DI-3-n-butylphthalide exerts neuroprotective effects by modulating hypoxia-inducible factor 1-alpha ubiquitination to attenuate oxidative stress-induced apoptosis

DI-3-n-butylphthalide is used to treat mild and moderate acute ischemic stroke.However,the precise underlying mechanism requires further investigation.In this study,we investigated the molecular mechanism of DI-3-n-butylphthalide action by various means.We used hydrogen peroxide to induce injury to PC12cells and RAW264.7 cells to mimic neuronal oxidative stress injury in stroke in vitro and examined the effects of DI-3-n-butylphthalide.We found that DI-3-nbutylphthalide pretreatment markedly inhibited the reduction in viability and reactive oxygen species production in PC12 cells caused by hydrogen peroxide and inhibited cell apoptosis.Furthermore,DI-3-n-butylphthalide pretreatment inhibited the expression of the pro-apoptotic genes Bax and Bnip3.DI-3-nbutylphthalide also promoted ubiquitination and degradation of hypoxia inducible factor 1α,the key transcription factor that regulates Bax and Bnip3 genes.These findings suggest that DI-3-n-butylphthalide exhibits a neuroprotective effect on stroke by promoting hypoxia inducible factor-1α ubiquitination and degradation and inhibiting cell apoptosis.

A pancreatic player in dementia:pathological role for islet amyloid polypeptide accumulation in the brain

Type 2 diabetes mellitus patients have a markedly higher risk of developing dementia.While multiple factors contribute to this predisposition,one of these involves the increased secretion of amylin,or islet amyloid polypeptide,that accompanies the pathophysiology of type 2 diabetes mellitus.Islet amyloid polypeptide accumulation has undoubtedly been implicated in various forms of dementia,including Alzheimer’s disease and vascular dementia,but the exact mechanisms underlying islet amyloid polypeptide’s causative role in dementia are unclear.In this review,we have summarized the literature supporting the various mechanisms by which islet amyloid polypeptide accumulation may cause neuronal damage,ultimately leading to the clinical symptoms of dementia.We discuss the evidence for islet amyloid polypeptide deposition in the brain,islet amyloid polypeptide interaction with other amyloids implicated in neurodegeneration,neuroinflammation caused by islet amyloid polypeptide deposition,vascular damage induced by islet amyloid polypeptide accumulation,and islet amyloid polypeptide-induced cytotoxicity.There are very few therapies approved for the treatment of dementia,and of these,clinical responses have been controversial at best.Therefore,investigating new,targetable pathways is vital for identifying novel therapeutic strategies for treating dementia.As such,we conclude this review by discussing islet amyloid polypeptide accumulation as a potential therapeutic target not only in treating type 2 diabetes mellitus but as a future target in treating or even preventing dementia associated with type 2 diabetes mellitus.

Diagnostic and classification value of immune-related lncRNAs in dilated cardiomyopathy

Background:Various physiological mechanisms are linked to dilated cardiomyopathy(DCM)development,including oxidative stress,immune irregularities,inflammation,fibrosis,and genetic changes.However,precise molecular drivers of DCM,especially regarding abnormal immune responses,remain unclear.This study investigates immune-related long non-coding RNAs(lncRNAs)in DCM’s diagnostic and therapeutic potential.Methods:GSE141910,GSE135055,and GSE165303 datasets were acquired from the GEO database.LASSO,SVM-RFE,and random forest algorithms identified DCM-associated immune-related lncRNAs.Diagnostic capabilities were assessed by Nomogram and receiver operating characteristic(ROC)curves.Multivariate linear regression explored lncRNA correlations with ejection fraction.Single-sample gene set enrichment analysis(ssGSEA)gauged immune cell infiltration/functions.Functional enrichment analyses were performed using Gene set variation analysis(GSVA),gene ontology(GO),and the Kyoto Encyclopedia of Genes and Genomes(KEGG).Consensus clustering categorized DCM cases.Results:Ten immune-related lncRNAs emerged:C10orf71-AS1,FHAD1-AS1,SCIRT,FNDC1-AS1,MELTFAS1,LOC101928834,GDNF-AS1,DCXR-DT,C3orf36,and LOC107985323.These lncRNAs,tied to immunomodulation,showed promising DCM diagnostic accuracy.Adjusted for confounders,they independently correlated with ejection fraction.Using lncRNA expression,DCM patients were grouped into subtypes.Subtype C1 displayed a higher level of immune cell infiltration and immune checkpoint expression compared to subtype C2,emphasizing the variations in the immune microenvironment.Conclusion:This study identifies ten immune-related lncRNAs for further exploration in DCM diagnosis and subtyping.Based on expression patterns,we propose two potential DCM subtypes.Notably,findings are preliminary and hypothesis-generating,demanding validation and further investigation.This research provides insights into DCM diagnosis and classification.

