In vivo single-molecule RNA structural profiling.

Cellular RNAs exhibit substantial heterogeneity in structure and function. Recently, Yang et al. developed an in vivo single-molecule RNA structure profiling methodology and revealed that individual isoforms of noncoding transcripts adopt multiple diverse and functionally relevant structural conformations, which change in abundance and structure in response to temperature conditions.

Genetically encoded RNA-based sensors with Pepper fluorogenic aptamer.

Sensors to measure the abundance and signaling of intracellular molecules are crucial for understanding their physiological functions. Although conventional fluorescent protein-based sensors have been designed, RNA-based sensors are promising imaging tools. Numerous RNA-based sensors have been developed. These sensors typically contain RNA G-quadruplex (RG4) motifs and thus may be suboptimal in living cells. Here we describe RNA-based sensors based on Pepper, a fluorogenic RNA without an RG4 motif. With Pepper, we engineered various sensors for metabolites, synthetic compounds, proteins and metal ions in vitro and in living cells. In addition, these sensors show high activation and selectivity, demonstrating their universality and robustness. In the case of sensors responding to S-adenosylmethionine (SAM), a metabolite produced by methionine adenosyltransferase (MATase), we showed that our sensors exhibited positively correlated fluorescence responding to different SAM levels. Importantly, we revealed the SAM biosynthesis pathway and monitored MATase activity and gene expression spatiotemporally in living individual human cells. Additionally, we constructed a ratiometric SAM sensor to determine the inhibition efficacy of a MATase inhibitor in living cells. Together, these sensors comprising Pepper provide a useful platform for imaging diverse cellular targets and their signaling pathway.

[Self-assembling peptide GFS-4 nanofiber scaffolds for three-dimensional cell cultures and myocardial infarction repair].

The purpose of this study is to investigate the effects of self-assembling peptide GFS-4 on three-dimen-sional myocardial cell culture and tissue repair of myocardial infarction. The circular dichroism (CD) spectrum was used to detect secondary structure of GFS-4, and atomic force microscope (AFM) was used to analyze the microstructure of self-assembly. The nanofiber scaffolds self-assembled by GFS-4 were used as the three-dimensional culture material to observe the growth effect of cardiomyocytes. The model of myocardial infarction was established and the effect of GFS-4 on myocardial infarction was studied. The results indicated that self-assembling peptide GFS-4 could form mainly ß-sheet structure that can form dense nanofiber scaffolds after 24 hours' self-assembling. The myocardial cells had a favorable growth status in GFS-4 nanofiber scaffold hydrogel when cells treated in three-dimen-sional cell culture. The experiment of repairing myocardial infarction in vitro proved that peptide GFS-4 hydrogel scaffold could alleviate tissue necrosis in a myocardial infarction area. As a new nanofiber scaffold material, self-assembling peptide GFS-4 can be used for three-dimensional cell culture and tissue repairing in myocardial infarction area.

Sustainability evaluation of sports tourism using a linguistic neutrosophic multi-criteria decision-making method.

Sports tourism represents a novel industrial manifestation of the profound integration between the tourism and sports sectors. The objective of this research is to examine an innovative multi-criteria decision-making (MCDM) method for the sustainability evaluation of sports tourism. The largest innovations are the expression and treatment of ambiguous data and interdependent evaluation criteria in the sports tourism sustainability evaluation process. On the one hand, intricate assessment data is represented using linguistic neutrosophic numbers (LNNs), which employ three linguistic variables to convey uncertainty and imprecision. On the other hand, to effectively capture the interrelationships among inputs, two novel aggregation operators are proposed. They are devised based on the Einstein operations and Heronian mean operators of LNNs. Subsequently, a linguistic neutrosophic evaluation method utilizing the aforementioned operators is presented. Comparative and sensitivity analyses conclude that great interdependence exists among five different dimensions of sustainability evaluation in sports tourism, and the proposed method can reflect the interrelations among inputs without redundant calculations.

Frequency of Vigorous physical activity and sleep difficulty in adolescents: A multiply-country cross-sectional study.

