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Sandip Patil,1– 3 Hongyu Chen,4 Chunna Guo,4 Xiaoli Zhang,3 Pei-Gen Ren,1 Ngiambudulu M Francisco,5 Feiqiu Wen2,3 1Shenzhen Institute of Advanced Technology, Shenzhen University Town, Shenzhen, Guangdong Province, People’s Republic of China; 2Department of Haematology and Oncology, Shenzhen Children’s Hospital, Shenzhen, Guangdong Province, People’s Republic of China; 3Paediatric Research Institute, Shenzhen Children’s Hospital, Shenzhen, Guangdong Province, People’s Republic of China; 4Department of Laboratory Medicine, Shenzhen Children’s Hospital, Shenzhen, Guangdong Province, People’s Republic of China; 5Grupo de Investigação Microbiana e Imunológica, Instituto Nacional de Investigação em Saúde (National Institute for Health Research), Luanda, AngolaCorrespondence: Feiqiu WenDepartment of Haematology and Oncology, Shenzhen Children’s Hospital, Shenzhen, Guangdong Province, People’s Republic of ChinaTel +86 18938690333Fax +86-755 83009888Email fwen62@163.comAim: We investigated the clonal diversity of carbapenemase-producing Klebsiella pneumoniae isolates from the Shenzhen Children’s Hospital, China, and drew conclusions on the clinical and public health impact of these isolates as multidrug-resistant.Methods: From January 2014 to December 2018, a total number of 36 unique carbapenemase-producing clinical isolates of Klebsiella pneumoniae were collected out of 900 clinical isolates in paediatric patients from the Shenzhen Children’s Hospital, China. After carbapenemase production confirmation, antimicrobial susceptibility, resistance determinants and phylogenetic relationship were determined.Results: The isolates showed resistance to ceftazidime, ertapenem, ampicillin, cefazolin, ceftriaxone, cefotetan, ticarcillin, cefaclor, cefpodoxime, azlocillin, cefcapene, mezlocillin and ampicillin-sulbactam. Of the 36 Klebsiella pneumoniae carbapenemase genes coding isolates, blaNDM was the mostly detected 50% (n=18) followed by blaKPC and blaIMP 19% (n=7), blaVIM 17% (n=6), blaOXA-48-like 8% (n=3) and blaSME 5% (n=2), whereas extended-spectrum β-lactamase (blaSHV) was predominantly detected 92% (n=33) followed by blaCTX-M 53% (n=19) and blaCMY 28% (n=10). Pulsed-field gel electrophoresis typing showed eight different patterns, and twenty-five distinct sequences types were observed with ST307 being predominantly identified 11% (n=4), followed by ST2407 8% (n=3). Plasmid replicon typing results indicated that IncFIS, IncHI2, IncFIC and IncFIA plasmids carry blaCTX-M, blaSHV and blaNDM genes.Conclusion: This study reports on the occurrence and spread of carbapenemase and extended-spectrum β-lactamase encoding genes co-existence in sporadic Klebsiella pneumoniae ST307 in paediatric patients from the Shenzhen Children’s Hospital, China.Keywords: Klebsiella pneumoniae, carbapenemase, ESBLs, antimicrobial susceptibility, molecular characterization

Sandip Patil,1,2 Hongyu Chen,2 Xiaoli Zhang,2 Ma Lian,2 Pei-Gen Ren,1 Feiqiu Wen2 1Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, Guangdong Province, People’s Republic of China; 2Shenzhen Children’s Hospital, Futian District, Shenzhen, Guangdong Province, People’s Republic of ChinaCorrespondence: Feiqiu WenShenzhen Children’s Hospital, 7019 Yitian Road, Futian District, Shenzhen, Guangdong Province 518038, People’s Republic of ChinaTel +86 18938690333Fax +86-755 83009888Email fwen62@163.comIntroduction: The emergence of multi-drug-resistant Gram-negative bacteria (GNB) is a concern in China and globally. This study investigated antimicrobial resistance traits and resistance determinant detection in GNB isolates from paediatric patients in China.Methods: In the present study, a total of 170 isolates of GNB including the most prevalent Escherichia coli, Klebsiella pneumoniae and Acinetobacter baumannii were collected from Shenzhen Children’s Hospital, China. ESBLs production was confirmed by using the combination disc diffusion method, and carbapenemase production was confirmed by using a carbapenem inactivation method followed by antimicrobial susceptibility. In addition, β-lactamase-encoding genes and co-existence of plasmid-borne colistin resistance mcr-1 gene were determined by PCR and sequencing.Results: Overall, 170 etiological agents (GNB) were recovered from 158 paediatric patients. The most prevalent species was E. coli 40% (n=68), followed by K. pneumoniae 17.64% (n=30), and Enterobacter cloacae 14.11% (n=24). Of 170 GNB, 71.76% (n=122) were multi-drug-resistant, 12.35% (n=21) extreme-drug resistant, and 7.64% (n=13) single-drug-resistant, while 8.23% (n=14) were sensitive to all of the studied antibiotics. The prevalence of ESBLs and carbapenemase producers were 60% and 17%, respectively. blaCTX-M was the most prevalent resistance gene (59.42%), followed by blaTEM (41.17%), blaSHV (34.270%), blaKPC (34.11%), blaOXA-48 (18.82%) and blaNDM-1 (17.64%).Conclusion: The present study provides insights into the linkage between the resistance patterns of GNB to commonly used antibiotics and their uses in China. The findings are useful for understanding the genetics of resistance traits and difficulty in tackling of GNB in paediatric patients.Keywords: Gram-negative bacteria, antimicrobial susceptibility, ESBLs, carbapenemase, molecular characterization

Endochondral ossification is the process by which cartilage is mineralized into bone, and is essential for the development of long bones. Osteocalcin (OCN), a protein abundant in bone matrix, also exhibits high expression in chondrocytes, especially hypertrophic chondrocytes, while its role in endochondral ossification remains unclear. Utilizing a new CRISPR/Cas9-mediated bglap-bglap2 deficiency (OCN em ) mouse model generated in our laboratory, we provide the first evidence of OCN's regulatory function in chondrocyte differentiation and endochondral ossification. The OCN em mice exhibited significant delays in primary and secondary ossification centers compared to wild-type mice, along with increased cartilage length in growth plates and hypertrophic zones during neonatal and adolescent stages. These anomalies indicated that OCN deficiency disturbed endochondral ossification during embryonic and postnatal periods. Mechanism wise, OCN deficiency was found to increase chondrocyte differentiation and postpone vascularization process. Furthermore, bone marrow mesenchymal stromal cells (BMSCs) from OCN em mice demonstrated an increased capacity for chondrogenic differentiation. Transcriptional network analysis implicated that BMP and TGF-β signaling pathways were highly affected in OCN em BMSCs, which is closely associated with cartilage development and maintenance. This elucidation of OCN's function in chondrocyte differentiation and endochondral ossification contributes to a more comprehensive understanding of its impact on skeletal development and homeostasis.

Obesity has become a global concern because of increasing the risk of many diseases. Alterations in human gut microbiota have been proven to be associated with obesity, yet the mechanism of how the microbiota are altered by high salt diet (HSD) remains obscure. In this study, the changes of Small Intestinal Microbiota (SIM) in obesity-T2DM mice were investigated. High-throughput sequencing was applied for the jejunum microbiota analysis. Results revealed that high salt intake (HS) could suppress the body weight (B.W.) in some extent. In addition, significant T2DM pathological features were revealed in high salt-high food diet (HS-HFD) group, despite of relatively lower food intake. High-throughput sequencing analysis indicated that the F/B ratio in HS intake groups increased significantly (P < 0.001), whereas beneficial bacteria, such as lactic acid or short chain fatty acid producing bacteria, were significantly decreased in HS-HFD group (P < 0.01 or P < 0.05). Furthermore, Halorubrum luteum were observed in small intestine for the first time. Above results preliminary suggested that in obesity-T2DM mice, high dietary salt could aggravate the imbalance of composition of SIM to unhealthy direction., (© 2023. The Author(s).)