In vivo tracking of human adipose-derived stem cells labeled with ferumoxytol in rats with middle cerebral artery occlusion by magnetic resonance imaging

Ferumoxytol, an iron replacement product, is a new type of superparamagnetic iron oxide ap- proved by the US Food and Drug Administration. Herein, we assessed the feasibility of tracking transplanted human adipose-derived stem cells labeled with ferumoxytol in middle cerebral artery occlusion-injured rats by 3.0 T MRI in vivo. 1 × 104 human adipose-derived stem cells labeled with ferumoxytol-heparin-protamine were transplanted into the brains of rats with middle cerebral artery occlusion. Neurologic impairment was scored at 1, 7, 14, and 28 days after transplantation. T2-weighted imaging and enhanced susceptibility-weighted angiography were used to observe transplanted cells. Results of imaging tests were compared with results of Prussian blue staining. The modified neurologic impairment scores were significantly lower in rats transplanted with cells at all time points except I day post-transplantation compared with rats without transplantation. Regions with hypointense signals on T2-weighted and enhanced susceptibility-weighted angiography images corresponded with areas stained by Prussian blue, suggesting the presence of superparamagnetic iron oxide particles within the engrafted cells. Enhanced susceptibility-weighted angiography image exhibited better sensitivity and contrast in tracing ferumoxytol-heparin-protamine-labeled human adipose-derived stem ceils compared with T2-weighted imaging in routine MRI.

Downregulation of caveolin-1 contributes to the synaptic plasticity deficit in the hippocampus of aged rats

Caveolin-1 is involved in the regulation of synaptic plasticity, but the relationship between its ex-pression and cognitive function during aging remains controversial. To explore the relationship be-tween synaptic plasticity in the aging process and changes in learning and memory, we examined caveolin-1 expression in the hippocampus, cortex and cerebellum of rats at different ages. We also examined the relationship between the expression of caveolin-1 and synaptophysin, a marker of synaptic plasticity. Hippocampal caveolin-1 and synaptophysin expression in aged （22-24 month old） rats was significantly lower than that in young （1 month old） and adult （4 months old） rats. Ex- pression levels of both proteins were significantly greater in the cortex of aged rats than in that of young or adult rats, and levels were similar between the three age groups in the cerebellum. Linear regression analysis revealed that hippocampal expression of synaptophysin was associated with memory and learning abilities. Moreover, synaptophysin expression correlated positively with caveolin-1 expression in the hippocampus, cortex and cerebellum. These results confirm that caveolin-1 has a regulatory effect on synaptic plasticity, and suggest that the downregulation of hippocampal caveolin-1 expression causes a decrease in synaptic plasticity during physiological aging.

Neuroregeneration and functional recovery after stroke: advancing neural stem cell therapy toward clinical application

Stroke is a main cause of death and disability worldwide.The ability of the brain to selfrepair in the acute and chronic phases after stroke is minimal;however,promising stem cell-based interventions are emerging that may give substantial and possibly complete recovery of brain function after stroke.Many animal models and clinical trials have demonstrated that neural stem cells(NSCs)in the central nervous system can orchestrate neurological repair through nerve regeneration,neuron polarization,axon pruning,neurite outgrowth,repair of myelin,and remodeling of the microenvironment and brain networks.Compared with other types of stem cells,NSCs have unique advantages in cell replacement,paracrine action,inflammatory regulation and neuroprotection.Our review summarizes NSC origins,characteristics,therapeutic mechanisms and repair processes,then highlights current research findings and clinical evidence for NSC therapy.These results may be helpful to inform the direction of future stroke research and to guide clinical decision-making.

