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Abstract Inflammation has a destructive effect on the homeostasis of the vascular wall, which is involved in the formation, growth, and rupture of human intracranial aneurysms (IAs) disease progression. However, inflammation-related markers have not been well studied in the risk stratification of unruptured IAs. The purpose of this study was to investigate the predictive value of serum inflammatory markers in the unstable progression of small saccular intracranial aneurysms (SIAs). This study retrospectively included 275 patients with small SIAs (aneurysm diameter less than or equal to 7 mm), to compare the level difference of serum inflammatory complex marker systemic immune-inflammatory index (SII), white blood cell to platelet ratio (WPR), and homocysteine (Hcy) in patients with stable (asymptomatic unruptured) and unstable (symptomatic unruptured, ruptured) small SIAs. 187 patients (68%) had aneurysm-related compression symptoms and rupture outcomes. In the multivariate logistic regression after adjusting for baseline differences, SII, WPR, and Hcy were independent risk factors for the instability of small SIAs, the prediction model combined with other risk factors (previous stroke history, aneurysm irregularity) showed good predictive ability for the instability of small SIAs, with an area under the curve of 0.905. In addition, correlation analysis showed that SII, WPR, and Hcy also had significant differences in patients with symptomatic unruptured and ruptured small SIAs, and higher inflammation levels often promoted the disease progression of small SIAs. Higher levels of SII, WPR and Hcy can be used as independent predictors of instability of small SIAs. As an economical and convenient biomarker, it is crucial for clinical treatment strategies of stable small SIAs.

Abstract Background MicroRNA (miRNA)-based therapies have shown great potential in myocardial repair following myocardial infarction (MI). MicroRNA-302 (miR302) has been reported to exert a protective effect on MI. However, miRNAs are easily degraded and ineffective in penetrating cells, which limit their clinical applications. Exosomes, which are small bioactive molecules, have been considered as an ideal vehicle for miRNAs delivery due to their cell penetration, low immunogenicity and excellent stability potential. Herein, we explored cardiomyocyte-targeting exosomes as vehicles for delivery of miR302 into cardiomyocyte to potentially treat MI. Methods To generate an efficient exosomal delivery system that can target cardiomyocytes, we engineered exosomes with cardiomyocyte specific peptide (CMP, WLSEAGPVVTVRALRGTGSW). Afterwards, the engineered exosomes were characterized and identified using transmission electron microscope (TEM) and Nanoparticle Tracking Analysis (NTA). Later on, the miR302 mimics were loaded into the engineered exosomes via electroporation technique. Subsequently, the effect of the engineered exosomes on myocardial ischemia and reperfusion (I/R) injury was evaluated in vitro and in vivo, including MTT, ELISA, real-time quantitative polymerase chain reaction (PCR), western blot, TUNNEL staining, echocardiogram and hematoxylin and eosin (HE) staining. Results Results of in vitro experimentation showed that DSPE-PEG-CMP-EXO could be more efficiently internalized by H9C2 cells than unmodified exosomes (blank‐exosomes). Importantly, compared with the DSPE-PEG-CMP-EXO group, DSPE-PEG-CMP-miR302-EXO significantly upregulated the expression of miR302, while exosomes loaded with miR302 could enhance proliferation of H9C2 cells. Western blot results showed that the DSPE-PEG-CMP-miR302-EXO significantly increased the protein level of Ki67 and Yap, which suggests that DSPE-PEG-CMP-miR302-EXO enhanced the activity of Yap, the principal downstream effector of Hippo pathway. In vivo, DSPE-PEG-CMP-miR302-EXO improved cardiac function, attenuated myocardial apoptosis and inflammatory response, as well as reduced infarct size significantly. Conclusion In conclusion, our findings suggest that CMP-engineered exosomes loaded with miR302 was internalized by H9C2 cells, an in vitro model for cardiomyocytes coupled with potential enhancement of the therapeutic effects on myocardial I/R injury.

Corneal melt is a sight-threatening complication of Boston type 1 keratoprosthesis (KPro). Severe corneal melt may result in hypotony, choroidal hemorrhage, and even spontaneous extrusion of the KPro, which may lead to a poor visual prognosis. Lamellar keratoplasty is one surgical option for the management of mild corneal melt, especially when a new KPro is not available. Herein, we present a new surgical technique application, intra-operative optical coherence tomography (iOCT) for the management of cornea graft melt after Boston type 1 KPro implantation. The visual acuity and the intra-ocular maintained stable at 6 months post-operatively, and the KPro remained in place without corneal melting, epithelial ingrowth, or infection. iOCT may prove to be a real-time, non-invasive, and accurate treatment for corneal lamellar dissection and suturing beneath the anterior plate of the KPro, which can effectively help the surgeon to make surgical decisions and reduce post-operative complications.

Both oxidative stress and the immune response are associated with heart failure (HF). In this study, our aim was to identify the hub genes associated with oxidative stress andimmune infiltration of HF by bioinformatics analysis and experimental verification. The expression profile of GSE36074 was obtained from the Gene Expression Omnibus (GEO) database. The differentially expressed genes (DEGs) were screened by GEO2R. The genes related to oxidative stress were extracted from GeneCards websites. Then, the functional enrichment analysis of oxidative stress-related DEGs (OSRDEGs) was performed using DAVID. In addition, we constructed a protein-protein interaction (PPI) network using the STRING database and screened for hub genes with Cytoscape software. We also used CIBERSORTx to analyze immune infiltration in mice heart tissues between the TAC and Sham groups and explored the correlation between immune cells and hub genes. Finally, the hub genes were carried out using reverse transcription quantitative PCR (RT-qPCR), immunohistochemistry (IHC) and western blot. A total of 136 OSRDEGs were found in GSE36074. Enrichment analysis revealed that these OSRDEGs were enriched in the mitochondrion, HIF-1, FoxO, MAPK and TNF signaling pathway. The five hub genes (Mapk14, Hif1a, Myc, Hsp90ab1, and Hsp90aa1) were screened by the cytoHubba plugin. The correlation analysis between immune cells and hub genes showed that Mapk14 was positively correlated with Th2 Cells, while Hif1a and Hsp90ab1exhibited a negative correlation with Th2 Cells; Myc exhibited a negative correlation with Monocytes; whereas, Hsp90aa1 was negatively correlated with NK Resting. Finally, five hub genes were validated by RT-qPCR, IHC and western blot. Mapk14, Hif1a, Myc, Hsp90ab1, and Hsp90aa1 are hub genes of HF and may play a critical role in the oxidative stress of HF. This study may provide new targets for the treatment of HF, and the potential immunotherapies are worthy of further study.

ObjectivesThe ratio of white blood cell to platelet count (WPR) is considered a promising biomarker in some diseases. However, its prediction of delayed cerebral ischemia (DCI) and prognosis after aneurysmal subarachnoid hemorrhage (aSAH) has not been studied. The primary objective of this study was to investigate the predictive value of WPR in DCI after aSAH and its impact on 90-day functional outcome.Materials and methodsThis study retrospectively analyzed the data of blood biochemical parameters in 447 patients with aSAH at early admission. Univariate and multivariate analyses were used to determine the risk factors for DCI. According to multivariate analysis results, a nomogram for predicting DCI is developed and verified by R software. The influence of WPR on 90-day modified Rankin score (mRS) was also analyzed.ResultsAmong 447 patients with aSAH, 117 (26.17%) developed DCI during hospitalization. Multivariate logistic regression analysis showed that WPR [OR = 1.236; 95%CI: 1.058–1.444; p = 0.007] was an independent risk factor for DCI. The receiver operating characteristic (ROC) curve analysis was used to evaluate the predictive ability of WPR for DCI, and the cut-off value of 5.26 (AUC 0.804, 95% CI: 0.757–0.851, p

Background: Chronic ocular graft-versus-host disease (oGVHD) is a common ocular complication following allogeneic hematopoietic stem cell transplantation (allo-HSCT), characterized by progressive inflammation of the ocular surface and refractory dry eye. In severe cases, sterile corneal perforation can occur, which poses a significant challenge, due to the low survival rate of grafts after corneal transplantation. Case Presentation: A 47-year-old female presented to our hospital with persistent dryness, foreign body sensation, and blurred vision in her left eye. Diagnosis of graft-versus-host disease with corneal descemetocele in the left eye was made after detailed history review and thorough examination. Multi-layer amniotic membrane transplantation was performed in the affected eye, resulting in amelioration of the patient’s symptoms. This amelioration of symptoms provided the patient with a level of comfort that permitted additional time while awaiting corneal transplantation. Conclusions: We report a successful case of multi-layer amniotic membrane transplantation for the management of corneal descemetocele following allo-HSCT.

