Prognositic value of anoikis and tumor immune microenvironment-related gene in the treatment of osteosarcoma

Objective:Osteosarcoma is a highly aggressive primary malignant bone tumor commonly seen in children and adolescents,with a poor prognosis.Anchorage-dependent cell death(anoikis)has been proven to be indispensable in tumor metastasis,regulating the migration and adhesion of tumor cells at the primary site.However,as a type of programmed cell death,anoikis is rarely studied in osteosarcoma,especially in the tumor immune microenvironment.This study aims to clarify prognostic value of anoikis and tumor immune microenvironment-related gene in the treatment of osteosarcoma.Methods:Anoikis-related genes(ANRGs)were obtained from GeneCards.Clinical information and ANRGs expression profiles of osteosarcoma patients were sourced from the therapeutically applicable research to generate effective therapies and Gene Expression Omnibus(GEO)databases.ANRGs highly associated with tumor immune microenvironment were identified by the estimate package and the weighted gene coexpression network analysis(WGCNA)algorithm.Machine learning algorithms were performed to construct long-term survival predictive strategy,each sample was divided into high-risk and low-risk subgroups,which was further verified in the GEO cohort.Finally,based on single-cell RNA-seq from the GEO database,analysis was done on the function of signature genes in the osteosarcoma tumor microenvironment.Results:A total of 51 hub ANRGs closely associated with the tumor microenvironment were identified,from which 3 genes(MERTK,BNIP3,S100A8)were selected to construct the prognostic model.Significant differences in immune cell activation and immune-related signaling pathways were observed between the high-risk and low-risk groups based on tumor microenvironment analysis(all P<0.05).Additionally,characteristic genes within the osteosarcoma microenvironment were identified in regulation of intercellular crosstalk through the GAS6-MERTK signaling pathway.Conclusion:The prognostic model based on ANRGs and tumor microenvironment demonstrate good predictive power and provide more personalized treatment options for patients with osteosarcoma.

In Situ Raman Monitoring of Trace Antibiotics in Different Harsh Water Environments

In situ surface-enhanced Raman scattering(SERS)is a widely used operando analytical technique,while facing numerous complex factors in applications under aqueous environment,such as low detection sensitivity,poor anti-interference capability,etc.,resulting in unreliable detectability.To address these issues,herein a new hydrophobic SERS strategy has been attempted.By comprehensively designing and researching a SERS-active structure of superhydrophobic ZnO/Ag nanowires,we demonstrate that hydrophobicity can not only draw analytes from water onto substrate,but also adjust"hottest spot"from the bottom of the nanowires to the top.As a result,the structure can simultaneously concentrate the dispersed molecules in water and the enhanced electric field in structure into a same zone,while perfecting its own anti-interference ability.The underwater in situ analytical enhancement factor of this platform is as high as 1.67×10^(11),and the operando limited of detection for metronidazole(MNZ)reaches to 10^(-9)M.Most importantly,we also successfully generalized this structure to various real in situ detection scenarios,including on-site detection of MNZ in corrosive urine,real-time warning of wrong dose of MNZ during intravenous therapy,in situ monitoring of MNZ in flowing wastewater with particulate interference,etc.,demonstrating the great application potential of this hydrophobic platform.This work realizes a synergistic promotion for in situ SERS performance under aqueous environment,and also provides a novel view for improving other in situ analytical techniques.

Preventing brain aging by the artificial enforcement of the unfolded protein response:future directions

As the life expectancy of the world’s population increases,age-related diseases are emerging as one of the greatest problems facing modern society.The onset of dementia and neurodegenerative diseases is strictly dependent on aging as a major risk factor and has a profound impact on various aspects of the lives of individuals and their families.

