Evaluation of appropriateness of antibiotic prescribing in primary healthcare institutions in China using proxy indicator.

Background: Our objectives were to develop a set of proxy indicators (PIs) suited for assessing antibiotic use appropriateness in China's primary healthcare institutions (PHIs), and assess performance scores of these PIs while exploring factors that influence the antibiotic appropriateness. Methods: We selected potential PIs for the PHIs through a RAND-modified Delphi procedure, and assessed clinimetric properties, focusing on measurability, applicability, and potential for improvement. PIs with favorable clinimetric properties were used to evaluate antibiotic prescription appropriateness by calculating performance scores of each PI. Institutions were categorized into three clusters representing different levels of appropriateness. We used the chi-square test and an ordinal logistic regression model at PHI level to explore factors influencing antibiotic appropriateness. Findings: Eighteen PIs were developed through two rounds of online surveys and one face-to-face meeting involving 20 stakeholders. All PIs met the clinimetric properties criteria and were used to analyze 209,662 antibiotic prescriptions across 269 PHIs. The percentage of PHIs meeting the target ranged from 3.1% to 69.3%, with 6 PIs below 10%. The appropriateness of antibiotic prescriptions was significantly associated with percentages of patients' gender of the PHIs. Interpretation: The varied and suboptimal performance of the PIs indicated the need for diverse efforts to enhance the rational antibiotic use at PHI level. It was necessary to devise distinct sets of PIs for diverse settings in future endeavors. Funding: This work was supported by the National Natural Science Foundation of China (grant numbers 72374009, 81973294).

Exploring flatfeet morphology in children aged 6-12 years: relationships with body mass and body height through footprints and three-dimensional measurements.

The aim of the study was to determine the relationship between flatfoot morphology and body mass and height in children aged 6-12 years. A total of 6471 Chinese children (mean age 9.0 ± 1.9 years, 41% female) were assessed for foot morphometry, body height, and body mass index. Foot morphology, including foot length, width, girth, arch height, hallux valgus angle, and rearfoot valgus angle, was measured using a 3D laser scanner. Flatfoot evaluations were conducted using the Sztriter-Godunov index (KY) from footprints. All measurements were analyzed by age and sex using the mean values of the left and right sides. Comparisons were performed between flatfoot groups, between body mass index (BMI) groups, and between body height groups. The study revealed a significant decrease in the incidence of bipedal flatfoot with age (p < 0.001), whereas the prevalence of obesity remained consistent (p > 0.05). Bipedal flatfoot was associated with distinct morphological changes, including lower arches, reduced instep height, diminished ankle heights and a greater rearfoot valgus angle (p < 0.05). When comparing the BMI groups, overweight children had larger and thicker feet (p < 0.05), but no differences were found in arch height and ankle height (p > 0.05). When comparing the body height groups, short-statured children had a shorter feet girth, shorter arches, and shorter ankle height (p < 0.05), but no differences were found in the rearfoot valgus angle (p > 0.05). CONCLUSION: The main characteristics of flat feet include lower arches and instep heights and ankle heights but higher rearfoot valgus angles. In general, overweight children's feet do not have the common features of flat feet. In contrast, short children had similar features of flatfoot except for rearfoot valgus. Assessment of posture, such as rearfoot valgus, can be critical in identifying children with flat feet. WHAT IS KNOWN: • The morphology of children's feet is associated with body growth, but the relationship between flatfeet and body mass and height remains controversial. WHAT IS NEW: • Three-dimensional foot measurement shows that body mass is generally not associated with flatfeet, while short children have lower arches but no rearfoot valgus.

Response of dissolved carbon dioxide and methane concentration to warming in shallow lakes.

