Design and evaluation of mRNA encoding recombinant neutralizing antibodies for botulinum neurotoxin type B intoxication prophylaxis.

Among all natural and synthetic toxins, botulinum neurotoxins (BoNTs), produced by Clostridium botulinum in an anaerobic environment, are the most toxic polymer proteins. Currently, the most effective modalities for botulism prevention and treatment are vaccination and antitoxin use, respectively. However, these modalities are associated with long response time for active immunization, side effects, and donor limitations. As such, the development of more promising botulism prevention and treatment modalities is warranted. Here, we designed an mRNA encoding B9-hFc - a heavy-chain antibody fused to VHH and human Fc that can neutralize BoNT serotype B (BoNT/B) effectively - and assessed its expression in vitro and in vivo. The results confirmed that our mRNA demonstrates good expression in vitro and in vivo. Moreover, a single mRNA lipid nanoparticle injection effectively prevents BoNT/B intoxication in vivo, with effects comparable to those of protein antibodies. In conclusion, we explored and clarified whether mRNA drugs encoding neutralizing antibodies prevent BoNT/B intoxication. Our results provide an efficient strategy for further research on the prevention and treatment of intoxication by botulinum toxin.

Shenqi Fuzheng injection hinders non-small cell lung cancer cell growth by regulating the Bax/Bcl-2 signaling pathway.

INTRODUCTION: Lung cancer (LC) is the most common solid tumor and is currently considered the primary cause of cancer-related deaths worldwide. In clinical efficacy studies, it was not difficult to find that the combination of SFI and chemotherapy could improve the general condition of patients, reduce the side effects of chemotherapy drugs, and have a cooperative antitumor effect in NSCLC patients. However, whether SFI can be used as a novel antitumor drug is still unknown. METHODS: First, meta-analysis aimed to explore the efficacy of SFI in NSCLC patients, and SFI was identified by ultra-performance liquid chromatography‒mass spectrometry (UPLC‒MS). Cell proliferation, migration, and invasion were explored by Cell Counting Kit-8 (CCK-8), scratch healing, and Transwell assays, respectively. Cell cycle and apoptosis assays were performed by flow cytometry. Transcriptome sequencing analysis was performed in four NSCLC cell lines. Differential expression analysis was used to identify potential targets. Apoptosis-related protein levels were detected by Western blotting assays. The effects of SFI in NSCLC were further investigated by mouse xenografts. RESULTS: SFI could markedly improve the chemotherapy efficacy of NSCLC patients. The main active ingredients include flavonoids and terpenoids, which can effectively exert antitumor effects. SFI could not only inhibit tumor cell proliferation and cell migration/invasion but also regulate the cell cycle and promote tumor cell apoptosis. In NSCLC, SFI could enhance the transcription level of the CHOP gene, thereby upregulating the expression of the proapoptotic proteins Bax and caspase 3, and inhibiting the expression of the antiapoptotic protein Bcl-2. SFI hindered the growth of mouse NSCLC xenografts in vivo. CONCLUSIONS: SFI hindered tumor progression and might promote apoptosis by increasing the expression of Bax, caspase 3 and decreasing the level of Bcl-2 in NSCLC.

Analysis of lumbar spine loading during walking in patients with chronic low back pain and healthy controls: An OpenSim-Based study.

Low back pain (LBP) is one of the most prevalent and disabling disease worldwide. However, the specific biomechanical changes due to LBP are still controversial. The purpose of this study was to estimate the lumbar and lower limb kinematics, lumbar moments and loads, muscle forces and activation during walking in healthy adults and LBP. A total of 18 healthy controls and 19 patients with chronic LBP were tested for walking at a comfortable speed. The kinematic and dynamic data of the subjects were collected by 3D motion capture system and force plates respectively, and then the motion simulation was performed by OpenSim. The OpenSim musculoskeletal model was used to calculate lumbar, hip, knee and ankle joint angle variations, lumbar moments and loads, muscle forces and activation of eight major lumbar muscles. In our results, significant lower lumbar axial rotation angle, lumbar flexion/extension and axial rotation moments, as well as the muscle forces of the four muscles and muscle activation of two muscles were found in patients with LBP than those of the healthy controls (p < 0.05). This study may help providing theoretical support for the evaluation and rehabilitation treatment intervention of patients with LBP.

Association between pain interference and motoric cognitive risk syndrome in older adults: a population-based cohort study.

