Mesenchymal stem cells-extracellular vesicles alleviate pulmonary fibrosis by regulating immunomodulators

BACKGROUND Pulmonary fibrosis(PF)is a chronic interstitial lung disease characterized by fibroblast proliferation and extracellular matrix formation,causing structural damage and lung failure.Stem cell therapy and mesenchymal stem cells-extracellular vesicles(MSC-EVs)offer new hope for PF treatment.AIM To investigate the therapeutic potential of MSC-EVs in alleviating fibrosis,oxidative stress,and immune inflammation in A549 cells and bleomycin(BLM)-induced mouse model.METHODS The effect of MSC-EVs on A549 cells was assessed by fibrosis markers[collagen I andα-smooth muscle actin(α-SMA),oxidative stress regulators[nuclear factor E2-related factor 2(Nrf2)and heme oxygenase-1(HO-1),and inflammatory regu-lators[nuclear factor-kappaB(NF-κB)p65,interleukin(IL)-1β,and IL-2].Similarly,they were assessed in the lungs of mice where PF was induced by BLM after MSC-EV transfection.MSC-EVs ion PF mice were detected by pathological staining and western blot.Single-cell RNA sequencing was performed to investigate the effects of the MSC-EVs on gene expression profiles of macrophages after modeling in mice.RESULTS Transforming growth factor(TGF)-β1 enhanced fibrosis in A549 cells,significantly increasing collagen I andα-SMA levels.Notably,treatment with MSC-EVs demonstrated a remarkable alleviation of these effects.Similarly,the expression of oxidative stress regulators,such as Nrf2 and HO-1,along with inflammatory regulators,including NF-κB p65 and IL-1β,were mitigated by MSC-EV treatment.Furthermore,in a parallel manner,MSC-EVs exhibited a downregulatory impact on collagen deposition,oxidative stress injuries,and inflammatory-related cytokines in the lungs of mice with PF.Additionally,the mRNA sequencing results suggested that BLM may induce PF in mice by upregulating pulmonary collagen fiber deposition and triggering an immune inflammatory response.The findings collectively highlight the potential therapeutic efficacy of MSC-EVs in ameliorating fibrotic processes,oxidative stress,and inflammatory responses associated with PF.CONCLUSION MSC-EVs could ameliorate fibrosis in vitro and in vivo by downregulating collagen deposition,oxidative stress,and immune-inflammatory responses.

Milk fat globule epidermal growth factor 8 alleviates liver injury in severe acute pancreatitis by restoring autophagy flux and inhibiting ferroptosis in hepatocytes

BACKGROUND Liver injury is common in severe acute pancreatitis(SAP).Excessive autophagy often leads to an imbalance of homeostasis in hepatocytes,which induces lipid peroxidation and mitochondrial iron deposition and ultimately leads to ferroptosis.Our previous study found that milk fat globule epidermal growth factor 8(MFG-E8)alleviates acinar cell damage during SAP via binding toαvβ3/5 integrins.MFG-E8 also seems to mitigate pancreatic fibrosis via inhibiting chaperone-mediated autophagy.AIM To speculate whether MFG-E8 could also alleviate SAP induced liver injury by restoring the abnormal autophagy flux.METHODS SAP was induced in mice by 2 hly intraperitoneal injections of 4.0 g/kg L-arginine or 7 hly injections of 50μg/kg cerulein plus lipopolysaccharide.mfge8-knockout mice were used to study the effect of MFG-E8 deficiency on SAPinduced liver injury.Cilengitide,a specificαvβ3/5 integrin inhibitor,was used to investigate the possible mechanism of MFG-E8.RESULTS The results showed that MFG-E8 deficiency aggravated SAP-induced liver injury in mice,enhanced autophagy flux in hepatocyte,and worsened the degree of ferroptosis.Exogenous MFG-E8 reduced SAP-induced liver injury in a dose-dependent manner.Mechanistically,MFG-E8 mitigated excessive autophagy and inhibited ferroptosis in liver cells.Cilengitide abolished MFG-E8’s beneficial effects in SAP-induced liver injury.CONCLUSION MFG-E8 acts as an endogenous protective mediator in SAP-induced liver injury.MFG-E8 alleviates the excessive autophagy and inhibits ferroptosis in hepatocytes by binding to integrinαVβ3/5.