Background: Osteoporosis (OP), a systemic skeletal disease common in aged population, is an important public health problem worldwide. Animal models are important tools for understanding OP. In ovariectomy (OVX) or orchiectomy (ORX) OP models, lumbar vertebrae are often used for evaluating of the OP progression. However, unlike the bipeds, the lumbar vertebrae are not weight loading bones in quadruped animal, but the head-bearing cervical vertebrae take much higher stress. So, we compared the murine cervical vertebrae with lumbar vertebrae for OP assessment., Methods: OVX and ORX mouse models were established on C57BL/6J mice. Serum estradiol, testosterone and bone related biomarkers were verified. Bone quantity and quality were determined using micro computed tomography (micro-CT) analysis. Hard tissue sections were prepared, stained for histomorphological analyzing, and micro-indentation measured for bone mechanical property evaluation., Results: In OVX and ORX mice, serum estradiol or testosterone levels reduced, bone resorption level and related biomarkers elevated, indicated the successful generation of the OP models. In the early stage, the trabecular bone mineral density (BMD) of cervical vertebrae was already reduced 16.1% (OVX) and 21.7% (ORX) one-month post-gonadectomy, respectively; while this decline in the fifth lumbar vertebra were only 5% and 7.4%, respectively. Six months post-gonadectomy, the reduction of BMD in cervical vertebrae and the fifth lumbar vertebra were 31.2% & 36.1% and 28.5% & 30.7% respectively. In biomechanical aspects, cervical spines showed worse Vickers hardness (HV) and elastic modulus than lumbar spine in six-month OVX and ORX mice., Conclusions: We provide a new OP early-stage evaluation mouse model based on the cervical spine. Through the radiographic, biological and biomechanical assessments, the mouse cervical spine is more suitable for bone remodeling evaluation in OP models than the conventional lumbar vertebrae, especially for early-stage OP study., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://qims.amegroups.com/article/view/10.21037/qims-22-717/coif). The authors have no conflicts of interest to declare., (2023 Quantitative Imaging in Medicine and Surgery. All rights reserved.)

The children's gut microbiota, associated with the development of obesity, is in maturation. The impact of obesity on the gut microbiota in childhood could have a more significant effect than on adulthood and eventually be lifelong lasting, but it has been rarely studied. Aimed to discover the difference in gut microbiota between children and adults with obesity, we collected published amplicon sequencing data from National Center for Biotechnology Information (NCBI) and re-analyzed them using a uniform bioinformatic pipeline, as well as predicted the obesity using gut microbiota based on the random forest model. Summarizing common points among these cohorts, we found that the gut microbiota had a significant difference between children with and without obesity, but this difference was not observed in adult cohorts. Based on the random forest model, it was more challenging to predict childhood obesity using gut microbiota than adulthood obesity. Our results suggest that gut microbiota in childhood is more easily affected than in adulthood. Early intervention for childhood obesity is essential to improve children's health and lifelong gut microbiota-related health., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2023 Yu, Yu, Zhao, Su and Ren.)

Type 2 diabetes (T2DM) is characterized by insulin secretion deficiencies and systemic insulin resistance (IR) in adipose tissue, skeletal muscle, and the liver. Although the mechanism of T2DM is not yet fully known, inflammation and insulin resistance play a central role in the pathogenesis of T2DM. G protein-coupled receptors (GPCRs) are involved in endocrine and metabolic processes as well as many other physiological processes. GPR50 (G protein-coupled receptor 50) is an orphan GPCR that shares the highest sequence homology with melatonin receptors. The aim of this study was to investigate the effect of GPR50 on inflammation and insulin resistance in 3T3-L1 preadipocytes. GPR50 expression was observed to be significantly increased in the adipose tissue of obese T2DM mice, while GPR50 deficiency increased inflammation in 3T3-L1 cells and induced the phosphorylation of AKT and insulin receptor substrate (IRS) 1. Furthermore, GPR50 knockout in the 3T3-L1 cell line suppressed PPAR-γ expression. These data suggest that GPR50 can attenuate inflammatory levels and regulate insulin signaling in adipocytes. Furthermore, the effects are mediated through the regulation of the IRS1/AKT signaling pathway and PPAR-γ expression., (© 2022 The Authors. FEBS Open Bio published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)

cAMP is a key second messenger that regulates diverse cellular functions including neural plasticity. However, the spatiotemporal dynamics of intracellular cAMP in intact organisms are largely unknown due to low sensitivity and/or brightness of current genetically encoded fluorescent cAMP indicators. Here, we report the development of the new circularly permuted GFP (cpGFP)-based cAMP indicator G-Flamp1, which exhibits a large fluorescence increase (a maximum ΔF/F 0 of 1100% in HEK293T cells), decent brightness, appropriate affinity (a K d of 2.17 μM) and fast response kinetics (an association and dissociation half-time of 0.20 and 0.087 s, respectively). Furthermore, the crystal structure of the cAMP-bound G-Flamp1 reveals one linker connecting the cAMP-binding domain to cpGFP adopts a distorted β-strand conformation that may serve as a fluorescence modulation switch. We demonstrate that G-Flamp1 enables sensitive monitoring of endogenous cAMP signals in brain regions that are implicated in learning and motor control in living organisms such as fruit flies and mice., (© 2022. The Author(s).)

High-calorie intake has become one of the most common causes of dietary obesity, which eventually develops into type 2 diabetes mellitus (T2DM). Microbiota, along with the length of the gastrointestinal tract, is related to metabolic disorders, but its shifts and following impact on metabolic disorders due to external perturbation are still unclear. To evaluate shifts of microbiota from the proximal to the distal intestine and their impact on metabolic disorders, we profiled jejunal and colonic microbiota with the perturbation using high salt (HS) and antibiotic-induced microbiota depletion (AIMD) in diet-induced obesity (DIO) mice and analyzed the association with parameters of both obesity and blood glucose. After ten weeks of feeding DIO mice with HS intake and AIMD, they failed to develop obesity. The DIO mice with HS intake had T2DM symptoms, whereas the AIMD DIO mice showed no significant difference in blood glucose parameters. We observed that the jejunal and colonic microbiota had shifted due to settled perturbation, and jejunal microbiota within a group were more dispersed than colonic microbiota. After further analyzing jejunal microbiota using quantified amplicon sequencing, we found that the absolute abundance of Colidextribacter  (R = 0.695, p = 0.001) and  Faecalibaculum  (R = 0.631, p = 0.005) in the jejunum was positively correlated with the changes in BW and FBG levels. The predicted pathway of glucose and metabolism of other substances significantly changed between groups (p <0.05). We demonstrated that the onset of obesity and T2DM in DIO mice is impeded when the gut microbiota is perturbed; thus, this pathogenesis depends on the gut microbiota., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Yu, Yu, Kerem and Ren.)

Due to the finding of severe side effects and low therapeutic efficacy with cancer chemotherapy, there still remains a great challenge to benefit patients with curative effect. In this work, we designed a self-powered drug delivery system comprising a current source derived from the disk TENG (D-TENG) and a pair of Au electrodes. Thus, cells seeded within the electrode gap could be stimulated by the current followed by D-TENG`s work. Under the rotation frequency of about 7.4 Hz, the peak output current and voltage of the D-TENG reached 3.7 μA and 135 V and achieved an average of 2.8 μA of output current. Furthermore, the D-TENG also showed its good stability to output steady current in a long-term condition. When applying the electric stimulation by this self-powered drug delivery system, a chemotherapy drug, doxorubicin (DOX), had significant uptake by cancer cells. Therefore, utilizing a novel TENG device as a part of chemotherapy would provide a new opportunity in future disease treatment., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Chen, Deng, He, Cheng, Ren and Xu.)

Cyclic adenosine 3',5'-monophosphate (cAMP) is an important intracellular second messenger molecule downstream of many G protein-coupled receptors (GPCRs). Fluorescence imaging with bright and sensitive cAMP indicators allows not only dissecting the spatiotemporal dynamics of intracellular cAMP, but also high-content screening of compounds against GPCRs. We previously reported the high-performance circularly permuted GFP (cpGFP)-based cAMP indicator G-Flamp1. Here, we developed improved G-Flamp1 variants G-Flamp2 and G-Flamp2b. Compared to G-Flamp1, G-Flamp2 exhibited increased baseline fluorescence (1.6-fold) and larger fluorescence change (ΔF/F 0 ) (1,300% vs. 1,100%) in HEK293T cells, while G-Flamp2b showed increased baseline fluorescence (3.1-fold) and smaller ΔF/F 0 (400% vs. 1,100%). Furthermore, live cell imaging of mitochondrial matrix-targeted G-Flamp2 confirmed cytosolic cAMP was able to enter the mitochondrial matrix. G-Flamp2 imaging also showed that adipose tissue extract activated the Gi protein-coupled orphan GPCR GPR50 in HEK293T cells. Taken together, our results showed that the high-performance of G-Flamp2 would facilitate sensitive intracellular cAMP imaging and activity measurement of compounds targeting GPCR-cAMP signaling pathway during early drug development., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Liu, Liu, Ren, Chu and Wang.)