Shallow lake ecosystems are highly sensitive to temperature fluctuation because of their high water surface-to-volume ratios. Shallow lakes have been increasingly identified as a hotspot of CO2 and CH4 emissions, but their response to temperature variation remains unclear. Here, we report from a 5-month outdoor mesocosm experiment where we investigated the impacts of a projected 3.5 °C future warming and monthly temperature changes on lake CO2 and CH4, as well as the key drivers affecting the lake carbon cycling. Our results show that CO2 and CH4 concentrations had a significantly positive correlation with monthly temperatures. CH4 concentration was primarily regulated by monthly temperature, while nutrients effects on CO2 concentration overrode climate warming and temporal temperature changes. These findings imply the varied roles that temperature and nutrient levels can play on CO2 and CH4 dynamics in shallow lake systems. The relationship between temperature and CO2 concentration was nonlinear, showing a threshold of approximately 9 °C, at which CO2 concentration could be strongly modified by nutrient level in the lake systems. Understanding this complex relationship between temperature with CO2 and CH4 concentrations in shallow lakes is crucial for effective lake management and efficient control of greenhouse gases (GHGs) emissions.

Exploring Facilitators and Barriers to Delayed Antibiotic Prescribing in Rural Northwest China: A Qualitative Study Using the Theoretical Domains Framework and Behavior Change Wheel.

BACKGROUND: Antimicrobial resistance, exacerbated by antibiotic misuse, poses a global threat. Though delayed antibiotic prescribing (DAP) can mitigate antibiotic overuse, its adoption in developing nations, such as China, is limited. This study probed barriers and facilitators to DAP in Xinjiang, characterized by extensive rural landscapes and primary care institutions (PCIs). METHODS: Adopting a qualitative methodology, we conducted key informant interviews with thirty participants across six county hospitals in Xinjiang using VooV Meeting. Employing a two-stage sampling method targeting economically diverse areas, our interviews spanned physicians, pharmacists, patients, and caregivers. We organized the data according to the Theoretical Domains Framework (TDF) and the Behavior Change Wheel (BCW), spotlighting behavioral and policy elements impacting DAP. RESULTS: Our research included thirty interviewees. Twelve physicians contemplated delayed prescriptions, while five adult patients and six caregivers encountered recommendations for delayed antibiotic prescriptions. Six patients sought pharmacists' advice on antibiotic necessity. Prominent TDF domains were memory, attention, and beliefs about consequences. Critical intervention functions included education and environmental restructuring, while vital policy categories encompassed communication/marketing and guidelines. CONCLUSIONS: Countering antibiotic misuse and resistance in China necessitates overcoming barriers through strategic resource distribution, comprehensive education, rigorous training, and consistent monitoring, thereby promoting DAP adoption. The adoption of DAP in rural healthcare settings in China has the potential to significantly reduce antibiotic misuse, thereby mitigating the global threat of antimicrobial resistance.

The Effects of Partial Substitution of Fertilizer Using Different Organic Materials on Soil Nutrient Condition, Aggregate Stability and Enzyme Activity in a Tea Plantation.

Fertilization plays a crucial role in enhancing tea production. However, it has been demonstrated that the long-term single application of chemical fertilizer will reduce soil nutrient content and the formation of soil aggregates, which is not conducive to the sustainable development of soil and agriculture. Many studies have shown that partial substitution of chemical fertilizer with organic fertilizer can improve soil physicochemical properties and soil nutrient content. This study compared the effects of different organic materials as substitutes for chemical fertilizer. We partially replaced chemical fertilizer with rabbit manure, wine lees and rapeseed cake, amounting to 30% of the total annual nitrogen application in the field experiment, and we set nine different fertilization methods to assess and analyze the soil nutrient condition, aggregate stability and enzyme activity. The results showed that the experimental soil aggregate mean weight diameter (MWD) and geometric mean diameter (GMD) were significantly increased compared with control (p < 0.05); the aforementioned fertilization methods also decreased the soil aggregate fractal dimension (D), disruption rate (PAD), average weight-specific surface area (MWSSA) and soil erodibility factor (K). The application of the fertilizer containing organic materials and microbial agent increased soil organic carbon (SOC) by 20.7% to 22.6% and total nitrogen (TN) by 34.6% to 38.1%; it also significantly promoted sucrase, urease and protease activities in all aggregate sizes (p < 0.05) and increased the 2-5 mm aggregate content. The correlation coefficients between the SOC and the enzyme activities were 0.18-0.95, and most of them showed an extremely significant positive correlation (p < 0.01). In conclusion, the application of fertilizers containing organic materials and microbial agents can improve soil aggregate stability, aggregate enzyme activity and soil structural stability.