Progress in clinical trials of stem cell therapy for cerebral palsy

Cerebral palsy is the most common disease in children associated with lifelong disability in many countries.Clinical research has demonstrated that traditional physiotherapy and rehabilitation therapies cannot alone cure cerebral palsy.Stem cell transplantation is an emerging therapy that has been applied in clinical trials for a variety of neurological diseases because of the regenerative and unlimited proliferative capacity of stem cells.In this review, we summarize the design schemes and results of these clinical trials.Our findings reveal great differences in population characteristics, stem cell types and doses, administration methods, and evaluation methods among the included clinical trials.Furthermore, we also assess the safety and efficacy of these clinical trials.We anticipate that our findings will advance the rational development of clinical trials of stem cell therapy for cerebral palsy and contribute to the clinical application of stem cells.

Transplantation of autologous peripheral blood mononuclear cells in the subarachnoid space for amyotrophic lateral sclerosis:a safety analysis of 14 patients

There is a small amount of clinical data regarding the safety and feasibility of autologous peripheral blood mononuclear cell transplantation into the subarachnoid space for the treatment of amyotrophic lateral sclerosis.The objectives of this retrospective study were to assess the safety and efficacy of peripheral blood mononuclear cell transplantation in 14 amyotrophic lateral sclerosis patients to provide more objective data for future clinical trials.After stem cell mobilization and collection,autologous peripheral blood mononuclear cells（1 × 109） were isolated and directly transplanted into the subarachnoid space of amyotrophic lateral sclerosis patients.The primary outcome measure was incidence of adverse events.Secondary outcome measures were electromyography 1 week before operation and 4 weeks after operation,Functional Independence Measurement,Berg Balance Scale,and Dysarthria Assessment Scale 1 week preoperatively and 1,2,4 and 12 weeks postoperatively.There was no immediate or delayed transplant-related cytotoxicity.The number of leukocytes,serum alanine aminotransferase and creatinine levels,and body temperature were within the normal ranges.Radiographic evaluation showed no serious transplant-related adverse events.Muscle strength grade,results of Functional Independence Measurement,Berg Balance Scale,and Dysarthria Assessment Scale were not significantly different before and after treatment.These findings suggest that peripheral blood mononuclear cell transplantation into the subarachnoid space for the treatment of amyotrophic lateral sclerosis is safe,but its therapeutic effect is not remarkable.Thus,a large-sample investigation is needed to assess its efficacy further.

Caveolin-1 downregulation promotes the dopaminergic neuron-like differentiation of human adipose-derived mesenchymal stem cells

Previous studies have shown that caveolin-1 is involved in regulating the differentiation of mesenchymal stem cells.However,its role in the differentiation of human adipose mesenchymal stem cells into dopaminergic neurons remains unclear.The aim of this study was to investigate whether caveolin-1 regulates the differentiation of human adipose mesenchymal stem cells into dopaminergic-like neurons.We also examined whether the expression of caveolin-1 could be modulated by RNA interference technology to promote the differentiation of human adipose mesenchymal stem cells into dopaminergic-like neurons.The differentiation of human adipose mesenchymal stem cells into dopaminergic neurons was evaluated morphologically and by examining expression of the markers tyrosine hydroxylase,Lmx1a and Nurr1.The analyses revealed that during the differentiation of human adipose mesenchymal stem cells into dopaminergic neurons,the expression of caveolin-1 is decreased.Notably,the downregulation of caveolin-1 promoted the differentiation of human adipose mesenchymal stem cells into dopaminergic-like neurons,and it increased the expression of tyrosine hydroxylase,Lmx1a and Nurr1.Together,our findings suggest that caveolin-1 plays a negative regulatory role in the differentiation of dopaminergic-like neurons from stem cells,and it may therefore be a potential molecular target for strategies for regulating the differentiation of these cells.This study was approved by the Medical Ethics Committee of the First Affiliated Hospital of Dalian Medical University of China(approval No.PJ-KS-KY-2020-54)on March 7,2017.