Sorafenib inhibits ossification of the posterior longitudinal ligament by blocking LOXL2-mediated vascularization

Ossification of the Posterior Longitudinal Ligament(OPLL)is a degenerative hyperostosis disease characterized by the transformation of the soft and elastic vertebral ligament into bone,resulting in limited spinal mobility and nerve compression.Employing both bulk and single-cell RNA sequencing,we elucidate the molecular characteristics,cellular components,and their evolution during the OPLL process at a single-cell resolution,and validate these findings in clinical samples.This study also uncovers the capability of ligament stem cells to exhibit endothelial cell-like phenotypes in vitro and in vivo.Notably,our study identifies LOXL2 as a key regulator in this process.Through gain-and loss-of-function studies,we elucidate the role of LOXL2 in the endothelial-like differentiation of ligament cells.It acts via the HIF1A pathway,promoting the secretion of downstream VEGFA and PDGF-BB.This function is not related to the enzymatic activity of LOXL2.Furthermore,we identify sorafenib,a broad-spectrum tyrosine kinase inhibitor,as an effective suppressor of LOXL2-mediated vascular morphogenesis.By disrupting the coupling between vascularization and osteogenesis,sorafenib demonstrates significant inhibition of OPLL progression in both BMP-induced and enpp1 deficiency-induced animal models while having no discernible effect on normal bone mass.These findings underscore the potential of sorafenib as a therapeutic intervention for OPLL.

Bioengineering breakthroughs:The impact of stem cell models on advanced therapy medicinal product development

The burgeoning field of bioengineering has witnessed significant strides due to the advent of stem cell models,particularly in their application in advanced therapy medicinal products(ATMPs).In this review,we examine the multifaceted impact of these developments,emphasizing the potential of stem cell models to enhance the sophistication of ATMPs and to offer alternatives to animal testing.Stem cell-derived tissues are particularly promising because they can reshape the preclinical landscape by providing more physiologically relevant and ethically sound platforms for drug screening and disease modelling.We also discuss the critical challenges of reproducibility and accuracy in measurements to ensure the integrity and utility of stem cell models in research and application.Moreover,this review highlights the imperative of stem cell models to align with regulatory standards,ensuring using stem cells in ATMPs translates into safe and effective clinical therapies.With regulatory approval serving as a gateway to clinical adoption,the collaborative efforts between scientists and regulators are vital for the progression of stem cell applications from bench to bedside.We advocate for a balanced approach that nurtures innovation within the framework of rigorous validation and regulatory compliance,ensuring that stem cell-base solutions are maximized to promote public trust and patient health in ATMPs.

Emerging and multifaceted potential contributions of polyphenols in the management of type 2 diabetes mellitus

Type 2 diabetes mellitus(T2DM)is recognized as a serious public health concern with a considerable impact on human life,long-term health expenditures,and substantial health losses.In this context,the use of dietary polyphenols to prevent and manage T2DM is widely documented.These dietary compounds exert their beneficial effects through several actions,including the protection of pancreatic islet β-cell,the antioxidant capacities of these molecules,their effects on insulin secretion and actions,the regulation of intestinal microbiota,and their contribution to ameliorate diabetic complications,particularly those of vascular origin.In the present review,we intend to highlight these multifaceted actions and the molecular mechanisms by which these plant-derived secondary metabolites exert their beneficial effects on type 2 diabetes patients.

Effect of nocturia in patients with different severity of obstructive sleep apnea on polysomnography: A retrospective observational study

Patients with suspected OSA were examined using PSG.They were divided into two groups based on the presence of nocturia.Nocturia was defined as a patient who needed to void at least once.Apneaehypopnea index(AHI)was employed to classify patients according to degrees of severity:AHI<5 events/h,5 events/hAHI<15 events/h,15 events/hAHI<30 events/h,and AHI30 events/h,defined as normal,mild OSA,moderate OSA,and severe OSA,respectively.Demographic variables,PSG parameters,International Prostate Symptom Scores(IPSSs),and quality of life scores due to urinary symptoms were analyzed.Results:In total 140 patients,114 patients had OSA(48 had mild OSA;34 had moderate OSA;and 32 had severe OSA)and 107 patients had nocturia.The total IPSS was significantly higher in nocturia patients in all groups except the group of severe OSA patients.With the increasing severity of OSA,more correlated factors related to nocturia were determined.In mild OSA patients,nocturia related to increased age(p=0.025),minimum arterial blood oxygenation saturation(p=0.046),and decreased AHI of non-rapid eye movement(p=0.047),AHI of total sleep time(p=0.010),and desaturation index(p=0.012).In moderate OSA patients,nocturia related to increased age(p<0.001),awake time(p=0.025),stage 1 sleep(p=0.033),and sleep latency(p=0.033),and decreased height(p=0.044),weight(p=0.025),and sleep efficiency(p=0.003).In severe OSA patients,nocturia related to increased weight(p=0.011),body mass index(p=0.009),awake time(p=0.008),stage 1 sleep(p=0.040),arousal number(p=0.030),arousal index(p=0.013),periodic limb movement number(p=0.013),and periodic limb movement index(p=0.004),and decreased baseline arterial blood oxygenation saturation(p=0.046).Conclusion:Our study revealed that there were more correlated factors related to nocturia with increasing severity of OSA.This study helps in clinical education and treatment for OSA patients with different severity.

