Naringin ameliorates H_(2)O_(2)-induced oxidative damage in cells and prolongs the lifespan of female Drosophila melanogaster via the insulin signaling pathway

Naringin exists in a wide range of Chinese herbal medicine and has proven to possess several pharmacological properties.In this study,PC12,HepG2 cells,and female Drosophila melanogaster were used to investigate the antioxidative and anti-aging effects of naringin and explore the underlying mechanisms.The results showed that naringin inhibited H_(2)O_(2)-induced decline in cell viability and decreased,the content of reactive oxygen species in cells.Meanwhile,naringin prolonged the lifespan of flies,enhanced the abilities of climbing and the resistance to stress,improved the activities of antioxidant enzymes,and decreased malondialdehyde content.Naringin also improved intestinal barrier dysfunction and reduced abnormal proliferation of intestinal stem cells.Moreover,naringin down-regulated the mRNA expressions of inr,chico,pi 3k,and akt-1,and up-regulated the mRNA expressions of dilp2,dilp3,dilp5,and foxo,thereby activating autophagy-related genes and increasing the number of lysosomes.Furthermore,the mutant stocks assays and computer molecular simulation results further indicated that naringin delayed aging by inhibiting the insulin signaling(IIS)pathway and activating the autophagy pathway,which was consistent with the result of network pharmacological predictions.

Molecular and prognostic heterogeneity of microsatellite-unstable colorectal cancer

Colorectal cancers (CRCs) with a high level of microsatellite instability (MSI-H) are clinicopathologically distinct tumors characterized by predominance in females, proximal colonic localization, poor differentiation, mucinous histology, tumor-infiltrating lymphocytes, a Crohn’s-like lymphoid reaction and a favorable prognosis. In terms of their molecular features, MSI-H CRCs are heterogeneous tumors associated with various genetic and epigenetic alterations, including DNA mismatch repair deficiency, target microsatellite mutations, BRAF mutations, a CpG island methylator phenotype-high (CIMP-H) status, and a low level of genomic hypomethylation. The molecular heterogeneity of MSI-H CRCs also depends on ethnic differences; for example, in Eastern Asian countries, relatively low frequencies of CIMP-H and BRAF mutations have been observed in MSI-H CRCs compared to Western countries. Although the prognostic features of MSI-H CRCs include a favorable survival of patients and low benefit of adjuvant chemotherapy, there may be prognostic differences based on the molecular heterogeneity of MSI-H CRCs. Here, we have reviewed and discussed the molecular and prognostic features of MSI-H CRCs, as well as several putative prognostic or predictive molecular markers, including HSP110 expression, beta2-microglobulin mutations, myosin 1a expression, CDX2/CK20 expression, SMAD4 expression, CIMP status and LINE-1 methylation levels.

Loss of CDX2 expression is associated with poor prognosis in colorectal cancer patients

AIM:To investigate the clinicopathologic characteristics and prognostic implications associated with loss of CDX2 expression in colorectal cancers(CRCs).METHODS:We immunohistochemically evaluated CDX2 expression in 713 CRCs and paired our findings to clinicopathologic and molecular characteristics of each individual.Endpoints included cytokeratin 7 and CK20 expression,microsatellite instability,Cp G island methylator phenotype,and KRAS and BRAF mutation statuses.Univariate and multivariate survival analysis was performed to reveal the prognostic value of CDX2 downregulation.RESULTS:CDX2 expression was lost in 42(5.9%) patients.Moreover,loss of CDX2 expression was associated with proximal location,infiltrative growth,advanced T,N,M and overall stage.On microscopic examination,loss of CDX2 expression was associated with poor differentiation,increased number of tumor-infiltrating lymphocytes,luminal serration and mucin production.Loss of CDX2 expression was also associated with increased CK7 expression,decreased CK20 expression,Cp G island methylator phenotype,microsatellite instability and BRAF mutation.In a univariate survival analysis,patients with loss of CDX2 expression showed worse overall survival(P < 0.001) and progression-free survival(P < 0.001).In a multivariate survival analysis,loss of CDX2 expression was an independent poor prognostic factor of overall survival [hazard ratio(HR) = 1.72,95%CI:1.04-2.85,P = 0.034] and progression-free survival(HR = 1.94,95%CI:1.22-3.07,P = 0.005).CONCLUSION:Loss of CDX2 expression is associated with aggressive clinical behavior and can be used as a prognostic marker in CRCs.