Nitrogen addition effect overrides warming effect on dissolved CO2 and phytoplankton structure in shallow lakes.

Shallow lakes are numerous in all climate zones, but our knowledge about their dissolved carbon dioxide (CO2) response to future climate change and nutrient enrichment is rather limited. Here we performed a mesocosm experiment with four treatments to investigate how warming and nitrogen addition will impact the partial pressure of CO2 (pCO2) and phytoplankton community individually and combined. We found that warming alone had no significant effect on pCO2, while nitrogen addition increased pCO2 significantly. The combined effects of nitrogen addition and warming on pCO2 level were prevalent, indicating that eutrophic shallow lakes would be double-jeopardized in the future climate. Warming and nitrogen addition together also showed to have changed the phytoplankton community structure, suggesting a potential shifting of biological system in shallow lakes under changing climate. These findings highlight the importance of reducing nitrogen pollution to shallow lake systems for sustainable development goal.

Artificial nerve graft constructed by coculture of activated Schwann cells and human hair keratin for repair of peripheral nerve defects.

Studies have shown that human hair keratin (HHK) has no antigenicity and excellent mechanical properties. Schwann cells, as unique glial cells in the peripheral nervous system, can be induced by interleukin-1ß to secrete nerve growth factor, which promotes neural regeneration. Therefore, HHK with Schwann cells may be a more effective approach to repair nerve defects than HHK without Schwann cells. In this study, we established an artificial nerve graft by loading an HHK skeleton with activated Schwann cells. We found that the longitudinal HHK microfilament structure provided adhesion medium, space and direction for Schwann cells, and promoted Schwann cell growth and nerve fiber regeneration. In addition, interleukin-1ß not only activates Schwann cells, but also strengthens their activity and increases the expression of nerve growth factors. Activated Schwann cells activate macrophages, and activated macrophages secrete interleukin-1ß, which maintains the activity of Schwann cells. Thus, a beneficial cycle forms and promotes nerve repair. Furthermore, our studies have found that the newly constructed artificial nerve graft promotes the improvements in nerve conduction function and motor function in rats with sciatic nerve injury, and increases the expression of nerve injury repair factors fibroblast growth factor 2 and human transforming growth factor B receptor 2. These findings suggest that this artificial nerve graft effectively repairs peripheral nerve injury.

Hedgehog Signalling Contributes to Trauma-Induced Tendon Heterotopic Ossification and Regulates Osteogenesis through Antioxidant Pathway in Tendon-Derived Stem Cells.

Heterotopic ossification (HO) is defined as the generation of pathological ectopic bony structures in soft tissues, but the molecular mechanisms of tendon HO are not fully revealed. Hedgehog (Hh) signalling is reportedly critical in hereditary HO. Our study focuses on the role of Hh signalling in the formation of trauma-induced tendon ossification. In this study, samples of healthy tendons and injured tendons from C57BL/6J female mice at 1, 4, 7, and 10 weeks after Achilles tenotomy were collected for quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemical analysis (IHC). At 1, 4, 7, and 10 weeks postinjury, tendon samples from the mice administered with vehicle, GANT58 (a GLI antagonist), or SAG (a smoothened agonist) were harvested for micro-CT, histological staining, qRT-PCR, and IHC. Rat tendon-derived stem cells (TDSCs) treated with vehicle, GANT58, or SAG were used to induce osteogenic and chondrogenic differentiation in vitro for qRT-PCR, alkaline phosphatase staining, Alcian blue staining, and reactive oxygen species (ROS) levels measurement. We found that Hh signalling is remarkably activated during the formation of trauma-induced tendon ossification in the model of Achilles tenotomy. The in vitro and in vivo assays both confirm that downregulation of Hh signalling significantly suppresses osteogenesis and chondrogenesis to inhibit tendon ossification, while upregulation of Hh signalling promotes this process. Under osteogenic induction, Hh signalling regulates antioxidant pathway and affects ROS generation of TDSCs. Collectively, Hh signalling contributes to trauma-induced tendon ossification and affects ROS generation through antioxidant pathway in osteogenic differentiation of TDSCs, indicating that targeting Hh signalling by GANT58 may be a potential treatment for trauma-induced tendon ossification.