Balloon expandable coronary stenting has revolutionized the field of interventional cardiology as a potential, minimally invasive modality for treating coronary artery disease. Even though stenting is successful compared to angioplasty (that leaves no stent in place), still there are many associated clinical complications. Bare metal stents are associated with in-stent restenosis caused mostly by neointimal hyperplasia, whereas success of drug-eluting stents comes at the expense of late-stent thrombosis and neoatherosclerosis. Even though innovative and promising, clinical trials with bioabsorbable stents reported thrombosis and a rapid pace of degradation without performing scaffolding action in several instances. It should be noted that a vast majority of these stents are based on a metallic platform which still holds the potential to mitigate major cardiovascular events and reduced economic burden to patients, alongside continuous improvement in stent technology and antiplatelet regimes. Hence, a systematic review was conducted following PRISMA guidelines to assess the clinically relevant material properties for a metallic stent material. From a materials perspective, the major causes identified for clinical failure of stents are inferior mechanical properties and blood-material interaction-related complications at the stent surface. In addition to these, the stent material should possess increased radiopacity for improved visibility and lower magnetic susceptibility values for artefact reduction. Moreover, the review provides an overview of future scope of percutaneous coronary interventional strategy. Most importantly, this review highlights the need for an interdisciplinary approach by clinicians, biomaterial scientists, and interventional cardiologists to collaborate in mitigating the impediments associated with cardiovascular stents for alleviating sufferings of millions of people worldwide., Competing Interests: Conflict of interestThe authors declare no competing interests., (© The Author(s), under exclusive licence to Springer Nature Switzerland AG 2022, corrected publication 2022.)

Increasing studies demonstrated that oncolytic activities of oHSV-1 are limited to the capacity of virus replicating in tumors. In order to potentiate the oHSV-1 oncolytic activity and expand the application of oHSV-1 treatment in multiple types of tumors, it is critical to explore the potential factors or mechanisms mediating tumor resistance to oHSV-1 infection. Here we evaluated the levels of oHSV-1 multiplication in various tumor cell lines and showed that glioblastoma cell line A172 had the lowest virus yields but intrinsically accumulated the highest levels of Mx2 protein. Subsequently we demonstrated that genetic depletion of Mx2 specifically enhanced oHSV-1 productive replication in A172 cells through promoting the nuclear translocation of uncoated viral genomic DNA and down-regulating innate antiviral response. In the further investigation, we found that Mx2 knockdown could alter the intrinsic mRNA accumulation of diverse sets innate immune genes in A172 cells, in particular DHX36 and MyD88. Mx2 depletion led to a decrease in mRNA levels of MyD88 and DHX36 in A172 cells and MyD88/DHX36 knockdown increased virus yield in A172 cells and decreased the production of IFNα, activation of IRF3 activity and NF-κB signaling in A172 cells. This shed new lights on understanding the roles of some intrinsic antiviral genes in oHSV-1 resistance, facilitating to offer potential targets to improve oHSV-1 oncolytic efficacy and develop candidates of biomarkers to predict the efficiency of oHSV-1 multiplication in tumors., (© 2021. The Author(s).)

Hyperandrogenism is a key pathological feature of polycystic ovarian syndrome (PCOS). Excess androgen can lead to PCOS-like cell hypertrophy in the ovaries and adipose tissue of rodents. Here, we established a dihydrotestosterone (DHT)-induced hyperandrogenic mouse model to analyze the differences in gene expression and signaling pathways of the ovaries and gonad fat pads of mice treated with or without DHT by RNA microarray analysis. From the results, we focused on the overlapping differentially expressed gene- Col6a5 -and the major differentially enriched signaling pathway-lipid metabolism. We employed DHT-induced mouse ovarian stromal cell, adipogenic 3T3-L1 cell and hepatic cell line NCTC1469 models to investigate whether androgens directly mediate lipid accumulation and hypertrophy. We found that DHT increased lipid droplet accumulation in ovarian stromal cells and adipogenic 3T3-L1 cells but not NCTC1469 cells. DHT significantly altered stromal cell cholesterol metabolism and steroidogenesis, as indicated by changes in cholesterol levels and the expression of related genes, but these effects were not observed in 3T3-L1 cells. Moreover, Col6a5 expression was significantly increased in ovaries and gonadal fat pads of DHT-treated mice, and Col6a5 inhibition alleviated DHT-induced excess lipid accumulation and hypertrophy of ovarian stromal cells and adipogenic 3T3-L1 cells, even improved lipid metabolism in overnourished NCTC1469 cells. Our results indicate that Col6a5 plays important roles in the pathogenesis of DHT-induced lipid metabolism disorder and the hypertrophy of ovarian stromal cells and adipocytes., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Sun, Yang, Wang, Zhao, Xiao, Li, Wang, Huang, Ren and Zhang.)

To effectively and selectively remove toxic anionic dyes which are heavily discharged and to promote them recovery, a sustainable cellulose nanofiber/chitosan (CNF/CS) composite film was elaborately designed through a facile procedure. Based on the strong supporting effect of CNF and excellent compatibility between CNF and CS, the composite film presents low swelling and acid-proof properties, which can prevent the adsorption process from the disintegration of adsorbent. Moreover, the positive electrical property of CNF/CS film increases the discrepancy in adsorption capacities for anionic and cationic dyes. The maximum adsorption capacity of anionic methyl orange (MO) on CNF/CS film reaches 655.23 mg/g with a desirable recyclability. The adsorption behavior attributed to a physico-chemical and monolayer adsorption process. This work opens a new route for the development of eco-friendly and highly efficient adsorbents on selective removal and recycling of anionic dyes from wastewater., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Osteoarthritis (OA) is a multifactorial joint disease with pathological changes that affect whole joint tissue. Obesity is acknowledged as the most influential risk factor for both the initiation and progression of OA in weight-bearing and non-weight-bearing joints. Obesity-induced OA is a newly defined phenotypic group in which chronic low-grade inflammation has a central role. Aside from persistent chronic inflammation, abnormal mechanical loading due to increased body weight on weight-bearing joints is accountable for the initiation and progression of obesity-induced OA. The current therapeutic approaches for OA are still evolving. Tissue-engineering-based strategy for cartilage regeneration is one of the most promising treatment breakthroughs in recent years. However, patients with obesity-induced OA are often excluded from cartilage repair attempts due to the abnormal mechanical demands, altered biomechanical and biochemical activities of cells, persistent chronic inflammation, and other obesity-associated factors. With the alarming increase in the number of obese populations globally, the need for an innovative therapeutic approach that could effectively repair and restore the damaged synovial joints is of significant importance for this sub-population of patients. In this review, we discuss the involvement of the systemic and localized inflammatory response in obesity-induced OA and the impact of altered mechanical loading on pathological changes in the synovial joint. Moreover, we examine the current strategies in cartilage tissue engineering and address the critical challenges of cell-based therapies for OA. Besides, we provide examples of innovative ways and potential strategies to overcome the obstacles in the treatment of obesity-induced OA., The Translational Potential of This Article: Altogether, this review delivers insight into obesity-induced OA and offers future research direction on the creation of tissue engineering-based therapies for obesity-induced OA., Competing Interests: The authors have no conflicts of interest to disclose in relation to this article., (© 2020 The Authors.)