Prognostic analysis of patients with combined hepatocellularcholangiocarcinoma after radical resection:A retrospective multicenter cohort study

BACKGROUND Combined hepatocellular-cholangiocarcinoma(cHCC-CCA)is a form of rare primary liver cancer that combines intrahepatic cholangiocarcinoma(ICC)and hepatocellular carcinoma.AIM To investigate overall survival(OS)and recurrence-free survival(RFS)after radical resection in patients with cHCC-CCA,and the clinicopathological factors affecting prognosis in two center hospitals of China.METHODS We reviewed consecutive patients with cHCC-CCA who received radical resection between January 2005 and September 2021 at Peking Union Medical College and the 5th Medical Center of the PLA General Hospital retrospectively.Regular follow-up and clinicopathological characteristics were systematic collected for baseline and prognostic analysis.RESULTS Our study included 95 patients who received radical resection.The majority of these patients were male and 82.7%of these patients were infected with HBV.The mean tumor size was 4.5 cm,and approximately 40%of patients had more than one lesion.The median OS was 26.8(95%CI:18.5-43.0)mo,and the median RFS was 7.27(95%CI:5.83-10.3)mo.Independent predictors of OS were CA19-9≥37 U/mL(HR=8.68,P=0.002),Child-Pugh score>5(HR=5.52,P=0.027),tumor number>1(HR=30.85,P=0.002),tumor size and transarterial chemoembolization(TACE)after surgery(HR=0.2,P=0.005).CONCLUSION The overall postoperative survival of cHCC-CCA patients is poor,and most patients experience relapse within a short period of time after surgery.Preoperative tumor biomarker(CA19-9,alphafetoprotein)levels,tumor size,and Child-Pugh score can significantly affect OS.Adjuvant TACE after surgery prolongs RFS,suggesting that TACE is a possible option for postoperative adjuvant therapy in patients with cHCC-CCA.

Astragaloside Ⅳ Improves Angiogenesis and Promotes Wound Healing in Diabetic Rats via the Activation of the SUMOylation Pathway

Radix astragali is the dried root of the legume Mongolian Astragalus or Astragalus membranaceus~([1]). Modern research has found that the main active ingredients in astragalus are astragalus polysaccharides, astragaloside, and astragalus isoflavones. Astragaloside IV(AS-IV) has been used internationally as a standard for evaluating the quality of astragalus medicinal materials.

Annexin A10 expression in colorectal cancers with emphasis on the serrated neoplasia pathway

AIM: To validate the utility of Annexin A10 as a surrogate marker of the serrated neoplasia pathway in invasive colorectal cancers(CRCs).METHODS: A total of 1133 primary CRC patients who underwent surgical resection at Seoul National University Hospital between January 2004 and December 2007 were enrolled.Expression of Annexin A10 was evaluated by immunohistochemistry using tissue microarray and paired to our findings on clinicopathologic and molecular characteristics of each individual.Cp G island methylator phenotype was determined by Methy Light assay and microsatellite instability was determined by high performance liquid chromatography.KRAS and BRAF mutation status was evaluated by direct sequencing and allele-specific PCR.Univariate and stage-specific survival analyses were performed to reveal the prognostic value of Annexin A10 expression.RESULTS: Annexin A10 expression was observed in 66(5.8%) of the 1133 patients.Annexin A10 expression was more commonly found in females and was associated with proximal location,ulcerative gross type,advanced T category,N category and TNM stage.CRCs with Annexin A10 expression showed an absence of luminal necrosis,luminal serration and mucin production.CRCs with Annexin A10 expression were associated with Cp G island methylator phenotype,microsatellite instability and BRAF mutation.In survival analysis,Annexin A10 expression was associated with poor overall survival and progression-free survival,especially in stage Ⅳ CRCs.CONCLUSION: Annexin A10 expression is associated with poor clinical behavior and can be used a supportive surrogate marker of the serrated neoplasia pathway in invasive CRCs.