OBJECTIVES: Motoric cognitive risk syndrome (MCR) is a pre-dementia condition characterized by subjective complaints in cognition and slow gait. Pain interference has previously been linked with cognitive deterioration; however, its specific relationship with MCR remains unclear. We aimed to examine how pain interference is associated with concurrent and incident MCR. METHODS: This study included older adults aged ≥ 65 years without dementia from the Health and Retirement Study. We combined participants with MCR information in 2006 and 2008 as baseline, and the participants were followed up 4 and 8 years later. The states of pain interference were divided into 3 categories: interfering pain, non-interfering pain, and no pain. Logistic regression analysis was done at baseline to examine the associations between pain interference and concurrent MCR. During the 8-year follow-up, Cox regression analysis was done to investigate the associations between pain interference and incident MCR. RESULTS: The study included 7120 older adults (74.6 ± 6.7 years; 56.8% females) at baseline. The baseline prevalence of MCR was 5.7%. Individuals with interfering pain had a significantly increased risk of MCR (OR = 1.51, 95% CI = 1.17-1.95; p = 0.001). The longitudinal analysis included 4605 participants, and there were 284 (6.2%) MCR cases on follow-up. Participants with interfering pain at baseline had a higher risk for MCR at 8 years of follow-up (HR = 2.02, 95% CI = 1.52-2.69; p < 0.001). CONCLUSIONS: Older adults with interfering pain had a higher risk for MCR versus those with non-interfering pain or without pain. Timely and adequate management of interfering pain may contribute to the prevention and treatment of MCR and its associated adverse outcomes.

Dual-channel deep graph convolutional neural networks.

The dual-channel graph convolutional neural networks based on hybrid features jointly model the different features of networks, so that the features can learn each other and improve the performance of various subsequent machine learning tasks. However, current dual-channel graph convolutional neural networks are limited by the number of convolution layers, which hinders the performance improvement of the models. Graph convolutional neural networks superimpose multi-layer graph convolution operations, which would occur in smoothing phenomena, resulting in performance decreasing as the increasing number of graph convolutional layers. Inspired by the success of residual connections on convolutional neural networks, this paper applies residual connections to dual-channel graph convolutional neural networks, and increases the depth of dual-channel graph convolutional neural networks. Thus, a dual-channel deep graph convolutional neural network (D2GCN) is proposed, which can effectively avoid over-smoothing and improve model performance. D2GCN is verified on CiteSeer, DBLP, and SDBLP datasets, the results show that D2GCN performs better than the comparison algorithms used in node classification tasks.

Identifying changes in dynamic plantar pressure associated with radiological knee osteoarthritis based on machine learning and wearable devices.

BACKGROUND: Knee osteoarthritis (KOA) is an irreversible degenerative disease that characterized by pain and abnormal gait. Radiography is typically used to detect KOA but has limitations. This study aimed to identify changes in plantar pressure that are associated with radiological knee osteoarthritis (ROA) and to validate them using machine learning algorithms. METHODS: This study included 92 participants with variable degrees of KOA. A modified Kellgren-Lawrence scale was used to classify participants into non-ROA and ROA groups. The total feature set included 210 dynamic plantar pressure features captured by a wearable in-shoe system as well as age, gender, height, weight, and body mass index. Filter and wrapper methods identified the optimal features, which were used to train five types of machine learning classification models for further validation: k-nearest neighbors (KNN), support vector machine (SVM), random forest (RF), AdaBoost, and eXtreme gradient boosting (XGBoost). RESULTS: Age, the standard deviation (SD) of the peak plantar pressure under the left lateral heel (f_L8PPP_std), the SD of the right second peak pressure (f_Rpeak2_std), and the SD of the variation in the anteroposterior displacement of center of pressure (COP) in the right foot (f_RYcopstd_std) were most associated with ROA. The RF model with an accuracy of 82.61% and F1 score of 0.8000 had the best generalization ability. CONCLUSION: Changes in dynamic plantar pressure are promising mechanical biomarkers that distinguish between non-ROA and ROA. Combining a wearable in-shoe system with machine learning enables dynamic monitoring of KOA, which could help guide treatment plans.

The effect of sense of security on job performance of medical staff: the mediating effect of psychological capital.