Application of patient-derived tumor models in anticancer drug development and individualized medicine

Malignant tumor is the second leading cause of death due to its high incidence, lack of effective treatment and poor prognosis. The evaluation of anticancer drugs used to based on NCI-60 cell line models, but the limited heterogeneity and the divorce from clinical practice of models lead to extremely low success rate of novel anticancer drugs during clinical trials (less than 10%). In recent years, because of the high heterogeneity and human derived tumor matrix, patient-derived tumor models have been gradually applied to the preclinical evaluation of various antitumor drugs, which shows certain advantages in predicting the clinical efficacy of antitumor drugs. Optimize the drug combination through patient-derived tumor models to achieve individualized medicine has gradually become an indispensable strategy in clinical cancer therapy. The current review summarized the development of patient-derived tumor models, characterized the application, advantages and challenges of them in preclinical antitumor drug evaluation and clinical precise medicine, which will provide a scientific basis and novel insights for further basic research, drug development and clinical application.

The Development of the “Monitoring Application for Inappropriate Expressions in Nursing Records”for PsyNACS©

Lengthening periods of hospitalization, increasing numbers of patients with age-related complications, and a shortage of nursing staff have been of great concern in medical psychiatry in Japan. Under these circumstances, countries such as Japan that face a super-aging society and a decline in the working-age population, have been recommended for use of advanced information and communications technology (ICT) to improve the efficiency of medical treatment and care. This study aims to develop the “Monitoring Application for Inappropriate Expressions in Nursing Records” for using PsyNACS©, which will enable psychiatric nursing plans to harness the advantages of ICT. The functions considered necessary are as follows: 1) identification of users who enter information;2) a necessary database for lists of inappropriate expressions;3) development of a matching function to recommend proper writing input and a warning function;and 4) a management function for an inappropriate expression list database. A demonstration experiment for developing the application in this study was conducted at a specialized psychiatric hospital. To introduce them to the application, nurses and nurse managers were informed about the system developed in this study, and a survey regarding their opinions on the functions of the application was conducted with ten nurse managers. The results were evaluated in terms of usefulness for nursing care, documentation, and the education of nurses from an ethical perspective. This study suggests that matching their input with an inappropriate expression database will allow nurses to record more appropriate expressions. From an ethical perspective, the ability to use appropriate expressions makes records more likely to withstand disclosure requests from patients and their families.