BACKGROUND: Sleep is an essential health behavior, and sleep difficulties are strongly associated with adolescent health, potentially leading to more severe sleep disorders. The beneficial effects of physical activity (PA) in alleviating sleep difficulties have been well-documented. Numerous investigations reveal influence in moderate to high-intensity physical activity (PA) positively influences sleep quality. Despite these findings, a gap in the literature exists, particularly regarding the association between frequency of vigorous-intensity physical activity (VPA) and sleep difficulties. AIM: This study aims to bridge the knowledge gap by exploring the link between sleep difficulty and frequency of VPA among adolescents. Insights are derived from analyzing data accumulated from the Health Behavior in School-aged Children (HBSC) project. METHODS: The analysis in this study utilized cross-sectional data from the HBSC (2017/2018). The study sample comprised a total of 171,233 respondents aged 11, 13, and 15 years, with males representing 51.1% of sample. Measurement instruments included a self-administered questionnaire, providing direct insight into sleep difficulty and frequency of VPA levels. Statistical analysis on the associaiton between frequency of VPA and sleep difficulties was conducted using Generalized Linear Models. RESULTS: 50.0% of adolescents reported no sleep difficulties, while 12.3% experienced sleep issues daily. Additionally, 17.1% of adolescents engaged in frequency of VPA on a daily basis, while 6.4% never participated in such activities. daily VPA was associated with fewer sleep difficulties (OR = 1.07 [1.00, 1.15]), 4-6 times a week (OR = 1.08 [1.01, 1.15]), and 2-3 times a week (OR = 1.08 [1.02, 1.16]). However, no significant association was found between sleep difficulties and frequency of VPA in girls. Furthermore, a negative association was observed between sleep difficulties and all frequencies of VPA (p < 0.05) in 11-year-old adolescents. For 13-year-olds, daily VPA was significantly associated with fewer sleep difficulties (OR = 1.10 [1.02, 1.19]), 4-6 times a week (OR = 1.15 [1.07, 1.24]), 2-3 times a week (OR = 1.19 [1.10, 1.27]), and once a week (OR = 1.13 [1.05, 1.22]). However, no significant association was found between sleep difficulties and frequency of VPA in 15-year-old adolescents. CONCLUSION: More participations in VPA would be an effective approach to reduce sleep difficulties in adolescents. Insights gleaned from this research illustrate a discernible link between sleep difficulty and frequency of VPA, particularly notable in male and 13-year-old participants. It is also imperative to underscore the variability in the connection between sleep difficulty and frequency of VPA, distinctly influenced by factors such as gender and age. Consequently, tailoring sleep intervention methodologies to align with the specific needs dictated by these variables emerges as a pivotal recommendation.

Evaluate teaching quality of physical education using a hybrid multi-criteria decision-making framework.

The teaching quality evaluation of physical education is an important measure to promote the professional development of physical teachers, improve the quality of school teaching and personnel training. It is helpful for students to achieve all-round development and better meet the needs of modern talents in the new era. This study aims to establish a novel MCDM (multi-criteria decision-making) framework for evaluating teaching quality of physical education. First, PFNs (picture fuzzy numbers) are suggested to reflect dissimilar attitudes or preferences of decision makers. Then, the typical SWARA (Step-wise Weight Assessment Ratio Analysis) model is modified with PFNs to calculate the weights of evaluation criteria. Considering that some criteria are non-compensatory during the evaluation process, the idea of ELECTRE (elimination and choice translating reality) is introduced to obtain the ranking results of alternatives. Specially, the MAIRCA (Multi-Attribute Ideal-Real Comparative Analysis) method is extended to construct the difference matrix in a picture fuzzy environment. Last, the hybrid MCDM model is utilized to assess the teaching quality of physical education. Its superiority is justified through comparison analyses. Results show that our approach is practicable and can provide instructions for the teaching quality evaluation of physical education.

Dynamic-stiffening collagen-coated substrate enhances osteogenic differentiation of mesenchymal stem cells through integrin α2ß1.