Objective: To investigate the current situation of sense of security, psychological capital and job performance of medical staff in Guangdong Province, and to explore the mediating role of psychological capital on the relationship between sense of security and job performance of medical staff. Methods: In this study, 969 health care workers were selected from February 2023 to April 2023 from 37 hospitals in Guangdong Province, China, using purposive sampling method. The Sense of Security Scale for Medical Staff (SSS-MS), psychological capital scale (PCS) in Chinese version and the Chinese version of job performance scale (JPS) were used in this study. We use SPSS 26.0 for statistical analysis and Amos 24.0 for structural equation modeling (SEM). The control variables entering SEM were selected by regression analysis. SEM analysis confirmed psychological capital scale's mediating function in the link between work performance scale and Sense of Security. Results: The overall SSS-MS, PCS, and JPS scores were 67.42 ± 16.136, 87.06 ± 15.04, and 77.87 ± 10.50, respectively. The results of Pearson's correlation analysis showed that there was a positive relationship between PCS and JPS (r = 0.722, P < 0.01), SSS-MS and JPS (r = 0.312, P < 0.01), and SSS-MS and PCS (r = 0.424, P < 0.01). PCS demonstrated a fully mediating influence on the link between medical workers' SSS-MS and JPS, according to structural equation modeling. Conclusion: The JPS of medical personnel in Guangdong Province is at a medium level, with much room for improvement. PCS is positively impacted by a sense of security. There is a supportive correlation between PCS, JPS, and SSS-MS. Furthermore, PCS fully mediates the relationship between medical staff members' JPS and their SSS-MS. The Job Diamond-Resource model and Conservation of Resource theory are further validated and supplemented by the findings of this study, which also gives managers a theoretical foundation for enhancing medical staff performance.

Promoting mental health in children and adolescents through digital technology: a systematic review and meta-analysis.

Background: The increasing prevalence of mental health issues among children and adolescents has prompted a growing number of researchers and practitioners to explore digital technology interventions, which offer convenience, diversity, and proven effectiveness in addressing such problems. However, the existing literature reveals a significant gap in comprehensive reviews that consolidate findings and discuss the potential of digital technologies in enhancing mental health. Methods: To clarify the latest research progress on digital technology to promote mental health in the past decade (2013-2023), we conducted two studies: a systematic review and meta-analysis. The systematic review is based on 59 empirical studies identified from three screening phases, with basic information, types of technologies, types of mental health issues as key points of analysis for synthesis and comparison. The meta-analysis is conducted with 10 qualified experimental studies to determine the overall effect size of digital technology interventions and possible moderating factors. Results: The results revealed that (1) there is an upward trend in relevant research, comprising mostly experimental and quasi-experimental designs; (2) the common mental health issues include depression, anxiety, bullying, lack of social emotional competence, and mental issues related to COVID-19; (3) among the various technological interventions, mobile applications (apps) have been used most frequently in the diagnosis and treatment of mental issues, followed by virtual reality, serious games, and telemedicine services; and (4) the meta-analysis results indicated that digital technology interventions have a moderate and significant effect size (g = 0.43) for promoting mental health. Conclusion: Based on these findings, this study provides guidance for future practice and research on the promotion of adolescent mental health through digital technology. Systematic review registration: https://inplasy.com/inplasy-2023-12-0004/, doi: 10.37766/inplasy2023.12.0004.

Protein-assisted synthesis of chitosan-coated minicells enhance dendritic cell recruitment for therapeutic immunomodulation within pulmonary tumors.

The efficacy of cancer therapies is significantly compromised by the immunosuppressive tumor milieu. Herein, we introduce a previously unidentified therapeutic strategy that harnesses the synergistic potential of chitosan-coated bacterial vesicles and a targeted chemotherapeutic agent to activate dendritic cells, thereby reshaping the immunosuppressive milieu for enhanced cancer therapy. Our study focuses on the protein-mediated modification of bacterium-derived minicells with chitosan molecules, facilitating the precise delivery of Doxorubicin to tumor sites guided by folate-mediated homing cues. These engineered minicells demonstrate remarkable specificity in targeting lung carcinomas, triggering immunogenic cell death and releasing tumor antigens and damage-associated molecular patterns, including calreticulin and high mobility group box 1. Additionally, the chitosan coating, coupled with bacterial DNA from the minicells, initiates the generation of reactive oxygen species and mitochondrial DNA release. These orchestrated events culminate in dendritic cell maturation via activation of the stimulator of interferon genes signaling pathway, resulting in the recruitment of CD4+ and CD8+ cytotoxic T cells and the secretion of interferon-ß, interferon-γ, and interleukin-12. Consequently, this integrated approach disrupts the immunosuppressive tumor microenvironment, impeding tumor progression. By leveraging bacterial vesicles as potent dendritic cell activators, our strategy presents a promising paradigm for synergistic cancer treatment, seamlessly integrating chemotherapy and immunotherapy.