Anticancer effect of linalool via cancer-specific hydroxyl radical generation in human colon cancer

AIM To investigate the anticancer mechanisms of the monoterpenoid alcohol linalool in human colon cancer cells.METHODS The cytotoxic effect of linalool on the human colon cancer cell lines and a human fibroblast cell line was examined using the WST-8 assay. The apoptosisinducing effect of linalool was measured using the terminal deoxynucleotidyl transferase d UTP nickend labeling assay and flow cytometry with Annexin V. Oxidative stress was investigated by staining for diphenyl-1-pyrenylphosphine, which is a cellular lipid peroxidation marker, and electron spin resonance spectroscopy. Sixteen SCID mice xenografted with human cancer cells were randomized into 3 groups for in vivo analysis: control and low-dose and high-dose linalool groups. The control group was administered tap water orally every 3 d. The linalool treatment groups were administered 100 or 200 μg/kg linalool solution orally for the same period. All mice were sacrificed under anesthesia 21 d after tumor inoculation, and tumors and organs were collected for immunohistochemistry using an anti-4-hydroxynonenal antibody. Tumor weights were measured and compared between groups. RESULTS Linalool induced apoptosis of cancer cells in vitro, following the cancer-specific induction of oxidative stress, which was measured based on spontaneous hydroxyl radical production and delayed lipid peroxidation. Mice in the high-dose linalool group exhibited a 55% reduction in mean xenograft tumor weight compared with mice in the control group(P < 0.05). In addition, tumor-specific lipid peroxidation was observed in the in vivo model.CONCLUSION Linalool exhibited an anticancer effect via cancerspecific oxidative stress, and this agent has potential for application in colon cancer therapy.

Human umbilical cord-derived mesenchymal stem cells promote repair of neonatal brain injury caused by hypoxia/ischemia in rats

Administration of human umbilical cord-derived mesenchymal stem cells(hUC-MSCs)is believed to be an effective method for treating neurodevelopmental disorde rs.In this study,we investigated the possibility of hUC-MSCs treatment of neonatal hypoxic/ischemic brain injury associated with maternal immune activation and the underlying mechanism.We established neonatal rat models of hypoxic/ischemic brain injury by exposing pregnant rats to lipopolysaccharide on day 16 or 17 of pregnancy.Rat offspring were intranasally administe red hUC-MSCs on postnatal day 14.We found that polypyrimidine tract-binding protein-1(PTBP-1)participated in the regulation of lipopolysaccharide-induced maternal immune activation,which led to neonatal hypoxic/ischemic brain injury.Intranasal delive ry of hUC-MSCs inhibited PTBP-1 expression,alleviated neonatal brain injury-related inflammation,and regulated the number and function of glial fibrillary acidic protein-positive astrocytes,there by promoting plastic regeneration of neurons and im p roving brain function.These findings suggest that hUC-MSCs can effectively promote the repair of neonatal hypoxic/ischemic brain injury related to maternal immune activation through inhibition of PTBP-1 expression and astrocyte activation.

Single nucleotide polymorphism of MYOC affected the severity of primary open angle glaucoma