We aimed to study the molecular mechanisms of chronic obstructive pulmonary disease (COPD) caused by cigarette smoke more comprehensively and systematically through different perspectives and aspects and to explore the role of protein acetylation modification in COPD. We established the COPD model by exposing C57BL/6J mice to cigarette smoke for 24 weeks, then analyzed the transcriptomics, proteomics, and acetylomics data of mouse lung tissue by RNA sequencing (RNA-seq) and liquid chromatography-tandem mass spectrometry (LC-MS/MS), and associated these omics data through unique algorithms. This study demonstrated that the differentially expressed proteins and acetylation modification in the lung tissue of COPD mice were co-enriched in pathways such as oxidative phosphorylation (OXPHOS) and fatty acid degradation. A total of 19 genes, namely, ENO3, PFKM, ALDOA, ACTN2, FGG, MYH1, MYH3, MYH8, MYL1, MYLPF, TTN, ACTA1, ATP2A1, CKM, CORO1A, EEF1A2, AKR1B8, MB, and STAT1, were significantly and differentially expressed at all the three levels of transcription, protein, and acetylation modification simultaneously. Then, we assessed the distribution and expression in different cell subpopulations of these 19 genes in the lung tissues of patients with COPD by analyzing data from single-cell RNA sequencing (scRNA-seq). Finally, we carried out the in vivo experimental verification using mouse lung tissue through quantitative real-time PCR (qRT-PCR), Western blotting (WB), immunofluorescence (IF), and immunoprecipitation (IP). The results showed that the differential acetylation modifications of mouse lung tissue are widely involved in cigarette smoke-induced COPD. ALDOA is significantly downregulated and hyperacetylated in the lung tissues of humans and mice with COPD, which might be a potential biomarker for the diagnosis and/or treatment of COPD.

ObjectiveTo evaluate the correlation of serum biological markers and related scales to the occurrence of delayed cerebral ischemia and clinical prognosis in patients with aneurysmal subarachnoid hemorrhage (aSAH) complicated with acute hydrocephalus before admission.MethodsThe clinical data of 227 patients with pre-admission aSAH complicated with acute hydrocephalus admitted to Henan Provincial People's Hospital from April 2017 to December 2020 were retrospectively analyzed. Patients were grouped according to the presence or absence of delayed cerebral ischemia (DCI) after surgery and the prognosis at 6 months after discharge. Univariate and multivariable logistic regression analysis were performed to analyze the relationship between serum biological indicators combined with aneurysm related clinical score scale and the occurrence and prognosis of delayed cerebral ischemia. ROC curves and nomogram were drawn.ResultsMultivariable Logistic regression analysis showed that high Hunt-Hess grade and surgical clipping were independent risk factors for postoperative DCI (P < 0.05). Older age, higher Hunt-Hess grade, higher CRP and neutrophil levels were independent risk factors for poor prognosis at 6 months after surgery (P < 0.05). ROC curve analysis showed that the area under the curve (AUC) of Hunt-Hess grade and surgical method for predicting DCI in patients with aSAH combined with hydrocephalus after surgery were 0.665 and 0.593. The combined AUC of Hunt-Hess grade and surgical method was 0.685, the sensitivity was 64.9%, and the specificity was 64.7%. The AUC of CRP, neutrophil, age and Hunt-Hess grade for predicting poor prognosis in patients with aSAH combined with hydrocephalus at 6 months after surgery were 0.804, 0.735, 0.596, 0.757, respectively. The combined AUC of CRP, neutrophil, age, Hunt-Hess grade was 0.879, the sensitivity was 79%, and the specificity was 84.5%. According to the correction curve, the predicted probability of the nomogram is basically consistent with the actual probability.ConclusionHunt-Hess grade and surgical method are independent predictors of postoperative DCI in patients with aSAH complicated with hydrocephalus. “CRP,” “neutrophil,” “age” and “Hunt-Hess grade” at admission are independent predictors of clinical prognosis in patients with aSAH complicated with hydrocephalus. The combination of the above indicators has high predictive value.

Abstract Long non-coding RNA (lncRNA) is receiving increasing attention in embryonic stem cells (ESCs) research. However, the roles of lncRNA in the differentiation of ESCs into pacemaker-like cells are still unclear. Therefore, the present study aims to explore the roles and mechanisms of lncRNA in the differentiation of ESCs into pacemaker-like cells. ESCs were cultured and induced differentiation to pacemaker-like cells. RNA sequencing was used to identify the differential expression lncRNAs during the differentiation of ESCs into pacemaker-like cells. Cell morphology observation, flow cytometry, quantitative real-time polymerase chain reaction, western blot, and immunofluorescence were used to detect the differentiation of ESCs into pacemaker-like cells. LncRNA and genes overexpression or knockdown through transfected adenovirus in the differentiation process. The fluorescence in situ hybridization (FISH) detected the lncRNA location in the differentiated ESCs. Luciferase reporter gene assay, methylation-specific PCR, chromatin immunoprecipitation assay, and RNA immunoprecipitation assay were performed to reveal the mechanism of lncRNA-regulating HCN4 expression. Rescue experiments were used to confirm that lncRNA regulates the differentiation of ESCs into pacemaker-like cells through HCN4. We cultured the ESCs and induced the differentiation of ESCs into pacemaker-like cells successfully. The expression of lncRNA RCPCD was significantly decreased in the differentiation of ESCs into pacemaker-like cells. Overexpression of RCPCD inhibited the differentiation of ESCs into pacemaker-like cells. RCPCD inhibited the expression of HCN4 by increasing HCN4 methylation at the promoter region through DNMT1, DNMT2, and DNMT3. RCPCD inhibited the differentiation of ESCs into pacemaker-like cells by inhibiting the expression of HCN4. Our results confirm the roles and mechanism of lncRNA RCPCD in the differentiation of ESCs into pacemaker-like cells, which could pave the path for the development of a cell-based biological pacemaker.

ObjectiveTo investigate the relationship between CLR and disease severity and clinical prognosis of aSAH.MethodsThe authors retrospectively analyzed the clinical data of 221 patients with aSAH, who were admitted to the intensive care unit from January 2017 to December 2020. The indicators of inflammatory factors in the first blood routine examination within 48 h of bleeding were obtained. The prognosis was evaluated by mRS score at discharge, mRS>2 was a poor outcome. Through the receiver operating characteristic (ROC) curve, the area under the curve was calculated and the predicted values of inflammatory factors (CLR, CRP, WBC, and neutrophils) were compared. Univariate and multivariable logistic regression analyses were used to evaluate the relationship between CLR and the clinical prognosis of patients. ROC curve analysis was performed to determine the optimal cut-off threshold, sensitivity, and specificity of CLR in predicting prognosis at admission.ResultsAccording to the mRS score at discharge, 139 (62.90%) patients were classified with poor outcomes (mRS>2). The inflammatory factor with the best predictive value was CLR, which had an optimal cut-off threshold of 10.81 and an area under the ROC curve of 0.840 (95%CI.788–0.892, P < 0.001). Multivariable Logistic regression analysis showed that the Modified Fisher grade, Hunt-Hess grade, and CLR at admission were independent risk factors for poor outcomes of patients with aSAH (P < 0.05). According to Hunt-Hess grade, patients were divided into a mild group (Hunt-Hess ≤ 3) and a severe group (Hunt-Hess > 3), and the CLR value was significantly higher in severe patients with aSAH than in mild patients. The optimal cut-off threshold of CLR in the severe group was 6.87, and the area under the ROC curve was 0.838 (95% CI.752–0.925, P < 0.001).ConclusionsThe CLR value at the admission of patients with aSAH was significantly associated with Hunt-Hess grade, The higher Hunt-Hess grade, the higher the CL R-value, and the worse the prognosis. Early CLR value can be considered as a feasible biomarker to predict the clinical prognosis of patients with aSAH.