MitoCur-1 induces ferroptosis to reverse vemurafenib resistance in melanoma through inhibition of USP14.

Melanoma is an aggressive malignant tumor with a poor prognosis. Vemurafenib (PLX4032, vem) is applied to specifically treat BRAF V600E-mutated melanoma patients. However, prolonged usage of vem makes patients resistant to the drug and finally leads to clinical failure. We previously tested the combination regimen of tubulin inhibitor VERU-111 with vem, as well as USP14 selective inhibitor b-AP15 in combination with vem, both of which have showed profound therapeutic effects in overcoming vem resistance in vitro and in vivo. Most importantly, we discovered that vem-resistant melanoma cell lines highly expressed E3 ligase SKP2 and DUB enzyme USP14, and we have demonstrated that USP14 directly interacts and stabilizes SKP2, which contributes to vem resistance. These works give us a clue that USP14 might be a promising target to overcome vem resistance in melanoma. MitoCur-1 is a curcumin derivative, which was originally designed to specifically target tumor mitochondria inducing redox imbalance, thereby promoting tumor cell death. In this study, we have demonstrated that it can work as a novel USP14 inhibitor, and thus bears great potential in providing an anti-tumor effect and sensitizing vem-resistant cells by inducing ferroptosis in melanoma. Application of MitoCur-1 dramatically induces USP14 inhibition and inactivation of GPX4 enzyme, meanwhile, increases the depletion of GSH and decreases SLC7A11 expression level. As a result, ferrous iron-dependent lipid ROS accumulated in the cell, inducing ferroptosis, thus sensitizes the vem-resistant melanoma cell. Interestingly, overexpression of USP14 antagonized all the ferroptosis cascade events induced by MitoCur-1, therefore, we conclude that MitoCur-1 induces ferroptosis through inhibition of USP14. We believe that by inhibition of USP14, vem resistance can be reversed and will finally benefit melanoma patients in future.

Transcriptome and metabolome analyses revealed the response mechanism of pepper roots to Phytophthora capsici infection.

BACKGROUND: Phytophthora root rot caused by the oomycete Phytophthora capsici is the most devastating disease in pepper production worldwide, and current management strategies have not been effective in preventing this disease. Therefore, the use of resistant varieties was regarded as an important part of disease management of P. capsici. However, our knowledge of the molecular mechanisms underlying the defense response of pepper roots to P. capsici infection is limited. METHODS: A comprehensive transcriptome and metabolome approaches were used to dissect the molecular response of pepper to P. capsici infection in the resistant genotype A204 and the susceptible genotype A198 at 0, 24 and 48 hours post-inoculation (hpi). RESULTS: More genes and metabolites were induced at 24 hpi in A204 than A198, suggesting the prompt activation of defense responses in the resistant genotype, which can attribute two proteases, subtilisin-like protease and xylem cysteine proteinase 1, involved in pathogen recognition and signal transduction in A204. Further analysis indicated that the resistant genotype responded to P. capsici with fine regulation by the Ca2+- and salicylic acid-mediated signaling pathways, and then activation of downstream defense responses, including cell wall reinforcement and defense-related genes expression and metabolites accumulation. Among them, differentially expressed genes and differentially accumulated metabolites involved in the flavonoid biosynthesis pathways were uniquely activated in the resistant genotype A204 at 24 hpi, indicating a significant role of the flavonoid biosynthesis pathways in pepper resistance to P. capsici. CONCLUSION: The candidate transcripts may provide genetic resources that may be useful in the improvement of Phytophthora root rot-resistant characters of pepper. In addition, the model proposed in this study provides new insight into the defense response against P. capsici in pepper, and enhance our current understanding of the interaction of pepper-P. capsici.

Cerebral Cortex Activation and Gait Performance between Healthy and Prefrail Older Adults during Cognitive and Walking Tasks.