AIM: To detect the mutations in two candidate genes, myocilin (MYOC ) and cytochrome P450 1B1 (CYP1B1 ), in a Chinese family with primary open angle glaucoma (POAG). ·METHODS:Thefamilywascomposedofthreemembers, the parents and a daughter. All members of the family underwent complete ophthalmologic examinations. Exons of MYOC and CYP1B1 genes were screened for sequence alterations by polymerase chain reaction (PCR) and direct DNA sequencing. ·RESULTS: The mother was the proband, she was diagnosed as POAG in both eyes. Her daughter was diagnosed as juvenile -onset POAG. The father was asymptomatic. One MYOC heterozygous mutation c.1150 G 】A (D384N) in exon 3 was identified in the mother, another MYOC heterozygous variation c.1058 C】T (T353I) in exon 3 was identified in the father, and the daughter inherited both of the variations. Meanwhile, three single nucleotide polymorphisms (SNPs) in CYP1B1 gene were found in the family. ·CONCLUSION: The D384N mutation of MYOC has been reported as one of disease-causing mutations in POAG, whereas T353I variation of MYOC was thought as a high risk factor for POAG. The two variations of MYOC were first reported in one juvenile -onset POAG patient whopresented with more severe clinical manifestations, suggesting that T353I polymorphism of MYOC may be associated with the severity of POAG.

Effect of Sonication on a Novel Designed Peptide

The reassembly of a half-sequence ionic self-complementarity peptide CH3CO-Pro-Ser-Phe- Cys-Phe-Lys-Phe-Glu-Pro-NH2 was reported, which could self-assemble into stable nanofibers, and formed hydrogel consisting of 〉99% water. In this study, the nanofiber scaffold was sonicated by an ultrasonic cell disruptor. The effects of sonication were detected by circular dichroism （CD）, atomic force microscopy （AFM）, and rheology. AFM image illustrated that the sonicated fragments could quickly reassemble into nanofibers, while the morphology was distinguishable from the original one. CD spectrum revealed that the conversion occurred mainly between regular β-strand structure and distorted β-strand structure. Rheological analyses showed that the storage modulus （G＇） of the peptide solution at the 7th day after sonication decreased by nearly 40% compared with the value of the solution before sonication. Finally, a plausible conversion model was proposed to interpret the reassembly process.

Dried Rehmannia root protects against glutamate-induced cytotoxity to PC12 cells through energy metabolism-related pathways

Rehmannia has been shown to be clinically effective in treating neurodegenerative diseases; however, the neuroprotective mechanisms remain unclear. In this study, we established a model of neurodegenerative disease using PC12 cytotoxic injury induced by glutamate. The cells were treated with 20 mM glutamate in the absence or presence of water extracts of dried Rehmannia root of varying concentrations（70%, 50% and 30%）. The different concentrations of Rehmannia water extract significantly increased the activity of glutamate-injured cells, reduced the release of lactate dehydrogenase, inhibited apoptosis, increased the concentrations of NADH, NAD and ATP in cells, ameliorated mitochondrial membrane potential, and reduced the levels of light chain 3. Taken together, our findings demonstrate that Rehmannia water extracts exert a cytoprotective effect against glutamate-induced PC12 cell injury via energy metabolism-related pathways.

Evaluation of direct intramural injection to the bladder wall as a method for developing orthotopic tumor models

Background:Bladder cancer poses a great burden on society and its high rate of recurrence and treatment failure necessitates use of appropriate animal models to study its pathogenesis and test novel treatments.Orthotopic models are superior to other types since they provide a normal microenvironment.Four methods are described for developing bladder cancer models inside the animal’s bladder.Direct intramural injection is one of these methods and is widely used.However,its efficacy in model development has not yet been studied.We aimed to evaluate the efficacy and success rate of the direct intramural injection method of developing an orthotopic model for the study of bladder cancer.Method:Tumor cell lines were prepared in four microtubes.Aliquots of 200×10^(3) cells were injected through a 27 gauge needle into the ventral wall of the bladders of 4male and 4 female BALB/c mice following a midline 1 cm laparotomy incision.In addition,1 million cells from each microtube were injected into the flanks of control mice.To prevent infection and alleviate pain,5 mg/kg enrofloxacin and 2.5 mg/kg flunixin meglumine,respectively,were injected subcutaneously.Results:Tumors formed in all mice,resulting in 100% take rate and zero post-operation mortality.Surgery time was≤15 min per mouse.In two mice,tumors were found in the peritoneal space as well.Conclusion:Direct intramural injection is a rapid,reliable,and reproducible method for developing orthotopic models of bladder cancer.It can be done on both male and female mice and only requires readily available surgical tools.However,needle track can result in cell spillage and peritoneal tumors.