Exosomal miR-30a-5p targets NLRP3 to suppress podocyte pyroptosis in diabetic nephropathy

Background:Mesenchymal stem cell(MSC)-derived exosomes are closely related to pyroptosis in diabetic nephropathy(DN).This study aimed to explore the protective effect of exosomal miR-30a-5p on podocyte pyroptosis in DN.Methods:Streptozotocin was used to establish the mouse model of DN.Human bone marrow MSC-derived exosomes were extracted and identified via transmission electron microscopy,nanoparticle tracking analysis,and western blotting.MiR-30a-5p mimics and non-control(NC)mimics were transfected into MSCs and podocytes,and exosomes were isolated from the MSCs.High glucose(HG)-induced podocyte model was established to determine the effect of exosomal miR-30a-5p on pyroptosis and inflammation in vitro.Results:MiR-30a-5p was expressed at low levels in DN models,while NLR family pyrin domain containing 3(NLRP3),caspase-1,gasdermin-N(GSDMD-N),and pro-inflammatory factors(tumor necrosis factor-alpha,interleukin(IL)-1beta,and IL-18)were augmented.In vitro,miR-30a-5p expression in the HG-damaged podocytes was down-regulated,while NLRP3 was up-regulated.Interestingly,miR-30a-5p overexpression diminished HG-induced podocyte injury,as proven by increased activity and decreased pyroptosis of podocytes.Concurrently,the up-regulation of miR-30a-5p could inhibit the expression of pro-inflammatory factors,caspase-1,GSDMD-N,and NLRP3 in HG-induced podocytes.MSC-derived exosomal miR-30a-5p treatment of HG-damaged cells has similar effects to miR-30a-5p mimics treatment.Overexpression of NLRP3 reversed the effect of miR-30a-5p mimics on HG-induced podocytes.Conclusion:This research confirmed that exosomal miR-30a-5p regulates pyroptosis via mediating NLRP3 in DN.

SUMO1 regulates post-infarct cardiac repair based on cellular heterogeneity

Small ubiquitin-related modifier(SUMOylation)is a dynamic post-translational modification that maintains cardiac function and can protect against a hypertrophic response to cardiac pressure overload.However,the function of SUMOylation after myocardial infarction(MI)and the molecular details of heart cell responses to SUMO1 deficiency have not been determined.In this study,we demonstrated that SUMO1 protein was inconsistently abundant in different cell types and heart regions after MI.However,SUMO1 knockout significantly exacerbated systolic dysfunction and infarct size after myocardial injury.Single-nucleus RNA sequencing revealed the differential role of SUMO1 in regulating heart cells.Among cardiomyocytes,SUMO1 deletion increased the Nppa^(+)Nppb^(+)Ankrd1^(+)cardiomyocyte subcluster pro-portion after MI.In addition,the conversion of fibroblasts to myofibroblasts subclusters was inhibited in SUMO1 knockout mice.Importantly,SUMO1 loss promoted proliferation of endothelial cell subsets with the ability to reconstitute neovascularization and expressed angiogenesis-related genes.Computational analysis of ligand/receptor interactions suggested putative pathways that mediate cardiomyocytes to endothelial cell communication in the myocardium.Mice preinjected with cardiomyocyte-specific AAV-SUMO1,but not the endothelial cell-specific form,and exhibited ameliorated cardiac remodeling following MI.Collectively,our results identified the role of SUMO1 in cardiomyocytes,fibroblasts,and endothelial cells after MI.These findings provide new insights into SUMO1 involvement in the patho-genesis of MI and reveal novel therapeutic targets.

