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Abstract Background Osteoporosis affects the mandible resulting in bone loss. Though impairments are not life threatening, they affect a person's quality-of-life particularly vulnerable elderly. MicroRNAs (miRNAs) are novel regulatory factors that play an important role in regulating bone metabolism. Autophagy is evolutionarily conserved intracellular self-degradation process and is vital in the maintenance of both miRNA and bone homeostasis. However, the role of autophagy in the pathogenesis of miRNA regulating osteoporosis remains unclear. Methods In the study, we established a rat osteoporosis model induced by ovariectomy (OVX) and isolated mesenchymal stem cells from mandible (MMSCs-M). Several miRNAs were identified to regulate osteoporosis in some studies. qRT-PCR was applied to examine the expression of miRNA, autophagy and osteogenic differentiation-related genes. Western blotting assays were performed to detect the expression of autophagy and osteogenic differentiation proteins. Immunofluorescence and transmission electron microscope were used to verify the autophagy activity. Transfecting technology was used to enhance or suppress the expression of miR-152-5p which enable us to observe the relationship between miR-152-5p, autophagy and osteogenic differentiation. Additionally, the measurement of reactive oxygen species was used to investigate the mechanism of autophagy affecting osteogenic differentiation. Results We found an upregulated expression of miR-152-5p in MMSCs-M in OVX group. Downregulated autophagy-related gene, proteins and autophagosome were detected in vitro of OVX group compared with sham group. Moreover, downregulation of miR-152-5p promoted osteogenic differentiation of MMSCs-M as well as enhanced autophagy-related proteins in OVX group. Conversely, overexpression of miR-152-5p showed opposite effect in sham group. Meanwhile, we found Atg14 (autophagy-related protein homolog 14) was identified to be a direct target of miR-152-5p theoretically and functionally. In other words, we confirmed inhibition of miR-152-5p promoted the osteogenic differentiation via promoting ATG14-mediated autophagy. Furthermore, miR-152-5p/ATG14-mediated autophagy regulated osteogenic differentiation by reducing the endogenous ROS accumulation and maintaining cellular redox homeostasis. Conclusion Our data suggest that miR-152-5p is the first identified to regulate osteogenic differentiation by directly targeting autophagy-related protein ATG14 and regulating oxidative stress and therapeutic inhibition of miR-152-5p may be an efficient anabolic strategy for osteoporosis.

Self‐repair of critical bone defects from periodontitis, tumor resection, and trauma is difficult. Osteoporosis is an abnormal bone metabolism disease characterized by increased bone resorption, decreased bone formation, and decreased activity of bone marrow mesenchymal stem cells (BMSCs), which further inhibits the repair of bone defects. Improving the osteogenic ability of BMSCs after osteoporosis is an important step in treating osteoporotic bone defects. Hydroxyapatite (HA) with good biocompatibility is widely used in bone material research. HA and strontium‐doped hydroxyapatite (Sr‐HA) microspheres are synthesized following a hydrothermal method, and the characteristics and the ability of these microspheres are explored to promote osteogenesis. Further, the role of the calcium‐sensing receptor (CaSR) and Janus kinase 2 gene/signal transducers and activators of transcription 3 (JAK2/STAT3) signaling pathway in this process are investigated. The results show that Sr‐HA, rather than HA alone, significantly promote the osteogenic differentiation of BMSCs in vitro and bone formation in vivo. Further, the effect of promoting osteogenic differentiation is realized by activating the CaSR‐JAK2/STAT3 signaling pathway. Therefore, the study demonstrates that Sr‐HA is the better choice for restoring critical bone defects.

