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In this paper, we will discuss the worship of fertility goddesses around the world and how these practices inform female figures as they appear in Western art. Aphrodite, or Venus, is a key figure in our discussion. In this study, we will focus on her origin, figures, and characteristics. We will also investigate female images from the later time when Europe became a fully patriarchal society. These images evidenced a shift from a muscular and curvaceous body to a soft and submissive-looking one. Once the feminist movement was underway, images from the ancient Greek and Roman period were revived and reemerged in new ways. In this paper, we’ll use Rosie the Riveter as a modern example for study. As the message and intent of artists throughout history makes clear: the ancient goddess’s expression of “Heroic Femininity” is still full of life in today’s context. Venus, the goddess of love, sex, justice, and fertility, created with a body to be desired by man, has become, in a new world setting – a beacon for women’s equal rights and freedom.

BACKGROUND: A number of studies have demonstrated that trichosanthin (TCS) can induce apoptosis in numerous types of tumor cell lines. However, whether TCS can induce pyroptosis has not yet been reported. This study aimed to investigate the role of TCS and its inhibitory effect on tumor growth by modulating pyroptosis in non-small cell lung cancer (NSCLC). METHODS: Effects of different concentrations of TCS on the cell viability, proliferation, migration and invasion of NSCLC were detected by Cell Counting Kit-8 (CCK-8), colony formation, migration, and invasion assays. Immunofluorescence was used to detect the effect of TCS on the expression of pyroptosis marker protein gasdermin-D (GSDMD)-N in A549 cells. A tumor xenograft animal model was established by injecting A549 cells into nude mice. RESULTS: In the present study, we found that TCS significantly inhibited the proliferation, migration, and invasion of A549 cells in a concentration-dependent manner. In addition, TCS at a high concentration (40 µg/mL) significantly promoted the expression of pyroptosis-related proteins [GSDMD-N, NLR family pyrin domain containing 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), caspase-1, and GSDMD], which showed an inhibitory effect on the pyroptosis of A549 cells. Additionally, we found that necrosulfonamide (NSA) significantly reversed the inhibitory effect of high concentrations of TCS on the pyroptosis of A549 cells. The in vivo experiments showed that TCS effectively reduced the tumor volume and inhibited the expression of Ki-67, whereas it increased the expression of GSDMD-N. CONCLUSIONS: Taken together, these results indicated that TCS could inhibit the progression of NSCLC by promoting pyroptosis. These findings provide further information on the possible underlying mechanism of TCS in the treatment of NSCLC.

Spinel LiMn2O4 (LMO) with a three-dimensional structure has become one of the cathode materials that has gained the most interest due to its safety, low price and abundant resources. However, the lithium ion transmission is limited by large particle size and particle agglomeration of LMO. Thus, reducing the particle size and agglomeration of LMO can effectively improve its lithium ion transmission. Here, we synthesized a LMO cathode material with a nanoscale crystal size using the flexible expanded graphite (EG) soft template and Pichini method. EG-controlled particle size and particle agglomeration of LMO is conducive to charge transfer and diffusion of lithium ions between LMO and the electrolyte, meanwhile, there are more redox sites on the nanosized LMO particles, which makes the redox reaction of LMO more thorough during the charge and discharge process, resulting in high capacitance performance. In order to obtain the considerably required lithium-ion capacitors (LICs) with high energy density and power density, we assembled aqueous LMO//activated carbon (AC) LICs with 5 M LiNO3 as the aqueous electrolytes, which are environmentally friendly, safe, low cost and have higher electrical conductivity than organic electrolytes. The optimal LIC has an energy density of 32.63 W h kg−1 at a power density of 500 W kg−1 and an energy density of 8.06 W h kg−1 at a power density of 10 000 W kg−1, which is higher than most of the LMO-based LICs in previous reports. After 2000 cycles, the specific capacitance retention rate was 75.9% at a current density of 3 A g−1. Therefore, our aqueous LMO//AC LICs synthesized by the soft template/Pichini method have wide prospects and are suitable for low-cost, high-safety and high-power applications.

