multivariable analysis,and P<0.05 was considered significant.Pearson’sχ^(2) test was used to compare the groups of patients followed up at the HNSC and those followed up at other sites.RESULTS The results were lower than those expected by the UNAIDS,with 94%of patients linked,91%retained,81%adhering to ART,and 84%in viral suppression.Age and site of follow-up were the variables with the highest statistical significance.A comparison showed that the cascade of patients from the HNSC had superior results than outpatients,with a significant difference in the last step of the cascade.CONCLUSION The specialized and continued care provided at the HNSC was associated with better results and was closer to the goals set by the UNAIDS.The development of the HIV cascade using local data allowed for the stratification and evaluation of risk factors associated with the losses occurring between each step of the cascade.

Phylogenetic Analyses Reveal a Unique Species of Elaphe(Serpentes,Colubridae)New to Science

The snakes comprising the monophyletic group referred to as ratsnakes are found throughout Asia,Europe and the New World.Recently,three snake samples likely belonging to the ratsnakes were collected in Zoige County,Sichuan Province,China.Species identity was difficult to delimit morphologically because the specimens were juveniles and partially damaged.Subsequently,a molecular phylogenetic approach was used.Portions of three mitochondrial genes(cyt b,ND4 and 12S rRNA) were sequenced and analyzed.The results showed that they were sister to the genus Elaphe.Very little genetic variation was found among the three samples.The minimum genetic distances between these samples and those within Elaphe were greater than any currently recognized species within the genus.We conclude that this likely represents a new species within the genus Elaphe.Adult specimens and a morphologic description are needed for further study.

Cloning and characterization of proliferating cell nuclear antigen gene of Alexandrium catenella (Dinoflagellate) with respect to cell growth

Harmful algal blooms （HABs） have been affecting negatively the shellfish and aquaculture industries around the world. Though a lot of efforts have been made to disclose the changes of environmental factors involved and their effects on the HABs events, the molecular mechanism of this process remains unclear. To address this problem, proliferating cell nuclear antigen gene （pcna） was isolated and characterized from Alexandrium catenella. It showed high homology to those of other dinoflagellates （89% and 91% homology to Pfiesteria piscicid and Pyrocystis lunula, respectively）, and also 42%–43% homology to those of plant and animals. The expression level of pcna revealed by quantitative real time PCR was the lowest at the late lagging cell growth phase, increased to the highest at the late exponential phase, and then decreased at the stationary phase. Though the cell growth rate was also changing, no positive correlation between pcna expression level and cell growth rate was displayed throughout the whole cell growth stages （r 2 =0.024 6）. However, the pcna expression level had the similar trend with the change of cell growth rate throughout the whole growing process, e.g., from increasing at the earlier cell growth stage to decreasing at the following stages, though slightly lagging to the latter.

The landscape of cognitive impairment in superoxide dismutase 1-amyotrophic lateral sclerosis

Although mutations in the superoxide dismutase 1 gene account for only a minority of total amyotrophic lateral sclerosis cases,the discovery of this gene has been crucial for amyotrophic lateral sclerosis research.Since the identification of superoxide dismutase 1 in 1993,the field of amyotrophic lateral sclerosis genetics has considerably widened,improving our understanding of the diverse pathogenic basis of amyotrophic lateral sclerosis.In this review,we focus on cognitive impairment in superoxide dismutase 1-amyotrophic lateral sclerosis patients.Literature has mostly reported that cognition remains intact in superoxide dismutase 1-amyotrophic lateral sclerosis patients,but recent reports highlight frontal lobe function frailty in patients carrying different superoxide dismutase 1-amyotrophic lateral sclerosis mutations.We thoroughly reviewed all the various mutations reported in the literature to contribute to a comprehensive database of superoxide dismutase 1-amyotrophic lateral sclerosis genotype-phenotype correlation.Such a resource could ultimately improve our mechanistic understanding of amyotrophic lateral sclerosis,enabling a more robust assessment of how the amyotrophic lateral sclerosis phenotype responds to different variants across genes,which is important for the therapeutic strategy targeting genetic mutations.Cognition in superoxide dismutase 1-amyotrophic lateral sclerosis deserves further longitudinal research since this peculiar frailty in patients with similar mutations can be conditioned by external factors,including environment and other unidentified agents including modifier genes.

Ardisia gigantifolia ethanolic extract inhibits cell proliferation and targets cancer stem cells in gastric cancer

Objective:To evaluate the effects of ethanol extract from Ardisia gigantifolia leaves on cell proliferation and cancer stem cell(CSC)number in gastric cancer.Methods:The inhibitory effect of Ardisia gigantifolia extract on the proliferation of MKN45 and MKN74 gastric cancer cells was assessed using 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide assay.Non-adherent culture(3D)model was used to evaluate the effect of the extract on tumorsphere size and number.Moreover,the expression of CD44,ALDH,and p21 was determined by immunofluorescence analysis.Flow cytometric analysis was performed to evaluate cell cycle arrest and the expression of gastric CSC markers CD44 and ALDH.Real-time PCR analysis was also carried out to assess the effect of the extract on the expression of cell cycle-regulated genes.Results:Ardisia gigantifolia extract effectively inhibited cell proliferation with an IC_(50)of 55.7μg/m L in MKN45 cells and 123.6μg/m L in MKN74 cells.The extract also arrested cell cycle in the G_(0)/G_(1)phase as well as significantly reduced the size and number of tumorspheres.The markedly increased expression of p21 was observed at both m RNA and protein levels in the extract-treated adherent cells and tumorspheres.In addition,Ardisia gigantifolia extract significantly reduced the number of CD44-and/or ALDH-expressing gastric CSC.Conclusions:The development of gastric CSC can be inhibited by the ethanol extract of Ardisia gigantifolia.