In vitromodel to study confined osteocyte networks exposed to flow-induced mechanical stimuli.

Osteocytes are considered the primary mechanical sensor in bone tissue and orchestrate the coupled bone remodeling activity of adjacent osteoblast and osteoclast cells.In vivoinvestigation of mechanically induced signal propagation through networks of interconnected osteocytes is confounded by their confinement within the mineralized bone matrix, which cannot be modeled in conventional culture systems. In this study, we developed a new model that mimics thisin vivoconfinement using gelatin methacrylate (GelMA) hydrogel or GelMA mineralized using osteoblast-like model cells. This model also enables real-time optical examination of osteocyte calcium (Ca2+) signaling dynamics in response to fluid shear stimuli cultured under confined conditions. Using this system, we discovered several distinct and previously undescribed patterns of Ca2+responses that vary across networks of interconnected osteocytes as a function of space, time and connectivity. Heterogeneity in Ca2+signaling may provide new insights into bone remodeling in response to mechanical loading. Overall, such a model can be extended to study signaling dynamics within cell networks exposed to flow-induced mechanical stimuli under confined conditions.

Electrospun aligned poly(ε-caprolactone) nanofiber yarns guiding 3D organization of tendon stem/progenitor cells in tenogenic differentiation and tendon repair.

Hierarchical anisotropy structure directing 3D cellular orientation plays a crucial role in designing tendon tissue engineering scaffolds. Despite recent development of fabrication technologies for controlling cellular organization and design of scaffolds that mimic the anisotropic structure of native tendon tissue, improvement of tenogenic differentiation remains challenging. Herein, we present 3D aligned poly (ε-caprolactone) nanofiber yarns (NFYs) of varying diameter, fabricated using a dry-wet electrospinning approach, that integrate with nano- and micro-scale structure to mimic the hierarchical structure of collagen fascicles and fibers in native tendon tissue. These aligned NFYs exhibited good in vitro biocompatibility, and their ability to induce 3D cellular alignment and elongation of tendon stem/progenitor cells was demonstrated. Significantly, the aligned NFYs with a diameter of 50 µm were able to promote the tenogenic differentiation of tendon stem/progenitor cells due to the integration of aligned nanofibrous structure and suitable yarn diameter. Rat tendon repair results further showed that bundled NFYs encouraged tendon repair in vivo by inducing neo-collagen organization and orientation. These data suggest that electrospun bundled NFYs formed by aligned nanofibers can mimic the aligned hierarchical structure of native tendon tissue, highlighting their potential as a biomimetic multi-scale scaffold for tendon tissue regeneration.

Construction and Validation of a Tumor Microenvironment-Based Scoring System to Evaluate Prognosis and Response to Immune Checkpoint Inhibitor Therapy in Lung Adenocarcinoma Patients.

BACKGROUND: Lung cancer is among the most dangerous malignant tumors to human health. Lung adenocarcinoma (LUAD) accounts for about 40% of all lung cancers. Accumulating evidence suggests that the tumor microenvironment (TME) is a crucial regulator of carcinogenesis and therapeutic efficacy in LUAD. However, the impact of tumor microenvironment-related signatures (TMERSs) representing the TME characteristics on the prognosis and therapeutic outcome of LUAD patients remains to be further explored. MATERIALS AND METHODS: Gene expression files and clinical information of 1630 LUAD samples and 275 samples with immunotherapy information from different databases such as The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO) and Cancer Research Institute (CRI) iAtlas were downloaded and analyzed. Three hundred tumor microenvironment-related signatures (TMERS) based on a comprehensive collection of marker genes were quantified by single sample gene set enrichment analysis (ssGSEA), and then eight significant signatures were selected to construct the tumor microenvironment-related signature score (TMERSscore) by performing Least Absolute Shrinkage and Selection Operator (LASSO)-Cox analysis. RESULTS: In this study, we constructed a TME-based prognostic stratification model for patients with LUAD and validated it in several external datasets. Furthermore, the TMERSscore was found to be positively correlated with tumor malignancy and a high TMERSscore predicted a poor prognosis. Moreover, the TMERSscore of responders treated with Immune Checkpoint Inhibitor (ICI) therapies was significantly lower than that of non-responders, and the TMERSscore was positively correlated with the tumor immune dysfunction and exclusion (TIDE) score, implying that a low TMERSscore predicts a better response to ICI treatment and may provide independent and incremental predictive value over current biomarkers. CONCLUSIONS: Overall, we constructed a TMERSscore that can be used for LUAD patient prognosis stratification as well as ICI therapeutic efficacy evaluation, supportive results from independent external validation sets showed its robustness and effectiveness.