Bioinformatics and in vitro study reveal the roles of microRNA-346 in high glucose-induced human retinal pigment epithelial cell damage

AIM:To study microRNAs(miRNAs)and their potential effects in high glucose-induced human retinal pigment epithelial cell damage.METHODS:We screened the GSE52233 miRNA expression dataset for differentially expressed miRNAs(DEMs).The target genes of the top 10 DEMs were predicted using miRWalk 2.0 database,followed by function enrichment and protein-protein interaction analysis.miRNA expression was determined in the human retinal pigment epithelial cell line ARPE-19 treated with high glucose(HG)by quantitative reverse transcription-polymerase chain reaction(qRT-PCR).Cell proliferation was determined using cell counting kit(CCK)-8 assay.Cell cycle,apoptosis,and reactive oxygen species(ROS)levels were determined by flow cytometry.The direct interaction between miRNA and targets was validated using dual-luciferase reporter assay.RESULTS:Thirty-nine DEMs were screened,and we predicted 125 miRNA-mRNA pairs for the top 10 DEMs,including 119 target genes of seven DEMs such as miR-346,which was upregulated in diabetic retinopathy(DR).miR-346 target genes were substantially enriched in the regulation of intracellular transport and retinoic acidinducible gene I(RIG-I)-like receptor signaling pathway.Expression of three upregulated and downregulated miRNAs were verified by qRT-PCR in HG-treated ARPE-19 cells.Expression of miR-346 was elevated in HG treated ARPE-19 cells in a dose-dependent manner.HG inhibited cell proliferation and induced apoptosis,which were partly reversed by transfecting an miR-346 inhibitor,which even decreased the ROS levels elevated due to HG.Argonaute 2(AGO2)was a target of miR-346.CONCLUSION:miR-346 is a key miRNA and plays an important role in HG-induced damage in human retinal pigment epithelial cells.

Expression of PDGFR-α in Patients with Valvular Atrial Fibrillation

Objective: To investigate the expression of Platelet-derived growth factor receptor alpha (PDGFR-α) in patients who have valvular atrial fibrillation. Methods: In this research, eighty-four patients with rheumatic heart disease who were going to undertake cardiac surgery were included. The subjects were divided into two groups: the AF group and the sinus rhythm group, the quantities are 39 and 45 respectively. Before the surgery, baseline demographics, physical examination, routine laboratory testing, echocardiography, echocardiographic data and additional clinical data were available for all patients. The right atrial tissue of the subjects was separated during surgery, with an area of approximately 0.3 - 0.5 mm3. Immunofluorescence staining was used to analyze the distribution of PDGFR-α of atrial tissue. mRNA of PDGFR-α in atrial tissue were determined by real-time quantitative PCR (Polymerase Chain Reaction);Western-Blot technique was used to measure the protein of PDGFR-α in atrial tissue. Results: There were no significant differences (P > 0.05) in sex ratio, age, blood pressure, blood biochemistry, and other aspects of medical history between the two groups. However, the right and left atrium diameters in the AF group were markedly larger than those in the SR group (P α from right atrial tissue were obviously higher in the AF group than that in the SR group (P Conclusion: The expression of PDGFR-α in the right atrial tissue of patients with atrial fibrillation was found to be significantly higher.