Machine learning insights concerning inflammatory and liver-related risk comorbidities in non-communicable and viral diseases

The liver is a key organ involved in a wide range of functions,whose damage can lead to chronic liver disease(CLD).CLD accounts for more than two million deaths worldwide,becoming a social and economic burden for most countries.Among the different factors that can cause CLD,alcohol abuse,viruses,drug treatments,and unhealthy dietary patterns top the list.These conditions prompt and perpetuate an inflammatory environment and oxidative stress imbalance that favor the development of hepatic fibrogenesis.High stages of fibrosis can eventually lead to cirrhosis or hepatocellular carcinoma(HCC).Despite the advances achieved in this field,new approaches are needed for the prevention,diagnosis,treatment,and prognosis of CLD.In this context,the scientific community is using machine learning(ML)algorithms to integrate and process vast amounts of data with unprecedented performance.ML techniques allow the integration of anthropometric,genetic,clinical,biochemical,dietary,lifestyle and omics data,giving new insights to tackle CLD and bringing personalized medicine a step closer.This review summarizes the investigations where ML techniques have been applied to study new approaches that could be used in inflammatoryrelated,hepatitis viruses-induced,and coronavirus disease 2019-induced liver damage and enlighten the factors involved in CLD development.

α-parvin controls chondrocyte column formation and regulates long bone development

Endochondral ossification requires proper control of chondrocyte proliferation,differentiation,survival,and organization.Here we show that knockout ofα-parvin,an integrin-associated focal adhesion protein,from murine limbs causes defects in endochondral ossification and dwarfism.The mutant long bones were shorter but wider,and the growth plates became disorganized,especially in the proliferative zone.With two-photon time-lapse imaging of bone explant culture,we provide direct evidence showing thatα-parvin regulates chondrocyte rotation,a process essential for chondrocytes to form columnar structure.Furthermore,loss ofα-parvin increased binucleation,elevated cell death,and caused dilation of the resting zones of mature growth plates.Single-cell RNA-seq analyses revealed alterations of transcriptome in all three zones(i.e.,resting,proliferative,and hypertrophic zones)of the growth plates.Our results demonstrate a crucial role ofα-parvin in long bone development and shed light on the cellular mechanism through whichα-parvin regulates the longitudinal growth of long bones.

A novel role of LncRNA in regulating tumor metabolism and angiogenesis under hypoxia

Tumor cells are characterized by metabolic reprogram-ming,a step that is necessary to support their high demand for energy and nutrients[1,2].Furthermore,once metastatic tumor cells are seeded,tumor microen-vironment remodeling is immediately initiated,leading to the formation of more suitable conditions for growth.Tumor cells secrete several types of growth factors,cytokines,and chemokines to induce angiogenesis and tumor-associated immune cell recruitment,all of which can promote proliferation and metastasis,as well as trig-ger immune tolerance and drug resistance[3].Exploring the synergistic effect between metabolic reprogramming and microenvironment remodeling will likely facili-tate the discovery of novel biomarkers and therapeutic targets.

Impacts of transmembrane serine protease 4 expression on susceptibility to severe acute respiratory syndrome coronavirus 2

To the Editor:Transmembrane serine protease 4(TMPRSS4;OMIM:606565),which encodes a member of the serine protease family,is involved in various biological processes related to diseases such as cancer and pediatric neurodegenerative disorders.

Microplastome visualizations:From spatiotemporal distribution to risk assessment

VECTOR EFFECT OF MICROPLASTIC PARTICLES.The microplastic pollution issue is infamous for its enormous multidimensionality and the intricate combination arising from plastics themselves and“plasticassociated chemicals.”Interactions between microplastic particles with multiple chemicals can affect biological health and alter the toxicity of the primary environmental pollutants.These complex combinations raise great challenges to experimental design for toxicity evaluation,going beyond the investigation of individual or dual contaminations.Systematic research and an integrated visualization strategy are needed to decipher these biochemical processes and molecular mechanisms.1,2.