Spinel LiMn

It is reported that olivine-type lithium iron phosphate (LFP) for Li-ion batteries is one of the most widely utilized cathode materials, but its high-power applications are limited due to its intrinsically poor ion transfer rate and conductivity. Therefore, it is highly desired to fabricate LFP Li-ion capacitors (LICs) with high power performance and excellent cyclic reversibility, especially in safe, low cost and environmentally friendly aqueous electrolytes. Here, we fabricate LFP/expanded graphite (EG) nanocomposites by a one-step process, in which polyethylene glycol (PEG) is used as the particle growth inhibitor combined with vacuum infiltration of the LFP precursor into EG as a conductive sub-phase, and further investigate their high-power performance in aqueous LICs. Embedding spherical LFP nanoparticles with well-controlled size and agglomeration into the pores of EG and wrapping LFP nanoparticles by EG films contribute to the rapid electron and ion diffusion in LFP/EG composites, resulting in excellent cyclic reversibility and rate performance of LFP/EG composites. The aqueous LFP/EG//active carbon (AC) LICs were assembled in LiNO3 electrolytes with LFP/EG composites and AC as the positive and negative electrodes, respectively. The optimal LIC shows a power density of 2367.9 W kg−1 at an energy density of 6.5 W h kg−1, dramatically favorable rate characteristics and excellent cycle life with 82.1% capacitance retention of its primary capacitance at 2 A g−1 after 6000 cycles, markedly higher than those of the commercial LFP LIC. The presented aqueous LFP/EG//AC LICs with excellent electrochemical performance are expected to have broad high-power appliances that are cost-sensitive and highly secure.

It is highly desired to simultaneously introduce active heteroatoms and abundant hierarchical pore structures for enhanced electrochemical performances on carbon materials. Herein, trehalose as a pore-forming agent was added into polyvinylpyrrolidone/melamine formaldedyde resin mixture with high concentrations of nitrogen and oxygen. Then a simple one-step carbonization/activation process was adopted and heteroatom-enriched carbon with hierarchical pores (HPC) was fabricated successfully. HPC/HPC symmetric supercapacitors were assembled using KOH electrolyte. It is clearly demonstrated that due to the pore-forming action of trehalose HPC shows the porous honeycomb, interconnected and worm-like pore structure, which is favorable to enhance the double-layer capacitance. It is confirmed that in our system the three active species of pyridinic nitrogen (N-6), pyrrolic nitrogen (N-5) and quinone type oxygen (O-I) are responsible for the pseudocapacitive behavior. Based on XPS, nitrogen adsorption/desorption isotherms and electrochemical impedance spectroscopy, it is deduced that the ratio-optimized HPC-T30 exhibits high concentration of three active species (8.17 at.%), increased specific area (351.26 m2 g−1) and tuned hierarchical pore structures with substantial micropores (micropore area of 321.68 m2 g−1) and a small amount of mesopores and macropores, which lead to decrease of charge transfer resistance, increase of transfer rate of electrolyte ions in the pores and excellent electrochemical performances. In cyclic voltammetry tests of three-electrode system and galvanostatic charge/discharge tests of two-electrode system, HPC-T30 displays high specific capacitance, 46% and 1.2-time enhancement compared to untreated HPC-T0, respectively. The optimized HPC-T30/HPC-T30 supercapacitor delivers the energy density of 6.69 W h kg−1 in 6 M KOH electrolyte. Furthermore, the supercapacitor shows a capacitance retention of 91.16% up to 6000 cycles and the coulombic efficiency reaches nearly 100% for each charged/discharge cycle, demonstrating its good cyclic stability.

Olivine-type LiFePO4 (LFP) is one of the most widely utilized cathode materials for high power Li-ion batteries (LIBs). In spite of rapidly growing popularity of LIBs, the rate performance of the highest power LFP cells is still insufficiently high for some high-power applications. In this work we demonstrate that vacuum-infiltration of LFP precursors into pores of low-cost expanded graphite (EG), an in-situ sol-gel process, followed by calcination, allows formation of LFP/EG nanocomposites that demonstrate remarkable performance in higher power Li-ion capacitor (LIC) applications. Such composites comprise spherical LFP particles embedded into EG pores and additionally wrapped by EG films, forming a highly efficient and stable conducting network. Such a morphology greatly accelerates Li-ion diffusion and improves Li-ion exchange between LFP and electrolyte. As a result, compared to commercial LFP particles of comparable size, the optimized LFP/EG nanocomposite shows significantly higher rate performance, dramatically better stability and higher specific capacitance of up to about 1200 F g−1. The use of environmentally friendly, safe and low-cost aqueous electrolyte is particularly advantageous for LIC applications that are cost-sensitive and require enhanced safety. Our results demonstrate a great promise of our approach, which is additionally applicable for a broad range of other intercalation chemistries.