Pre-frailty is a transitional stage between health and frailty. Previous studies have demonstrated that individuals with pre-frailty experience declines in cognitive and gait performances compared with healthy individuals. However, the basic neural mechanism underlying this needs to be clarified. In this cross-sectional study, twenty-one healthy older adults and fifteen with pre-frailty underwent three conditions, including a single cognitive task (SC), single walking task (SW), and dual-task (DT), while cortical hemodynamic reactions were measured using functional near-infrared spectroscopy (fNIRS). The prefrail group (PG) showed a significantly lower activation of the left dorsolateral prefrontal cortex (L-DLPFC) than the healthy group (HG) when performing SC (p < 0.05). The PG showed a significantly lower Timed Up and Go test and step speed than the HG during SW (p < 0.05). The coefficient of variation (CV) of the step length of the PG was significantly higher than that of the HG when performing DT (p < 0.05). No significant correlation in cerebral cortex activation and gait parameters in the HG when performing SW and DT was noted (p > 0.05). Participants of the PG with a higher oxygenated area in the left anterior prefrontal cortex (L-APFC) had a lower step frequency during SW (r = -0.533, p = 0.041), and so did the following indicators of the PG during DT: L-APFC and step speed (r = -0.557, p = 0.031); right anterior prefrontal cortex and step speed (r = -0.610, p = 0.016); left motor cortex and step speed (r = -0.674, p = 0.006); step frequency (r = -0.656, p = 0.008); and step length (r = -0.535, p = 0.040). The negative correlations between the cerebral cortex and gait parameters of the PG indicated a neural compensatory effect of pre-frailty. Therefore, older adults with pre-frailty promote prefrontal activation to compensate for the impaired sensorimotor systems.

Effects of inspiratory muscle training in patients with hypertension: a meta-analysis.

Objective: To explore the effects of inspiratory muscle training (IMT) on hypertension and provide guidance for its clinical application as an auxiliary approach. Methods: Articles published prior to July 2022 were searched in Cochrane Library, Web of Science, PubMed, Embase, CNKI, and Wanfang databases. Included were randomized controlled studies that used IMT to treat individuals with hypertension. The mean difference (MD) was computed using the Revman 5.4 software. In individuals with hypertension, the effects of IMT on systolic blood pressure (SBP), diastolic blood pressure (DBP), heart rate (HR), and pulse pressure (PP) were compared and studied. Results: There were found to be eight randomized controlled trials totaling 215 patients. According to a meta-analysis, the IMT reduced the SBP (MD: -12.55 mmHg, 95% CI: -15.78, -9.33), DBP (MD: -4.77 mmHg, 95% CI: -6.00, -3.54), HR (MD: -5.92 bpm, 95% CI: -8.72, -3.12), and PP (MD: -8.92 mmHg, 95% CI: -12.08, -5.76) in patients with hypertension. In subgroup analyses, low-intensity IMT showed a better reduction in SBP (MD: -14.47 mmHg, 95% CI: -17.60, -11.34), DBP (MD: -7.70 mmHg, 95% CI: -10.21, -5.18). Conclusion: IMT may become an auxiliary means to improve the four hemodynamic indexes (SBP, DBP, HR and PP) in patients with hypertension. In subgroup analyses, low-intensity IMT was more effective in regulating blood pressure than medium-high-intensity IMT. Systematic Review Registration: https://www.crd.york.ac.uk/prospero/, identifier: CRD42022300908.

The relationship between ethylene-induced autophagy and reactive oxygen species in Arabidopsis root cells during the early stages of waterlogging stress.

The response of plants to waterlogging stress is a complex process, with ethylene playing a crucial role as a signaling molecule. However, it remains unclear how ethylene is initially triggered in response to waterlogging stress when plants are continuously waterlogged for less than 12 hours. Here, we have shown that ethylene-induced autophagy leads to the degradation of damaged mitochondria (the main organelles producing reactive oxygen species (ROS)) to reduce ROS production during oxidative stress in Arabidopsis thaliana, which improves the survival rate of root cells in the early stages of waterlogging stress. Waterlogging stress activated ethylene-related genes, including ACO2, ACS2, ERF72, ERF73, and EIN3, and ethylene content of plants increased significantly within 24 h of continuous waterlogging. As stress duration increased, increased amounts of ROS accumulated in Arabidopsis thaliana roots, and the activity of antioxidant enzymes initially increased and then decreased. Concurrently, the level of ethylene-induced autophagy, which participates in antioxidant defense, is higher in wild-type plants than in the octuple acs mutant cs16651 (acs2-1/acs4-1/acs5-2/acs6-1/acs7-1/acs9-1/amiRacs8acs11). Exogenous application of 1-aminocyclopropanecarboxylic acid (ACC), resulted in a more pronounced manifestation of autophagy in the stele of Arabidopsis roots. Compared with the waterlogging treatment group or the ACC treatment group, the waterlogging + ACC treatment can induce autophagy to occur earlier and expand the autophagic range to the epidermis of Arabidopsis thaliana roots. Overall, our results provide insight into the important role of ethylene-induced autophagy in enhancing the antioxidative capacity of Arabidopsis thaliana during the early stages of waterlogging stress. Furthermore, we suggest ethylene as a potential candidate for mitigating the deleterious effects caused by waterlogging in Arabidopsis thaliana.