Matrix stiffness dynamically increases during the bone formation process. Enhancement of the osteogenic differentiation of mesenchymal stem cells (MSCs) by the dynamic stiffening of the substrate has been reported in previous research. However, the mechanism by which the dynamic stiffening of the matrix effects the osteogenic differentiation of MSCs remains quite unknown. A previously reported dynamic hydrogel system with dynamic matrix stiffening was used in this study to investigate the mechanical transduction mechanism of MSCs. The integrin α2ß1 and phosphorylation focal adhesion kinase levels were evaluated. The results indicated that dynamic stiffening of the matrix mediated the activation of integrin α2ß1, and further influenced the focal adhesion kinase (FAK) phosphorylation level of MSCs. In addition, integrin α2 is a probable integrin subunit that causes integrin ß1 activation during the matrix dynamic stiffening process. The integrin ß1 is the main integrin subunit regulating the osteogenic differentiation of MSCs induced by FAK phosphorylation. Overall, the results suggested that the dynamic stiffness facilitated the osteogenic differentiation process of the MSCs by regulating the integrin-α2ß1-mediated mechanical transduction pathway, which implied that integrin α2ß1 played a crucial role in the physical biological coupling in the dynamic matrix microenvironment.

Zonally Stratified Decalcified Bone Scaffold with Different Stiffness Modified by Fibrinogen for Osteochondral Regeneration of Knee Joint Defect.

Articular cartilage is generally known to be a complex tissue with multiple layers. Each layer has different composition, structure, and mechanical properties, making regeneration after knee joint defects a troubling clinical problem. A novel integrated stratified decalcified bone matrix (SDBM) scaffold with different stiffness to mimic the mechanical properties of articular cartilage is presented herein. This SDBM scaffold was modified using fibrinogen (Fg) (Fg + SDBM) to enhance its vascularization ability and improve its repair efficiency for osteochondral defects of knee joints. A Fg + SDBM scaffold with different elastic modulus in each layer (high-stiffness DBM (HDBM) layer, 174.208 ± 44.330 MPa (Fg + HDBM); medium-stiffness DBM (MDBM) layer, 21.214 ± 6.922 MPa (Fg + MDBM); and low-stiffness DBM (LDBM) layer, 0.678 ± 0.269 MPa (Fg + LDBM)) was constructed by controlling the stratified decalcification time with layered embedding paraffin (0, 3, and 5 days). The low- and medium-stiffness layers of the Fg + SDBM scaffold remarkably promoted the cartilage differentiation of bone marrow mesenchymal stem cells in vitro. Subcutaneous transplantation and rabbit knee joint osteochondral defect repair experiments revealed that the low- and medium-stiffness layers of the Fg + SDBM scaffold exhibited wonderful cartilage capacity, whereas the high-stiffness layer of Fg + SDBM scaffold exhibited good osteogenesis ability. Furthermore, this scaffold could promote blood vessel formation in subchondral bone area. This study presents a feasible strategy for osteochondral regeneration of defective knee joints, which is of great clinical value for tissue repair.

Exercise Intervention as a Therapy in Patients with Diabetes Mellitus and Sarcopenia: A Meta-Analysis.

INTRODUCTION: Sarcopenia is defined as a progressive and generalized muscle disorder that involves accelerated loss of muscle mass and impaired function. It is believed to influence the ability to carry out daily activities, muscle strength, and physical capacity in the elderly. Studies have shown that sarcopenia has been implicated as both a cause and a consequence of diabetes mellitus. In this analysis, we aimed to systematically show the impact of exercise intervention as a therapy for patients with diabetes mellitus and sarcopenia. METHODS: Electronic databases, including PubMed, EMBASE, Web of Science, and the Cochrane database, were searched from November to December 2021 for publications based on exercise intervention in patients with sarcopenia. After the selection of studies for this analysis, patients with diabetes mellitus were retrieved. Since dichotomous data including mean and standard deviation were reported, weighted mean difference (MD) with 95% confidence intervals (CI) were used to represent the data following analysis. RESULTS: A total of 431 participants with diabetes mellitus and sarcopenia were included in this meta-analysis. A statistical analysis was carried out on patients with diabetes mellitus who were assigned to the exercise intervention group. Our analysis showed that "sit-to-stand test" and "timed up and go" were significantly in favor of exercise intervention: MD -1.57, 95% confidence interval (CI) -2.26 to -0.87 (P = 0.0001) versus MD -0.61, 95% CI -1.21 to -0.01 (P = 0.05), respectively. Handgrip strength, walking speed and leg strength were also assessed. Another statistical analysis was carried out, this time on patients with diabetes mellitus and sarcopenia who were not assigned to an exercise intervention. The results showed no significant difference among sit-to-stand test, timed up and go, handgrip strength, and leg strength. CONCLUSION: Exercise intervention significantly improved the time taken to stand up from a sitting position, and to "stand up and go" in patients with diabetes mellitus and sarcopenia. Therefore, exercise intervention should be considered a relevant therapy for such patients.