Myocyte enhancer factor 2D (MEF2D) is involved in many aspects of cancer progression, including cell proliferation, invasion, and migration. However, little is known about the role of MEF2D in tumor angiogenesis. Using clinical specimens, colorectal cancer (CRC) cell lines and a mouse model in the present study, we found that MEF2D expression was positively correlated with CD31-positive microvascular density in CRC tissues. MEF2D promoted tumor angiogenesis in vitro and in vivo and induced the expression of proangiogenic cytokines in CRC cells. MEF2D was found to be a downstream effector of hypoxia-inducible factor (HIF)-1α in the induction of tumor angiogenesis. HIF-1α transactivates MEF2D expression by binding to the MEF2D gene promoter. These results demonstrate that the HIF-1α/MEF2D axis can serve as a therapeutic target for the treatment of CRC.

Cerium oxide nanoparticles (CNPs) have been intensively explored for biomedical applications in recent few years due to the versatile enzyme mimetic activities of the nanoparticles. However, the control of CNPs quality through the optimization of synthesis conditions remains largely unexplored as most of the previous studies only focus on utilizing the catalytic activities of the nanoparticles. In the present study, CNPs with size about 5 nm were synthesized by thermal decomposition method using traditional convective heating and recently developed microwave irradiation as heating source. The quality of CNPs synthesized by the two heating manner was evaluated. The CNPs synthesized by convective heating were slightly smaller than that synthesized by microwave irradiation heating. The cores of the CNPs synthesized by the two heating manner have similar crystal structure. While the surface subtle structures of the CNPs synthesized by two heating manner were different. The CNPs synthesized by microwave irradiation have more surface reactive hot spot than that synthesized by convective heating as the nanoparticles responded more actively to the redox environment variation. This difference resulted in the higher superoxide dismutase (SOD) mimetic activity of CNPs synthesized by microwave irradiation heating than that of the convective heating. Preliminary experiments indicated that the CNPs synthesized by microwave irradiation heating could better protect cells from oxidative stress due to the higher SOD mimetic activity of the nanoparticles.

It is highly desired to introduce active nitrogen atoms and abundant pore structures for enhanced electrochemical performances of carbon materials for supercapacitors. Hence a facile one-step carbonization/activation method was adopted to treat different mass ratios of polyvinylpyrrolidone/melamine formaldehyde resin (PVP/MF) and ammonium carbonate in order to obtain porous N-doped carbon (PNC). Symmetric supercapacitors with PNC as electrodes were assembled using KOH aqueous electrolyte. It is clearly demonstrated that due to the introduction of ammonium carbonate the resulting PNC displays increased specific surface areas, enriched pore structures and increased active nitrogen concentration. The ratio-optimized PNC with 7 wt% ammonium carbonate exhibits the highest proportion of negatively charged N-6 and N-5 with the pseudocapacitance and the highest micropore volume with the electrical double-layer capacitance, which also lead to the decrease of charge transfer resistance and the increase of transfer rate of electrolyte ions in the pores, and these are beneficial effects for the improvement of the overall capacitive performances. The ratio-optimized PNC displays an enhanced specific capacitance, which is about 32% higher than that of the PNC prepared without ammonium carbonate. And the symmetric supercapacitor assembled with the ratio-optimized PNC shows superior cycling performance and its capacitance retention is 94.43% after 1500 cycles at 20 mA cm−2.

Chondrocyte hypertrophy is a physiological process in endochondral ossification. However, the hypertrophic-like alterations of chondrocytes at the articular surface may result in osteoarthritis (OA). In addition, the generation of fibrocartilage with a decreased biological function in tissue engineered cartilage, has been attributed to chondrocyte hypertrophy. Therefore, suppressing chondrocyte hypertrophy in OA and the associated regeneration of non-active cartilage is of primary concern. The present study examined the effects of xanthotoxin (XAT), which is classified as a furanocoumarin, on chondrocyte hypertrophic differentiation of mesenchymal stem cells. Following XAT treatment, the expression levels of genes associated with chondrocyte hypertrophy were detected via immunohistochemistry, western blotting and reverse transcription-quantitative polymerase chain reaction. The results revealed that XAT inhibited the expression of various chondrocyte hypertrophic markers, including runt related transcription factor 2 (Runx2), matrix metalloproteinase 13 and collagen type X α1 chain. Further exploration indicated that XAT reduced the activation of p38-mitogen activated protein kinase and then increased the expression of histone deacetylase 4 to suppress Runx2. The findings indicated that XAT maintained the chondrocyte phenotype in regenerated cartilage and therefore may exhibit promise as a potential drug for the treatment of OA in the future.