Integrated Microbiome and Metabolomic Analysis Reveal Responses of Rhizosphere Bacterial Communities and Root exudate Composition to Drought and Genotype in Rice (Oryza sativa L.).

BACKGROUND: As climate change events become more frequent, drought is an increasing threat to agricultural production and food security. Crop rhizosphere microbiome and root exudates are critical regulators for drought adaptation, yet our understanding on the rhizosphere bacterial communities and root exudate composition as affected by drought stress is far from complete. In this study, we performed 16S rRNA gene amplicon sequencing and widely targeted metabolomic analysis of rhizosphere soil and root exudates from two contrasting rice genotypes (Nipponbare and Luodao 998) exposed to drought stress. RESULTS: A reduction in plant phenotypes was observed under drought, and the inhibition was greater for roots than for shoots. Additionally, drought exerted a negligible effect on the alpha diversity of rhizosphere bacterial communities, but obviously altered their composition. In particular, drought led to a significant enrichment of Actinobacteria but a decrease in Firmicutes. We also found that abscisic acid in root exudates was clearly higher under drought, whereas lower jasmonic acid and L-cystine concentrations. As for plant genotypes, variations in plant traits of the drought-tolerant genotype Luodao 998 after drought were smaller than those of Nipponbare. Interestingly, drought triggered an increase in Bacillus, as well as an upregulation of most organic acids and a downregulation of all amino acids in Luodao 998. Notably, both Procrustes analysis and Mantel test demonstrated that rhizosphere microbiome and root exudate metabolomic profiles were highly correlated. A number of differentially abundant genera responded to drought and genotype, including Streptomyces, Bacillus and some members of Actinobacteria, were significantly associated with organic acid and amino acid contents in root exudates. Further soil incubation experiments showed that Streptomyces was regulated by abscisic acid and jasmonic acid under drought. CONCLUSIONS: Our results reveal that both drought and genotype drive changes in the compositions of rice rhizosphere bacterial communities and root exudates under the greenhouse condition, and that organic acid exudation and suppression of amino acid exudation to select specific rhizosphere bacterial communities may be an important strategy for rice to cope with drought. These findings have important implications for improving the adaptability of rice to drought from the perspective of plant-microbe interactions.

Effect of Technology Acceptance on Blended Learning Satisfaction: The Serial Mediation of Emotional Experience, Social Belonging, and Higher-Order Thinking.

This study explored the relationship between technology acceptance and learning satisfaction in the context of blended learning, with a particular focus on the mediating effects of online behaviors, emotional experience, social belonging, and higher-order thinking. A total of 110 Chinese university students participated in this study and completed a questionnaire at the end of 11 weeks of blended learning. The results demonstrate that technology acceptance directly and indirectly relates to blended learning satisfaction. The mediation analysis further revealed two significant mediating pathways from technology acceptance to blended learning satisfaction: one through higher-order thinking, and the other through serial mediation of emotional experience, social belonging, and higher-order thinking. Moreover, there was no significant mediating effect of online learning behaviors on blended learning satisfaction. Based on these results, we have proposed practical implications for improving blended learning practice to promote learner satisfaction. These results contribute to our understanding of blended learning as an integrated construct under the triadic interplay of technical environment, learning behaviors, and individual perceptions.

The opening of mitochondrial permeability transition pore (mPTP) and the inhibition of electron transfer chain (ETC) induce mitophagy in wheat roots under waterlogging stress.