[Cardiotoxicity of Anti-PD-L1 Antibody and the Effect of Levothyroxine 
in Attenuating the Related Mortality in Mice].

BACKGROUND: Immune checkpoint inhibitors (ICIs) such as antibodies against programmed cell death 1 (PD-1) and programmed cell death ligand 1 (PD-L1), have shown remarkable efficacies in many subtypes of cancers. However, ICIs may also cause severe immune-related adverse events in the recipient patients. Recently, ICI-associated myocarditis have been reported in hundreds of patients worldwide, with a mortality rate of approximately 50% in these cases. This study aims to recapitulate the cardiotoxicity and explore the detoxicifying approaches to attenuate mortality caused by PD-1/PD-L1 inhibitors in healthy mice. METHODS: Six to eight-week-old C57BL/6 mice were inoculated with anti-PD-1 antibody (12.5 µg/g every 5 days for 6 injections), anti-PD-L1 antibody (10 µg/g once a week for 6 weeks), anti-PD-L1 antibody (with the same dosage described above) in combination with levothyroxine (0.25 µg/g, intraperitoneally injected half an hour before anti-PD-L1 antibody injection), or isotype control immunoglobulin IgG (10 µg/g once a week for 6 weeks). The ejection function of the hearts was detected by echocardiography, body temperature and blood pressure were detected by Mouse MonitorTM and non-invassive blood pressure minotor, and serum free thyroxine concentration was detected by The enzyme linked immunosorbent assay (ELISA). RESULTS: PD-L1 was expressed at different levels by the cardiomyocytes of the mice. The isotype control immunoglobulin and anti-PD-1 antibody did not cause death of the mice. The 12 mice receiving 3-6 injections of anti-PD-L1 antibody showed a significant increase in the heart-to-tibial ratio and cardiomyoctye degeneration, hyalinization and extravascular inflammatory cell infiltration. In addition, the serum thyroxine was mardedly decreased to 1/3 of that in the control group mice, and the blood pressure and body temperature were abnormally decreased in mice upon treatment with PD-L1 blockade. Eight of the 12 (66.7%) mice died from multiple intravenous injection of anti-PD-L1 antibody.Intraperitoneal injection of levothyroxine 30 min before the injection of anti-PD-L1 antibody significantly attenuated the mortality rate of the anti-PD-L1 antibody-treated mice. CONCLUSIONS: The anti-PD-L1 antibody is cardiotoxic and lethal, and levothyroxine is able to rescue the mice from this immune checkpoint inhibitor-caused mortality.

The Therapeutic Effects of Mild to Moderate Intensity Aerobic Exercise on Glycemic Control in Patients with Type 2 Diabetes Mellitus: A Meta-Analysis of Randomized Trials.