NK cell-based cancer immunotherapy: from basic biology to clinical application

Natural killer(NK) cells, which recognize and kill target cells independent of antigen specificity and major histocompatibility complex(MHC) matching, play pivotal roles in immune defence against tumors. However, tumor cells often acquire the ability to escape NK cell-mediated immune surveillance. Thus, understanding mechanisms underlying regulation of NK cell phenotype and function within the tumor environment is instrumental for designing new approaches to improve the current cell-based immunotherapy. In this review, we elaborate the main biological features and molecular mechanisms of NK cells that pertain to regulation of NK cell-mediated anti-tumor activity. We further overview current clinical approaches regarding NK cell-based cancer therapy, including cytokine infusion, adoptive transfer of autologous or allogeneic NK cells, applications of chimeric antigen receptor(CAR)-expressing NK cells and adoptive transfer of memory-like NK cells. With these promising clinical outcomes and fuller understanding the basic questions raised in this review, we foresee that NK cell-based approaches may hold great potential for future cancer immunotherapy.

Mutant p53 promotes cell spreading and migration via ARHGAP44

The tumor suppressor p53 protein is either lost or mutated in about half of all human cancers.Loss of p53 function is well known to influence cell spreading,migration and invasion.While expression of mutant p53 is not equivalent to p53 loss,mutant p53 can acquire new functions to drive cell spreading and migration via different mechanisms.In our study,we found that mutant p53 significantly increased cell spreading and migration when comparing with p53-null cells.RNA-Seq analysis suggested that Rho GTPase activating protein 44(ARHGAP44) is a new target of mutant p53,which suppressed AKHGAP44 transcription.ARHGAP44 has GAP activity and catalyze GTP hydrolysis on Cdc42.Higher level of GTP-Cdc42 was correlated with increase expression of mutant p53 and reduced ARHGAP44.Importantly,wt-ARHGAP44 but not mutant ARHGAP44(R291A) suppressed mutant p53 mediated cell spreading and migration.Bioinformatics analysis indicated lower expression of ARHGAP44 in lung carcinoma compared with normal tissues,which was verified by RT-qPCR using specimens from patients.More interestingly,ARHGAP44 mRNA level was lower in tumors with mutant p53 than those with normal p53.Collectively,our results disclose a new mechanism by which mutant p53 stimulates cell spreading and migration.

Inducible CRISPRa screen identifies putative enhancers

Enhancers are critical cis-regulatory elements that regulate spatiotemporal gene expression and control cell fates.However,the identification of enhancers in native cellular contexts still remains a challenge.Here,we develop an inducible CRISPR activation(CRISPRa)system by transgenic expression of doxycycline(Dox)-inducible dCas9-VPR in mouse embryonic stem cells(iVPR ESC).With this line,a simple introduction of specific guide RNAs targeting promoters or enhancers allows us to realize the effect of CRISPRa in an inducible,reversible,and Dox concentration-dependent manner.Taking advantage of this system,we induce tiled CRISPRa across genomic regions(105 kilobases)surrounding T(Brachyury),one of the key mesodermal development regulator genes.Moreover,we identify several CRISPRa-responsive elements with chromatin features of putative enhancers,including a region the homologous sequence in which humans harbors a body height risk variant.Genetic deletion of this region in ESC does affect subsequent T gene activation and osteogenic differentiation.Therefore,our inducible CRISPRa ESC line provides a convenient platform for high-throughput screens of putative enhancers.

DNMT3A reads and connects histone H3K36me2 to DNA methylation

Dear Editor,DNA methylation at the 5-position of cytosine(5mC)is a crucial epigenetic mark in regulating biological processes including gene silencing,gene imprinting,and X chromo-some inactivation(Jaenisch and Bird,2003;Smith and Meissner,2013).Human genome encodes three DNA methyltransferases,DNMT1,DNMT3A and DNMT3B to catalyze 5mC.Although not tightly restricted,DNMT1 is thought to maintain the established pattern of 5mC throughout DNA replication,while DNMT3A and DNMT3B are largely responsible for the de novo establishment of 5mC.It has long been questioned how de novo DNA 5mC patterns are established in different genomic regions and whether histone modifications crosstalk to the process.Until recently,it was reported that through recognition of histone H3K36me3 mark,DNMT3B plays a dominant role in medi-ating DNA 5mC in the genic region undergoing active tran-scription(Baubec et al.,2015;Neri et al,2017).However,5mC occurs at both intergenic and genic regions,while H3K36me3 is largely absent in the intergenic regions,indi-cating that the intergenic 5mC may be mediated through diferent mechanisms.

