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Circular RNAs (circRNAs) in controlling gene expression have been highlighted by increasing evidence, and their dysregulation has been linked to various diseases. However, the limited role of circRNAs in the adipogenesis of bone marrow-derived mesenchymal stem cells (BMSCs) has been explored. High-throughput sequencing of circRNA was carried out on BMSCs and AD induction 7d BMSCs. Then a substantial upregulation of circNDUFA13 was detected among circRNAs in AD induction 7d BMSCs. We found that the adipogenic differentiation of BMSCs was positively linked with circNDUFA13 expression levels. Adipogenesis in BMSCs was effectively inhibited by circNDUFA13 knockdown, whereas overexpression of circNDUFA13 promoted adipogenesis. It was noted that circNDUFA13 regulated the adipogenic differentiation of BMSCs by directly interacting with the signal transducer and activator of transcription 3 (STAT3), which activates CEBPß transcription. The in vitro model also validated the in vivo findings. our results suggest that circNDUFA13 controlled the adipogenic differentiation of BMSCs by targeting STAT3 and CEBPß activation.

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The process of bone remodeling occurs and is regulated through interactions between osteoclasts, which resorb bone, and osteoblasts, which generate bone tissue. When the homeostatic balance between these two cell types is dysregulated, this can contribute to abnormal bone remodeling resulting in a loss of bone mass as is observed in osteoporosis (OP) and other forms of degenerative bone metabolic diseases. At present, details of molecular mechanism underlying the development of bone metabolic diseases such as OP remain to be elucidated. Circular RNAs (circRNAs) are small non-coding RNA molecules with a closed-loop structure that can regulate the differentiation of osteoclasts and osteoblasts. The present review provides a systematic overview of recent literature on the processes through which circRNAs regulate the dynamic balance between osteoblasts and osteoclasts that ultimately preserve bone homeostasis. It will also give insight that can shape current understanding of the pathogenesis of OP and other bone metabolic diseases to better guide diagnostic and treatment strategies for affected patients.

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The significant morphological differences and abundant germplasm resources of Chinese indigenous dog breeds can be attributed to the diverse geographical environment, including plateaus, mountains, and a long history of raising dogs. The combination of both natural and artificial selection during the past several thousand years has led to hundreds of dog breeds with distinct morphological traits and environmental adaptations. China is one of the earliest countries to domesticate dogs and there are more than 50 ancient indigenous dog breeds. In this study, the run of homozygosity (ROH) and proportion of the autosomal genome covered by ROHs (FROH) were calculated for 10 dog breeds that are the most representative Chinese indigenous dogs based on 170K SNP microarray. The results of FROH showed that the Chuandong hound dogs (HCSSC) have the highest level of inbreeding among the tested breeds. The inbreeding in HCSSC occurred more recently than the Liangshan dogs (SCLSQ) dogs because of more numbers of long ROHs in HCSSC dogs, and the former also have higher inbreeding degree. In addition, there are significant differences in the inbreeding degree among different subpopulations of the same breed, such as the Thin dogs from Shaanxi and Shandong province. To explore genome-wide selection signatures among different breeds, including coat color, ear shape, and altitude adaptability, we performed genome selection analyses of FST and cross population extended haplotype homozygosity (XP-EHH). For the coat color, the FST analysis between Xiasi dogs (XSGZ) and HCSSC dogs was performed and identified multiple genes involved in coat color, hair follicle, and bone development, including MC1R, KITLG, SOX5, RSPO2, and TBX15. For the plateau adaptability, we performed FST and XP-EHH analyses between dogs from Tibet (Tibetan Mastiffs and Nyingchi dogs) and plain regions (Guangxi Biwei dogs GXBWQ and Guandong Sharpei dogs). The results showed the EPAS1 gene in dogs from Tibet undergo strong selection. Multiple genes identified for selection signals based on different usage of dogs. Furthermore, the results of ear shape analyses showed that MSRB3 was likely to be the main gene causing the drop ear of domestic dogs. Our study provides new insights into further understanding of Chinese indigenous dogs.

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The delicate equilibrium between osteoblast and adipocyte differentiation of MSCs is highly regulated. We screened for early-stage osteogenesis- or adipogenesis-based MSCs protein expression profiles using TMT-based quantitative proteomic analysis to identify novel participating molecules. Protein annotation, hierarchical clustering, functional stratification, and protein-protein association assessments were performed. Moreover, two upregulated proteins, namely, FBLN2 and NPR3, were validated to participate in the osteogenic differentiation process of MSCs. After that, we independently downregulated FBLN2 and NPR3 over seven days of osteogenic differentiation, and we performed quantitative proteomics analysis to determine how different proteins were regulated in knockdown vs. control cells. Based on gene ontology (GO) and network analyses, FBLN2 deficiency induced functional alterations associated with biological regulation and stimulus-response, whereas NPR3 deficiency induced functional alterations related to cellular and metabolic processes, and so on. These findings suggested that proteomics remains a useful method for an in-depth study of the MSCs differentiation process. This will assist in comprehensively evaluating its role in osteoporosis and provide additional approaches for identifying as-yet-unidentified effector molecules.