Mitochondria are crucial for the regulation of intracellular energy metabolism, biosynthesis, and cell survival. And studies have demonstrated the role of mitochondria in oxidative stress-induced autophagy in plants. Previous studies found that waterlogging stress can induce the opening of mitochondrial permeability transition pore (mPTP) and the release of cytochrome c in endosperm cells, which proved that mPTP plays an important role in the programmed cell death of endosperm cells under waterlogging stress. This study investigated the effects of the opening of mPTP and the inhibition of ETC on mitophagy in wheat roots under waterlogging stress. The results showed that autophagy related genes in the mitochondria of wheat root cells could respond to waterlogging stress; waterlogging stress led to the degradation of the characteristic proteins cytochrome c and COXII in the mitochondria of root cells. With the prolongation of waterlogging time, the protein degradation degree and the occurrence of mitophagy gradually increased. Under waterlogging stress, exogenous mPTP opening inhibitor CsA inhibited mitophagy in root cells and alleviated mitophagy induced by flooding stress, while exogenous mPTP opening inducer CCCP induced mitophagy in root cells; exogenous mPTP opening inducer CCCP induced mitophagy in root cells. The electron transfer chain inhibitor antimycin A induces mitophagy in wheat root cells and exacerbates mitochondrial degradation. In conclusion, waterlogging stress led to the degradation of mitochondrial characteristic proteins and the occurrence of mitophagy in wheat root cells, and the opening of mPTP and the inhibition of ETC induced the occurrence of mitophagy.

Efficacy and safety of whole-body vibration therapy for post-stroke spasticity: A systematic review and meta-analysis.

Background: One of the main objectives of stroke rehabilitation is to alleviate post-stroke spasticity. Over the recent years, many studies have explored the potential benefits of whole-body vibration (WBV) treatment for post-stroke spasticity, but it is still controversial. Objective: The current study aims to assess the efficacy and safety of WBV for post-stroke spasticity and determine the appropriate application situation. Methods: From their establishment until August 2022, the following databases were searched: PubMed, Web of Science, Embase, Cochrane Central Register of Controlled Trials (CENTRAL), Medline, China National Knowledge Infrastructure (CNKI), and Wanfang. Only randomized controlled trials (RCTs) that were published in either English or Chinese were taken into consideration. We independently filtered the research, gathered the data from the studies, and evaluated the research quality (Cochrane RoB tool) and the overall evidence quality (GRADE). Rev Man 5.4 software was utilized to conduct statistical analysis. Results: In this analysis, 11 RCTs with 475 patients that reported on the effectiveness of WBV therapy for post-stroke spasticity were taken into account. Compared to the control groups, the results revealed that WBV combined with conventional rehabilitation at a vibration frequency lower than 20 Hz (SMD = -0.58, 95% CI: -0.98 to -0.19, P = 0.004) was more effective in relieving upper (SMD = -0.53, 95% CI: -1.04 to 0.03, P = 0.03) and lower limb spasticity (SMD = -0.21, 95% CI: -0.40 to -0.01, P = 0.04); similarly, it was superior for patients aged under 60 years (SMD = -0.41, 95% CI: -0.66 to -0.17, P = 0.0008) with acute and subacute stroke (SMD = -0.39, 95% CI: -0.68 to -0.09, P = 0.01). The valid vibration for reducing spasticity was found to last for 10 min (SMD = -0.41, 95% CI: -0.75 to -0.07, P = 0.02). None of the included studies revealed any serious adverse impact. Conclusion: Moderate-quality evidence demonstrated when WBV was used as an adjuvant, vibration <20 Hz for 10 min was effective and secure in treating upper and lower limb spasticity in patients with acute and subacute stroke under the age of 60 years. Systematic review registration: https://www.crd.york.ac.uk/PROSPERO/, identifier: CRD42022293951.

Efficacy of non-invasive brain stimulation on cognitive and motor functions in multiple sclerosis: A systematic review and meta-analysis.