Inducing human induced pluripotent stem cell differentiation through embryoid bodies:A practical and stable approach

Human induced pluripotent stem cells(hiPSCs)are invaluable resources for producing high-quality differentiated cells in unlimited quantities for both basic research and clinical use.They are particularly useful for studying human disease mechanisms in vitro by making it possible to circumvent the ethical issues of human embryonic stem cell research.However,significant limitations exist when using conventional flat culturing methods especially concerning cell expansion,differentiation efficiency,stability maintenance and multicellular 3D structure establishment,differentiation prediction.Embryoid bodies(EBs),the multicellular aggregates spontaneously generated from iPSCs in the suspension system,might help to address these issues.Due to the unique microenvironment and cell communication in EB structure that a 2D culture system cannot achieve,EBs have been widely applied in hiPSC-derived differentiation and show significant advantages especially in scaling up culturing,differentiation efficiency enhancement,ex vivo simulation,and organoid establishment.EBs can potentially also be used in early prediction of iPSC differentiation capability.To improve the stability and feasibility of EB-mediated differentiation and generate high quality EBs,critical factors including iPSC pluripotency maintenance,generation of uniform morphology using micro-pattern 3D culture systems,proper cellular density inoculation,and EB size control are discussed on the basis of both published data and our own laboratory experiences.Collectively,the production of a large quantity of homogeneous EBs with high quality is important for the stability and feasibility of many PSCs related studies.

Oral microbiome and pancreatic cancer

BACKGROUND Microbiota profiles differ between patients with pancreatic cancer and healthy people,and understanding these differences may help in early detection of pancreatic cancer.Saliva sampling is an easy and cost-effective way to determine microbiota profiles compared to fecal and tissue sample collection.AIM To investigate the saliva microbiome distribution in patients with pancreatic adenocarcinoma(PDAC)and the role of oral microbiota profiles in detection and risk prediction of pancreatic cancer.METHODS We conducted a prospective study of patients with pancreatic cancer(n=41)and healthy individuals(n=69).Bacterial taxa were identified by 16S ribosomal ribonucleic acid gene sequencing,and a linear discriminant analysis effect size algorithm was used to identify differences in taxa.Operational taxonomic unit values of all selected taxa were converted into a normalized Z-score,and logistic regressions were used to calculate risk prediction of pancreatic cancer.RESULTS Compared with the healthy control group,carriage of Streptococcus and Leptotrichina(z-score)was associated with a higher risk of PDAC[odds ratio(OR)=5.344,95%confidence interval(CI):1.282-22.282,P=0.021 and OR=6.886,95%CI:1.423-33.337,P=0.016,respectively].Veillonella and Neisseria(z-score)were considered a protective microbe that decreased the risk of PDAC(OR=0.187,95%CI:0.055-0.631,P=0.007 and OR=0.309,95%CI:0.100-0.952,P=0.041,respectively).Among the patients with PDAC,patients reporting bloating have a higher abundance of Porphyromonas(P=0.039),Fusobacterium(P=0.024),and Alloprevotella(P=0.041);while patients reporting jaundice had a higher amount of Prevotella(P=0.008);patients reporting dark brown urine had a higher amount of Veillonella(P=0.035).Patients reporting diarrhea had a lower amount of Neisseria and Campylobacter(P=0.024 and P=0.034),and patients reporting vomiting had decreased Alloprevotella(P=0.036).CONCLUSION Saliva microbiome was able to distinguish patients with pancreatic cancer and healthy individuals.Leptotrichia may be specific for patients living in Sichuan Province,southwest China.Symptomatic patients had different bacteria profiles than asymptomatic patients.Combined symptom and microbiome evaluation may help in the early detection of pancreatic cancer.