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INTRODUCTION: Adipogenesis, a highly coordinated process regulated by numerous effectors, is largely responsible for the quantity and size of adipocytes. Attenuation of adipocyte differentiation has been proposed as a viable technique for reducing obesity and its associated diseases. MicroRNAs play an important role in human bone marrow mesenchymal stem cells (hMSCs) adipogenic differentiation. However, there is a lack of clarity regarding the role of miR-328-5p in adipogenesis. MATERIAL AND METHODS: Using the lentiviral vectors to overexpress fatty acid synthase (FASN) and miR-328-5p, RT-qPCR and Western blotting were carried out to assess RNA expression and protein levels of FASN and adipogenic marker factors. Meanwhile, Oil red O staining and lipid quantification was performed to evaluate the accumulation of intracellular lipid droplets. Additionally, the validity of FASN as a potential target gene for miR-328-5p was carried out using a luciferase reporter assay. RESULTS: Our data showed that hMSCs adipogenic differentiation was associaed with the reduced miR-328-5p expression, while an elevated expression of the underlined miRNA attenuated adipogenesis and the expression of adipogenic marker genes. Luciferase reporter assay validated FASN as a target gene of miR-328-5p, and an elevated FASN expression reversed the anti-adipogenic effects of miR-328-5p. CONCLUSIONS: The results revealed that miR-328-5p inhibits hMSCs adipogenic differentiation by targeting FASN. These findings contribute to our understanding of obesity-related disease development.

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MicroRNAs play an important role in the adipogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs). How miR-100-3p influences such adipogenesis, however, remains uncertain. In this study, hMSC adipogenic differentiation was associated with miR-100-3p downregulation, and overexpressing this miRNA inhibited adipogenesis and the expression of adipogenic marker genes. Through bioinformatics approaches, miR-100-3p can bind the 3'-untranslated region (3'-UTR) of the mRNA encoding phosphoinositide 3-kinase regulatory subunit 1 (PIK3R1) such that miR-100-3p overexpression resulted in significant reductions in PIK3R1 expression. Importantly, overexpressing PIK3R1 was sufficient to reverse the anti-adipogenic effects of miR-100-3p overexpression. PIK3R1 is a critical component of the PI3K/AKT signaling pathway, and miR-100-3p overexpression resulted in reduced AKT phosphorylation in the context of adipogenesis. In addition, the adipogenic differentiation of hMSCs in which miR-100-3p was overexpressed was further enhanced upon treatment with the PI3K/AKT agonist 740Y-P relative to miR-100-3p overexpression alone. Taken together, these findings provide evidence that miR-100-3p inhibits the adipogenic differentiation of hMSCs by targeting PIK3R1 via the PI3K/AKT signaling pathway.

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BACKGROUND: Transfer RNA-derived small RNAs (tsRNAs) are a recently discovered form of non-coding RNA capable of regulating myriad physiological processes. The role of tsRNAs in hMSC adipogenic differentiation, however, remains incompletely understood. The purpose of this study was to identify the novel tsRNA-16902 as a regulator of hMSC adipogenic differentiation. METHODS: In this study, we conducted transcriptomic sequencing of hMSCs after inducing their adipogenic differentiation, and we were thereby able to clarify the molecular mechanism underlying the role of tsRNA-16902 in this context via a series of molecular biology methods. RESULTS: When we knocked down tsRNA-16902 expression, this impaired hMSC adipogenic differentiation and associated marker gene expression. Bioinformatics analyses further revealed tsRNA-16902 to target retinoic acid receptor γ (RARγ). Luciferase reporter assays also confirmed the ability of tsRNA-16902 to bind to the RARγ 3'-untranslated region. Consistent with this, RARγ overexpression led to impaired hMSC adipogenesis. Further analyses revealed that Smad2/3 phosphorylation was increased in cells that either overexpressed RARγ or in which tsRNA-16902 had been knocked down. We also assessed the adipogenic differentiation of hMSCs in which tsRNA-16902 was knocked down and at the same time a Smad2/3 inhibitor was added to disrupt Smad2/3 phosphorylation. The adipogenic differentiation of hMSCs in which tsRNA-16902 was knocked down was further enhanced upon the addition of a Smad2/3 signaling inhibitor relative to tsRNA-16902 knockdown alone. CONCLUSIONS: Through a comprehensive profiling analysis of tsRNAs that were differentially expressed in the context of hMSC adipogenic differentiation, we were able to identify tsRNA-16902 as a previously uncharacterized regulator of adipogenesis. tsRNA-16902 is able to regulate hMSC adipogenic differentiation by targeting RARγ via the Smad2/3 signaling pathway. Together, our results may thus highlight novel strategies of value for treating obesity.