Background: Periodontitis is a chronic and progressive disease accompanied by bone loss. It is still a challenge to restore the bone structure. The osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) plays a decisive role in bone restoration and regeneration. Marine natural products (MNPs) have multiple biological activities, including anti-tumor and anti-inflammatory properties. However, the exploration of MNPs in osteogenesis is far from sufficient.Methods: We obtained a series of derivatives through structural optimization from 4-phenyl-3,4-dihydroquinolin-2(1H)-one alkaloid isolated from Scopulariopsis sp. Some preliminary cytological experiments showed that CHNQD-00603, obtained by adding a methoxy group to the position C3 and a hydroxyl group to the position C4 of 4-phenyl-3,4-dihydroquinolin-2(1H)-one, might promote the osteogenic differentiation of BMSCs. To further investigate the effects of CHNQD-00603 on BMSCs, we performed a CCK-8 assay and qRT-PCR, alkaline phosphatase staining (ALP), and alizarin red S staining to assess the cytotoxicity and the ability of osteogenic differentiation of CHNQD-00603. The autophagy level was assessed and validated by WB, qRT-PCR, and transmission electron microscopy. Then, 3-methyladenine (3-MA) was added to further examine the role of autophagy. Based on the expression of autophagy-related genes, we predicted and examined the potential miRNAs by bioinformatics.Results: CCK-8 assay showed that CHNQD-00603 at 1 µg/ml did not influence BMSCs activity. However, the proliferation rate decreased from the seventh day. qRT-PCR, ALP staining, ALP activity assay, and Alizarin red S staining showed that the best concentration of CHNQD-00603 to promote osteogenic differentiation was 1 µg/ml. Further investigations indicated that CHNQD-00603 activated autophagy, and the inhibition of autophagy by 3-MA attenuated CHNQD-00603-enhanced osteogenic differentiation. Subsequently, the findings from bioinformatics and qRT-PCR indicated that miR-452-3p might be a regulator of autophagy and osteogenesis. Furthermore, we transfected BMSCs with miR-452-3p NC and mimics separately to further determine the function of miR-452-3p. The data showed that the overexpression of miR-452-3p moderated the level of autophagy and osteogenic differentiation of CHNQD-00603-treated BMSCs.Conclusion: Our data suggested that CHNQD-00603 promoted the osteogenic differentiation of BMSCs by enhancing autophagy. Meanwhile, miR-452-3p played a regulatory role in this process.

Abstract Cloud model similarity algorithm is an important part of cloud modelling theory. Most of the existing cloud model similarity algorithms suffer from poor discriminability, poor classification, unstable results, and low time efficiency. In this paper, a new similarity algorithm is proposed that considers the triangular cloud model distance and shape. First, according to the $${{D}}_{\text{T}}$$ D T distance formula, a new exponential closeness measure is defined, with which the distance similarity of cloud models is characterized. Then, the shape similarity is calculated according to the variance of the cloud model cloud drops. Finally, the two similarities are synthesized to define a similarity algorithm for determining the distance from the $${{D}}_{\text{T}}$$ D T distance formula and shape based on the triangular cloud model (DDTSTCM). In this paper, discriminability, stability, efficiency and theoretical interpretability are taken as the evaluation indices. Equipment security system capability evaluation experiment, cloud model differentiation simulation experiment and time series classification accuracy experiment are set up to verify the effectiveness of the algorithm in terms of the four above aspects. The experimental results show that DDTSTCM has good differentiation and excellent classification effects. In the classification experiment for the time series, the average classification accuracy of DDTSTCM reaches 91.78%, which is at least 2.78% higher than those of the other seven commonly used algorithms. The CPU running efficiency of DDTSTCM is also extremely high, and the average CPU running time of group training is always on the order of milliseconds, which effectively reduces the time cost. Finally, a case study is conducted to analyse a risk assessment problem for China’s island microgrid industry, and the evaluation results based on DDTSTCM are in line with human cognition and have good value for engineering applications. Graphical abstract

Plums (Prunus salicina and Prunus domestica) are prevalent in southwestern China, and have attracted interest owing to their delectable taste and exceptional nutritional properties. Therefore, this study aimed to investigate the nutritional and flavor properties of plum to improve its nutritional utilization. Specifically, we determined the soluble sugars, organic acids, and phenolic components in 86 accessions using high-performance liquid chromatography. Notably, glucose, fructose, malic, and quinic acids were the predominant sweetness and acidity in plums, with sucrose contributing more to the sweetness of the flesh than the peel. Moreover, The peel contains 5.5 fold more phenolics than flesh, epicatechin, gallic acid, and proanthocyanidins C1 and B2 were the primary sources of astringency. Correlation and principal component analyses showed eight core factors for plum flavor rating, and a specific rating criterion was established. Conclusively, these findings provide information on the integrated flavor evaluation criteria and for enhancing optimal breeding of plums.