Background & Aims: Circulating peptides and G protein-coupled receptors (GPCRs) have gained much attention because of their biofunctions in metabolic disorders including obesity and non-alcoholic fatty liver disease (NAFLD). Herein, we aimed to characterize the role and therapeutic potential of a newly identified peptide hormone in NAFLD., Methods: Using bioinformatics, we identified a murine circulating pentadecapeptide flanked by potential convertase cleavage sites of osteocalcin (OCN), which we named 'metabolitin (MTL)'. We used ligand-receptor binding, receptor internalization, bioluminescence resonance energy transfer and Nano isothermal titration calorimetry assays to study the binding relationship between MTL and GPRC6A. For in vivo biological studies, wild-type mice kept on a high-fat diet (HFD) were injected or gavaged with MTL to study its function in NAFLD., Results: We confirmed that MTL binds to GPRC6A and OCN interacts with GPRC6A using in vitro biological studies. Both intraperitoneal and oral administration of MTL greatly improved NAFLD and insulin resistance in a mouse model. Interacting with GPRC6A expressed in intestines, MTL can significantly inhibit intestinal neurotensin secretion, which in turn inhibits triglyceride but not cholesterol gut absorption, mediated by the 5'AMP-activated protein kinase pathway. In addition, glucagon like peptide-1 secretion was induced by MTL treatment., Conclusions: Oral or intraperitoneal MTL significantly improves the symptoms of NAFLD by inhibiting lipid absorption and insulin resistance. MTL could be a potential therapeutic candidate for the treatment of NAFLD., Lay Summary: A novel murine peptide hormone, herein named 'metabolitin', inhibits fatty acid absorption and improves systemic insulin resistance in a murine model of obesity and non-alcoholic fatty liver disease. Thus, metabolitin has therapeutic potential for the treatment of patients with non-alcoholic fatty liver disease., Competing Interests: Conflict of interest The authors declare no conflicts of interest that pertain to this work. Please refer to the accompanying ICMJE disclosure forms for further details., (Copyright © 2020 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)

The pressure sensor with high sensitivity and a broad pressure sensing range is highly desired for flexible electronics. Here, a high-performance pressure sensor based on a hybrid structure was facilely fabricated using the glass template method, which consists of polyurethane (PU) mesodomes embedded with gradient-distributed silver nanowire (AgNW). Such a novel hybrid architecture enables the as-prepared PU/AgNW pressure sensor to have high sensitivity as well as a wide detection range. Moreover, the obtained PU/AgNW pressure sensors have a fast response time (20 ms), good cycling stability, and excellent flexibility. The pressure sensor, benefiting from its outstanding comprehensive sensing performance, can be used for expression recognition and human activity monitoring, showing tremendous application potential in wearable devices. The proposed architecture and developed methodology in this work is promising for future flexible electronic applications.

Objective: The aim of this study was to understand more about long noncoding RNAs (lncRNAs) as potential prediction biomarkers or therapeutic targets for obesity and type 2 diabetes mellitus (T2DM). This study aimed to find more lncRNA candidates related to obesity and T2DM., Methods: In this study, a high-fat diet (HFD)-induced obesity-T2DM mouse model was used, and a mRNA and lncRNA expression map was drawn up in adipose tissue by microarray technology. Then Gene Ontology analysis and Kyoto Encyclopedia of Genes and Genomes pathway analysis were performed and revealed that the most associated genes and pathways were metabolism-related ones. The candidate lncRNA expression was further validated in adipose tissue from HFD-induced mice by quantitative real-time polymerase chain reaction analysis., Results: Transcriptome analyses were performed to show expression profiles of mRNAs and lncRNAs in epididymal adipose tissue in the obesity-T2DM mice. A total of 124 lncRNAs and 1,606 mRNAs were differentially expressed between the chow and HFD groups. Then, an mRNA-lncRNA coexpression network was constructed. Based on a series of analyses, 15 candidate lncRNAs were screened, and their expression was further validated by quantitative real-time polymerase chain reaction analysis., Conclusions: The results reveal significant differences between the transcriptomes of the HFD and control groups in adipose tissue that provide clues to the molecular mechanisms of diet-induced metabolic disorders as well as biomarkers of risk for these disorders., (© 2019 The Obesity Society.)

Obesity is a chronic multifactorial disease prevalent in many areas of the world and is a major cause of morbidity and mortality. In women, obesity increases the risks of both metabolic and reproductive diseases, such as diabetes and infertility. The mechanisms underlying these effects, especially in young women, are largely unknown. To explore these mechanisms, we established a high-fat diet (HFD) model of obesity in immature female mice. Microarray analysis of gene expression in ovaries and white adipose tissue identified a large number of differentially expressed genes (>1.3-fold change) in both tissues. In ovaries of the HFD group, there were 208 differentially expressed messenger RNAs (mRNAs), including 98 upregulated and 110 downregulated, and 295 differentially expressed lncRNAs (long non coding RNAs), including 63 upregulated and 232 downregulated. In white adipose tissue, there were 625 differentially expressed mRNAs, including 220 upregulated and 605 downregulated in the HFD group, and 1595 differentially expressed lncRNAs, including 1320 and 275 downregulated in the HFD group. Our results reveal significant differences between the transcriptomes of the HFD and control groups in both ovaries and white adipose tissue that provide clues to the molecular mechanisms of diet-induced female reproductive dysfunction and metabolic disorders, as well as biomarkers of risk for these disorders.

An efficient electromagnetic interference (EMI) shielding paper with excellent water repellency and mechanical flexibility has been developed, by assembling silver nanowires (AgNWs) and hydrophobic inorganic ceramic on the cellulose paper, via a facile dip-coating preparation. Scanning electron microscope (SEM) observations confirmed that AgNWs were interconnected and densely coated on both sides of the cellulose fiber, which endows the as-prepared paper with high conductivity (33.69 S/cm in-plane direction) at a low AgNW area density of 0.13 mg/cm 2 . Owing to multiple reflections and scattering between the two outer highly conductive surfaces, the obtained composite presented a high EMI shielding effectiveness (EMI SE) of up to 46 dB against the X band, and ultrahigh specific EMI SE of 271.2 dB mm -1 . Moreover, the prepared hydrophobic AgNW/cellulose (H-AgNW/cellulose) composite paper could also maintain high EMI SE and extraordinary waterproofness (water contact angle > 140°) by suffering dozens of bending tests or one thousand peeling tests. Overall, such a multifunctional paper might have practical applications in packaging conductive components and can be used as EMI shielding elements in advanced application areas, even under harsh conditions.

Flexible strain sensors have attracted a great amount of attention for promising applications in next-generation artificially intelligent devices. However, it is difficult for conventional planar strain sensors to meet the requirements of miniature size and light weight for flexible electronics. Herein, a highly sensitive and stretchable fiber strain sensor with a millimeter diameter was innovatively fabricated by the capillary tube method to integrate silver nanowires (AgNWs) in polyurethane (PU) fibers. Scanning electron microscopy results demonstrate that AgNWs were embedded into the surface layer of PU fibers and formed completely conductive networks. The unique AgNW networks endow the PU/AgNW fibers with superior electrical conductivity of 3.1 S/cm, high elongation at break of 265%, wide response range of 43%, high gauge factor of 87.6 up to 22% strain, fast response time of 49 ms, and excellent reliability and stability. Such satisfactory stretchability and sensitivity is attributed to the combination of the highly stretchable PU matrix and the embedded architecture of the AgNW conductive network. Moreover, PU/AgNW fibers can be employed as wearable devices to detect various human motions and to drive light-emitting diodes at a lower voltage (2.7 V).

Regenerated cellulose (RC) films exhibit poor water barrier performance, which seriously restricts its applications. To address this issue, an impermeable and hydrophobic graphene oxide modified by chemically grafting octadecylamine (GO-ODA) was utilized to enhance the water vapor barrier performance of RC nanocomposite films. Compared to the neat RC film, more than 20% decrease in the coefficient of water vapor permeability ( P H 2 O ) was achieved by loading only 2.0 wt % GO-ODA. The promising hydrophobicity of GO-ODA effectively retarded the formation of hydrogen bonding at the relatively weakened interface between GO and RC, compensating for the diffusion of water vapor molecules at the interface; on the other hand, the fully exfoliated GO-ODA nanosheets were inclined to align with the surface of the as-prepared RC nanocomposite films during hot-pressure drying, creating a much more tortuous pathway for diffusion of water molecules. The new insights could be valuable for widening application of cellulose such as packaging., Competing Interests: The authors declare no competing financial interest.