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Understanding the mechanisms of adipogenic differentiation may lead to the discovery of novel therapeutic targets for obesity. The natural plant polyphenol compound curcumin can improve obesity-associated inflammation and diabetes in obese mice. The role of curcumin in adipogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs) is still unclear. We used hMSCs to investigate the details of the mechanism underlying the adipogenic effects of curcumin. At different time points (i.e., 5 days and 10 days) of hMSC adipocyte differentiation, an accumulation of large lipid droplets was analyzed in Oil Red O-stained cultured cells in two curcumin (5 µM and 10 µM) groups and the control group. The cells were also harvested for the detection of mRNA and protein expressions by quantitative real-time polymerase chain reaction and Western blot analysis. The results showed that curcumin can suppresses adipocyte differentiation in a dose-dependent manner and inhibited the expression of PPARγ, C/EBPα, and FABP4. Importantly, curcumin can also suppress the expression of Kruppel-like factor 15, which may bind to the PPARγ promoter, resulting in downregulation of PPARγ expression to inhibit the adipogenic differentiation of hMSCs.

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MicroRNAs are members of the family of non-coding small RNAs that regulate gene expression either by inhibiting mRNA translation or by promoting mRNA degradation at the post-transcriptional level. They play an important role in the differentiation of human bone marrow mesenchymal stem cells (hMSCs) into adipocytes. However, the role of microRNAs in this process remains to be poorly understood. Here, we observed that miR-377-3p expression was markedly decreased during adipogenic differentiation of hMSCs. Overexpression of miR-377-3p decreased adipocyte differentiation and downregulated the expression of adipogenic markers. Meanwhile, bioinformatics-based studies suggested that LIFR is a target of miR-377-3p. Further analysis confirmed that expression of LIFR present markedly increased during adipogenic differentiation of hMSCs. In addition, downregulation expression of LIFR significantly inhibited the process of adipocyte differentiation. To confirm the relation between miR-377-3p and LIFR, luciferase reporter assays were carried out. The results indicated that miR-377-3p bound directly to the 3'-untranslated region of LIFR. These data indicate that miR-377-3p suppressed adipogenesis of hMSCs by targeting LIFR, which provides novel insights into the molecular mechanism of miRNA-mediated cellular differentiation.

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The present study aimed to screen several differentially expressed genes (DEGs) and differentially expressed microRNAs (miRNAs) for two types of mesenchymal stem cell (MSC) differentiation. Bone morphogenetic protein 6 (BMP­6) and dexamethasone were used to induce MSCs towards osteoblastic differentiation or adipocytic differentiation. The t­test in the Bioconductor bioinformatics software tool was used to screen DEGs and differentially expressed miRNAs in the two samples. Subsequent gene ontology (GO) and pathway analyses on the DEGs were performed using the GO and Kyoto Encyclopedia of Genes and Genomes databases, respectively; potential target genes for the screened miRNAs were predicted using the TargetScan database. In addition, an interaction network between the DEGs and miRNAs was constructed. Numerous DEGs and miRNAs were screened during osteoblastic and adipocytic differentiation of MSCs. Important pathways, such as glutathione metabolism, pathogenic Escherichia coli infection and Parkinson's disease, and GO terms, including cytoskeletal protein binding and phospholipase inhibitor activity, were enriched in the screened DEGs from MSCs undergoing osteogenic differentiation and adipocytic differentiation. miRNAs, including miRNA (miR)­382 and miR­203, and DEGs, including neuronal growth regulator 1 (NEGR1), phosphatidic acid phosphatase 2B (PPAP2B), platelet­derived growth factor receptor alpha (PDGFRA), interleukin 6 signal transducer (IL6ST) and sortilin 1 (SORT1), were demonstrated to be involved in osteoblastic differentiation. In addition, the downregulated miRNAs (including miR­495, miR­376a and miR­543), the upregulated miR­106a, the upregulated DEGs, including enabled homolog (ENAH), polypeptide N­acetylgalactosaminyltransferase 1 and acyl­CoA synthetase long­chain family member 1, and the downregulated repulsive guidance molecule family member B and semaphorin SEMA7A were demonstrated to be involved in adipocytic differentiation. The results of the present study suggested that miRNAs (miR­203 and miR­382) and DEGs (NEGR1, PPAP2B, PDGFRA, IL6ST and SORT1) may serve pivotal functions in the osteoblastic differentiation of MSCs, whereas miR­495, which is also involved in osteoblast differentiation and had four targets, including NEGR1, miR­376a, miR­543 and ENAH may have crucial roles in adipocytic differentiation of MSCs.