IntroductionExogenous melatonin (MT) can promote horticultural crops growth under stress conditions.MethodsIn this study, the effects of exogenous MT on the accumulation of selenium (Se) in grape were studied under Se stress.Results and discussionUnder Se stress, exogenous MT increased the biomass, content of photosynthetic pigments and antioxidant enzyme activity of grapevines. Compared with Se treatment, MT increased the root biomass, shoot biomass, chlorophyll a content, chlorophyll b content, carotenoids, superoxide dismutase activity, and peroxidase activity by 18.11%, 7.71%, 25.70%, 25.00%, 25.93%, 5.73%, and 9.41%, respectively. Additionally, MT increased the contents of gibberellin, auxin, and MT in grapevines under Se stress, while it decreased the content of abscisic acid. MT increased the contents of total Se, organic Se and inorganic Se in grapevines. Compared with Se treatment, MT increased the contents of total Se in the roots and shoots by 48.82% and 135.66%, respectively. A transcriptome sequencing analysis revealed that MT primarily regulated the cellular, metabolic, and bioregulatory processes of grapevine under Se stress, and the differentially expressed genes (DEGs) were primarily enriched in pathways, such as aminoacyl-tRNA biosynthesis, spliceosome, and flavonoid biosynthesis. These involved nine DEGs and nine metabolic pathways in total. Moreover, a field experiment showed that MT increased the content of Se in grapes and improved their quality. Therefore, MT can alleviate the stress of Se in grapevines and promote their growth and the accumulation of Se.

Intestinal microbial community plays an important part in maintaining health and skeletal muscle development in livestock. This study is the first of its kind in the world. In order to better understand the relationship between gut microbiota and gene expression in skeletal muscle of rabbits, caecum contents and longissimus dorsi tissues of rabbits at 0 d (S1), 35 d (S2) and 70d (S3) were collected and subjected for 16S rRNA sequencing and transcriptome sequencing. Our results showed that, among three groups of rabbits, Firmicutes and Bacteroidetes were the dominant phyla at the phylum level, while Akmansia, Bacteroides and Ruminobacter were the dominant genera at the genus level, and the relative abundance of Akmansia and Bacteroides increased firstly and then decreased from 0 d to 70 d. By analyzing the transcriptome sequencing data, we identified 2866, 2446 and 4541 differentially expressed genes (DEGs) in S1 vs S2, S2 vs S3 and S1 vs S3 groups, respectively. Finally, we performed correlation analysis between gut microbiota and the expression levels of muscle development-related genes of rabbits at 0 d and 70 d. Compared with 0 day old rabbits, in 70 day old rabbits Acinetobacter and Cronbacter with decreased abundance, and Ruminococcaceae_UCG-014 and Ruminococcus_1 with increase abundance is beneficial to caecum health in rabbits. These results will lay a foundation for further re-searches about the relationship between caecum microflora and muscle development in rabbits.

Abstract The purpose of this research is to examine the correlation between blood urea nitrogen (BUN) and severe abdominal aortic calcification (AAC) among American adults aged 40 years and older. A total of 2757 participants in the NHANES from 2013 to 2014 were included in the final analysis. BUN was measured by means of the enzymatic conductivity rate method. AAC scores were quantified by the Kauppila scoring system, and severe AAC was defined as an AAC score ≥ 6. Multivariable logistic regression and restricted cubic splines were used in the analyses. In the multivariable logistic regression model, the highest BUN level (log 2-transformed) was associated with an increased risk of severe AAC [odds ratio (OR) = 1.77, 95% CI 1.17, 2.71]. The restricted cubic spline plot displayed a reverse l-shaped association between BUN (log2-transformed) and severe AAC (p for nonlinearity

Abstract Under the background of automobile intelligence, cockpit comfort is receiving increasing attention, and intelligent cockpit comfort evaluation is especially important. To study the intelligent cockpit comfort evaluation model, this paper divides the intelligent cockpit comfort influencing factors into four factors and influencing indices: acoustic environment, optical environment, thermal environment, and human–computer interaction environment. The subjective and objective evaluation methods are used to obtain the subjective weights and objective weights of each index by the analytic hierarchy process and the improved entropy weight method, respectively. On this basis, the weights are combined by using the game theory viewpoint to obtain a comprehensive evaluation model of the intelligent automobile cockpit comfort. Then, the cloud algorithm was used to generate the rank comprehensive cloud model of each index for comparison. The research results found that among the four main factors affecting the intelligent automobile cockpit comfort, human–computer interaction has the greatest impact on it, followed by the thermal environment, acoustic environment, and optical environment. The results of the study can be used in intelligent cockpit design to make intelligent cockpits provide better services for people.