Chinese physicians' perceptions of fecal microbiota transplantation

AIM: To explore Chinese physicians' perceptions towards fecal microbiota transplantation(FMT) and to provide information and an assessment of FMT development in China.METHODS: A self-administered questionnaire was developed according to the FMT practice guidelines and was distributed to physicians in hospitals via Internet Research Electronic Data Capture(REDcap) software and electronic mails to assess their attitudes toward and knowledge of FMT. The questionnaire included a brief introduction of FMT that was followed by 20 questions. The participants were required to respond voluntarily, under the condition of anonymity and without compensation. Except for the fill-in-the-blank questions, all of the other questions were required in the REDcap data collection systems, and the emailed questionnaires were completed based on eligibility.RESULTS: Up to December 9, 2014, 844 eligible questionnaires were received out of the 980 distributed questionnaires, with a response rate of 86.1%. Among the participants, 87.3% were from tertiary hospitals, and there were 647(76.7%) gastroenterologists and 197(23.3%) physicians in other departments(nongastroenterologists). Gastroenterologists' awareness of FMT prior to the survey was much higher than non-gastroenterologists'(54.3 vs 16.5%, P < 0.001); however, acceptance of FMT was not statistically different(92.4 vs 87.1%, P = 0.1603). Major concerns of FMT included the following: acceptability to patients(79.2%), absence of guidelines(56.9%), and administration and ethics(46.5%). On the basis of understanding, the FMT indications preferred byphysicians were recurrent Clostridium difficile infection(86.7%), inflammatory bowel disease combined with Clostridium difficile infection(78.6%), refractory ulcerative colitis(70.9%), ulcerative colitis(65.4%), Crohn's disease(59.4%), chronic constipation(43.7%), irritable bowel syndrome(39.1%), obesity(28.1%) and type 2 diabetes(23.9%). For donor selection, the majority of physicians preferred individuals with a similar gut flora environment to the recipients. 76.6% of physicians chose lower gastrointestinal tract as the administration approach. 69.2% of physicians considered FMT a safe treatment. CONCLUSION: Chinese physicians have awareness and a high acceptance of FMT, especially gastroenterologists, which provides the grounds and conditions for the development of this novel treatment in China. Physicians' greatest concerns were patient acceptability and absence of guidelines.

Construction of targeted plasmid vector pcDNA3.1-Egr.1p-p16 and its expression in pancreatic cancer JF305 cells induced by radiation in vitro

To construct pcDNA3.1-Egr.1p-p16 recombinant plasmid and investigate the expression of p16 in pancreatic cancer JF305 cells induced by radiation and the feasibility of gene radiotherapy for pancreatic carcinoma.

Platelet-Derived Growth Factor-A Overexpression Correlates with Atrial Fibrosis in the Patients with Atrial Fibrillation Secondary to Rheumatic Valvular Disease

Objective: To investigate the relationship between platelet-derived growth factor-A (PDGF-A) and atrial fibrosis in patients who have developed atrial fibrillation (AF) secondary to rheumatic valvular disease. Methods: 84 selected patients participated in the current study who have developed rheumatic heart disease and were going to have a cardiac surgical operation. In the current study, whole subjects were divided into two group, they were atrial fibrillation (AF) group (the quantity is thirty-nine) and sinus rhythm (SR) group (the quantity is forty-five). Before the operation, complete clinical data was available for the whole patients. During the operation, the right atrial tissue (0.3 - 0.5 mm3) was disserted from every patient. Right atrial fibrosis was observed by Masson staining and the distribution of PDGF-A in right atrium specimen was observed by immunohistochemistry. RT-PCR techniques were applied to admeasure the mRNA expressions of PDGF-A in patients’ atrial tissue. At the same time, western-Blot techniques were employed to admeasure the protein expressions of PDGF-A. Results: In baseline clinical characteristics, in both AF group and SR group, there was no apparently difference between them (P > 0.05);compared with SR group, the diameters of left atrium and right atrium in AF group were apparently increased (P Conclusion: Atrial remodeling plays an important role in patients with valvular atrial fibrillation;PDGF-A in patients with AF was highly expressed in the right atrial, and was closely related with atrial fibrosis.