STEAM contests play a pivotal role in maintaining students' interest and motivation in STEAM education. Finding factors that are related to students' continuance intention (CI) to participate in STEAM contests is important for promoting STEAM education. Previous studies mostly focused on exploring the relationships between students' behavioural attitude, positive affect and their CI for a STEAM contest. However, few studies have explored students' CI from the perspective of help-seeking attitude with the belief-attitude-behaviour model. Thus, this study investigated the relationships among students' hands-on making self-efficacy (HMSE), incremental belief of intelligence (IBI), help-seeking attitudes including instrumental help-seeking (IHS) and executive help-seeking (EHS), and their CI for STEAM contests based on the belief-attitude-behaviour model. A total of 405 elementary and junior high school students who took part in the 2022 STEAM PowerTech contest participated in this study. Structural equation modelling was used for confirmatory analysis to validate the hypotheses. Results indicated that EHS exhibited a negative association with CI, whereas IHS exhibited a positive association with CI. HMSE and IBI were negatively associated with EHS, and positively associated with IHS. This study implies that students with a high level of HMSE and IBI are inclined to exhibit IHS and thus have a strong CI for STEAM contests. The findings provide suggestions on enhancing students' CI for STEAM contests.

Abstract Background Temozolomide (TMZ) resistance limits its application in glioma. Exosome can carry circular RNAs (circRNAs) to regulate drug resistance via sponging microRNAs (miRNAs). miRNAs can control mRNA expression by regulate the interaction with 3’UTR and methylation. Nanog homeobox (NANOG) is an important biomarker for TMZ resistance. Hitherto, it is unknown about the role of exosomal hsa_circ_0072083 (circ_0072083) in TMZ resistance in glioma, and whether it is associated with NANOG via regulating miRNA sponge and methylation. Methods TMZ-resistant (n = 36) and sensitive (n = 33) patients were recruited. The sensitive cells and constructed resistant cells were cultured and exposed to TMZ. circ_0072083, miR-1252-5p, AlkB homolog H5 (ALKBH5) and NANOG levels were examined via quantitative reverse transcription polymerase chain reaction and western blot. The half maximal inhibitory concentration (IC50) of TMZ, cell proliferation, apoptosis, migration and invasion were analyzed via Cell Counting Kit-8, colony formation, flow cytometry, wound healing and transwell assays. The in vivo function was assessed using xenograft model. The N6-methyladenosine (m6A) level was analyzed via methylated RNA immunoprecipitation (MeRIP). Target relationship was investigated via dual-luciferase reporter assay and RNA immunoprecipitation. Warburg effect was investigated via lactate production, glucose uptake and key enzymes expression. Exosome was isolated and confirmed via transmission electron microscopy and specific protein expression. Results circ_0072083 expression was increased in TMZ-resistant glioma tissues and cells. circ_0072083 knockdown restrained the resistance of resistant cells via decreasing IC50 of TMZ, proliferation, migration, invasion and xenograft tumor growth and increasing apoptosis. circ_0072083 silence reduced NANOG expression via blocking ALKBH5-mediated demethylation. circ_0072083 could regulate NANOG and ALKBH5 via targeting miR-1252-5p to control TMZ resistance. Warburg effect promoted the release of exosomal circ_0072083 in resistant cells. Exosomal circ_0072083 from resistant cells increased the resistance of sensitive cells to TMZ in vitro and xenograft model. Exosomal circ_0072083 level was enhanced in resistant patients, and it had a diagnostic value and indicated a lower overall survival in glioma. Conclusion Exosomal circ_0072083 promoted TMZ resistance via increasing NANOG via regulating miR-1252-5p-mediated degradation and demethylation in glioma.

Background Intimal hyperplasia is a common pathological process of restenosis following angioplasty, atherosclerosis, pulmonary hypertension, vein graft stenosis, and other proliferative diseases. This study aims to screen for potential novel gene targets and mechanisms related to vascular intimal hyperplasia through an integrated microarray analysis of the Gene Expression Omnibus Database (GEO) database. Material and Methods The gene expression profile of the GSE56143 dataset was downloaded from the Gene Expression Omnibus database. Functional enrichment analysis, protein-protein interaction (PPI) network analysis, and the transcription factor (TF)-target gene regulatory network were used to reveal the biological functions of differential genes (DEGs). Furthermore, the expression levels of the top 10 key DEGs were verified at the mRNA and protein level in the carotid artery 7 days after ligation. Results A total of 373 DEGs (199 upregulated DEGs and 174 downregulated DEGs) were screened. These DEGs were significantly enriched in biological processes, including immune system process, cell adhesion, and several pathways, which were mainly associated with cell adhesion molecules and the regulation of the actin cytoskeleton. The top 10 key DEGs (Ptprc, Fn1, Tyrobp, Emr1, Itgb2, Itgax, CD44, Ctss, Ly86, and Aif1) acted as key genes in the PPI network. The verification of these key DEGs at the mRNA and protein levels was consistent with the results of the above-mentioned bioinformatics analysis. Conclusion The present study identified key genes and pathways involved in intimal hyperplasia induced by carotid artery ligation. These results improved our understanding of the mechanisms underlying the development of intimal hyperplasia and provided candidate targets.

Students’ learning engagement is recognized as one of the main components of effective instruction and a necessary prerequisite for learning, but students’ learning engagement in flipped classroom poses some pedagogical challenges. This study aimed to promote students’ learning engagement via the flipped classroom approach. Design-based research (DBR) was adopted in this study to conduct an experiment involving three iterations in a Modern Educational Technology (MET) course in a Chinese university. The participants included 36 third-year pre-service teacher undergraduates. Classroom observations and a learning engagement questionnaire were used to measure the effectiveness of the flipped instruction in terms of students’ learning engagement. Data analysis applied descriptive statistics, ANOVA, and paired samples t tests. The results showed that after three rounds of iterative experiments, students’ learning engagement (behavioral, cognitive, and emotional) significantly improved. Several principles are provided as guidelines for instructors to implement flipped classroom approach to promote students’ learning engagement.

Purpose. The aim of this study is to report the clinical characteristics, visual outcomes, and antibiotic susceptibilities of patients with Pseudomonas aeruginosa endophthalmitis. Methods. The medical records of patients with culture-proven Pseudomonas aeruginosa endophthalmitis treated from June 2013 to December 2019 were reviewed. Results. This study included 36 eyes of 36 patients. The clinical settings included ocular trauma (15/36), corneal ulcer (9/36), postoperative endophthalmitis (5/36), endogenous (3/36), and unknown (4/36). Sixteen patients underwent evisceration, 13 patients underwent pars plana vitrectomy (PPV), 2 patients were treated with only intravitreal antibiotics, and 5 patients did not undergo surgery. Only one patient achieved a visual acuity of 20/400, and the others had all counting fingers or below. The cultured Pseudomonas aeruginosa was 100% sensitive to gentamicin, tobramycin, amikacin, ciprofloxacin, and levofloxacin and, approximately 95% sensitive to meropenem, imipenem, and aztreonam. Conclusion. The visual outcomes of Pseudomonas aeruginosa endophthalmitis were very poor, and the evisceration rate remained high. Pseudomonas aeruginosa has good susceptibility to gentamicin, tobramycin, amikacin, ciprofloxacin, and levofloxacin.

The recurrence of glioma is a difficult problem in clinical treatment. The molecular markers of primary tumors after resection cannot fully represent the characteristics of recurrent tumors. Here, abundant tumor DNA was detected in tumor in situ fluid (TISF). We report that TISF-derived tumor DNA (TISF-DNA) can detect genomic changes in recurrent tumors and facilitate recurrence risk analysis, providing valuable information for diagnosis and prognosis. The tumor DNA in TISF is more representative and sensitive than that in cerebrospinal fluid. It reveals the mutational landscape of minimal residual disease after glioma surgery and the risk of early recurrence, contributing to the clinical management and clinical research of glioma patients.

Subarachnoid hemorrhage (SAH) is one of the common clinical neurological emergencies. Its incidence accounts for about 5–9% of cerebral stroke patients. Even surviving patients often suffer from severe adverse prognoses such as hemiplegia, aphasia, cognitive dysfunction and even death. Inflammatory response plays an important role during early nerve injury in SAH. Toll-like receptors (TLRs), pattern recognition receptors, are important components of the body’s innate immune system, and they are usually activated by damage-associated molecular pattern molecules. Studies have shown that with TLR 4 as an essential member of the TLRs family, the inflammatory transduction pathway mediated by it plays a vital role in brain injury after SAH. After SAH occurrence, large amounts of blood enter the subarachnoid space. This can produce massive damage-associated molecular pattern molecules that bind to TLR4, which activates inflammatory response and causes early brain injury, thus resulting in serious adverse prognoses. In this paper, the process in research on TLR4-mediated inflammatory response mechanism in brain injury after SAH was reviewed to provide a new thought for clinical treatment.