Study of tumor necrosis factor receptor in the inflammatory bowel disease

Ulcerative colitis(UC)and Crohn’s disease(CD)are part of Inflammatory Bowel Diseases(IBD)and have pathophysiological processes such as bowel necrosis and enteric neurons and enteric glial cells.In addition,the main inflammatory mediator is related to the tumor necrosis factor-alpha(TNF-α).TNF-αis a mediator of the intestinal inflammatory processes,thus being one of the main cytokines involved in the pathogenesis of IBD,however,its levels,when measured,are present in the serum of patients with IBD.In addition,TNF-αplays an important role in promoting inflammation,such as the production of interleukins(IL),for instance IL-1βand IL-6.There are two receptors for TNF as following:The tumor necrosis factor 1 receptor(TNFR1);and the tumor necrosis factor 2 receptor(TNFR2).They are involved in the pathogenesis of IBD and their receptors have been detected in IBD and their expression is correlated with disease activity.The soluble TNF form binds to the TNFR1 receptor with,and its activation results in a signaling cascade effects such as apoptosis,cell proliferation and cytokine secretion.In contrast,the transmembrane TNF form can bind both to TNFR1 and TNFR2.Recent studies have suggested that TNF-αis one of the main pro-inflammatory cytokines involved in the pathogenesis of IBD,since TNF levels are present in the serum of both patients with UC and CD.Intravenous and subcutaneous biologics targeting TNF-αhave revolutionized the treatment of IBD,thus becoming the best available agents to induce and maintain IBD remission.The application of antibodies aimed at neutralizing TNF-αin patients with IBD that induce a satisfactory clinical response in up to 60%of patients,and also induced long-term maintenance of disease remission in most patients.It has been suggested that anti-TNF-αagents inactivate the pro-inflammatory cytokine TNF-αby direct neutralization,i.e.,resulting in suppression of inflammation.However,anti-TNF-αantibodies perform more complex functions than a simple blockade.

Contributions of the receptor for advanced glycation end products axis activation in gastric cancer

Compelling shreds of evidence derived from both clinical and experimental research have demonstrated the crucial contribution of receptor for advanced glycation end products(RAGE)axis activation in the development of neoplasms,including gastric cancer(GC).This new actor in tumor biology plays an important role in the onset of a crucial and long-lasting inflammatory milieu,not only by supporting phenotypic changes favoring growth and dissemination of tumor cells,but also by functioning as a pattern-recognition receptor in the inflammatory response to Helicobacter pylori infection.In the present review,we aim to highlight how the overexpression and activation of the RAGE axis contributes to the proliferation and survival of GC cells as and their acquisition of more invasive phenotypes that promote dissemination and metastasis.Finally,the contribution of some single nucleotide polymorphisms in the RAGE gene as susceptibility or poor prognosis factors is also discussed.

Metabolic regulation of innate immunity in cancer immunotherapy

Innate immunity,originally recognized as the primary defense mechanism against pathogenic infections,has also been shown to have an important role in anti-tumor immunity.Host cells recognize cytosolic DNA and RNA,which triggers a cascade of signaling events via nucleic-acid sensing receptors,including endosomal Toll-like receptors(TLRs),cytoplasmic cyclic GMP-AMP synthase(cGAS)for double-stranded DNA sensing,and cytoplasmic retinoic acid-inducible gene I(RIG-I)for double-stranded RNA detection.

Assessment of functional prognosis of anterior cruciate ligament reconstruction in athletes based on a body shape index

BACKGROUND A healthy body shape is essential to maintain athletes’sports level.At present,little is known about the effect of athletes’body shape on anterior cruciate ligament reconstruction(ACLR).Moreover,the relationship between body shape and variables such as knee joint function after operation and return to the field has not been well studied.AIM To verify the relationship between a body shape index(ABSI)and the functional prognosis of the knee after ACLR in athletes with ACL injuries.METHODS We reviewed 76 athletes with unilateral ACL ruptures who underwent ACLR surgery in the First Hospital of Shanxi Medical University between 2017 and 2020,with a follow-up period of more than 24 mo.First,all populations were divided into a High-ABSI group(ABSI>0.835,n=38)and a Low-ABSI group(ABSI<0.835,n=38)based on the arithmetic median(0.835)of ABSI values.The primary exposure factor was ABSI,and the outcome indicators were knee function scores as well as postoperative complications.The correlation between ABSI and postoperative knee function scores and postoperative complications after ACLR were analyzed using multifactorial logistic regression.RESULTS The preoperative knee function scores of the two groups were similar.The surgery and postoperative rehabilitation exercises,range of motion(ROM)compliance rate,Lysholm score,and Knee Injury and Osteoarthritis Outcome Score of the two groups gradually increased,whereas the quadriceps atrophy index gradually decreased.The knee function scores were higher in the Low-ABSI group than in the High-ABSI group at the 24-mo postoperative follow-up(P<0.05).In multifactorial logistic regression,ABSI was a risk factor of low knee joint function score after surgery,specifically low ROM scores(odds ratio[OR]=1.31,95%confidence interval[CI][1.10-1.44];P<0.001),low quadriceps atrophy index(OR=1.11,95%CI[0.97-1.29];P<0.05),low Lysholm scores(OR=2.34,95%CI[1.78-2.94];P<0.001),low symptoms(OR=1.14,95%CI[1.02-1.34];P<0.05),low activity of daily living(OR=1.34,95%CI[1.18-1.65];P<0.05),low sports(OR=2.47,95%CI[1.78-2.84];P<0.001),and low quality of life(OR=3.34,95%CI[2.88-3.94];P<0.001).ABSI was also a risk factor for deep vein thrombosis of the lower limb(OR=2.14,95%CI[1.88-2.36],P<0.05]and ACL recurrent rupture(OR=1.24,95%CI[0.98-1.44],P<0.05)after ACLR.CONCLUSION ABSI is a risk factor for the poor prognosis of knee function in ACL athletes after ACLR,and the risk of poor knee function after ACLR,deep vein thrombosis of lower limb,and ACL recurrent rupture gradually increases with the rise of ABSI.