White Fat tissue’s Browning:Gaps in Knowledge and Current Clinical Significance

There are three types of fat tissue in mammals:white fat tissue(WAT),classic brown fat tissue(cBAT),and inducible brown fat tissue(iBAT)or beige fat tissue.The brown fat tissue in the human body is more similar to the induced brown fat tissue in mice.The main function of WAT is to store energy in the form of triacylglycerols and to regulate endocrine function.The main function of cBAT and iBAT is to generate heat and maintain the energy balance of the body.A cold environment and exercise can stimulate the browning of white fat tissue.The browning of white fat tissue is a complex process involving many factors,with PRDM16,PPARγand PGC-1αbeing the core transcription factors related to the process.Previous studies showed that white adipose browning is beneficial for the treatment of obesity,diabetes,fatty liver and other diseases.On the other hand,it can promote the progression of malignant tumors.Under normal conditions,the different types of fat tissue maintain a dynamic balance in the human body.Therefore,obtaining a better understanding of this balance and determining how to intervene at the early stages of metabolic imbalance are necessary.

Expression and Clinical Significance of miRNA-495 in the Peripheral Blood of Acute Myeloid Leukemia Patients

Objective:To analyze the expression and clinical significance of miRNA-495 in the peripheral blood of patients with acute myeloid leukemia.Methods:Fifty-six patients with acute myeloid leukemia and 56 healthy controls were selected.Fasting venous blood was drawn and centrifuged,and the plasma was collected.The target miRNA was directly amplified and reverse transcribed into cDNA.The expression of plasma miR-495 was detected by qRT-PCR.Results:The expression level of miRNA-495 in newly diagnosed acute myeloid leukemia(AML[ND])and relapsed/refractory acute myeloid leukemia(AML[RR])was significantly lower than that in complete remission(AML[CR])and normal control group(Control)(p<0.0001).There was no significant difference between AML(ND)group and AML(RR)group(p>0.05).The area under the ROC curve(AUC)of miRNA-495 was 0.9503,the 95%confidence interval was 0.9113–0.9892(p<0.0001),the standard error was 0.020,the sensitivity and specificity were 91.1%and 92.9%,respectively,and the Jordan index was 0.857.There was no significant difference between the expression level of miRNA-495 and gender,age,leukocyte count,hemoglobin,and platelet count(p>0.05).However,it was found related to the proportion of primitive bone marrow cells in patients(p=0.017).Conclusion:The decreased expression of plasma miR-495 in AML patients can be used as a new indicator for the diagnosis and prognosis of AML.

Reference values of carotid intima-media thickness and arterial stiffness in Chinese adults based on ultrasound radio frequency signal:A nationwide,multicenter study

Background:Carotid intima-media thickness(IMT)and diameter,stiffness,and wave reflections,are independent and important clinical biomarkers and risk predictors for cardiovascular diseases.The purpose of the present study was to establish nationwide reference values of carotid properties for healthy Chinese adults and to explore potential clinical determinants.Methods:A total of 3053 healthy Han Chinese adults(1922 women)aged 18-79 years were enrolled at 28 collaborating tertiary centers throughout China between April 2021 and July 2022.The real-time tracking of common carotid artery walls was achieved by the radio frequency(RF)ultrasound system.The IMT,diameter,compliance coefficient,βstiffness,local pulse wave velocity(PWV),local systolic blood pressure,augmented pressure(AP),and augmentation index(AIx)were then automatically measured and reported.Data were stratified by age groups and sex.The relationships between age and carotid property parameters were analyzed by Jonckheere-Terpstra test and simple linear regressions.The major clinical determinants of carotid properties were identified by Pearson’s correlation,multiple linear regression,and analyses of covariance.Results:All the parameters of carotid properties demonstrated significantly age-related trajectories.Women showed thinner IMT,smaller carotid diameter,larger AP,and AIx than men.Theβstiffness and PWV were significantly higher in men than women before forties,but the differences reversed after that.The increase rate of carotid IMT(5.5µm/year in women and 5.8µm/year in men)and diameter(0.03 mm/year in both men and women)were similar between men and women.For the stiffness and wave reflections,women showed significantly larger age-related variations than men as demonstrated by steeper regression slopes(all P for age by sex interaction<0.05).The blood pressures,body mass index(BMI),and triglyceride levels were identified as major clinical determinants of carotid properties with adjustment of age and sex.Conclusions:The age-and sex-specific reference values of carotid properties measured by RF ultrasound for healthy Chinese adults were established.The blood pressures,BMI,and triglyceride levels should be considered for clinical application of corresponding reference values.