Catch bond-inspired hydrogels with repeatable and loading rate-sensitive specific adhesion

Biological receptor-ligand adhesion governed by mammalian cells involves a series of mechanochemical pro-cesses that can realize reversible,loading rate-dependent specific interfacial bonding,and even exhibit a counterintuitive behavior called catch bonds that tend to have much longer lifetimes when larger pulling forces are applied.Inspired by these catch bonds,we designed a hydrogen bonding-meditated hydrogel made from acrylic acid-N-acryloyl glycinamide(AA-NAGA)copolymers and tannic acids(TA),which formed repeatable specific adhesion to polar surfaces in an ultra-fast and robust way,but hardly adhered to nonpolar materials.It demonstrated up to five-fold increase in shear adhesive strength and interfacial adhesive toughness with external loading rates varying from 5 to 500 mm min^(-1).With a mechanochemical coupling model based on Monte Carlo simulations,we quantitatively revealed the nonlinear dependence of rate-sensitive interfacial adhesion on external loading,which was in good agreement with the experimental data.Likewise,the developed hydrogels were biocompatible,possessed antioxidant and antibacterial properties and promoted wound healing.This work not only reports a stimuli-responsive hydrogel adhesive suitable for multiple biomedical applications,but also offers an innovative strategy for bionic designs of smart hydrogels with loading rate-sensitive specific adhesion for various emerging areas including flexible electronics and soft robotics.

Correction to:DNMT3A reads and connects histone H3K36me2 to DNA methylation

The author would like to add the below information in this correction.A similar study from Chao Lu group was published online on 5 September 2019 in Nature,entitled“The histone mark H3K36me2 recruits DNMT3A and shapes the intergenic DNA methylation landscape”(Weinberg et al.,2019).Although both studies reported the preferential recognition of H3K36me2 by DNMT3A PWWP,ours in addition uncovered a stimulation function by such interaction on the activity of DNMT3A.On the disease connections,we used a NSD2 gain-of-function model which led to the discovery of potential therapeutic implication of DNA inhibitors in the related cancers,while the other study only used NSD1 and DNMT3A loss-of-function models.

Biomimetic hydrogels with mesoscale collagen architecture for patient-derived tumor organoids culture

Patient-derived tumor organoids(PDTOs)shows great potential as a preclinical model.However,the current methods for establishing PDTOs primarily focus on modulating local properties,such as sub-micrometer topographies.Nevertheless,they neglect to capture the global millimeter or intermediate mesoscale architecture that have been demonstrated to influence tumor response to therapeutic treatment and tumor progression.In this study,we present a rapid technique for generating collagen bundles with an average length of 90±27μm and a mean diameter of 5±1.5μm from tumor tissue debris that underwent mechanical agitation following enzymatic digestion.The collagen bundles were subsequently utilized for the fabrication of biomimetic hydrogels,incorporating microbial transglutaminase(mTG)crosslinked gelatin.These biomimetic hydrogels,referred to as MC-gel,were specifically designed for patient-derived tumor organoids.The lung cancer organoids cultured in MC-gel exhibited larger diameters and higher cell viability compared to those cultured in gels lacking the mesoscale collagen bundle;moreover,their irregular morphology more closely resembled that observed in vivo.The MC-gel-based lung cancer organoids effectively replicated the histology and mutational landscapes observed in the original donor patient’s tumor tissue.Additionally,these lung cancer organoids showed a remarkable similarity in their gene expression and drug response across different matrices.This recently developed model holds great potential for investigating the occurrence,progression,metastasis,and management of tumors,thereby offering opportunities for personalized medicine and customized treatment options.