Inflammatory/immune-suppressive microenvironment characteristics of pancreatic cancer patients with Shi-Re syndrome identified by extracellular vesicle long RNA profiling

Background:Numerous long RNAs were detected in extracellular vesicles(EVs),some of which were related with the tissue origins and immune cell types.This study examined the molecular basis of different traditional Chinese medicine syndrome diagnoses(also called syndrome differentiation)in pancreatic ductal adenocarcinoma.Methods:128 pancreatic ductal adenocarcinoma patients with different syndrome diagnoses were retrospectively reviewed in this study.Long RNA sequencing was conducted to analyze the EV long RNA profile of plasma samples.Differentially regulated EV long RNAs were annotated and assessed for Gene Ontology pathway enrichment using DAVID.The online program xCell were used to perform the cell-type enrichment analysis.Results:An average of 15,000 annotated genes,mainly including messenger RNAs,were stably detected per sample.Different syndrome diagnoses exhibited unique EV mRNA expression profiles and therefore different enriched pathways.Gene Set Enrichment Analysis discovered transforming growth factor-βand kirsten rat sarcoma viral oncogene homolog signaling activation as the hallmarks of cancer with Shi-Re syndrome.Cell-type enrichment analysis also revealed a varied inflammation/immune cell type distribution among patients with or without Shi-Re diagnosis.Mast cells,platelets and Tregs were significantly enriched but basophils,common lymphoid progenitors,dendritic cells,and conventional dendritic cells were decreased in patients with Shi-Re diagnosis compared with patients without Shi-Re diagnosis.Conclusion:We identified the hallmarks of cancer with different syndrome diagnoses based on plasma EV long RNA sequencing.In particular,transforming growth factor-βand kirsten rat sarcoma viral oncogene homolog signaling activation were the hallmarks of Shi-Re syndrome,which contribute to shape an inflammatory/immune-suppressive tumor microenvironment in pancreatic ductal adenocarcinoma.

Diagnostic use of superb microvascular imaging in evaluating septic arthritis of the manubriosternal joint:A case report

BACKGROUND Septic arthritis of the manubriosternal joint is a diagnostic challenge due to its rarity and anatomical characteristic.Conventional ultrasound,plain radiographs,and computed tomography are not able to confirm or even suspect arthritis early.Superb microvascular imaging is a new advanced Doppler technique in evaluating low-flow microvascular patterns.The higher sensitivity for increased perisynovial vascularity helps to suspect septic arthritis and forms a methodical approach to using magnetic resonance imaging(MRI).CASE SUMMARY A 34-year-old immunocompetent woman presented with a fever and a dull ache in the chest radiating to the right arm.Traumatic injury and the most common respiratory and cardiac disorders were ruled out.Blood cultures came back positive for Staphylococcus aureus,and sepsis was confirmed.A small lump was noted on the chest during the first week of hospitalization.Superb microvascular imaging was performed and septic arthritis of the manubriosternal joint was detected.MRI confirmed the diagnosis and showed septic arthritis of the manubriosternal joint with several localized abscesses behind the sternum.The patient was treated for three weeks with intravenous antibiotics and the outcome was favorable:Inflammatory markers became normal,and the lump disappeared.Three months later,the patient was examined for a new episode of mild pain in the sternum and was diagnosed with persistent perichondritis by ultrasound in comparison with MRI.CONCLUSION Superb microvascular imaging is a useful tool for the early diagnosis of septic arthritis of the manubriosternal joint and following-up.