It is important to be able to measure the similarity between two uncertain concepts for many real-life AI applications, such as image retrieval, collaborative filtering, risk assessment, and data clustering. Cloud models are important cognitive computing models that show promise in measuring the similarity of uncertain concepts. Here, we aim to address the shortcomings of existing cloud model similarity measurement algorithms, such as poor discrimination ability and unstable measurement results. We propose an EPTCM algorithm based on the triangular fuzzy number EW-type closeness and cloud drop variance, considering the shape and distance similarities of existing cloud models. The experimental results show that the EPTCM algorithm has good recognition and classification accuracy and is more accurate than the existing Likeness comparing method (LICM), overlap-based expectation curve (OECM), fuzzy distance-based similarity (FDCM) and multidimensional similarity cloud model (MSCM) methods. The experimental results also demonstrate that the EPTCM algorithm has successfully overcome the shortcomings of existing algorithms. In summary, the EPTCM method proposed here is effective and feasible to implement.

Aiming at the comfort evaluation of automobile intelligent cockpit, an evaluation model based on improved combination weighting-cloud model is established. By consulting relevant literature, 4 first-class indexes and 15 second-class indexes, including noise and vibration, light environment, thermal environment and human-computer interaction, are selected to establish a comfort evaluation system. Later the subjective and objective weights obtained by improved Analytic Hierarchy Process (AHP) and Technique for Order Preference by Similarity to an Ideal Solution (TOPSIS) are combined by Game Theory. Considering the fuzziness and randomness of the index system, the combination weights obtained by Game Theory are combined with the cloud model. The floating cloud algorithms is used to determine the first-class and second-class index clouds and the comprehensive evaluation cloud parameters. Improvements were made in two commonly used similarity calculation methods, the expectation curve method (ECM) and the maximum boundary curve method (MCM). A new similarity calculation method is defined to optimize the evaluation results and determine the final comfort evaluation grade. Lastly, a 2021 Audi intelligent car under a certain working condition was selected to verify the correctness and rationality of the model using the fuzzy evaluation method. The results show that the cockpit comfort evaluation model based on the improved combination weighting-cloud model can better reflect the comprehensive comfort of automobile cockpit.

Abstract Background Bluetongue virus (BTV) causes a disease among wild and domesticated ruminants which is not contagious, but which is transmitted by biting midges of the Culicoides species. BTV can induce an intense cytopathic effect (CPE) in mammalian cells after infection, although Culicoides- or mosquito-derived cell cultures cause non-lytic infection with BTV without CPE. However, little is known about the transcriptome changes in Aedes albopictus cells infected with BTV. Methods Transcriptome sequencing was used to identify the expression pattern of mRNA transcripts in A. albopictus cells infected with BTV, given the absence of the Culicoides genome sequence. Bioinformatics analyses were performed to examine the biological functions of the differentially expressed genes. Subsequently, quantitative reverse transcription–polymerase chain reaction was utilized to validate the sequencing data. Results In total, 51,850,205 raw reads were generated from the BTV infection group and 51,852,293 from the control group. A total of 5769 unigenes were common to both groups; only 779 unigenes existed exclusively in the infection group and 607 in the control group. In total, 380 differentially expressed genes were identified, 362 of which were up-regulated and 18 of which were down-regulated. Bioinformatics analyses revealed that the differentially expressed genes mainly participated in endocytosis, FoxO, MAPK, dorso-ventral axis formation, insulin resistance, Hippo, and JAK-STAT signaling pathways. Conclusion This study represents the first attempt to investigate transcriptome-wide dysregulation in A. albopictus cells infected with BTV. The understanding of BTV pathogenesis and virus–vector interaction will be improved by global transcriptome profiling.

Abstract Background Tubby-like proteins (TLPs), characterized by a signature tubby domain, are widespread in plants and animals. To date, only plant TLPs involved in multifarious stress responses and male gametophyte development have been identified. However, studies on the molecular functions of plant TLPs are largely unknown. Results In this investigation, the roles of a TLP from Malus domestica (MdTLP7) in response to abiotic stresses were characterized by expressing it in Arabidopsis. The expression of wild-type full-length MdTLP7 (FL) significantly increased the stress tolerance of Arabidopsis seedlings to osmotic, salt, cold and heat stress, while the expression of truncated MdTLP7 containing only the tubby domain (Tub) also showed some function. Located on a central α helix surrounded by 12 anti-parallel β strands in the tubby domain, the K190/R192 site may be involved in fixation to the plasma membrane, as shown by 3D homology modelling with animal TLPs. This site might play a crucial role in anti-stress functions since site-directed mutagenesis of MdTLP7 reduced stress tolerance. Subcellular localization showed that MdTLP7 was mainly localized in the plasma membrane in plant cells, suggesting that it might participate in the transduction of stress signals. Conclusions The results of this study showed that MdTLP7 could improve abiotic stress tolerance not only in bacteria but also in plants. The K190/R192 residues in the tubby domain were not only the plasma membrane binding site of MdTLP7 but also played a key role in stress tolerance. These results may provide a basis for further exploring the mechanism of anti-stress functioning and downstream target genes of plant TLPs.