The COVID-19 pandemic has caused great disruption in education systems around the world. Schools have in some cases ended or limited on-site teaching, and have shifted toward home online teaching. This situation is likely to cause increased uncertainty and anxiety for parents who on one hand may question the quality of home online learning yet, on the other, may not feel sufficiently confident or competent to guide their children's home online learning. Resulting anxiety is expected to be most evident in competitive educational contexts, such as those found throughout much of East Asia. Therefore, using China as the setting, and social comparison theory as the framework, this study examined how variation in parent social comparison relates to parent tutoring anxiety and, in turn, to the types of guided strategies parents use to promote their children's home online learning. The results indicated a positive relationship between parental upward social comparison and parental tutoring anxiety, but a negative relationship between parent downward social comparison and parental anxiety. Parental tutoring anxiety is positively related to the confirmation and structure types, but negatively related to the discovery type of guided approaches. The implication of this study is that parents who resist tendencies of competitive upward social comparison are likely to adopt more effective approaches to guiding their children's home online learning.

Abstract Introduction Lung large cell neuroendocrine carcinoma (L‐LCNEC) is a rare, aggressive tumor, for which the optimal treatment strategies for LCNEC have not yet been established. In order to explore how to improve the outcome of prognosis for patients with LCNEC, this study investigated the effect of different treatments based on the data obtained from the Surveillance, Epidemiology, and End Results (SEER) database. Methods A total of 2594 LCNEC cases with conditional information were extracted from SEER database. Propensity Score Matching (PSM) method was conducted to reduce possible bias between groups. One‐way ANOVA was used to test the differences of characteristics between groups. Univariate and multivariate Cox proportional hazard models were applied to identify prognostic factors. Results Clinicopathologic characteristics including gender, age, TNM stage, T stage, N stage, and M stage were all identified as independent prognostic factors. Surgery benefited stage I, II, and III LCNEC patients’ prognoses. The combination treatment that surgery combining with chemotherapy was the optimal treatment for stage I, II, and III LCENC patients. Compared with palliative treatment, stage IV patients obtained better prognoses with the treatment of radiation, chemotherapy, or chemoradiation. When comparing the effect of the three treatments (radiation, chemotherapy, and chemoradiation) in achieving better prognosis for stage IV patients, chemotherapy alone was better than the other treatments. Conclusion Surgery combining with chemotherapy was the optimal treatment for stage I, II, and III LCNEC patients; chemotherapy alone achieves more benefit than the other treatments for stage IV patients.

Abstract Introduction The purpose of this study was to describe the results using the Nd:YAG laser to reopen blocked glaucoma tube shunts in three ocular chemical burn patients with Boston keratoprostesis type I (KPro) implantation. Methods The medical records of the three patients at Zhongshan Ophthalmic Centre were reviewed. Results Patient 1, who had glaucoma secondary to KPro implantation, had undergone Ahmed glaucoma valve (AGV) implantation to control an elevated intraocular pressure (IOP). One day after surgery, the tube was observed to be embedded in the residual lens capsule. The capsule was opened by one 1.5 mJ laser pulse, with a subsequent drop in the IOP. In patient 2, the AGV and KPro had been implanted simultaneously. One month after surgery, the IOP increased to 35 mmHg, estimated by palpation, and a vitreous gel was seen blocking the tube. A Nd:YAG laser pulse was used to open the occluded tube. In patient 3, the tube was blocked by iris tissue; Nd:YAG laser treatment opened the tube. Conclusions The cases described here indicate that Nd:YAG laser treatment seems to be a valuable option for opening an occluded AGV tube in patients with KPro implantation. The correct location of the tube tip, visualized through the KPro optic, is essential for laser treatment.

Objectives: To evaluate the feasibility, efficiency, and safety of the transradial approach (TRA) for cerebral angiography versus the transfemoral approach (TFA) in patients. Methods: In this trial, 2314 patients underwent cerebral angiography, with 1085 patients undergoing the procedure via radial access and 1229 via transfemoral access. The arterial puncture time, operation success rate, oppression time on puncture points, local vascular complication incidence (including bleeding, hematoma, and pseudoaneurysm), deep venous thrombosis of lower limbs (DVT), and bradycardia/hypotension were observed and compared between the two groups. Results: Of the patients who underwent cerebral angiography via radial access, the procedure was successful in 1070 patients; compared with 1219 patients with transfemoral access, there was no significant difference (P > 0.05) in the success rate or the arterial puncture time. Radial access patients were less likely to present with oppression time on puncture points, local vascular complications, DVT, and bradycardia/hypotension compared with femoral access patients. Conclusions: For patients undergoing cerebral angiography, radial and femoral approaches are both safe and effective. However, the lower rate of local vascular complications may be a reason to use the radial approach. Keywords: Cerebral angiography, Transradial approach, Transfemoral approach, Intervention

TiO2 films exhibiting structural colors were successfully prepared using one-step electrochemical oxidation. Results of theoretical analyses and digital simulations revealed that the structural color of a TiO2 thin film could be regulated by adjusting oxidation voltage and oxidation time with different oxidation voltages leading to changes in structural color annulus number. At a low oxidation voltage, each thin film exhibited a single structural color, while thin films with different structural colors were obtained by varying the oxidation time. By contrast, at a higher oxidation voltage, each film exhibited iridescent and circular structural color patterns associated with symmetrical decreases in surface oxidation current density along radial lines emanating from the film center to its outer edges. TiO2 films exhibiting iridescent structural colorations have broad application prospects in industrial fields related to photocatalysis and photovoltaic cells.

Background: Although independent association of socioeconomic status (SES) or physical activity (PA) with obesity has been well-documented in urban settings, their independent and joint associations on obesity measures are limited in rural regions.Methods: Almost 38,000 (n = 37,922) individuals were included from the Henan Rural Cohort Study. The International Physical Activity Questionnaire (IPAQ) was used to evaluate PA. Obesity was reflected by body mass index (BMI), waist circumference (WC), waist-to-hip ratio (WHR), waist-to-height ratio (WHtR), body fat percentage (BFP), and visceral fat index (VFI). The independent and interactive effects of SES and PA on obesity were analyzed by logistic regression models and generalized linear regression models, respectively.Results: Compared with high education level, the OR (95%CI) of obesity defined by BMI with low education level was 1.466 (1.337, 1.608), 1.064 (0.924, 1.225), and 1.853 (1.625, 2.114) in total population, men and women, respectively. Besides, the OR (95%CI) of obesity defined by BMI associated with per capita monthly income were 1.089 (1.015, 1.170), 1.192 (1.055, 1.347), 1.038 (0.951, 1.133) in total population, men and women, respectively. Similar results had been observed in other obesity measures. Negative interactive association of low education level and PA on obesity measures were observed only in women (all P < 0.05).Conclusions: This study suggests that women are more susceptible to obesity concerning low SES and that adequate PA may be a potential target for mitigating the negative effect of low SES on obesity in women.Clinical Trial Registration: The Henan Rural Cohort Study has been registered at Chinese Clinical Trial Register (Registration number: ChiCTR-OOC-15006699) http://www.chictr.org.cn/showproj.aspx?proj=11375.

Meningioma (MEN) is a common central nervous system disease. Accumulating evidence indicated that long non-coding RNA maternally expressed gene 3 (MEG3) participated in the progression of MEN. However, the potential mechanisms of MEG3 in altering the aggressive phenotypes of MEN need further exploration. Levels of MEG3, microRNA (miR)-29c, and A-kinase anchor protein 12 (AKAP12) were determined using quantitative real-time Polymerase Chain Reaction (qRT-PCR) assay. Dual-luciferase reporter and RNA immunoprecipitation (RIP) assays were performed to verify the relationship between miR-29c and MEG3 or AKAP12. The protein level of AKAP12 was detected by western blot. Moreover, cell-cycle arrest, migration, invasion, and proliferation were assessed by flow cytometry, wound healing, transwell assays, and CCK-8 assay, respectively. Levels of MEG3 and AKAP12 were downregulated, while miR-29c was effectively increased in MEN tissues and cell line. Mechanically, MEG3 was a sponge of miR-29c to regulate the expression of AKAP12. Functionally, increase of MEG3 diminished cell-cycle, migration, invasion, and proliferation in MEN cells, and reintroduction of miR-29c could eliminate these effects. In addition, AKAP12 depletion overturned the inhibitory effects of miR-29c absence on cell-cycle, migration, invasion, and proliferation in vitro. Also, AKAP12 was co-regulated by MEG3/miR-29c axis. MEG3 mediated the aggressive behaviors of MEN cells via miR-29c/AKAP12 axis, supporting that MEG3 served as a promising biomarker for the diagnosis and treatment of human MEN.