The Application and Efficacy Evaluation of Autologous Fat Transplantation in Antiaging of the Face: Systematic Review and Meta-Analysis.

Objective: To evaluate the therapy of autologous fat fine-grain transplantation in the application of antiaging facial by metaelaboration method. Methods: In Chinese and English databases, such as Wanfang, Weipu, CNKI, PubMed, Cochrane Library, and EMBASE, the literature on the use of autologous fat fines graft in antiaging facial was collected, covering randomized comparative trials, nonrandomized comparative studies, comparative case studies, cohort studies, case reports, and so forth. Meta-analyses were also conducted on complication rates, beauty seeker satisfaction, and one-shot success rates. Results: This study included 23 studies that held metaelaborations, including a total of 2852 beauty seekers. According to metaelaboration, the complication rate was 0.02 (95% CI: 0.01, 0.04), the satisfaction rate of beauty seekers was 0.95 (95% CI: 0.91, 0.97), and the success rate of one injection is 0.79 (95% CI: 0.73, 0.84). Conclusion: Autologous fat fine-grain transplantation is obvious and stable in the use of antiaging facial, and it is necessary to popularize the use in clinical practice.

Perceived Overqualification and Intensive Smartphone Use: A Moderated Mediation Model.

Previous studies only considered the impact of personal or environmental factors on intensive smartphone use separately, while largely ignoring the impact of person-environment (P-E) fit on it. Drawing on the P-E fit theory, we proposed that perceived overqualification (POQ), an indicator of person-job misfit, positively affects intensive smartphone use via job boredom, and affective commitment moderates this indirect effect. We examined our hypotheses using four-wave time-lag data of 450 workers from 62 teams. The results revealed that POQ raised job boredom of an individual and thus increased their intensive smartphone use. In addition, when the affective commitment was high, the indirect effect from POQ to intensive smartphone use via job boredom was weaker. The implications, limitations, and future directions of this research were discussed.

Humble leadership and its outcomes: A meta-analysis.

The importance of humble leadership has garnered attention from both researchers and practitioners. Unfortunately, despite the accumulation of recent findings on the effects of leader humility, a quantitative review remains scant. In addressing this void, this study is among the first to conduct a meta-analytic review of humble leadership and its outcomes. Eighty-four correlations (N = 16,534) from 53 independent studies are synthesized. The authors found that: (a) humble leadership is positively related to affective commitment (ρ = 0.56), affective trust (ρ = 0.62), creativity (ρ = 0.39), engagement (ρ = 0.40), leader-member exchange (LMX) (ρ = 0.58), job satisfaction (ρ = 0.51), organizational identification (ρ = 0.48), psychological empowerment (ρ = 0.33), self-efficacy (ρ = 0.24), task performance (ρ = 0.33), and voice (ρ = 0.34); and that (b) humble leadership contributes a significant incremental variance beyond transformational, servant, and ethical leadership in several crucial criterion variables, providing solid evidence for the construct's uniqueness. However, humble leadership does not explain incremental variance in some criterion variables, indicating that future studies should control for the influence of some positive leadership (e.g., transformational and servant leadership). Age, gender, study design, country, and year partially moderate the correlations of interest. We discuss our findings with caution and propose future research directions.

Inhibition of glucose use improves structural recovery of injured Achilles tendon in mice.