AbstractTubby-like proteins (TLPs), which have a highly conserved β-barrel tubby domain, have been found to be associated with some animal-specific characteristics. In the plant kingdom, more than 10 TLP family members were identified in Arabidopsis, rice and maize, and they were found to be involved in responses to stress. The publication of the apple genome makes it feasible to systematically study the TLP family in apple. In this investigation, 9 TLP encoding genes (TLPs for short) were identified. When combined with the TLPs from other plant species, the TLPs were divided into three groups (group A, B and C). Most plant TLP members in group A contained an additional F-box domain at the N-terminus. However, no common domain was identified other than tubby domain either in group B or in group C. An analysis of the tubby domains of MdTLPs identified three types of conserved motifs. Motif 1 and 2, the signature motifs in the confirmed TLPs, were always present in MdTLPs, while motif 3 was absent from group B. Homology modeling indicated that the tubby domain of most MdTLPs had a closed β barrel, as in animal tubby domains. Expression profiling revealed that the MdTLP genes were expressed in multiple organs and were abundant in roots, stems, and leaves but low in flowers. An analysis of cis-acting elements showed that elements related to the stress response were prevalent in the promoter sequences of MdTLPs. Expression profiling by qRT-PCR indicated that almost all MdTLPs were up-regulated at some extent under abiotic stress, exogenous ABA and H2O2 treatments in leaves and roots, though different MdTLP members exhibited differently in leaves and roots. The results and information above may provide a basis for further investigation of TLP function in plants.

Introduction. The purpose of this study was to investigate the changes of chitinase 3-like 1 (CHI3L1) in the aorta of patients with coronary atherosclerosis and to determine whether inhibition of CHI3L1 by lentivirus-mediated RNA interference could stabilize atherosclerotic plaques in apolipoprotein E-knockout (ApoE−/−) mice. Methods. We collected discarded aortic specimens from patients undergoing coronary artery bypass graft surgery and renal arterial tissues from kidney donors. A lentivirus carrying small interfering RNA targeting the expression of CHI3L1 was constructed. Fifty ApoE−/− mice were divided into control group and CHI3L1 gene silenced group. A constrictive collar was placed around carotid artery to induce plaques formation. Then lentivirus was transfected into carotid plaques. Results. We found that CHI3L1 was overexpressed in aorta of patients with atherosclerosis and its expression was correlated with the atherosclerotic risk factors. After lentivirus transduction, mRNA and protein expression of CHI3L1 were attenuated in carotid plaques, leading to reduced plaque content of lipids and macrophages, and increased plaque content of collagen and smooth muscle cells. Moreover, CHI3L1 gene silencing downregulated the expression of local proinflammatory mediators. Conclusions. CHI3L1 is overexpressed in aorta from patients with atherosclerosis and the lentivirus-mediated CHI3L1 gene silencing could represent a new strategy to inhibit plaques progression.

The myelodysplastic syndromes (MDS) comprise a heterogeneous group of malignant neoplasms with distinctive clinicopathological features. Currently, there is no specific approach for the treatment of MDS. Here, we report that bortezomib (BTZ), a proteasome inhibitor that has been used to treat plasma cell myeloma, induced G2/M phase cycle arrest in the MDS cell line SKM-1 through upregulation of Wee1, a negative regulator of G2/M phase transition. Treatment by BTZ led to reduced SKM-1 cell viability as well as increased apoptosis and autophagy. The BTZ-induced cell death was associated with reduced expression of p-ERK. To elucidate the implications of downregulation of p-ERK, we established the BTZ resistant cell line SKM-1R. Our data show that resistance to BTZ-induced apoptosis could be reversed by the MEK inhibitors U0126 or PD98059. Our results suggest that MAPK pathway may play an important role in mediating BTZ resistance.