PurposeThe aim of this study was to explore the correlation and clinical significance of preoperative fibrinogen and neutrophil-lymphocyte ratio (F-NLR) scoring system with 3-year progression-free survival (PFS) of patients with atypical meningioma.Materials and MethodsClinical, pathological, radiological, and laboratory variables were collected to analyze their correlation with 3-year PFS in the training set with 163 patients. Patients were classified by different F-NLR scores (0, 1, or 2). External validation for the predictive value of F-NLR scoring system was performed in the validation set with 105 patients.ResultsOverall, 37.3% (100 of 268) of the enrolled patients were male. The scoring system showed good performance in predicting 3-year PFS (AUC = 0.872, 95%CI = 0.811–0.919, sensitivity = 66.1%, specificity = 93.3%, and Youden index = 0.594). DeLong’s test indicated that the AUC of F-NLR scoring system was significantly greater than that of fibrinogen level and NLR (Z = 2.929, P = 0.003; Z = 3.376, P < 0.001). Multivariate Cox analysis revealed that tumor size (HR = 1.39, 95%CI = 1.10–1.76, P = 0.007), tumor location (HR = 3.11, 95%CI = 1.60–6.95, P = 0.001), and F-NLR score (score of 1: HR = 12.78, 95%CI = 3.78–43.08, P < 0.001; score of 2: HR = 44.58, 95%CI = 13.02–152.65, P < 0.001) remained significantly associated with 3-year PFS. The good predictive performance of F-NLR scoring system was also demonstrated in the validation set (AUC = 0.824, 95%CI = 0.738–0.891, sensitivity = 62.5%, specificity = 87.9%, and Youden index = 0.504).ConclusionOur study confirmed the correlation and clinical significance of preoperative F-NLR scoring system with 3-year PFS of patients with atypical meningioma. A prospective and large-scale study is required to validate our findings.

Abstract Background The neuroglobin (Ngb) is well recognized as a potential biomarker for the hypoxic‐ischemic brain injury. However, connection between Ngb and delayed cerebral ischemia (DCI) following aneurysmal subarachnoid hemorrhage (aSAH) is still unclear. Objective To investigate the relationship between early stage Ngb level of aSAH patient and the occurrence of DCI. Methods We evaluated 126 aSAH patients who were enrolled into a prospective observational cohort study. Serum Ngb level on days 1, 2, 3, 5, and 7 after aSAH were determined using a commercial enzyme‐linked immunosorbent assay kit. The relationship between Ngb level and DCI was analyzed. Results Forty‐six (36.5%) aSAH patients experienced DCI. Patients with DCI had significantly higher Ngb levels than those without (p 8.4 ng/ml has a significantly worse DCI survival rate than those having day 3 Ngb level

Poly(vinylphosphonic acid) (PVPA) and polyethylenepolyamine (PEPA) are used as novel intumescent flame retardants to improve the properties of MXene (2D Ti3C2Tx)/poly(vinyl alcohol) (PVA) nanocomposites. We investigated the flame-retardant properties, thermal stability, and mechanical properties of MXene/PVA nanocomposites. The results show that MXene was homogeneously dispersed in the PVA matrix containing PVPA and PEPA. PVPA and PEPA effectively improved the flame-retardant properties of MXene/PVA nanocomposites and they did not obviously change the thermal degradation of the MXene/PVA nanocomposites. Moreover, MXene improved the thermal stability of the PVA matrix. The elongation at break of MXene/PVA nanocomposites reached its maximum when the MXene mass fraction was 1.0 wt.%, regardless of whether PVPA and PEPA were present in the PVA matrix, whereas the tensile strength and Young’s modulus of MXene/PVA nanocomposites increased with the increase in MXene content in the PVA matrix.

Segmentation of carotid intima-media (IM) borders from ultrasound images is of great importance for predicting cardiovascular risks. In this paper, we have developed a fully automatic approach to sequentially segment the carotid IM borders in each image throughout ultrasound sequences. First, the first frame of an ultrasound sequence is automatically segmented using edge detectors and dynamic programming, and then the rest frames are segmented successively under the state-space framework. Under this framework, we developed a variant of the snake method for a precise measurement. The evaluation of our segmentation result is done by comparison with average manual delineations of three physicians on a total of 65 sequences. The accuracy of our method is high. (Segmentation error is 32.1 ± 37.5 μm for LI and 35.0 ± 41.5 μm for MA.) The BA plot and the linear regression also demonstrate that our method is in agreement with the ground truth. This paper strengthens the potential of the state-space and snake-based approach in segmenting IM borders for clinical diagnosis by demonstrating a fully automatic scheme.

Purpose:To evaluate complications and anatomical retention when using autologous ear cartilage as a carrier in type 1 Boston keratoprosthesis (Kpro) surgery. Methods: In New Zealand White rabbits, the Kpro was surgically implanted using autologous cartilage (n = 5) or ipsilateral autologous cornea (n = 1, control) as carrier. Eyes were followed up using slit-lamp photography, anterior segment optical coherence tomography, and histological analysis for 19 days to 24 months. Results: Kpros were retained in all eyes. One eye in the cartilage group had an uncomplicated course; 3 developed granulation tissue over the Kpros optic, 1 developed a retroprosthetic membrane, and 3 developed glaucoma. Two eyes had lacunae. The gap between the Kpros flange and cartilage carrier filled with fibrovascular tissue over time. The control eye developed corneal tissue thinning and an enlarged gap. Conclusion: Auricular cartilage carriers may encourage Kpro-graft adhesion in a rabbit model and may be useful in Kpro recipients who cannot use bandage contact lenses.

Arts design has been proven to enable designers to have a meaningful learning of STEAM, but research on a checklist for assessing Arts design based on evaluating entries in a STEAM contest is still rare. Therefore, this study developed a checklist for assessing Arts design for evaluating miniature robots in a STEAM contest. Using an example of STEAM contest, named PowerTech, which has been held 22 years in Taiwan, we developed five dimensions of the checklist are: aesthetics, material usage, bionic, performance, and additional devices. There are two sub-indices in each aspect to be considered as the identification of form characteristics. Eight international experts were invited to review the content validity of the Arts design scale, and 30 completed checklists were used to test the reliability and validity though Kendall's coefficient of concordance. The results showed that the Arts design scale had high reliability and good validity, and so rubric index could be used to measure and assess Arts design with miniature robots in that integrated STEAM contest. Based on this, it is suggested that educational authorities can refer this study and for encourage students to create Arts in any integrated STEAM contests.

This study reports an investigation of the teaching engineering self-efficacy among Chinese high school general technology teachers. The teaching engineering self-efficacy scale (TESS) was the measurement tool. 557 qualified questionnaires were analyzed, and the results showed that: (1) TESS can be used to assess teaching engineering self-efficacy in the Chinese context after confirmatory factor analysis. (2) Technology teachers don't really believe that they are capable of teaching engineering. Teachers are especially pessimistic about their ability in engineering pedagogical content knowledge. (3) There are school area differences in teaching engineering self-efficacy. Teachers' teaching engineering self-efficacy in the town is the lowest. (4) There are training frequency differences in teaching engineering self-efficacy. Teachers who participated in more than 6 training events have higher teaching engineering self-efficacy than other teachers, and teachers with no training have the lowest teaching engineering self-efficacy. (5) There are educational level differences in teaching engineering self-efficacy. Teachers with a less than bachelor's degree have the lowest teaching engineering self-efficacy, and teachers with a master's degree have higher teaching engineering self-efficacy than teachers with a bachelor's degree.

Creativity and problem-solving are 21st-century skills that students must have to respond to the complex world dominated by technology. Teaching activities integrating Design Thinking (DT) have the potential to cultivate individual skills. As future in-service teachers, pre-service teachers need to understand DT so as to be able to use it in their future classrooms. In line with this, the present study aimed to explore the effects of the Stanford DT model on pre-service teachers' creativity self-efficacy, inventive problem-solving skills, and technology-related motivation. Quasi-experimental research with a pre- and post-test control group design was applied in the current study. A total of 70 pre-service teachers participated in this study, consisting of 36 experimental group (EG) students who learned with the DT model approach and 34 control group (CG) students who learned with a regular instructional approach. Design Thinking Activities were completed in 12 weeks, with three activities in the Modern Educational Technology (MET) course. The creativity self-efficacy, inventive problem-solving skills, and technology-related motivation scale were used to collect data before and after the experiment. An independent t test, Mann--Whitney U test, and Wilcoxon Test were used to analyze the data. The results indicated that there were significant differences in technology-related motivation, creative self-efficacy, and inventive problem-solving skills between the CG and EG students. The Stanford DT model integrated with the MET course was conducive to enhancing pre-service teachers' creativity self-efficacy, inventive problem-solving skills, and technology-related motivation in the context of the technological literacy course. Implications for the findings of this study are discussed.