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INTRODUCTION: Adipogenesis comprises multiple processes by which mesenchymal stem cells differentiate into adipocytes. To increase our knowledge of the mechanism underlying adipogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs), we performed full-genome gene expression microarray and gene ontology analyses of induced differentiation of hMSCs. MATERIAL AND METHODS: Adipogenic differentiation of hMSCs was induced by an adipogenic medium, and total RNA was extracted from undifferentiated hMSCs (day 0) and differentiated adipocytes (day 14). Then microarray hybridization of RNA samples was performed. The GeneChip Operating Software was used to analyze the hybridization data to identify differentially expressed genes, which were performed Gene Ontology categorization and pathway analysis. Pathway-act-network and genes-act-network were built according to the Kyoto Encyclopedia of Genes and Genomes database. Some differentially expressed genes were subjected to qRT-PCR to verify the microarray data. RESULTS: We detected a total of 3,821 differentially expressed genes, of which 753 were upregulated and 3,068 downregulated. These genes were well represented in a variety of functional categories, including collagen fibril organization, brown fat cell differentiation, cell division, and S phase of mitotic cell cycle. Subsequently, pathway analysis was conducted, and significant pathways (from top 50) were selected for pathway-act-network analysis, which indicated that the mitogen-activated protein kinase (MAPK) pathway and cell cycle were of high degrees (> 10). Gene-act-network analysis showed that insulin-like growth factor 1 receptor (IGF1R), histone deacetylase 1 (HDAC1), HDAC2, MAPK13, MAPK8, phosphoinositide-3-kinase regulatory subunit 1 (PI3KR1), and PI3KR2 also had high degrees (> 18). CONCLUSIONS: Collectively, these data provide novel information and could serve as a basis for future study to clarify the mechanisms underlying adipocyte differentiation of hMSCs.

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OBJECTIVE: To screen the differentially expressed miRNAs and their target genes in adipogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs) to better understand the mechanism for regulating the balance between osteoblast and adipocyte differentiation. METHODS: Cultured hMSCs were induced for adipogenic differentiation, and at 0, 7, 14, and 21 days of induction, the cells were examined for miRNA and mRNA expression profiles using miRNA chip and transcriptome sequencing (RNA-seq) techniques. Correlation analysis was carried out for the miRNAs and mRNAs of potential interest. The databases including TargetScan, PicTar and miRanda were used to predict the target genes of the differentially expressed miRNA. RESULTS: The expression of miR-140-5p was down-regulated and leukemia inhibitory factor receptor (LIFR) expression increased progressively during adipogenic differentiation of hMSCs, showing a negative correlation between them. Target gene prediction using the 3 databases identified LIFR as the target gene of miR-140-5p. CONCLUSION: miRNA-140-5p may play an important role by regulating its target gene LIFR during adipogenic differentiation of hMSCs.

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OBJECTIVE: To verify whether insulin promotes the differentiation of skeletal myoblasts into myocytes and whether wortmannin and U0126 inhibit the promoting effect with an attempt to explore the molecular mechanisms of inducing the differentiation of muscular cells in rats. METHODS: Primary skeletal myoblasts were separated and cultured from rats, and then were treated in DMEM containing various concentrations of insulin. The morphology of cells was monitored under a phase-contrast microscope. And the expression of myogenin was detected by immunocytochemistry and Western blotting. The change in the effect of insulin on the differentiation of myoblasts was observed after the intervention of a phosphatidylinositide 3-kinase (PI3K) inhibitor wortmannin and a specific MEK inhibitor U0126. RESULTS: Insulin markedly promoted myotube formation of myoblasts. Two days after insulin treatment, myotubes started to form; later, more and more myotubes appeared, and to the peak at 7 days. Insulin increased the expression of myogenin in a concentration-dependent manner. However, wortmannin and U0126 inhibited the effect of insulin on the differentiation of skeletal myoblasts in rats. CONCLUSION: Wortmannin and U0126 can suppress the promoting effect of insulin on the differentiation of skeletal myoblasts into myocytes in rats and decrease the formation of myotubes and the expression of myogenin.