The adipokine Chemerin and its receptor, chemokine-like receptor 1 (CMKLR1), are associated with osteoblastogenic differentiation of mesenchymal stem cells (MSCs) and osteoclastogenic differentiation of osteoclast precursors in vitro, suggesting that CMKLR1 would affect the bone mineral density (BMD). However, the role of CMKLR1 on BMD in vivo remains unknown. Here, using CMKLR1 knockout mouse model, we unveiled that CMKLR1 effected the amount of Leydig cells in testis and regulated androgen-dependent bone maintenance in male mice, which exhibited lower serum testosterone levels, thereby reducing the trabecular bone mass. Correspondingly, the mRNA expression of testosterone synthesis enzymes in testis decreased. The bone tissue also showed decreased mRNAs expression of osteogenic markers and increased mRNA levels for osteoclast markers. Furthermore, by in vitro differentiation models, we found CMKLR1-deficiency could break the balance between osteoblastogenesis and osteoclastogenesis that caused a shift from osteogenic to adipogenic differentiation in MSCs and enhanced osteoclast formation. In addition, bone mass increase in CMKLR1 KO male mice can be promoted by treatment with 5α-dihydrotestosterone (DHT), and the inactivation of CMKLR1 in male wild-type (WT) mice with antagonist treatment can lead to low bone mass. Taken together, these data indicate that CMKLR1 positively regulates bone metabolism through mediating testosterone production and the balance between osteoblast and osteoclast formation.

Polypropylene random co-polymer (PPR)/styrene-ethylene-butylene-styrene (SBS) block copolymer blends with high toughness and favorable tensile properties were successfully obtained by blending with traces of multi-wall carbon nanotubes (MWCNTs) and β-nucleating agents (β-NAs). β-NAs can effectively induce the ductile β-form crystal in the PPR matrix. Although the addition of MWCNTs was reported to be only benefit for the tensile strength of PPR and relatively disadvantageous for the toughness, the obviously synergistic toughening effect in PPR/SBS blends was found when MWCNTs and β-NAs coexisted. The notched izod impact strength of PPR/30 wt % SBS blend with MWCNTs and β-NAs increased from 11.3 to 58.9 kJ/m²; more than 5-fold increment compared with pure PPR. Meanwhile, the tensile strength retention of this PPR blend is still above 72.2%. The micro-morphology indicated that the MWCNTs can act as bridges between SBS particle and PPR matrix, effectively transferring the stress and absorbing impact energy among SBS particles.

Composite aerogels consisting of graphene oxide (GO) and regenerated cellulose (RCE) were prepared via a solution mixing-regeneration and freeze-drying process. The prepared RCE/GO composites aerogel exhibited 3D network thin-walled pore structure with large specific surface area, also favorable compression recovery capability, outstanding dye elimination efficiency and reusability after the addition of GO. With addition of only 0.5 wt% GO, the methylene blue (MB) elimination efficiency of RCE/GO aerogel reached 99.0% and still remained at 90.5% after five-time reused under oscillation adsorption. In addition, our research indicates that the excellent MB adsorption of RCE/GO composite aerogel was driven by electrostatic interactions and followed a pseudo-second-order adsorption kinetic and monolayer Langmuir adsorption isotherm. This investigation provides the guidance for the development of green environmental adsorbents to remove organic dye from sewage water., (Copyright © 2018 Elsevier Inc. All rights reserved.)

In this work, we propose novel layer-structured polymer composites (PCs) for manipulating the electromagnetic (EM) wave transport, which holds unique electromagnetic interference (EMI) shielding features. The as-prepared PCs with a multilayered structure exhibits significant improvement in overall EMI shielding effectiveness (EMI SE) by adjusting the contents and distribution of electrical and magnetic loss fillers. The layer-structured PCs with low nanofiller content (5 wt % graphene nanosheets (GNSs) and 15 wt % Fe₃O₄) and a thickness of only 2 mm exhibited ultrahigh electrical conductivity and excellent EMI SE, reaching up to 2000 S/m and 45.7 dB in the X-band, respectively. The increased EMI SE of the layer-structured PCs was mainly based on the improved absorption rather than the reflection of electromagnetic waves, which was attributed to the "absorb-reflect-reabsorb" process for the incident electromagnetic waves. This work may provide a simple and effective approach to achieve new EMI shielding materials, especially for absorption-dominated EMI shielding.

Background: Polycystic ovary syndrome (PCOS) is a complex genetic disease with multifarious phenotypes. Many researches use dehydroepiandrosterone (DHEA) to induce PCOS in pubertal mouse models. The aim of this study was to investigate the role of GPR1 in dehydroepiandrosterone (DHEA)-induced hyperandrogenized mice., Methods: Prepubertal C57BL/6 mice (25 days of age) and Gpr1-deficient mice were each divided into two groups and injected daily with sesame oil with or without DHEA (6 mg/100 g) for 21 consecutive days. Hematoxylin and eosin (H&E) staining was performed to determine the characteristics of the DHEA-treated ovaries. Real-time PCR was used to examine steroid synthesis enzymes gene expression. Granulosa cell was cultured to explore the mechanism of DHEA-induced, GPR1-mediated estradiol secretion., Results: DHEA treatment induced some aspects of PCOS in wild-type mice, such as increased body weight, elevated serum testosterone, increased number of small, cystic, atretic follicles, and absence of corpus luteum in ovaries. However, Gpr1 deficiency significantly attenuated the DHEA-induced weight gain and ovarian phenotype, improving steroidogenesis in ovaries and estradiol synthesis in cultured granulosa cells, partially through mTOR signaling., Conclusions: In conclusion, Gpr1 deficiency leads to the improvement of steroid synthesis in mice hyperandrogenized with DHEA, indicating that GPR1 may be a therapeutic target for DHEA-induced hyperandrogenism.

Carbon nanotube/high density polyethylene (CNT/HDPE) foam composites with high electrical conductivity and electromagnetic interference (EMI) shielding performance were developed by means of compression molding plus salt-leaching. The uniform porous structure and interconnected CNT networks throughout the cell backbones endowed the as-prepared foam composites with a significantly lower electrical percolation threshold (0.22 vol%) than that of the solid composites (0.84 vol%). Owing to the multiple reflections and scattering between the cell-matrix interfaces, the foam composites presented a superior specific EMI shielding effectiveness (EMI SE) of 104.3 dB cm 3 g -1 , 2.2 times higher than that of their solid counterpart. Besides this, the pore sizes of the CNT/HDPE foam composites could be easily tuned by controlling the particle size of the porogen. Also, the electrical conductivity and specific EMI SE increased with an increase in the cell diameter, which was attributed to the formation of a more perfect conductive network in the cell backbones. Our approach provides a novel idea for fabricating new lightweight EMI shielding materials, especially for aircraft and spacecraft applications., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Low-dose irradiation (LDI) has been used in clinics to treat human diseases, including chronic inflammation. This study assessed the effects of LDI on the inflammatory response of activated mouse primary peritoneal macrophages, and the underlying signal pathways. Primary peritoneal macrophages were isolated from mice and then incubated with lipopolysaccharide (LPS)-coated Ti microparticles (Ti-positive control) with or without brief exposure to LDI (X-ray, 0.5 Gy) 1 h later (Ti-LDI group) or left untreated in culture medium (Ti-negative control). The macrophages were then subjected to qRT-PCR, Western blot, cell viability CCK-8 assay, and ELISA. qRT-PCR analysis revealed the Ti-LDI group expressed significantly lower levels of IL-1β, IL-6, and TNF-α mRNA than those of the Ti-positive control group, while the ELISA data showed that Ti-LDI group had significantly lower secretion of IL-1β, IL-6, and TNF-α proteins. The most significant reduction associated with LDI was the secretion TNF-α protein, which barely increased from 13 to 25 h after treatment. Western blot data demonstrated that phosphorylation of p65 and ERK was much lower in the Ti-LDI group than in the controls. The data from the current study suggests that LDI of activated mouse macrophages was associated with significantly lower inflammation responses, compared with non-exposed activated macrophages, which was possibly through inhibition of the NF-κB and ERK pathways.