Purpose. To explore the probability and variation in biomechanical measurements of rabbit cornea by a modified Scheimpflug device. Methods. A modified Scheimpflug device was developed by imaging anterior segment of the model imitating the intact eye at various posterior pressures. The eight isolated rabbit corneas were mounted on the Barron artificial chamber and images of the anterior segment were taken at posterior pressures of 15, 30, 45, 60, and 75 mmHg by the device. The repeatability and reliability of the parameters including CCT, ACD, ACV, and CV were evaluated at each posterior pressure. All the variations of the parameters at the different posterior pressures were calculated. Results. All parameters showed good intraobserver reliability (Cronbach’s alpha; intraclass correlation coefficient, α, ICC > 0.96) and repeatability in the modified Scheimpflug device. With the increase of posterior pressures, the ratio of CCT decreased linearly and the bulk modulus gradually reduced to a platform. The increase of ACD was almost linear with the posterior pressures elevated. Conclusions. The modified Scheimpflug device was a valuable tool to investigate the biomechanics of the cornea. The posterior pressure 15–75 mmHg range produced small viscoelastic deformations and nearly linear pressure-deformation response in the rabbit cornea.

The performance of community health service centers (CHSCs) has not been well monitored and analysed since China's latest community health reforms in 2009. The aim of the current investigation was to evaluate the performing trends of the CHSCs and to analyze the main factors that could affect the performance in Pudong new district of Shanghai, China.A regional performance assessment indicator system was applied to the evaluation of Pudong CHSCs' performance from 2011 to 2013. All of the data were sorted out by a panel, and analyzed using descriptive statistics and a generalized estimating equation model.We found that the overall performance increased annually, with a growing number of CHSCs achieving high scores. Significant differences were observed in institutional management, public health services, basic medical services and comprehensive satisfaction during the period of three years. However, we found no differences in the service scores of Chinese traditional medicine (CTM). The investigation also demonstrated that the key factors affecting performance were the location, information system level, family GP program and medical association program rather than the size of the center. However, the medical association participation appeared to have a significant negative effect on performance.It can be concluded from the three-year investigation that the overall performance was improved, but that it could have been further enhanced, especially in institutional management and basic medical service; therefore, it is imperative that CHSCs undertake approaches such as optimizing the resource allocation and utilization, reinforcing the establishment of the information system level, extending the family GP program to more local communities, and promoting the medical association initiative.

To investigate the activation of three unfolded protein response (UPR) pathways in the lenses of age-related, high myopia-related and congenital cataracts.Lens specimens were collected from patients during small incision cataract surgery. Lenses from young cadaver eyes were collected as normal controls. Real-time PCR and Western blotting were performed to detect the expression of GRP78, p-eIF2α, spliced XBP1, ATF6, ATF4 and p-IRE1α in the lenses of normal human subjects and patients with age-related, myopia-related or congenital cataracts.In the lenses of the age-related and high myopia-related cataract groups, the protein levels of ATF6, p-eIF2α and p-IRE1α and the gene expression levels of spliced XBP1, GRP78, ATF6 and ATF4 were greatly increased. Additionally, in the congenital cataract group, the protein levels of p-eIF2α and p-IRE1α and the gene expression levels of spliced XBP1, GRP78 and ATF4 were greatly increased. However, the protein and gene expression levels of ATF6 were not up-regulated in the congenital cataract group compared with the normal control group.The UPR is activated via different pathways in the lenses of age-related, high myopia-related and congenital cataracts. UPR activation via distinct pathways might play important roles in cataractogenesis mechanisms in different types of cataracts.

To investigate the expression of αA- and αB-crystallin and the unfolded protein response in the lens epithelium of patients with high myopia-related cataracts.The central portion of the human anterior lens capsule together with the adhering epithelial cells, approximately 5 mm in diameter, were harvested and processed within two hours after cataract surgery from high myopia-related (spherical equivalent ≥-10.00 diopters) and age-related cataract patients or from high myopia but non-cataractous patients (tissue were collected from ocular trauma patients with high myopia and lens trauma). Anterior lens samples from fresh cadaver normal human eyes were used as normal control (collected within 6 hours from death). Real-time PCR was performed to detect the mRNA levels of α-crystallins as well as unfolded protein response (UPR)-related GRP78, spliced-XBP1, ATF4 and ATF6. Western blot analysis was used to determine the protein level of α-crystallin, GRP78, p-IRE1α, p-eIF2α and ATF6.In the lens epithelium of the high myopia-related cataract group and the age related cataract group, the mRNA and soluble protein expression of αA- and αB-crystallin were both decreased; additionally, the protein levels of ATF6, p-eIF2α and p-IRE1α and the gene expression levels of spliced XBP1, GRP78, ATF6 and ATF4 were greatly increased relative to the normal control.These results suggest the significant loss of soluble α-crystallin and the activation of the UPR in the lens epithelium of patients with high myopia-related cataract, which may be associated with the cataractogenesis of high myopia-related cataract.

Abstract The battery energy storage system (BESS) combines backup and load regulation functions, making it a potential alternative to the diesel generator (DG) as the backup power source for data centers. Some studies have been conducted on the reliability and cost–benefit of equipping data centers with BESS, but the impact of the reliability of external utility power and the expected outage losses have not been considered. Therefore, this study established a power supply reliability model that included the external utility power reliability and the electrical equipment reliability, and a cost–benefit model that took into account the BESS construction and replacement cost, the outage loss reduction, and the load regulation benefits. A case study was conducted on a data center in Beijing. The results indicate that: (a) the reliability and cost–benefit of BESS significantly vary with the reliability of the external utility power; (b) based on the 2022 utility power reliability data from various regions in China, in over 48% of the regions, the objective function value of BESS, the annual net income considering power outage losses, and the investment payback period are all superior to those of DG; and in over 27% of regions, the BESS–DG parallel system can serve as a suboptimal alternative to DG; (c) in regions with reliable utility power, BESS exhibits the potential as a feasible substitute for DG in providing backup and load regulation power to data centers.

Abstract Black soldier fly larvae (BSFL) could convert a variety of organic wastes, including spent mushroom substrate (SMS) and wet distiller’s grains (WDG). Nevertheless, little is known about the conversion of these wastes by BSFL. Thus, this study investigates the conversion of SMS and WDG in five different proportions by BSFL. This study demonstrates that BSFL can convert SMS, WDG, and their mixtures. It can also encourage the humification of the substrate, increasing the amount of element in the residues. It is evident that there were differences in the carbon and nitrogen element fractionation mode as well as the microbial community present in the residue. The microbial community of the substrate and the physiochemical parameters are intimately related to this. Although the mixture treated with BSFL helps to generate a residue with more humus, it might not be stable.

Abstract Background Umbilical cord blood (UCB) is a rich source of multifunctional stem cells characterized by low immunogenicity. Recent research in the fields of aging and regenerative medicine has revealed the potential of human umbilical cord blood-derived exosomes (UCB-Exos) in promoting wound healing, anti-aging, and regeneration. However, their role in neurodegenerative diseases, specifically Parkinson’s disease (PD), remains unexplored. This study investigates the potential therapeutic effects and underlying mechanisms of UCB-Exos on PD. Methods Large extracellular vesicles (LEv), Exos, and soluble fractions (SF) of human UCB plasma were extracted to investigate their effects on motor dysfunction of the MPTP-induced PD mouse model and identify the key components that improve PD symptoms. UCB-Exos were administered by the caudal vein to prevent or treat the PD mouse model. The motor function and pathological markers were detected. Differentially expressed gene and KEGG enrichment pathways were screened by transcriptome sequence. MN9D and SH-SY5Y cells were cultured and evaluated for cell viability, oxidative stress, cell cycle, and aging-related indexes by qRT-PCR, western blot, immunofluorescence, and flow cytometry. The protein expression level of the MAPK p38 and ERK1/2 signaling pathway was detected by western blot. Results We observed that LEv, Exos, and SF all exhibited potential in ameliorating motor dysfunction in MPTP-induced PD model mice, with UCB-Exos demonstrating the most significant effect. UCB-Exos showed comparable efficacy in preventing and treating motor dysfunction, cognitive decline, and substantia nigra pathological damage in PD mice. Further investigations revealed that UCB-Exos could potentially alleviate oxidative damage, aging and degeneration, and energy metabolism disorders in neurons. Transcriptome sequencing results corroborated that genes differentially expressed due to UCB-Exos were primarily enriched in the neuroactive ligand-receptor interaction, Dopaminergic synapse, and MAPK signaling pathway. We also observed that UCB-Exos significantly inhibited the hyperphosphorylation of the MAPK p38 and ERK1/2 signaling pathways both in vitro and in vivo. Conclusions Our study provides a comprehensive evaluation of UCB-Exos on the neuroprotective effects and suggests that inhibition of hyperphosphorylation of MAPK p38 and ERK 1/2 signaling pathways by regulating transcription levels of HspB1 and Ppef2 may be the key mechanism for UCB-Exos to improve PD-related pathological features. Graphical Abstract