Injured tendons do not regain their native structure except at fetal or very young ages. Healing tendons often show mucoid degeneration involving accumulation of sulfated glycosaminoglycans (GAGs), but its etiology and molecular base have not been studied substantially. We hypothesized that quality and quantity of gene expression involving the synthesis of proteoglycans having sulfated GAGs are altered in injured tendons and that a reduction in synthesis of sulfated GAGs improves structural and functional recovery of injured tendons. C57BL6/j mice were subjected to Achilles tendon tenotomy surgery. The injured tendons accumulated sulfate proteoglycans as early as 1-week postsurgery and continued so by 4-week postsurgery. Transcriptome analysis revealed upregulation of a wide range of proteoglycan genes that have sulfated GAGs in the injured tendons 1 and 3 weeks postsurgery. Genes critical for enzymatic reaction of initiation and elongation of chondroitin sulfate GAG chains were also upregulated. After the surgery, mice were treated with the 2-deoxy-d-glucose (2DG) that inhibits conversion of glucose to glucose-6-phosphate, an initial step of glucose metabolism as an energy source and precursors of monosaccharides of GAGs. The 2DG treatment reduced accumulation of sulfated proteoglycans, improved collagen fiber alignment, and reduced the cross-sectional area of the injured tendons. The modulus of the 2DG-treated groups was higher than that in the vehicle group, but not of statistical significance. Our findings suggest that mucoid degeneration in injured tendons may result from the upregulated expression of genes involved the synthesis of sulfate proteoglycans and can be inhibited by reduction of glucose utilization.

Corrigendum: Vitamin B12 Enhances Nerve Repair and Improves Functional Recovery After Traumatic Brain Injury by Inhibiting ER Stress-Induced Neuron Injury.

[This corrects the article DOI: 10.3389/fphar.2019.00406.].

Perfusion-based co-culture model system for bone tissue engineering.

In this work, we report on a perfusion-based co-culture system that could be used for bone tissue engineering applications. The model system is created using a combination of Primary Human Umbilical Vein Endothelial Cells (HUVECs) and osteoblast-like Saos-2 cells encapsulated within a Gelatin Methacrylate (GelMA)-collagen hydrogel blend contained within 3D printed, perfusable constructs. The constructs contain dual channels, within a custom-built bioreactor, that were perfused with osteogenic media for up to two weeks in order to induce mineral deposition. Mineral deposition in constructs containing only HUVECs, only Saos-2 cells, or a combination thereof was quantified by microCT to determine if the combination of endothelial cells and bone-like cells increased mineral deposition. Histological and fluorescent staining was used to verify mineral deposition and cellular function both along and between the perfused channels. While there was not a quantifiable difference in the amount of mineral deposited in Saos-2 only versus Saos-2 plus HUVEC samples, the location of the deposited mineral differed dramatically between the groups and indicated that the addition of HUVECs within the GelMA matrix allowed Saos-2 cells, in diffusion limited regions of the construct, to deposit bone mineral. This work serves as a model on how to create perfusable bone tissue engineering constructs using a combination of 3D printing and cellular co-cultures.

Stabilization of HIF-1α alleviates osteoarthritis via enhancing mitophagy.

Mitochondrial dysfunction leads to osteoarthritis (OA) and disc degeneration. Hypoxia inducible factor-1α (HIF-1α) mediated mitophagy has a protective role in several diseases. However, the underlying mechanism of HIF-1α mediated mitophagy in OA remains largely unknown. This current study was performed to determine the effect of HIF-1α mediated mitophagy on OA. Therefore, X-ray and tissue staining including HE staining, safranin O-fast green (S-O) and Alcian Blue were used to assess imageology and histomorphology differences of mouse knee joint. Transcriptional analysis was used to find the possible targets in osteoarthritis. Western blot analysis, RT-qPCR and immunofluorescence staining were used to detect the changes in gene and protein levels in the vitro experiment. The expression of HIF-1α was increased in human and mouse OA cartilage. HIF-1α knockdown by siRNA further impair the hypoxia-induced mitochondrial dysfunction; In contrast, HIF-1α mediated protective role was reinforced by prolylhydroxylase (PHD) inhibitor dimethyloxalylglycine (DMOG). In addition, HIF-1α stabilization could alleviate apoptosis and senescence via mitophagy in chondrocytes under hypoxia condition, which could also ameliorate surgery-induced cartilage degradation in mice OA model. In conclusion, HIF-1α mediated mitophagy could alleviate OA, which may serve as a promising strategy for OA treatment.