Objective: In this study, we aimed to investigate the effects of non-invasive brain stimulation (NIBS) on cognitive and motor functions in patients with multiple sclerosis (pwMS). Methods: A literature search was performed in the Cochrane Library, Embase, PubMed, Web of Science, Medline, CNKI, and Wan fang. The time interval used for database construction was up to December 2022, and the language was not limited. The collected trials were subsequently screened, the data were extracted, the quality was evaluated, and the effect sizes were computed using STATA/MP Version 13 for outcome analysis. Standard mean difference (SMD) and 95% confidence interval (CI) were calculated for domain of interest. Results: In total, 17 articles that examined 364 patients with multiple sclerosis were included in this analysis. Non-invasive brain stimulation did not improve the overall cognitive function [SMD = 0.18, 95% CI (-0.32, 0.69), P = 0.475] but helped improve motor function in patients [SMD = 0.52, 95% CI (0.19, 0.85), P = 0.002]. Moreover, this study specifically indicated that non-invasive brain stimulation improved alerting [SMD = 0.68, 95% CI (0.09, 1.26), P = 0.02], whereas non-invasive brain stimulation intervention improved motor function in patients aged <45 years [SMD = 0.67, 95% CI (0.23, 1.10), P = 0.003] and in patients with expanded disability status scale scores (EDSS) <3.5 [SMD = 0.82, 95% CI (0.22, 1.42), P = 0.007]. In particular, NIBS contributed to the improvement of spasticity in pwMS [SMD = 0.68, 95% CI (0.13, 1.23), P = 0.015]. Conclusion: These results of this present study provide evidence that non-invasive brain stimulation could improve alertness in pwMS. Furthermore, NIBS may help pwMS with motor function and those who are under 45 years of age or with EDSS < 3.5 improve their motor function. For the therapeutic use of NIBS, we recommend applying transcranial magnetic stimulation as an intervention and located on the motor cortex M1 according to the subgroup analysis of motor function. These findings warrant verification. Systematic review registration: https://www.crd.york.ac.uk/PROSPERO/, identifier CRD42022301012.

Inhibition of USP14 enhances anti-tumor effect in vemurafenib-resistant melanoma by regulation of Skp2.

BACKGROUND: The mutation of BRAF V600E often occurred in melanoma and results in tumorigenesis. BRAF mutation drives hyperactivation of the RAF-MAPK-ERK pathway. The acquired drug resistance upon prolonged use of BRAF inhibitors (such as vemurafenib) still remains the main obstacle. Previously, we have found that E3 ligase Skp2 over-expresses vemurafenib-resistant melanoma cells, and knockdown of Skp2 enhances the anti-tumor effect of vemurafenib. Interestingly, the literature has reported that the selective USP14/UCHL5 inhibitor b-AP15 displays great potential in melanoma therapy; however, the molecular mechanism still remains unknown. METHODS: In vitro, the effect of the combination regimen of vemurafenib (Vem, PLX4032) and b-AP15 on vem-sensitive and vem-resistant melanoma has been investigated by wound healing, colony formation, transwell invasion assay, flow cytometry, lysosome staining, and ROS detection. In vivo, the combination effect on vem-resistant melanoma has been evaluated with a nude mice xenograft tumor model. GST-pulldown and co-immunoprecipitation (co-IP) assays have been applied to investigate the interactions between USP14, UCHL5, and Skp2. Cycloheximide (CHX) assay and ubiquitination assays have been used to explore the effect of USP14 on Skp2 protein half-life and ubiquitination status. RESULTS: In the present study, we have revealed that repression of USP14 sensitizes vemurafenib resistance in melanoma through a previously unappreciated mechanism that USP14 but not UCHL5 stabilizes Skp2, blocking its ubiquitination. K119 on Skp2 is required for USP14-mediated deubiquitination and stabilization of Skp2. Furthermore, the mutated catalytic activity amino acid cysteine (C) 114 on USP14 abrogates stabilization of Skp2. Stabilization of Skp2 is required for USP14 to negatively regulate autophagy. The combination regimen of Skp2 inhibitor vemurafenib and USP14/UCHL5 inhibitor b-AP15 dramatically inhibits cell viability, migration, invasion, and colony formation in vemurafenib-sensitive and vemurafenib-resistant melanoma. Vemurafenib and b-AP15 hold cells in the S phase thus leading to apoptosis as well as the formation of the autophagic vacuole in vemurafenib-resistant SKMEL28 cells. The enhanced proliferation effect of USP14 and Skp2 is mainly due to a more effective reduction of cell apoptosis and autophagy. Further evaluation of various protein alterations has revealed that the increased expression of cleaved-PARP, LC3, and decreased Ki67 are more obvious in the combination of vemurafenib and b-AP15 treatment than those in single-drug treatment. Moreover, the co-treatment of vemurafenib and b-AP15 dramatically inhibits the growth of vemurafenib-resistant melanoma xenograft in vivo. Collectively, our findings have demonstrated that the combination of Skp2 inhibitor and USP14 inhibitor provides a new solution for the treatment of BRAF inhibitor resistance melanoma.