Erratum to“3D collagen matrices modulate the transcriptional trajectory of bone marrow hematopoietic progenitors into macrophage lineage commitment”[Bioact.Mater.10C(April 2022)255-268]

The publisher regrets the omission of the below footnote for the author Dr.Nu Zhang.The publisher would like to apologise for any inconvenience caused.

3D collagen matrices modulate the transcriptional trajectory of bone marrow hematopoietic progenitors into macrophage lineage commitment

Physical signals provided by the extracellular matrix(ECM)are key microenvironmental parameters for the fate decision of hematopoietic stem and progenitor cells(HSPC)in bone marrow.Insights into cell-ECM interactions are critical for advancing HSC-based tissue engineering.Herein,we employed collagen hydrogels and collagen-alginate hydrogels of defined stiffness to study the behaviors of hematopoietic progenitor cells(HPCs).Three-dimensional(3D)collagen hydrogels with a stiffness of 45 Pa were found to promote HPC maintenance and colony formation of monocyte/macrophage progenitors.Using single-cell RNA sequencing(scRNA-seq),we also characterized the comprehensive transcriptional profiles of cells randomly selected from two-dimensional(2D)and 3D hydrogels.A distinct maturation trajectory from HPCs into macrophages within the 3D microenvironment was revealed by these results.3D-derived macrophages expressed high levels of various cytokines and chemokines,such as Saa3,Cxcl2,Socs3 and Tnf.Furthermore,enhanced communication between 3D-macrophages and other hematopoietic clusters based on ligand-repair interactions was demonstrated through bioinformatic analyses.Our research underlines the regulatory role of matrix-dimensionality in HPC differentiation and therefore probably be applied to the generation of specialized macrophages.

The DIRECT consortium and the REST-meta-MDD project:towards neuroimaging biomarkers of major depressive disorder

Despite a growing neuroimaging literature on the pathophysiology of major depressive disorder(MDD),repro-ducible findings are lacking,probably reflecting mostly small sample sizes and heterogeneity in analytic approaches.To address these issues,the Depression Imaging REsearch ConsorTium(DIRECT)was launched.The REST-meta-MDD project,pooling 2428 functional brain images processed with a standardized pipeline across all participating sites,has been the first effort from DIRECT.In this review,we present an overview of the moti-vations,rationale,and principal findings of the studies so far from the REST-meta-MDD project.Findings from the first round of analyses of the pooled repository have included alterations in functional connectivity within the default mode network,in whole-brain topological properties,in dynamic features,and in functional lat-eralization.These well-powered exploratory observations have also provided the basis for future longitudinal hypothesis-driven research.Following these fruitful explorations,DIRECT has proceeded to its second stage of data sharing that seeks to examine ethnicity in brain alterations in MDD by extending the exclusive Chinese original sample to other ethnic groups through international collaborations.A state-of-the-art,surface-based preprocessing pipeline has also been introduced to improve sensitivity.Functional images from patients with bipolar disorder and schizophrenia will be included to identify shared and unique abnormalities across diag-nosis boundaries.In addition,large-scale longitudinal studies targeting brain network alterations following antidepressant treatment,aggregation of diffusion tensor images,and the development of functional magnetic resonance imaging-guided neuromodulation approaches are underway.Through these endeavours,we hope to accelerate the translation of functional neuroimaging findings to clinical use,such as evaluating longitudinal effects of antidepressant medications and developing individualized neuromodulation targets,while building an open repository for the scientific community.