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Schizophrenia is a serious neuropsychiatric disease of uncertain etiology, which causes human mental disorder and affects about 1% of the population. In recently years, some studies showed that some cases of schizophrenia may be associated with Toxoplasma gondii infection. In order to investigate a potential association between Toxoplasma infection and schizophrenia, we investigated the relative clinical symptom of schizophrenia such as learning and memory capability, depression and stereotypy to find some useful information by behavioral test in mouse models. Our results demonstrated that mice from Toxoplasma infection and MK-801 administration (as the model of schizophrenia) were impaired in learning and memory capability, and they had more serious depression and stereotypy compared with the control mice, especially the mice from congenital Toxoplasma infection. In addition, our results clearly showed that the number of cysts in brain tissue of congenital Toxoplasma infection mice was significantly low than in acquired Toxoplasma infected mice. Collectively, these results suggested a potential association between Toxoplasma infection and schizophrenia.

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OBJECTIVE: Using papain to prepare polypeptide from Eupolyphaga sinensis, then study the immune function of polypeptide from Eupolyphaga sinensis in vivo. METHODS: Used hydrolysis degree as index, pH value, enzyme dosage, thermometer reaction time were optimized. Studied the influence of polyeptide on the mice immune functions through mice immune organs index, phagocytic function and the level of IL-2. RESULTS: The optimum enzymolysis condition was as follows: pH 8.0, enzyme 1%, temperature 55 degrees C, reaction time 4. 5 h. In vivo test of mice demonstraed that, Eupolyphaga sinensis could elevate index of thymus and spleen, enhance the phagocytic function of macrophage and promote the level of IL-2 in serum. CONCLUSION: Eupolyphaga sinensis has immunoregulatory effect.

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The aim of this study was to investigate the influence of neonatal isolation stress on hyperlocomotion in complexin II knockout mouse (Cplx2(-/-)). The mice were randomly divided into 4 groups: Cplx2(-/-) with stress, Cplx2(+/+) with stress, Cplx2(-/-) without stress and Cplx2(+/+) without stress. Isolation stress was employed on the pups of stress groups from the 2nd day after the postnatal to the 21st day. The PCR was used to determine the gene type and the hyperlocomotion test was employed to detect the change of animal behavior after methamphetamine or saline injection (i.p.). The results showed that the animals of all groups increased their movement after injection of 0.2 mg/kg methamphetamine in different levels (P < 0.01), compared with those injected with saline. The Cplx2(-/-) mouse with stress revealed a significant increase in the distance of free movement after injection of 0.2 mg/kg methamphetamine compared with the knockout mouse without stress (P < 0.001). When Cplx2(-/-) mouse with stress was compared with wild type with stress, Cplx2(-/-) mouse with stress had more movement (P < 0.001), indicating that Cplx2 has effect on the hyperlocomotion as well. These results suggest an involvement of stress and Cplx2 in the movement behavior of mice.

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OBJECTIVE: To obtain the cDNA sequence encoding fibrinolytic enzyme from Eupolyphaga sinensis and express it in prokaryotic and eukaryotic expression system. METHOD: The primers were designed according to the cDNA of other animals'fibrinolytic enzyme. The cDNA sequence was cloned by RT-PCR and 3 RACE. RESULT: Sequence analysis revealed that the length of the cDNA fragment was 672 bp and encoded a protein of 224 amino acid residues, the N end amino acid sequence residues was IVGG in accordance with other fibrinolytic enzyme. The cDNA sequence was expressed in E. coli, inactive protein was obtained. While expressed in Pichia pastoris, recombinant protein had fibrinolytic activity. CONCLUSION: The cDNA sequence of fibrinolytic enzyme from E. sinensis Walker was cloned and expressed for the first time and it proved a good basis for further functional study of the enzyme.

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OBJECTIVE: To obtain polysaccharide from Scolopendra subspinipes mutilans and study its partial properties. METHODS: A raw polysaccharide was isolated from Chinese medicine Scolopendra subspinipes mutilans, and three groups were isolated by DEAE-52. The thin layer chromatography and gel filtration chromatography were used to detect the main monosaccharide composition and molecular weight of the group I. The inhibitory effect of the group I on tumor cells was detected by MTT assay. RESULTS: The molecular weight of the group I was 33.1 kD. The inhibitory effect of the group I on Hela cells was obvious, as its inhibitive rate was 60.8% on Hela cells when the polysaccharide's concentration was 3.13 microg/mL, but it had no effect on the Eca 109 cells. CONCLUSION: The group I has specific effects on different tumor cells.

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