It is reported that olivine-type lithium iron phosphate (LFP) for Li-ion batteries is one of the most widely utilized cathode materials, but its high-power applications are limited due to its intrinsically poor ion transfer rate and conductivity. Therefore, it is highly desired to fabricate LFP Li-ion capacitors (LICs) with high power performance and excellent cyclic reversibility, especially in safe, low cost and environmentally friendly aqueous electrolytes. Here, we fabricate LFP/expanded graphite (EG) nanocomposites by a one-step process, in which polyethylene glycol (PEG) is used as the particle growth inhibitor combined with vacuum infiltration of the LFP precursor into EG as a conductive sub-phase, and further investigate their high-power performance in aqueous LICs. Embedding spherical LFP nanoparticles with well-controlled size and agglomeration into the pores of EG and wrapping LFP nanoparticles by EG films contribute to the rapid electron and ion diffusion in LFP/EG composites, resulting in excellent cyclic reversibility and rate performance of LFP/EG composites. The aqueous LFP/EG//active carbon (AC) LICs were assembled in LiNO

Background & Aims Immune checkpoint inhibitors have some efficacy in the treatment of hepatocellular carcinoma (HCC). Programmed cell death 1 ligand 1 (PD-L1), expressed on some cancer cells, binds to the receptor programmed cell death 1 (PDCD1, also called PD1) on T cells to prevent their proliferation and reduce the antigen-tumor immune response. Immune cells that infiltrate some types of HCCs secrete interferon gamma (IFNG). Some HCC cells express myocyte enhancer factor 2D (MEF2D), which has been associated with shorter survival times of patients. We studied whether HCC cell expression of MEF2D regulates expression of PD-L1 in response to IFNG. Methods We analyzed immune cells from 20 fresh HCC tissues by flow cytometry. We analyzed 225 fixed HCC tissues (from 2 cohorts) from patients in China by immunohistochemistry and obtained survival data. We created mice with liver-specific knockout of MEF2D (MEF2DLPC-KO mice). We knocked out or knocked down MEF2D, E1A binding protein p300 (p300), or sirtuin 7 (SIRT7) in SMMC-7721, Huh7, H22, and Hepa1-6 HCC cell lines, some incubated with IFNG. We analyzed liver tissues from mice and cell lines by RNA sequencing, immunoblot, dual luciferase reporter, and chromatin precipitation assays. MEF2D protein acetylation and proteins that interact with MEF2D were identified by coimmunoprecipitation and pull-down assays. H22 cells, with MEF2D knockout or without (controls), were transplanted into BALB/c mice, and some mice were given antibodies to deplete T cells. Mice bearing orthotopic tumors grown from HCC cells, with or without knockout of SIRT7, were given injections of an antibody against PD1. Growth of tumors was measured, and tumors were analyzed by immunohistochemistry and flow cytometry. Results In human HCC specimens, we found an inverse correlation between level of MEF2D and numbers of CD4+ and CD8+ T cells; level of MEF2D correlated with percentages of PD1-positive or TIM3-positive CD8+ T cells. Knockout of MEF2D from H22 cells reduced their growth as allograft tumors in immune-competent mice but not in immune-deficient mice or mice with depletion of CD8+ T cells. When MEF2D-knockout cells were injected into immune-competent mice, they formed smaller tumors that had increased infiltration and activation of T cells compared with control HCC cells. In human and mouse HCC cells, MEF2D knockdown or knockout reduced expression of PD-L1. MEF2D bound the promoter region of the CD274 gene (encodes PD-L1) and activated its transcription. Overexpression of p300 in HCC cells, or knockout of SIRT7, promoted acetylation of MEF2D and increased its binding, along with acetylated histones, to the promoter region of CD274. Exposure of HCC cells to IFNG induced expression of p300 and its binding MEF2D, which reduced the interaction between MEF2D and SIRT7. MEF2D-induced expression of PD-L1 upon IFNG exposure was independent of interferon-regulatory factors 1 or 9. In HCC cells not exposed to IFNG, SIRT7 formed a complex with MEF2D that attenuated expression of PD-L1. Knockout of SIRT7 reduced proliferation of HCC cells and growth of tumors in immune-deficient mice. Compared with allograft tumors grown from control HCC cells, in immune-competent mice, tumors grown from SIRT7-knockout HCC cells expressed higher levels of PD-L1 and had reduced infiltration and activation of T cells. In immune-competent mice given antibodies to PD1, allograft tumors grew more slowly from SIRT7-knockout HCC cells than from control HCC cells. Conclusions Expression of MEF2D by HCC cells increases their expression of PD-L1, which prevents CD8+ T-cell–mediated antitumor immunity. When HCC cells are exposed to IFNG, p300 acetylates MEF2D, causing it to bind the CD274 gene promoter and up-regulate PD-L1 expression. In addition to promoting HCC cell proliferation, SIRT7 reduced acetylation of MEF2D and expression of PD-L1 in HCC cells not exposed to IFNG. Strategies to manipulate this pathway might increase the efficacy of immune therapies for HCC.