Characteristics of Sleep and Autonomic Activity in Active Older Adults Based on Metabolic Age: A Comparative Case Study

Background: The balance of autonomic nervous system activity and its relationship with body composition, sleep quality, and activities of daily living among older people is still unclear. Purpose: This comparative case study examined the characteristics of body composition, sleep quality, and autonomic nerve activity in active older adults with a younger body age-calculated from age trends in body composition and basal metabolic rate. Methods: We selected two cases with a metabolic age younger than their actual age. They had good sleep quality, no sarcopenia, strong muscle and grip strength, and balanced autonomic nervous system activity. They were compared with two other age- and gender-matched cases, who had poor sleep quality, unbalanced autonomic nervous system activity, and had a physical age closer to their actual age. Results: Older adults with more muscle mass and higher basal metabolism were younger than their actual age, had a better sleep status, and had a good balance of autonomic nervous activity during exercise stimulation. They also had lower percentages of body and visceral fat and higher percentages of body water. Conclusion: Two cases had a metabolic age younger than their actual age were found to be much younger than their actual age. However, the older adults with normal muscle mass and basal metabolic rate had poor sleep status and no sympathetic hyperactivity during exercise simulation.

Analysis of Emotions Using Multimodal Data: A Case Study

In this case study, we hypothesized that sympathetic nerve activity would be higher during conversation with PALRO robot, and that conversation would result in an increase in cerebral blood flow near the Broca’s area. The facial expressions of a human subject were recorded, and cerebral blood flow and heart rate variability were measured during interactions with the humanoid robot. These multimodal data were time-synchronized to quantitatively verify the change from the resting baseline by testing facial expression analysis, cerebral blood flow, and heart rate variability. In conclusion, this subject indicated that sympathetic nervous activity was dominant, suggesting that the subject may have enjoyed and been excited while talking to the robot (normalized High Frequency < normalized Low Frequency: 0.22 ± 0.16 < 0.78 ± 0.16). Cerebral blood flow values were higher during conversation and in the resting state after the experiment than in the resting state before the experiment. Talking increased cerebral blood flow in the frontal region. As the subject was left-handed, it was confirmed that the right side of the brain, where the Broca’s area is located, was particularly activated (Left < right: 0.15 ± 0.21 < 1.25 ± 0.17). In the sections where a “happy” facial emotion was recognized, the examiner-judged “happy” faces and the MTCNN “happy” results were also generally consistent.

Echinacoside attenuates glandular fibrosis in benign prostatic hyperplasia via inhibiting MKK6/MK2 signaling pathway

Background:Lower urinary tract symptoms commonly occur in the elderly population and seriously constrain the quality of life.Glandular fibrosis is an important pathobiological process in benign prostatic hyperplasia and is also a main inducing factor for benign prostatic hyperplasia-associated lower urinary tract symptoms.Cistanches species is an important herbal medicine resource and is traditionally used in ameliorating renal and prostatic defects.Methods:This study was to investigate the potential protective function of echinacoside(a bioactive compound from Cistanches)against prostatic fibrosis in mice and human benign prostatic hyperplasia epithelial-1 cell models.Results:It was found that echinacoside attenuated testosterone-induced prostatic hyperplasia and collagen deposition in mice,relieved prostate local inflammation and oxidative damage,and ameliorated prostatic epithelial-mesenchymal transition.Additionally,echinacoside inhibited the activation of the MKK6/MK2 signaling pathway both in vivo and in vitro.Conclusion:This study added new evidence for the anti-fibrotic function of echinacoside on the prostate and provided new insights for understanding its possible pharmacological mechanisms.