Mechanoresponse of stem cells for vascular repair

Stem cells have shown great potential in vascular repair.Numerous evidence indicates that mechanical forces such as shear stress and cyclic strain can regulate the adhesion,proliferation,migration,and differentiation of stem cells via serious signaling pathways.The enrichment and differentiation of stem cells play an important role in the angiogenesis and maintenance of vascular homeostasis.In normal tissues,blood flow directly affects the microenvironment of vascular endothelial cells(ECs);in pathological status,the abnormal interactions between blood flow and vessels contribute to the injury of vessels.Next,the altered mechanical forces are transduced into cells by mechanosensors to trigger the reformation of vessels.This process occurs when signaling pathways related to EC differentiation are initiated.Hence,a deep understanding of the responses of stem cells to mechanical stresses and the underlying mechanisms involved in this process is essential for clinical translation.In this the review,we provide an overview of the role of stem cells in vascular repair,outline the performance of stem cells under the mechanical stress stimulation,and describe the related signaling pathways.

Intratracheal administration of umbilical cord-derived mesenchymal stem cells attenuates hyperoxia-induced multi-organ injury via heme oxygenase-1 and JAK/STAT pathways

BACKGROUND Bronchopulmonary dysplasia(BPD)is not merely a chronic lung disease,but a systemic condition with multiple organs implications predominantly associated with hyperoxia exposure.Despite advances in current management strategies,limited progress has been made in reducing the BPD-related systemic damage.Meanwhile,although the protective effects of human umbilical cord-derived mesenchymal stem cells(hUC-MSCs)or their exosomes on hyperoxia-induced lung injury have been explored by many researchers,the underlying mechanism has not been addressed in detail,and few studies have focused on the therapeutic effect on systemic multiple organ injury.AIM To investigate whether hUC-MSC intratracheal administration could attenuate hyperoxia-induced lung,heart,and kidney injuries and the underlying regulatory mechanisms.METHODS Neonatal rats were exposed to hyperoxia(80%O_(2)),treated with hUC-MSCs intratracheal(iT)or intraperitoneal(iP)on postnatal day 7,and harvested on postnatal day 21.The tissue sections of the lung,heart,and kidney were analyzed morphometrically.Protein contents of the bronchoalveolar lavage fluid(BALF),myeloper oxidase(MPO)expression,and malondialdehyde(MDA)levels were examined.Pulmonary inflammatory cytokines were measured via enzyme-linked immunosorbent assay.A comparative transcriptomic analysis of differentially expressed genes(DEGs)in lung tissue was conducted via RNA-sequencing.Subsequently,we performed reverse transcription-quantitative polymerase chain reaction and western blot analysis to explore the expression of target mRNA and proteins related to inflammatory and oxidative responses.RESULTS iT hUC-MSCs administration improved pulmonary alveolarization and angiogenesis(P<0.01,P<0.01,P<0.001,and P<0.05 for mean linear intercept,septal counts,vascular medial thickness index,and microvessel density respectively).Meanwhile,treatment with hUC-MSCs iT ameliorated right ventricular hypertrophy(for Fulton’s index,P<0.01),and relieved reduced nephrogenic zone width(P<0.01)and glomerular diameter(P<0.001)in kidneys.Among the beneficial effects,a reduction of BALF protein,MPO,and MDA was observed in hUC-MSCs groups(P<0.01,P<0.001,and P<0.05 respectively).Increased pro-inflammatory cytokines tumor necrosis factor-alpha,interleukin(IL)-1β,and IL-6 expression observed in the hyperoxia group were significantly attenuated by hUC-MSCs administration(P<0.01,P<0.001,and P<0.05 respectively).In addition,we observed an increase in anti-inflammatory cytokine IL-10 expression in rats that received hUC-MSCs iT compared with rats reared in hyperoxia(P<0.05).Transcriptomic analysis showed that the DEGs in lung tissues induced by hyperoxia were enriched in pathways related to inflammatory responses,epithelial cell proliferation,and vasculature development.hUC-MSCs administration blunted these hyperoxia-induced dysregulated genes and resulted in a shift in the gene expression pattern toward the normoxia group.hUC-MSCs increased heme oxygenase-1(HO-1),JAK2,and STAT3 expression,and their phosphorylation in the lung,heart,and kidney(P<0.05).Remarkably,no significant difference was observed between the iT and iP administration.CONCLUSION iT hUC-MSCs administration ameliorates hyperoxia-induced lung,heart,and kidney injuries by activating HO-1 expression and JAK/STAT signaling.The therapeutic benefits of local iT and iP administration are equivalent.