This study was aimed to investigate the effect of Forkhead Box G1 (FOXG1) on the epithelial-mesenchymal transition (EMT) of colorectal cancer (CRC) cells and the underlying mechanism. For this purpose, FOXG1 lentiviral interference (shRNA) plasmid and expression plasmid were constructed. Western blotting was used to analyze the expression of FOXG1 protein in five CRC cells, namely RKO, SW480, SW620, LoVo and DLD-1. The shRNA fragment of FOXG1 (shFOXG1) was designed and synthesized. Recombinant plasmids were obtained with the aid of DNA recombination technique. Double digestion and sequencing were used to identify the recombinant plasmids, and then lentivirus packaging, purification and stable transfection were carried out. Additionally, stable CRC cell lines were screened out. The changes of FOXG1 knockdown and overexpression efficiency, E-cadherin, Vimentin, Fibronectin, Snail, Twist mRNA and protein were investigated respectively by Western blotting and qRT-PCR analysis. Furthermore, the changes of cell morphology after knockdown and cell migration ability were evaluated respectively with optical microscopy, scratch test and Transwell assay. FOXG1 had the highest protein expression in RKO and the lowest in DLD-1 among the five CRC cells. Compared with those of the control group, the cell morphology in FOXG1 knockdown RKO group was changed from spindle into round or polygonal shape, cell polarization was enhanced and tight junction assembly was acclerated while cell migration distance was noticeably decreased. Moreover, the number of cells invaded and migrated through chambers was significantly reduced. Among these key factors of EMT, the expression of E-cadherin was increased while the expressions of Vimentin, Fibronectin, Snail and Twist were decreased. The opposite was the case in the overexpressed FOXG1 group. The overexpression of FOXG1 in CRC promoted the invasion and metastasis of CRC cells and played a crucial role in regulating the EMT. Thus, FOXG1 might be a novel therapeutic target in CRC treatment.构建叉头框G1 (Forkhead box G1, FOXG1) 的慢病毒干扰 (shRNA) 质粒及表达质粒，通过敲低和过表达FOXG1 探讨其对结直肠癌细胞上皮-间质转化EMT 的作用及其机制。应用Western blotting 检测FOXG1 在RKO、SW480、SW620、LOVO、DLD-1 五种结直肠癌细胞中蛋白的表达水平，设计并合成FOXG1 的shRNA 片段 (shFOXG1)，运用DNA 重组技术获得重组质粒，经双酶切技术及测序方法鉴定后进行慢病毒的包装、纯化及稳定转染，经筛选后获得稳定的结直肠癌细胞株，通过Western blotting 和qRT-PCR 技术检测FOXG1 敲低和过表达效率及EMT 关键因子E-cadherin、Vimentin、Fibronectin、Snail、Twist mRNA 和蛋白的变化，光学显微镜观察敲低后细胞形态学变化，通过划痕实验检测迁移能力变化，Transwell 检测侵袭迁移能力的变化。5 种结直肠癌细胞中，FOXG1 在RKO 细胞中蛋白表达量最高，而在DLD-1 细胞中表达量最低，与对照组相比较，在RKO细胞中敲低FOXG1，细胞形态由长梭型变成了类圆形或者多边形，细胞极性和紧密连接增加，细胞迁移距离明显降低，侵袭转移穿过小室的细胞数也明显减少，EMT 关键因子E-cadherin 表达增高，Vimentin、Fibronectin、Snail、Twist 表达降低，过表达FOXG1 组则相反。FOXG1 在结直肠癌中高表达，这种基因的高表达能够促进结直肠癌细胞的侵袭和转移，对结直肠癌细胞的EMT 起着重要的调控作用。.