Heat shock proteins (HSPs), also known as heat stress proteins, can be divided into five main families, including HSP100, HSP90, HSP70, HSP60, and small heat shock proteins (sHSPs), according to their molecular weight. Further studies have shown that HSPs represent a class of highly conserved proteins that widely exist in the subcellular compartments of prokaryotic and eukaryotic cells. They have many functions, such as protein folding, repair, and immunity, and play important roles in organism growth and development. For example, Dmhsp70-A and Dmhsp70-B showed similar trends in the development of asexual embryos in Daphnia magna. However, they exhibited different trends in the development of sexual embryos despite high expression in both embryo types. Penaeus japonicus is one of the most important commercial shrimp species in China, Japan, and Southeast Asian countries. Relevant studies have shown that hsp genes widely exist in P. japonicus and play important roles in growth and development. For example, hsp10 and hsp60 were expressed in all tissues of P. japonicus, and the differential expression of hsp60 was identified in the early development stages. However, previous studies were based on single genes, and more systematic studies based on the role of the hsp gene family in the growth and development of P. japonicus have not been published. Our laboratory has completed the genome sequence of P. japonicus, which provides basic data for analyzing the molecular mechanism of hsp in the growth and development of P. japonicus. To further study the characteristics of the hsp gene family in P. japonicus and the expression characteristics of hsp genes during different development stages, the hsp gene family of P. japonicus was identified and analyzed using bioinformatics methods, and variation in its expression during the development of P. japonicus was measured using reverse transcription and quantitative real-time (RT-qPCR). Fifteen members of the hsp gene family were identified from the whole genome of P. japonicus via bioinformatics analysis, and their gene structures, motif compositions, chromosome localizations, and phylogenetic characteristics were analyzed. The expression of hsp genes in maternal-to-zygotic cells, blastocysts, gastrulae, limb buds, intramembranous nauplii, stage 1 nauplii, stage 1 zoeae, stage 1 myses, and stage 1 post-larval juveniles of P. japonicus was measured using qPCR. A total of 15 hsp genes were identified, including 1 hsp10, 2 hsp20, 2 hsp40, 1 hsp60, 6 hsp70, and 3 hsp90 genes. The physicochemical properties of seven proteins were stable, with instability coefficients less than 40. Subcellular localization results showed that most of the hsp gene family members were located in the cytoplasm, as seven hsp genes were localized to the cytoplasm, two to the nucleus, two to the extracellular region, two to the endoplasmic reticulum, and two to the mitochondria. Isoelectric point (pI) is related to the number and proportion of acidic and basic amino acids contained in a protein. Most HSPs exhibited a pI < 7, suggesting they were acidic proteins. The 15 hsp genes were located on 11 chromosomes. Phylogenetic tree analysis showed that most of the HSP family members were first clustered with HSP family members from species such as Penaeus monodon, Litopenaeus vannamei, and Fenneropenaeus chinensis and then with HSP family members of other species, which was consistent with the traditional taxonomic status. A few HSP family members were clustered first with HSP family members from Eriocheir sinensis, D. melanogaster, and other insects, and then with HSP family members from other species. Motif analysis showed that the hsp gene family was highly conserved; however, the functions of different families were quite different. Structural domain analysis showed that the HSP family included cpn10-, metazoan ACD-, DnaJ bact superfamily-, GroEL-, PTZ00009 superfamily-, dnaK-, HSP70 superfamily-, HSP90 superfamily-, PRK14083 superfamily-, and HATpase_HSP90-like domains. Each gene family contained corresponding domains and was important in molecular chaperone processes. hsp genes were expressed in different developmental stages of P. japonicus and played important roles in growth and development. The trend of Pjhsp10-19.517 was similar to that of Pjhsp60-19.518, which was highly expressed during embryonic development and significantly downregulated during larval development. This is consistent with the conclusion of previous studies that HSP60 and HSP10 are molecular chaperones for each other to ensure the correct folding of the target protein. The hsp genes such as Pjhsp40-15.349, Pjhsp70-39.287, Pjhsp70-1.298, and Pjhsp90 were highly expressed in the embryonic period; Pjhsp20, Pjhsp70-3.662, Pjhsp70-15.369, Pjhsp70-32.916, and Pjhsp90-12.759 were highly expressed during the larval period. The hsp genes expressed at high levels during the embryonic period satisfy the high protein requirements of embryos by playing the role of molecular chaperone and regulating the transcription of vitellogenin to meet the embryonic need for vitellin. The hsp genes expressed at high levels during the larval period protect the body from environmental pressure, pathogens, and xenobiotics through a synergistic immune effect. The authors believe that hsp genes play important roles in the growth and development of P. japonicus, and the specific pathways and mechanisms should be further studied. This study provides basic data for further investigations of the role of hsp genes in the growth and development of P. japonicus.

Abstract. Background:. We previously reported that activation of the cell cycle in human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) enhances their remuscularization capacity after human cardiac muscle patch transplantation in infarcted mouse hearts. Herein, we sought to identify the effect of magnesium lithospermate B (MLB) on hiPSC-CMs during myocardial repair using a myocardial infarction (MI) mouse model. Methods:. In C57BL/6 mice, MI was surgically induced by ligating the left anterior descending coronary artery. The mice were randomly divided into five groups (n = 10 per group); a MI group (treated with phosphate-buffered saline only), a hiPSC-CMs group, a MLB group, a hiPSC-CMs + MLB group, and a Sham operation group. Cardiac function and MLB therapeutic efficacy were evaluated by echocardiography and histochemical staining 4 weeks after surgery. To identify the associated mechanism, nuclear factor (NF)-κB p65 and intercellular cell adhesion molecule-1 (ICAM1) signals, cell adhesion ability, generation of reactive oxygen species, and rates of apoptosis were detected in human umbilical vein endothelial cells (HUVECs) and hiPSC-CMs. Results:. After 4 weeks of transplantation, the number of cells that engrafted in the hiPSC-CMs + MLB group was about five times higher than those in the hiPSC-CMs group. Additionally, MLB treatment significantly reduced tohoku hospital pediatrics-1 (THP-1) cell adhesion, ICAM1 expression, NF-κB nuclear translocation, reactive oxygen species production, NF-κB p65 phosphorylation, and cell apoptosis in HUVECs cultured under hypoxia. Similarly, treatment with MLB significantly inhibited the apoptosis of hiPSC-CMs via enhancing signal transducer and activator of transcription 3 (STAT3) phosphorylation and B-cell lymphoma-2 (BCL2) expression, promoting STAT3 nuclear translocation, and downregulating BCL2-Associated X, dual specificity phosphatase 2 (DUSP2), and cleaved-caspase-3 expression under hypoxia. Furthermore, MLB significantly suppressed the production of malondialdehyde and lactate dehydrogenase and the reduction in glutathione content induced by hypoxia in both HUVECs and hiPSC-CMs in vitro. Conclusions:. MLB significantly enhanced the potential of hiPSC-CMs in repairing injured myocardium by improving endothelial cell function via the NF-κB/ICAM1 pathway and inhibiting hiPSC-CMs apoptosis via the DUSP2/STAT3 pathway.

Critical size bone defects represent a significant challenge worldwide, often leading to persistent pain and physical disability that profoundly impact patients’ quality of life and mental well-being. To address the intricate and complex repair processes involved in these defects, we performed single-cell RNA sequencing and revealed notable shifts in cellular populations within regenerative tissue. Specifically, we observed a decrease in progenitor lineage cells and endothelial cells, coupled with an increase in fibrotic lineage cells and pro-inflammatory cells within regenerative tissue. Furthermore, our analysis of differentially expressed genes and associated signaling pathway at the single-cell level highlighted impaired angiogenesis as a central pathway in critical size bone defects, notably influenced by reduction of Spp1 and Cxcl12 expression. This deficiency was particularly pronounced in progenitor lineage cells and myeloid lineage cells, underscoring its significance in the regeneration process. In response to these findings, we developed an innovative approach to enhance bone regeneration in critical size bone defects. Our fabrication process involves the integration of electrospun PCL fibers with electrosprayed PLGA microspheres carrying Spp1 and Cxcl12. This design allows for the gradual release of Spp1 and Cxcl12 in vitro and in vivo. To evaluate the efficacy of our approach, we locally applied PCL scaffolds loaded with Spp1 and Cxcl12 in a murine model of critical size bone defects. Our results demonstrated restored angiogenesis, accelerated bone regeneration, alleviated pain responses and improved mobility in treated mice.