RPLP0/TBP are the most stable reference genes for human dental pulp stem cells under osteogenic differentiation

BACKGROUND Validation of the reference gene(RG)stability during experimental analyses is essential for correct quantitative real-time polymerase chain reaction(RT-qPCR)data normalisation.Commonly,in an unreliable way,several studies use genes involved in essential cellular functions[glyceraldehyde-3-phosphate dehydro-genase(GAPDH),18S rRNA,andβ-actin]without paying attention to whether they are suitable for such experimental conditions or the reason for choosing such genes.Furthermore,such studies use only one gene when Minimum Information for Publication of Quantitative Real-Time PCR Experiments guidelines recom-mend two or more genes.It impacts the credibility of these studies and causes dis-tortions in the gene expression findings.For tissue engineering,the accuracy of gene expression drives the best experimental or therapeutical approaches.We cultivated DPSCs under two conditions:Undifferentiated and osteogenic dif-ferentiation,both for 35 d.We evaluated the gene expression of 10 candidates for RGs[ribosomal protein,large,P0(RPLP0),TATA-binding protein(TBP),GAPDH,actin beta(ACTB),tubulin(TUB),aminolevulinic acid synthase 1(ALAS1),tyro-sine 3-monooxygenase/tryptophan 5-monooxygenase activation protein,zeta(YWHAZ),eukaryotic translational elongation factor 1 alpha(EF1a),succinate dehydrogenase complex,subunit A,flavoprotein(SDHA),and beta-2-micro-globulin(B2M)]every 7 d(1,7,14,21,28,and 35 d)by RT-qPCR.The data were analysed by the four main algorithms,ΔCt method,geNorm,NormFinder,and BestKeeper and ranked by the RefFinder method.We subdivided the samples into eight subgroups.RESULTS All of the data sets from clonogenic and osteogenic samples were analysed using the RefFinder algorithm.The final ranking showed RPLP0/TBP as the two most stable RGs and TUB/B2M as the two least stable RGs.Either theΔCt method or NormFinder analysis showed TBP/RPLP0 as the two most stable genes.However,geNorm analysis showed RPLP0/EF1αin the first place.These algorithms’two least stable RGs were B2M/GAPDH.For BestKeeper,ALAS1 was ranked as the most stable RG,and SDHA as the least stable RG.The pair RPLP0/TBP was detected in most subgroups as the most stable RGs,following the RefFinfer ranking.CONCLUSION For the first time,we show that RPLP0/TBP are the most stable RGs,whereas TUB/B2M are unstable RGs for long-term osteogenic differentiation of human DPSCs in traditional monolayers.

Self‑Healing Dynamic Hydrogel Microparticles with Structural Color for Wound Management

Chronic diabetic wounds confront a significant medical challenge because of increasing prevalence and difficult-healing circumstances.It is vital to develop multifunctional hydrogel dressings,with well-designed morphology and structure to enhance flexibility and effectiveness in wound management.To achieve these,we propose a self-healing hydrogel dressing based on structural color microspheres for wound management.The microsphere comprised a photothermal-responsive inverse opal framework,which was constructed by hyaluronic acid methacryloyl,silk fibroin methacryloyl and black phosphorus quantum dots(BPQDs),and was further re-filled with a dynamic hydrogel.The dynamic hydrogel filler was formed by Knoevenagel condensation reaction between cyanoacetate and benzaldehyde-functionalized dextran(DEX-CA and DEX-BA).Notably,the composite microspheres can be applied arbitrarily,and they can adhere together upon near-infrared irradiation by leveraging the BPQDs-mediated photothermal effect and the thermoreversible stiffness change of dynamic hydrogel.Additionally,eumenitin and vascular endothelial growth factor were co-loaded in the microspheres and their release behavior can be regulated by the same mechanism.Moreover,effective monitoring of the drug release process can be achieved through visual color variations.The microsphere system has demonstrated desired capabilities of controllable drug release and efficient wound management.These characteristics suggest broad prospects for the proposed composite microspheres in clinical applications.