The reconstruction of critically sized bone defects remains a serious clinical problem because of poor angiogenesis within tissue-engineered scaffolds during repair, which gives rise to a lack of sufficient blood supply and causes necrosis of the new tissues. Rapid vascularization is a vital prerequisite for new tissue survival and integration with existing host tissue. The de novo generation of vasculature in scaffolds is one of the most important steps in making bone regeneration more efficient, allowing repairing tissue to grow into a scaffold. To tackle this problem, the genetic modification of a biomaterial scaffold is used to accelerate angiogenesis and osteogenesis. However, visualizing and tracking in vivo blood vessel formation in real-time and in three-dimensional (3D) scaffolds or new bone tissue is still an obstacle for bone tissue engineering. Multiphoton microscopy (MPM) is a novel bio-imaging modality that can acquire volumetric data from biological structures in a high-resolution and minimally-invasive manner. The objective of this study was to visualize angiogenesis with multiphoton microscopy in vivo in a genetically modified 3D-PLGA/nHAp scaffold for calvarial critical bone defect repair. PLGA/nHAp scaffolds were functionalized for the sustained delivery of a growth factor pdgf-b gene carrying lentiviral vectors (LV-pdgfb) in order to facilitate angiogenesis and to enhance bone regeneration. In a scaffold-implanted calvarial critical bone defect mouse model, the blood vessel areas (BVAs) in PHp scaffolds were significantly higher than in PH scaffolds. Additionally, the expression of pdgf-b and angiogenesis-related genes, vWF and VEGFR2, increased correspondingly. MicroCT analysis indicated that the new bone formation in the PHp group dramatically improved compared to the other groups. To our knowledge, this is the first time multiphoton microscopy was used in bone tissue-engineering to investigate angiogenesis in a 3D bio-degradable scaffold in vivo and in real-time.

Maintenance of healthy bone quality and quantity requires a well-coordinated balance between bone formation by osteoblasts and bone resorption by osteoclasts. Chemerin is a novel adipokine with known functions such as regulating immunity and energy homeostasis through activation of chemokine-like receptor 1 (CMKLR1). G protein-coupled receptor 1 (GPR1) is the second mammalian chemerin receptor with similar binding affinity as CMKLR1. In male GPR1 -/- mice, a phenotype with significantly low bone mineral density was observed. We hypothesise that GPR1 might participate the process of bone remodelling. In this study, we investigated the role of GPR1 in regulating bone mass maintenance in male mice, and for the first time, revealed that GPR1 -/- male mice manifested seriously trabecular bone loss and lower serum testosterone levels compared to the wild type animals. Accordingly, the mRNA expression of biomarkers related to both osteoblast [collagen type I alpha 2 ( Col1A2 ), osteocalcin ( OCN )] and osteoclast [tartrate-resistant acid phosphatase ( TRAP ), Cathepsin K , NFATc1 ] were significantly decreased or increased in GPR1 -/- mice relative to the wild type, respectively. However, other osteogenic markers, Osterix and ALP levels, were increased. Microcomputed tomography scanning and histological analyses proved that there was a myriad of trabecular bone loss in GPR1 -/- mice. In the meantime, GPR1 -/- mice presented a significant decrease in serum testosterone level. Taken together, these findings suggested that chemerin-GPR1 signalling might be directly or indirectly communicated with testosterone synthesis on bone turnover regulation. Further detailed studies are required to unveil how chemerin-GPR1 participates in bone metabolism. The translational potential of this article: More studies and knowledge about GPR1 regulating function in bone turnover might supply a novel therapeutic target for osteoporosis in the future.

Low-dose irradiation (LDI) exhibits a positive effect on osteoblasts and inhibitory effect of inflammation. Here, we test the hypothesis that LDI can promote osseointegration and inhibit the inflammatory membrane formation in the presence of titanium (Ti) particles. Endotoxin-free titanium particles were injected into rabbit, prior to the insertion of a Ti6-Al-4-V sticks pre-coated with hydroxyapatite. Two days after operation, both distal femurs of the animal were exposed to 0.5 Gy X-ray irradiation. All ani-mals were euthanized 8 weeks after the operation. The PINP concentration was determined at day 0, 2, 4, and 8 weeks after operation. Trabecular morphology around the implants 8 weeks after operation was assessed using micro-CT, then the maximum push out force of simples was assessed using biomechanics test. Five samples in each group were chosen for bone histomorphology study without decalcification 8 weeks after operation. The results confirmed that the LDI can significantly improve ingrowth of bone into the prosthetic interface and stability of the prosthesis when there was no wear particles. Although promotion effects for bone formation induced by LDI can be counteracted by wear particles, LDI can significantly inhibit the interface membrane formation around the implant induced by wear particles. Based on these results, we conclude that LDI may be useful for enhancing the stability of prosthesis when there are no wear particles and for inhibiting the interface membrane formation during the early stage of aseptic loosening in the presence of wear particles. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:1688-1696, 2016., (© 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.)

Unlabelled: Reconstruction of critical size bone defects remains a major clinical challenge because of poor bone regeneration, which is usually due to poor angiogenesis during repair. Satisfactory vascularization is a prerequisite for the survival of grafts and the integration of new tissue with existing tissue. In this work, we investigated angiogenesis in 3D scaffolds by in vivo multiphoton microscopy during bone formation in a murine calvarial critical bone defect model and evaluated bone regeneration 8weeks post-implantation. The continuous release of bioactive lentiviral vectors (LV-pdgfb) from the scaffolds could be detected for 5days in vitro. In vivo, the released LV-pdgfb transfected adjacent cells and expressed PDGF-BB, facilitating angiogenesis and enhancing bone regeneration. The expression of both pdgfb and the angiogenesis-related genes vWF and VEGFR2 was significantly increased in the pdgfb gene-carrying scaffold (PHp) group. In addition, microCT scanning and histomorphology results proved that there was more new bone ingrowth in the PHp group than in the PLGA/nHA (PH) and control groups. MicroCT parameters, including BMD, BV/TV, Tb.Sp, and Tb.N indicated that there was significantly more new bone formation in the PHp group than in the other groups. With regard to neovascularization, 8weeks post-implantation, blood vessel areas (BVAs) were 9428±944μm(2), 4090±680.3μm(2), and none in the PHp, PH, and control groups, respectively. At each time point, BVAs in the PHp scaffolds were significantly higher than in the PH scaffolds. To our knowledge, this is the first use of multiphoton microscopy in bone tissue-engineering to investigate angiogenesis in scaffolds in vivo. This method represents a valuable tool for investigating neovascularization in bone scaffolds to determine if a certain scaffold is beneficial to neovascularization. We also proved that delivery of the pdgfb gene alone can improve both angiogenesis and bone regeneration Acronyms., Statement of Significance: Reconstruction of critical size bone defects remains a major clinical challenge because of poor bone regeneration, which is usually due to poor angiogenesis during repair. Satisfactory vascularization is a prerequisite for the survival of grafts and the integration of new tissue with existing tissue. In this work, we investigated angiogenesis in 3D scaffolds by in vivo multiphoton microscopy during bone formation in a murine calvarial critical bone defect model and evaluated bone regeneration 8weeks post-implantation. To verify that pdgfb-expressing vectors carried by the scaffolds can promote angiogenesis in 3D-printed scaffolds in vivo, we monitored angiogenesis within the implants by multiphoton microscopy. To our knowledge, this is the first study to dynamically investigate angiogenesis in bone tissue engineering scaffolds in vivo., (Copyright © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Magnesium (Mg) is a promising biodegradable metal offering many potential advantages over current scaffold technologies. Many studies have reported on the corrosion characteristics the Mg and its bioeffects in vitro and in vivo, but there are few studies on the biological effects of the corrosive products of Mg - the micron-size Mg particles (MgMPs). In this study, the effects of size-selected commercial MgMPs on bone turnover and macrophages were investigated in vivo and in vitro. We found that MgMPs were susceptible to engulfment by macrophages, leading to cell lysis, likely resulting from H2 gas production. We also found that the inflammatory cytokines IL-1, IL-6, and TNF-α were induced more strongly by titanium particles (TiMPs) group than by either MgMPs or control. Examination of the expression of bone remodeling markers revealed that MgMPs are beneficial for bone regeneration. Micro-CT scanning indicated that, 30 days postimplantation, unlike TiMPs, MgMPs had no adverse effect on either bone quality or quantity. We have investigated the bioeffects of micron-size MgMPs in vivo and in vitro, and our results indicate that MgMPs may promote bone regeneration without inducing inflammation. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 923-931, 2016., (© 2015 Wiley Periodicals, Inc.)