High cholesterol induces apoptosis and autophagy through the ROS-activated AKT/FOXO1 pathway in tendon-derived stem cells.

BACKGROUND: Hypercholesterolemia increases the risk of tendon pain and tendon rupture. Tendon-derived stem cells (TDSCs) play a vital role in the development of tendinopathy. Our previous research found that high cholesterol inhibits tendon-related gene expression in TDSCs. Whether high cholesterol has other biological effects on TDSCs remains unknown. METHODS: TDSCs isolated from female SD rats were exposed to 10 mg/dL cholesterol for 24 h. Then, cell apoptosis was assessed using flow cytometry and fluorescence microscope. RFP-GFP-LC3 adenovirus transfection was used for measuring autophagy. Signaling transduction was measured by immunofluorescence and immunoblotting. In addition, Achilles tendons from ApoE -/- mice fed with a high-fat diet were histologically assessed using HE staining and immunohistochemistry. RESULTS: In this work, we verified that 10 mg/dL cholesterol suppressed cell proliferation and migration and induced G0/G1 phase arrest. Additionally, cholesterol induced apoptosis and autophagy simultaneously in TDSCs. Apoptosis induction was related to increased expression of cleaved caspase-3 and BAX and decreased expression of Bcl-xL. The occurrence of autophagic flux and accumulation of LC3-II demonstrated the induction of autophagy by cholesterol. Compared with the effects of cholesterol treatment alone, the autophagy inhibitor 3-methyladenine (3-MA) enhanced apoptosis, while the apoptosis inhibitor Z-VAD-FMK diminished cholesterol-induced autophagy. Moreover, cholesterol triggered reactive oxygen species (ROS) generation and activated the AKT/FOXO1 pathway, while the ROS scavenger NAC blocked cholesterol-induced activation of the AKT/FOXO1 pathway. NAC and the FOXO1 inhibitor AS1842856 rescued the apoptosis and autophagy induced by cholesterol. Finally, high cholesterol elevated the expression of cleaved caspase-3, Bax, LC3-II, and FOXO1 in vivo. CONCLUSION: The present study indicated that high cholesterol induced apoptosis and autophagy through ROS-activated AKT/FOXO1 signaling in TDSCs, providing new insights into the mechanism of hypercholesterolemia-induced tendinopathy. High cholesterol induces apoptosis and autophagy through the ROS-activated AKT/FOXO1 pathway in tendon-derived stem cells.

Calcium ion assisted fluorescence determination of microRNA-167 using carbon dots-labeled probe DNA and polydopamine-coated Fe3O4 nanoparticles.

A selective and sensitive fluorescence biosensor is described for determination of microRNA-167 using fluorescent resonant energy transfer (FRET) strategy. The FRET system comprises carbon dots (CDs, donor) labeled with probe DNA (pDNA) and polydopamine (PDA)-coated Fe3O4 nanoparticles (Fe3O4@PDA NPs, acceptor). The CDs-pDNA can be absorbed onto the surface of Fe3O4@PDA NPs because of the strong π interaction between pDNA and PDA. With the enhanced adsorption ability of Fe3O4@PDA NPs by Ca2+, the fluorescence intensity of CDs at 445 nm (excitation at 360 nm) is quenched. In presence of microRNA-167, the hybridized complex of CDs-pDNA-microRNA-167 will be released from the surface of Fe3O4@PDA NPs due to the weak π interaction of the complex and PDA. This results in the fluorescence recovery of CDs. By application of twice-magnetic separation, the biosensor shows a wide linear range of 0.5-100 nM to microRNA-167 with a 76 pM detection limit. The method was applied to the determination of microRNA-167 in samples of total microRNA extractions from A. thaliana seedlings, and the recoveries ranged from 96.4 to 98.3%.