Insufficient blood perfusion is one of the critical problems that hamper the clinical application of tissue engineering bone (TEB). Current methods for improving blood vessel distribution in TEB mainly rely on delivering exogenous angiogenic factors to promote the proliferation, migration, differentiation, and vessel formation of endothelial cells (ECs) and/or endothelial progenitor cells (EPCs). However, obstacles including limited activity preservation, difficulty in controlled release, and high cost obstructed the practical application of this strategy. In this study, TEB scaffold were modified with cerium oxide nanoparticles (CNPs) and the effects of CNPs existed at the scaffold surface on the growth and paracrine behavior of mesenchymal stem cells (MSCs) were investigated. The CNPs could improve the proliferation and inhibit the apoptosis of MSCs. Meanwhile, the interaction between the cell membrane and the nanoparticle surface could activate the calcium channel of MSCs leading to the rise of intracellular free Ca(2+) level, which subsequently augments the stability of HIF-1α. These chain reactions finally resulted in high expression of angiogenic factor VEGF. The improved paracrine of VEGF could thereby promote the proliferation, differentiation, and tube formation ability of EPCs. Most importantly, in vivo ectopic bone formation experiment demonstrated this method could significantly improve the blood vessel distribution inside of TEB.

Mesenchymal stem cells (MSCs) are widely used in cartilage tissue engineering to repair articular cartilage defects. However, hypertrophy of chondrocytes derived from MSCs might hinder the stabilization of hyaline cartilage. Thus, it is very important to find a suitable way to maintain the chondrogenic phenotype of chondrocytes. It has been reported that cordycepin has anti-inflammatory and anti-tumor functions. However, the role of cordycepin in chondrocyte hypertrophy remains unclear. Therefore, the objective of this study was to determine the effect of cordycepin on chondrogenesis and chondrocyte hypertrophy in MSCs and ATDC5 cells. Cordycepin upregulated chondrogenic markers including Sox9 and collagen type II while down-regulated hypertrophic markers including Runx2 and collagen type X. Further exploration showed that cordycepin promoted chondrogenesis through inhibiting Nrf2 while activating BMP signaling. Besides, cordycepin suppressed chondrocyte hypertrophy through PI3K/Bapx1 pathway and Notch signaling. Our results indicated cordycepin had the potential to maintain chondrocyte phenotype and reconstruct engineered cartilage. [BMB Reports 2016; 49(10): 548-553].

Bone homeostasis is maintained by the balance between osteoblastic bone formation and osteoclastic bone resorption. Osteoclasts are multinucleated cells derived from hematopoietic stem cells (HSCs) or monocyte/macrophage progenitor cells and formed by osteoclasts precursors (OCPs) fusion. Cyanidin is an anthocyanin widely distributed in food diet with novel antioxidant activity. However, the effect of cyanidin on osteoclasts is still unknown. We investigated the effect of cyanidin on RANKL-induced osteoclasts differentiation and cell fusion. The results showed that cyanidin had a dual effect on RANKL-induced osteoclastogenesis. Lower dosage of cyanidin ( 10 µg/ml) has an inhibitory effect. Fusogenic genes like CD9, ATP6v0d2, DC-STAMP, OC-STAMP, and osteoclasts related genes like NFATc1, mitf, and c-fos were all regulated by cyanidin consistent to its dual effect. Further exploration showed that low concentration of cyanidin could increase osteoclasts fusion whereas higher dosage of cyanidin lead to the increase of LXR-β expression and activation which is suppressive to osteoclasts differentiaton. All these results showed that cyanidin exhibits therapeutic potential in prevention of osteoclasts related bone disorders.