Elevated serum chemerin levels correlate with increased severity of polycystic ovary syndrome (PCOS). However, the role of CMKLR1 signaling in ovarian biology under conditions of excess DHT remains unclear. In this study we compared the effects of continuous 90-day high dose DHT exposure (83.3 □g/day) on wild type and CMKLR1-deficient mice. DHT induced PCOS-like clinical signs in wild type mice as well as significant changes in the expression of hormone receptors, steroid synthesis enzymes, and BMPs and their receptors. In contrast, CMKLR1-deficient mice significantly attenuated DHT-induced clinical signs of PCOS and alterations in ovarian gene expression. To determine whether the BMP4 signaling pathway was involved in the pathogenic effects of CMKLR1 signaling in DHT-induced ovarian steroidogenesis, antral follicles were isolated from wild type and CMKLR1 knockout (KO) mice and treated in vitro with combinations of hCG, DHT, and BMP4 inhibitors. BMP4 inhibition attenuated the induction effects of hCG and DHT on estrogen and progesterone secretion in CMKLR1 KO mice, but not in WT mice, implicating the BMP4 signaling pathway in the CMKLR1-dependent response to DHT. In conclusion, CMKLR1 gene deletion attenuates the effects of chronic DHT treatment on ovarian function in experimental PCOS, likely via BMP4 signaling.

An in situ thermally reduced graphene/polyethylene conductive composite with a segregated structure was fabricated, which achieved a high electromagnetic interference shielding effectiveness of up to 28.3-32.4 dB at an ultralow graphene loading of 0.660 vol.%. Our work suggests a new way of effectively using graphene.

Wear particles and by-products from joint replacements and other orthopaedic implants may result in a local chronic inflammatory and foreign body reaction. This may lead to persistent synovitis resulting in joint pain and swelling, periprosthetic osteolysis, implant loosening and pathologic fracture. Strategies to modulate the adverse effects of wear debris may improve the function and longevity of joint replacements and other orthopaedic implants, potentially delaying or avoiding complex revision surgical procedures. Three novel biological strategies to mitigate the chronic inflammatory reaction to orthopaedic wear particles are reported. These include (i) interference with systemic macrophage trafficking to the local implant site, (ii) modulation of macrophages from an M1 (pro-inflammatory) to an M2 (anti-inflammatory, pro-tissue healing) phenotype in the periprosthetic tissues, and (iii) local inhibition of the transcription factor nuclear factor kappa B (NF-κB) by delivery of an NF-κB decoy oligodeoxynucleotide, thereby interfering with the production of pro-inflammatory mediators. These three approaches have been shown to be viable strategies for mitigating the undesirable effects of wear particles in preclinical studies. Targeted local delivery of specific biologics may potentially extend the lifetime of orthopaedic implants.

The novel adipokine chemerin plays a role in the regulation of lipid and carbohydrate metabolism, and recent reports of elevated chemerin levels in polycystic ovarian syndrome and preeclampsia have pointed to an emerging role of chemerin in reproduction. We hypothesised that chemerin, like other adipokines, may function to regulate male gonadal steroidogenesis. In this study, we show that chemerin and its three receptors chemokine-like receptor 1 (CMKLR1), G-protein-coupled receptor 1 (GPR1) and chemokine (C-C motif) receptor-like 2 were expressed in male reproductive tracts, liver and white adipose tissue. CMKLR1 and GPR1 proteins were localised specifically in the Leydig cells of human and rat testes by immunohistochemistry. The expression of chemerin and its receptors in rat testes was developmentally regulated and highly expressed in Leydig cells. In vitro treatment with chemerin suppressed the human chorionic gonadotropin (hCG)-induced testosterone production from primary Leydig cells, which was accompanied by the inhibition of 3β-hydroxysteroid dehydrogenase gene and protein expression. The hCG-activated p44/42 MAPK (Erk1/2) pathway in Leydig cells was also inhibited by chemerin cotreatment. Together, these data suggest that chemerin is a novel regulator of male gonadal steroidogenesis.

Sclerostin is a glycoprotein secreted by osteocytes and inhibits osteoblastogenesis via inhibition of Wnt signaling. We hypothesized that sclerostin antibody (Scl-AbIII) would accelerate the healing of a murine femoral non-critical size bone defect model. A unilateral and unicortical 0.8 mm-sized drill hole was made in the proximal femoral shaft of adult female nude mice. One group of mice received subcutaneous injections of Scl-AbIII and a second group received vehicle only. Reporter MC3T3 osteoprogenitor cells were injected via the tail vein 3 days after surgery to monitor systemic trafficking of exogenous osteoprogenitors. Bioluminescence imaging (BLI), microcomputed tomography (microCT), micropositron emission tomography (microPET) and histological analysis were used to compare the bone healing responses to Scl-AbIII treatment. Bone mineral density (BMD) significantly increased at the defect site after week 1, and was significantly higher in the treatment compared with the control group at all time points. This finding was also confirmed on histological analysis by increased deposition of new woven bone. MicroPET scanning showed a trend for greater activity in the control group at day 21 compared with the Scl-AbIII group, indicating early bone maturation following treatment with Scl-AbIII. Whereas the BLI signals derived from the injected osteoprogenitor cells showed no differences between vehicle and Scl-AbIII treated groups, systemic migration of MC3T3 cells to the bone defect was clearly identified in both groups using immunohistochemistry. Systemic administration of Scl-AbIII resulted in earlier healing and maturation of a non-critical size bone defect. These findings underscore the potential use of Scl-AbIII for treatment of complicated fractures, non-unions, and other clinical scenarios., (Copyright © 2012 Orthopaedic Research Society.)

Based on the bioinformatic prediction, Zhang and colleagues discovered obestatin, a new 23-amino acid hormone from rat stomach extract encoded by the ghrelin gene. Obestatin is present not only in the gastrointestinal tract, but also in the spleen, mammary gland, breast milk, and plasma. Obestatin appears to function as part of a complex gut-brain network whereby hormones and substances from the stomach, intestine and the brain about satiety or hunger. Given the current research regarding the effects of obestatin and its possible cognate receptor(s), this chapter provides the latest review of the physiological and pathological characteristics of this hormone and its possible receptor(s) in energy homeostasis and obesity., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Understanding how relevant cell types respond to wear particles will reveal new avenues for treating osteolysis following joint replacements. In this study, we investigate the effects of ultrahigh molecular weight polyethylene (UHMWPE) particles on preosteoblast migration and function. We infused UHMWPE particles or saline into the left femur of mice and injected luciferase-expressing preosteoblasts (MC3T3 cells) into each left ventricle. Bioluminescence imaging (BLI) confirmed systemic administration of MC3T3 cells. BLI throughout the 28-day experiment showed greater MC3T3 migration to the site of particle infusion than to the site of saline infusion, with significant differences on days 0, 4, and 6 (p≤0.055). Immunostaining revealed a greater number of osteoblasts and osteoclasts in the particle-infused femora, indicating greater bone turnover. The bone mineralization of the particle-infused femora increased significantly when compared to saline-infused femora (an increase of 146.4±27.9 vs. 12.8±8.7 mg/mL, p=0.008). These results show that infused preosteoblasts can migrate to the site of wear particles. Additionally, as the migrated cells were associated with increased bone mineralization in spite of the presence of particles, increasing osteoblast recruitment is a potential strategy for combating bone loss due to increased osteoclast/macrophage number and decreased osteoblast function.