Prior transfusion of umbilical cord mesenchymal stem cells can effectively alleviate symptoms of motion sickness in mice through interleukin 10 secretion

BACKGROUND Motion sickness(MS)is a disease that occurs during unbalanced movement,characterized by gastrointestinal symptoms and autonomic nervous system activation.Current clinical treatments for MS are limited.Recent evidence indicates that the levels of pro-inflammatory cytokines increase during MS and are associated with an inner ear immune imbalance.In the present study,mesenchymal stem cells(MSCs)have been shown to exert strong immunosuppressive effects.AIM To explore whether umbilical cord-derived mesenchymal stem cells(UC-MSCs)can prevent the occurrence of MS,and the underlying mechanism regulated by MSCs in a mouse model of MS.METHODS A total of 144(equal numbers of males and females)5wkold BALB/c mice were randomly divided into five groups:Normal group(n=16),MS group(n=32),MSCs group(n=32),MS+MSCs group(n=32),and MS+AS101/MSCs group(n=32).The MSCs group(n=32),MS+MSCs group(n=32),and MS+AS101/MSCs group(n=32)were preventively transplanted with UC-MSCs or AS101-treated UC-MSCs(1×106 cells/mouse).Mice in the MS(n=32),MS+MSCs,and MS+AS101/MSCs groups were subjected to rotation on a centrifuge for 10 min at 8×g/min for MS model establishment on days 3,5,8,and 10 after UC-MSCs injection.The Morris water maze(MWM)test was used to observe the symptom of dizziness.Enzyme-linked immunosorbent assay(ELISA)and reverse transcription-quantitative polymerase chain reaction(RT-qPCR)were used to detect the levels of inflammatory cytokines in mice peripheral blood and the petrous part of the temporal bone samples.Western blot analysis was performed to analyze the JAK2/STAT3 signaling pathway in the cochlear tissues.Histological examination was performed by hematoxylin and eosin(HE)staining for conventional morphological evaluation in the petrous part of temporal bone samples.RESULTS The MWM test demonstrated that UC-MSCs improved the symptoms of MS.The MS+MSCs group was faster than the MS group on days 3 and 5(P=0.036 and P=0.002,respectively).ELISA and RT-qPCR showed that the serum and mRNA levels of interleukin-10(IL-10)in the cochlear tissues were increased after transplantation with UC-MSCs(MS+MSCs group vs MS group at 3 and 5 d,P=0.002 and cP<0.001,respectively).RT-qPCR results confirmed a significant increase in IL-10 levels at four time points(MS+MSCs group vs MS group,P=0.009,P=0.009,P=0.048,and P=0.049,respectively).This suggested that UCMSCs reduced the sensitivity of the vestibular microenvironment by secreting IL-10.Moreover,Western blot analysis showed that the MSCs activated the JAK2/STAT3 signaling pathway in the cochlear tissues.The levels of IL-10,IL-10RA,JAK2,STAT3,and phosphorylated JAK2 and STAT3 in the MS+MSCs group were increased compared to those of the MS group(P<0.05).The morphological changes in the four groups showed no significant differences.The role of IL-10 secretion on the ability of UC-MSCs to successfully improve the symptoms of MS was confirmed by the diminished therapeutic effects associated with treatment with the IL-10 inhibitor ammonium trichloro(dioxoethylene-o,o′)tellurate(AS101).CONCLUSION Prophylactic transplantation of UC-MSCs can alleviate the clinical symptoms of MS in mice,particularly at 3-5 d after preventive transplantation.The mechanism for UC-MSCs to reduce the sensitivity of vestibular cortex imbalance may be the secretion of IL-10.The next step is to demonstrate the possibility of curing MS in the vestibular environment by intermittent transplantation of MSCs.Above all,MSCs are expected to become a new method for the clinical prevention and treatment of MS.