Bone is a dynamic organ continuously undergoing shaping, repairing and remodeling. The homeostasis of bone is maintained by the balance between osteoblastic bone formation and osteoclastic bone resorption. Osteoclasts (OCs) are specialized multinucleated cells derived from hematopoietic stem cells (HSCs) or monocytes/macrophage progenitor cells. There are different stages during osteoclastogenesis, and one of the most important steps to form functional osteoclasts is realized by cell-cell fusion. In our study, microarray was performed to detect the expression profiles of lncRNA, mRNA, circRNA and miRNA at different stages during osteoclastogenesis of RAW264.7 cells. Often changed RNAs were selected and clustered among the four groups with Venn analysis. The results revealed that expressions of 518 lncRNAs, 207 mRNAs, 24 circRNAs and 37 miRNAs were often altered at each stage during OC differentiation. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) biological pathway analysis were performed to predict the functions of differentially expressed lncRNAs and co-expressed potential targeting genes. Co-expression networks of lncRNA-mRNA and circRNA-miRNA were constructed based on the correlation analysis between the differentially expressed RNAs. The present study provided a systematic perspective on the potential function of non-coding RNAs (ncRNAs) during osteoclastogenesis.

DC-STAMP is a key regulating molecule of osteoclastogenesis and osteoclast precursor (OCP) fusion. Emerging lines of evidence showed that microRNAs play crucial roles in bone metabolism and osteoclast differentiation, but no microRNA has yet been reported to be directly related to OCPs fusion. Through a microarray, we found that the expression of miR-7b in RAW264.7 cells was significantly decreased after induction with M-CSF and RANKL. The overexpression of miR-7b in RAW264.7 cells attenuated the number of TRAP-positive cells number and the formation of multinucleated cells, whereas the inhibition of miR-7b enhanced osteoclastogenesis. Through a dual luciferase reporter assay, we confirmed that miR-7b directly targets DC-STAMP. Other fusogenic molecules, such as CD47, ATP6v0d2, and OC-STAMP, were detected to be down-regulated in accordance with the inhibition of DC-STAMP. Because DC-STAMP also participates in osteoclast differentiation through the ITAM-ITIM network, multiple osteoclast-specific genes in the ITAM-ITIM network were detected to identify how DC-STAMP is involved in this process. The results showed that molecules associated with the ITAM-ITIM network, such as NFATc1 and OSCAR, which are crucial in osteoclastogenesis, were consistently altered due to DC-STAMP inhibition. These findings suggest that miR-7b inhibits osteoclastogenesis and cell-cell fusion by directly targeting DC-STAMP. In addition, the inhibition of DC-STAMP and its downstream signals changed the expression of other fusogenic genes and key regulating genes, such as Nfatc1, c-fos, Akt, Irf8, Mapk1, and Traf6. In conclusion, our findings indicate that miR-7b may be a potential therapeutic target for the treatment of osteoclast-related bone disorders.

Bone homeostasis is maintained by the balance between osteoblastic bone formation and osteoclastic bone resorption. Osteoclasts are multinucleated cells derived from hematopoietic stem cells (HSCs) or monocyte/macrophage progenitor cells and formed by osteoclasts precursors (OCPs) fusion. Cyanidin is an anthocyanin widely distributed in food diet with novel antioxidant activity. However, the effect of cyanidin on osteoclasts is still unknown. We investigated the effect of cyanidin on RANKL-induced osteoclasts differentiation and cell fusion. The results showed that cyanidin had a dual effect on RANKL-induced osteoclastogenesis. Lower dosage of cyanidin ( 1 µg/ml) has a promoting effect on osteoclastogenesis while higher dosage of cyanidin ( 10 µg/ml) has an inhibitory effect. Fusogenic genes like CD9, ATP6v0d2, DC-STAMP, OC-STAMP, and osteoclasts related genes like NFATc1, mitf, and c-fos were all regulated by cyanidin consistent to its dual effect. Further exploration showed that low concentration of cyanidin could increase osteoclasts fusion whereas higher dosage of cyanidin lead to the increase of LXR-β expression and activation which is suppressive to osteoclasts differentiaton. All these results showed that cyanidin exhibits therapeutic potential in prevention of osteoclasts related bone disorders.