INTRODUCTION: It has been recommended that physical activity be a part of treatment and management regimens of type 2 diabetes mellitus (T2DM), and research has shown that regular physical exercise facilitates glycemic control in these patients. In this analysis, our aim was to systematically show the therapeutic effects of mild to moderate intensity aerobic exercise on glycemic control in patients with T2DM. METHODS: From February to April 2021, we searched the https://www.clinicaltrials.gov , EMBASE, MEDLINE, Cochrane Central Register of Controlled Trials (CENTRAL), Web of Science and Google Scholar databases for trials that showed the effects of aerobic exercise on glycemic control in patients with T2DM. Glycated hemoglobin (HbA1c) was the endpoint in the analysis. The RevMan version 5.4 statistical program was used for statistical analysis, and the mean difference (MD) and 95% confidence intervals (CI) used to represent the data following analysis. RESULTS: Eighteen trials involving 972 participants with T2DM were included in this meta-analysis, of whom 523 were assigned to an exercise group and 449 were assigned to a control group. A comparison pre- versus post-aerobic exercise showed that aerobic exercise significantly improved glycemic control (HbA1c) (MD 0.35, 95% CI 0.23-0.48; P = 0.00001) in these patients with T2DM. A second comparison, T2DM participants in the experimental group post-exercise versus T2DM participants from the control group at the end of the follow-up, also showed that aerobic exercise significantly improved glycemic control (MD - 0.46, 95% CI - 0.69 to - 0.22; P = 0.0001). However, a comparison of HbA1c of T2DM participants in the control group at the beginning of the study compared to those at the end of follow-up did not show any significant improvement in glycemic control (MD 0.08, 95% CI - 0.05 to 0.21; P = 0.21). CONCLUSION: The current analysis showed that mild to moderate intensity aerobic exercise significantly improved glycemic control in patients with T2DM. Patients with T2DM who regularly participated in aerobic exercise activities had a better control of their disease than those who were not on a regular aerobic exercise regimen. These results lead to the recommendation that at least mild to moderate intensity aerobic exercise should be included in the treatment and management regimens of patients with T2DM.

Degradation of SARS-CoV-2 receptor ACE2 by the E3 ubiquitin ligase Skp2 in lung epithelial cells.

An unexpected observation among the COVID-19 pandemic is that smokers constituted only 1.4%-18.5% of hospitalized adults, calling for an urgent investigation to determine the role of smoking in SARS-CoV-2 infection. Here, we show that cigarette smoke extract (CSE) and carcinogen benzo(a)pyrene (BaP) increase ACE2 mRNA but trigger ACE2 protein catabolism. BaP induces an aryl hydrocarbon receptor (AhR)-dependent upregulation of the ubiquitin E3 ligase Skp2 for ACE2 ubiquitination. ACE2 in lung tissues of non-smokers is higher than in smokers, consistent with the findings that tobacco carcinogens downregulate ACE2 in mice. Tobacco carcinogens inhibit SARS-CoV-2 spike protein pseudovirions infection of the cells. Given that tobacco smoke accounts for 8 million deaths including 2.1 million cancer deaths annually and Skp2 is an oncoprotein, tobacco use should not be recommended and cessation plan should be prepared for smokers in COVID-19 pandemic.

Autophagy inhibition enhances the inhibitory effects of ursolic acid on lung cancer cells.

The aim of the present study was to identify natural compounds that bear significant anti­tumor activity. Thus, the effects of 63 small molecules that were isolated from traditional Chinese medicinal herbs on A549 human non­small cell lung cancer (NSCLC) and MCF­7 breast cancer cells were examined. It was found that ursolic acid (UA), a natural pentacyclic triterpenoid, exerted significant inhibitory effect on these cells. Further experiments revealed that UA inhibited the proliferation of various lung cancer cells, including the NSCLC cells, H460, H1975, A549, H1299 and H520, the human small cell lung cancer (SCLC) cells, H82 and H446, and murine Lewis lung carcinoma (LLC) cells. UA induced the apoptosis and autophagy of NSCLC cells. The inhibition of the mammalian target of rapamycin (mTOR) signaling pathway, but not the activation of the extracellular signal­regulated kinase 1/2 (ERK1/2) signaling pathway contributed to the UA­induced autophagy of NSCLC cells. Moreover, the inhibition of autophagy by chloroquine (CQ) or siRNA for autophagy­related gene 5 (ATG5) enhanced the UA­induced inhibition of cell proliferation and promotion of apoptosis, indicating that UA­induced autophagy is a pro­survival mechanism in NSCLC cells. On the whole, these findings suggest that combination treatment with autophagy inhibitors may be a novel strategy with which enhance the antitumor activity of UA in lung cancer.