In this study, lotus seed starch was used as the raw material for preparing RS3 resistant starch (EHP-LRS3) through thermoacidophilic type Ⅲ pullulan hydrolase synergistic high-pressure heat treatments. The parameters for this preparation process were optimized using a single-factor experimental design and response surface methodology. The morphological and crystal structures of EHP-LRS3 were observed and analyzed using scanning electron microscopy (SEM) and X-ray diffraction (XRD), respectively, and its pro-proliferative effect on Bifidobacterium longum was investigated. According to the results, the optimal conditions for preparing EHP-LRS3 with TK-PUL enzymatic hydrolysis temperature of 80 ℃ were as follows: Treatment of 35.32% lotus seed starch slurry (pH5.00) with 25.00 U/g (lotus seed starch) TK-PUL at 80 ℃ for 12.70 h, followed by high-pressure heat treatment of the mixture at 121 ℃ for 10 min and retrogradation at 4 ℃ for 24 h. An EHP-LRS3 yield of 58.46% was achieved under these optimal conditions. The SEM analysis revealed that the prepared EHP-LRS3 possessed an irregular ravine-like structure. The XRD analysis showed that the resistant starch had a B-type crystal structure, which differed from the A-type crystal structure of lotus seed starch. The pro-proliferative effect of EHP-LRS3 on B. longum was superior to that of RS3 resistant starch prepared using high-pressure heating alone (HP-LRS3). In summary, a high EHP-LRS3 yield was obtained using both TK-PUL and high-pressure heat treatments. The superior pro-proliferative effect of EHP-LRS3 on B. longum was verified. The results provide a theoretical basis for the high-value utilization of lotus seed starch.

This study aimed to increase the expression and secretion of type III pullulan hydrolase (TK-PUL) by Brevibacillus choshinensis through the co-expression of molecular chaperone proteins and the optimization of fermentation conditions. By constructing multiple recombinant B. choshinensis co-expressing TK-PUL and molecular chaperone proteins, and screening them using extracellular enzyme activity as an indicator, the most favorable molecular chaperone protein and corresponding recombinant B. choshinensis for TK-PUL secretion and expression were determined. On this basis, single factor experiments and response surface methodology were used to optimize the fermentation conditions of recombinant B. choshinensis. Results showed that, the extracellular enzyme activity of the recombinant B. choshinensis strain co-expressing the molecular chaperone protein PrsABa reached 98.79 U/mL, representing a 0.31-fold increase. The optimal medium for this recombinant B. choshinensis was composed of 19.65 g/L of glucose, 21.46 g/L of yeast extract, 12.01 g/L of MgCl2·6H2O, 9.02 g/L of proline, 0.01 g/L of FeSO4·7H2O, 0.01 g/L of MnSO4·4H2O, and 0.001 g/L of ZnSO4·7H2O. Culturing the optimized recombinant B. choshinensis in the above-mentioned optimized medium for 66 hours at 35 °C and an initial pH of 7.0 increased the extracellular TK-PUL activity to 192.68 U/mL, which represented a 1.56-fold increase. Efficient secretion of TK-PUL in B. choshinensis was achieved through the co-expression of molecular chaperone proteins and the optimization of fermentation conditions. This study provides a foundation for exploring the industrial-scale application of TK-PUL.

It is well known that calcium oxide (CaO) has better catalytic efficiency than most heterogeneous catalysts in many transesterification reactions. However, the gradual deactivation problem prevents its large-scale application in industry. In this paper, the deactivation mechanism of CaO in a fixed-bed reactor is investigated based on the transesterification reaction of propylene carbonate and methanol. The leaching amount of CaO during the reaction was estimated by the concentration of Ca in the products. The pretreated and recovered catalysts were characterized by FT-IR, XRD, TG-MS and SEM-EDS. It is evident from experiments and characterization that the deactivation process of CaO is accompanied by the leaching of calcium species and the generation of CaCO3, which are also verified by DFT calculations. At high temperature and high weight hourly space velocity, the deactivation was attributed to the formation of dense CaCO3 shell, which prevents the contact between the feedstock and the active species inside.

This study investigated the effect of incorporating black soldier fly (BSF) larvae meal and oil on laying hens' performance, egg quality, serum profile, intestinal structure, and gut health. A total of 378 Lohmann laying hens (age 48 wk) were randomly assigned to 6 treatments with 3 replicates of 21 hens each. Following 7 d acclimation, the trail was conducted for 8 weeks. The dietary groups include: basal corn-soybean meal diet (S) without BSF (BSO) oil (S+BSO 0), S with BSF oil (S+BSO 100), BSF meal (9 %) without BSF oil (BSF 9+BSO 0), BSF meal (9 %) with BSF oil (BSF 9+BSO 100), BSF meal (18 %) without BSF oil (BSF 18+BSO 0), and BSF meal (18 %) with BSF oil (BSF 18+BSO 100). The results showed that the BSF 18 + BSO 100 diet significantly reduced egg weight (P < 0.001) compared to other dietary treatments. The addition of BSF meal reduced feed intake (P < 0.001) and the Haugh units (P < 0.05) in hens fed 18 % BSF meal with and without BSO. The jejunum villus area, crypt depth, and intestinal wall thickness increased with the increase in the inclusion of BSF larvae meal (P < 0.001). The ileum villus height, crypt depth and intestinal wall thickness increased (P < 0.001) at 9 % BSF meal and then decreased at 18 % BSF meal with and without BSF oil. The bacteria genera Ruminococcus, Clostridiales, Bacteroidales, Ruminococcus torques, and Intestinimonas were positively associated with the dietary treatments, while Prevotellaceae UCG-001, Clostridium, and Faecalibacterium were negatively associated with the dietary treatments. The inclusion of BSF meal and oil enriched the functional network of several pathways, including ascorbate and aldarate metabolism, D-arginine and D-ornithine metabolism, and fatty acid metabolism, highlighting the positive effects of BSF larvae meal and oil on the chicken gut microbiota. In conclusion, BSF meal at 9 % with BSF oil and BSF meal at 18 % without BSF oil can be incorporated into the diet without impairing the performance and gut health of laying hens.

ABSTRACT: Black soldier fly (BSF) larvae convert wastes into protein, playing a vital role in addressing the challenge of sustainable poultry production. These larvae accumulate toxic substances, posing a risk to feed and food safety. This study investigates the effects of substituting soybean meal with different levels of BSF larvae meal on laying performance, egg quality, serum analysis, and the deposition of various metals in eggs, meat, and excreta. A total of 1,008 Lohmann hens (age 48 wk) were randomly assigned to 4 treatments. The treatments consisted of corn-soybean meal (CK) diet replaced with 7% (BSF7), 14% (BSF14), and 21% (BSF21) BSF larvae meal. Each treatment consisted of 6 replicates with 42 hens each and the trial lasted for 8 wk. Dietary BSF larvae meal treatments increased (linear, P < 0.05) the laying rate (1.52 to 1.95%) and decreased (linear, P < 0.01) the feed intake (3.64–3.86 g) during the entire experiment. During 48 to 52 wk, egg weight was decreased (P < 0.001) 0.93 g for the BSF21 group compared to CK group, however, during 52-56 wk, no differences in egg weight were observed among treatments. The addition of BSF larvae meal enhanced the eggshell strength (linear, P < 0.05), with no effect on the albumen height and yolk weight (P > 0.05). Low transfer of arsenic, lead, and cadmium concentration was observed in the egg yolk and egg white across different treatments (P > 0.05). Conversely, the concentrations of these metals and iron showed an increase, while Zinc exhibited a decrease in excreta as the dietary intake of BSF larvae meal increased. The chromium and iron increased linearly (P < 0.001) in the egg white with the inclusion of BSF larvae meal. Egg white iron and chromium showed a strong positive correlation with the dietary zinc and copper. Taken together, BSF larvae can replace soybean meal completely in laying diet, however, careful attention requires to elevated metal levels in diet and excreta.