The biological mechanisms leading to periprosthetic osteolysis involve both chemokines and the monocyte/macrophage cell lineage. Whether MCP-1 plays a major role in macrophage recruitment in the presence of wear particles is unknown. We tested two hypotheses: (1) that exogenous local delivery of MCP-1 induces systematic macrophage recruitment and (2) that blockade of the MCP-1 ligand-receptor axis decreases macrophage recruitment and osteolysis in the presence of ultra high molecular weight polyethylene (UHMWPE) particles. Six groups of nude mice were used. We used non-invasive imaging to assay macrophage recruitment and osteolysis. A murine macrophage cell line and primary wild type and CCR2 knockout murine macrophages were used as the reporter cells. Particles were infused into the femoral canal. Bioluminescence and immunohistochemical staining were used to confirm the migration of reporter cells. Locally infused MCP-1 induced systemic macrophage trafficking to bone. Injection of MCP-1 receptor antagonist significantly decreased reporter cell recruitment to bone infused with UHMWPE particles and decreased osteolysis. Systemic migration of reporter cells to infused particles was decreased when the reporter cells were deficient in the CCR2 receptor. Interruption of the MCP-1 ligand-receptor axis appears to be a viable strategy to mitigate trafficking of macrophages and osteolysis due to UHMWPE particles., (Copyright © 2011 Orthopaedic Research Society.)

Graphene oxide (GO) was successfully prepared by a modified Hummer's method. The reduction effect and mechanism of the as-prepared GO reduced with hydrazine hydrate at different temperatures and time were characterized by x-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR), elemental analysis (EA), x-ray diffractions (XRD), Raman spectroscopy and thermo-gravimetric analysis (TGA). The results showed that the reduction effect of GO mainly depended on treatment temperature instead of treatment time. Desirable reduction of GO can only be obtained at high treatment temperature. Reduced at 95 °C for 3 h, the C/O atomic ratio of GO increased from 3.1 to 15.1, which was impossible to obtain at low temperatures, such as 80, 60 or 15 °C, even for longer reduction time. XPS, 13C NMR and FTIR results show that most of the epoxide groups bonded to graphite during the oxidation were removed from GO and form the sp(2) structure after being reduced by hydrazine hydrate at high temperature (>60 °C), leading to the electric conductivity of GO increasing from 1.5 × 10(-6) to 5 S cm(-1), while the hydroxyls on the surface of GO were not removed by hydrazine hydrate even at high temperature. Additionally, the FTIR, XRD and Raman spectrum indicate that the GO reduced by hydrazine hydrate can not be entirely restored to the pristine graphite structures. XPS and FTIR data also suggest that carbonyl and carboxyl groups can be reduced by hydrazine hydrate and possibly form hydrazone, but not a C = C structure.

Background: Aseptic loosening and periprosthetic osteolysis resulting from wear debris are major complications of total joint arthroplasty. Monocyte/macrophages are the key cells related to osteolysis at the bone-implant interface of joint arthroplasties. Whether the monocyte/macrophages found at the implant interface in the presence of polyethylene particles are locally or systemically derived is unknown., Questions/purposes: We therefore asked (1) whether macrophages associated with polyethylene particle-induced chronic inflammation are recruited locally or systemically and (2) whether the recruited macrophages are associated with enhanced osteolysis locally., Methods: Noninvasive in vivo imaging techniques (bioluminescence and microCT) were used to investigate initial macrophage migration systemically from a remote injection site to polyethylene wear particles continuously infused into the femoral canal. We used histologic and immunohistologic staining to confirm localization of migrated macrophages to the polyethylene particle-treated femoral canals and monitor cellular markers of bone remodeling., Results: The values for bioluminescence were increased for animals receiving UHMWPE particles compared with the group in which the carrier saline was infused. At Day 8, the ratio of bioluminescence (operated femur divided by nonoperated contralateral femur of each animal) for the UHMWPE group was 13.95 ± 5.65, whereas the ratio for the saline group was 2.60 ± 1.14. Immunohistologic analysis demonstrated the presence of reporter macrophages in the UHMWPE particle-implanted femora only. MicroCT scans showed the bone mineral density for the group with both UHMWPE particles and macrophage was lower than the control groups., Conclusions: Infusion of clinically relevant polyethylene particles, similar to the human scenario, stimulated systemic migration of remotely injected macrophages and local net bone resorption.

Wear particles generated from total joint arthroplasty (TJA) stimulate macrophages to release chemokines. The role of chemokines released from wear particle-stimulated macrophages on the migration of macrophages and osteoprogenitor cells in vitro has not been elucidated. In this study, we challenged murine macrophages (RAW 264.7) with clinically relevant polymethyl methacrylate (PMMA, 1-10 microm) and ultra high molecular weight polyethylene (UHMWPE, 2-3 microm) particles. The chemotactic effects of the conditioned media (CM) were tested in vitro using human macrophages (THP-1) and human mesenchymal stem cells (MSCs) as the migrating cells. CM collected from both particle types had a chemotactic effect on human macrophages, which could be eliminated by monocyte chemotactic protein-1 (MCP-1) neutralizing antibody. Blocking the CCR1 receptor eliminated the chemotactic effect, while CCR2 antibody only partially decreased THP-1 cell migration. CM from PMMA but not UHMWPE-exposed macrophages led to chemotaxis of MSCs; this effect could be eliminated by macrophage inflammatory protein-1 alpha (MIP-1alpha) neutralizing antibody. Neither CCR1 nor CCR2 blocking antibodies showed an effect on the migration of MSCs. Chemokines released by macrophages stimulated by wear particles can have an effect on the migration of macrophages and MSCs. This effect seems to be dependent on the particle type, and may be modulated by MCP-1 and MIP-1alpha, however, more than one chemokine may be necessary for chemotaxis., ((c) 2010 Wiley Periodicals, Inc.)

Growth factors control the proliferation and differentiation of osteoprogenitor cells. This study explores the effects of modulating growth factors (VEGF, IGF-1, FGF-2 and BMP-2) on osteogenesis of mesenchymal stem cells (MSCs) in vitro. Constant and profiled delivery protocols, in accordance with protein expression in vitro, were applied to deliver or neutralize growth factors. Cell number, alkaline phosphatase (ALP-2) and osteocalcin (OC) expression, and mineralization were measured as outcome variables. Profiled addition of VEGF increased MSC proliferation. Constant and profiled application of FGF-2 and neutralization of IGF-1 and BMP-2 decreased ALP-2 levels. Profiled addition of BMP-2 vastly increased OC release from MSCs, but constant addition of IGF-1, constant and profiled neutralization of IGF-1 and FGF-2 reduced OC levels. Constant addition of IGF-1 and FGF-2, as well as profiled loading of FGF-2 decreased mineralization of MSCs. This study indicated that endogenous IGF-1 and FGF-2 are essential to osteogenesis; excess IGF-1 and FGF-2 were inhibitory to bone formation. Selective, temporally specific addition of growth factors, such as BMP-2 and VEGF appears to be an important strategy to enhance osteogenesis., (Copyright 2010 Elsevier Ltd. All rights reserved.)

Macrophages constitute a major part of the cell response to wear particles produced at articulating and nonarticulating interfaces of joint replacements. This foreign body reaction can result in periprosthetic osteolysis and implant loosening. We demonstrate that ultra-high molecular weight polyethylene (UHMWPE) particles induce systemic trafficking of macrophages by noninvasive in vivo imaging and immunohistochemistry. The distal femora of nude mice were injected with 60 mg/mL UHMWPE suspension or saline alone. Reporter RAW264.7 macrophages that stably expressed the bioluminescent reporter gene and the fluorescence reporter gene were injected intravenously. Bioluminescence imaging was performed using an in vivo imaging system immediately after macrophage injection and at 2-day intervals. Compared with the nonoperated contralateral femora, at day 4, 6, and 8, the bioluminescent signal of femora containing UHMWPE suspension increased 1.30 +/- 0.09-, 2.36 +/- 0.92-, and 10.32 +/- 7.61-fold, respectively. The values at same time points for saline-injected control group were 1.08 +/- 0.07-, 1.14 +/- 0.27-, and 1.14 +/- 0.35-fold, respectively. The relative bioluminescence of the UHMWPE group was higher at all postinjection days and significantly greater than the saline group at day 8 (p < 0.05). Histological analysis confirmed the presence of reporter macrophages within the medullary canal of mice with implanted UHMWPE particles. The presence of UHMWPE particles induced enhanced bone remodeling activity. Clinically relevant UHMWPE particles stimulated the systemic recruitment of macrophages during an early time course using the murine femoral implant model. Interference with systemic macrophage trafficking may potentially mitigate UHMWPE particle-induced periprosthetic osteolysis., ((c) 2010 Wiley Periodicals, Inc.)