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Programmed cell death plays a critical role in the progression of spinal cord injury (SCI). Autophagy is a protective factor for controlling neuronal damage, while necroptosis promotes further cell death and neuroinflammation after SCI. DADLE (d-Ala2, d-Leu5) is a selective agonist for delta opioid receptor (DOR) and has been identified as a promising drug for its neuroprotective effects. Our present work aims to investigate the therapeutic effect of DADLE on locomotive function recovery following SCI and its concrete mechanism. By establishing a mouse model of spinal cord contusion injury and using functional behavioural assessment, our results showed that DADLE promoted functional recovery after SCI. Through experimental methods such as western blotting and immunofluorescence, we found that DADLE promoted autophagic flux and inhibited necroptosis. Then, analysis of the enzyme activity of NAG and related protein expression of CTSD and CTSB in lysosomes and cytoplasm revealed that DADLE decreased lysosomal membrane permeabilization (LMP). The autophagy inhibitor CQ reversed the protective effect of inhibiting necroptosis. Further analysis identified that DADLE decreased phosphorylated cPLA2, and network pharmacology analysis revealed that the AMPK (Adenosine monophosphate-activated protein kinase) signalling pathway may be involved in the therapeutic effect of DADLE. Finally, blocking the interaction between DOR and DADLE by using naltrindole abolished the anti-phosphorylation effect of DADLE on cPLA2 and p38, resulting in a decrease in autophagic markers and an increase in necroptosis and LMP markers. Altogether, our study indicated that DADLE promotes autophagic flux and inhibits necroptosis by decreasing LMP by interacting with DOR and then activating the AMPK/SIRT1/P38/cPLA2 pathway after SCI, which may have potential clinical application value in the future.

The ternary component NiCoAl-layered double hydroxide (NiCoAl-LDH) and carbon nanotube (CNT) nano-composite (CNT/NiCoAl-LDH) were successfully prepared by a simple hydrothermal method. The NiCoAl-LDH nanosheets were effectively and uniformly grown on CNTs, forming a cross-linked conductive network structure, and stainless steel (SS) mesh was used as the base to load CNT/NiCoAl-LDH for microbial fuel cell (MFC) cathode. X-ray diffraction (XRD) results presented that the CNT/NiCoAl-LDH hybrid exhibited the (003), (006), (012), (015), (018), (110) and (113) crystal planes of hydrotalcite reflection. The surface functional groups C-O, C=O, C-H, C-N and M-O of the hybrid were confirmed. The cross-linked network structure of the hybrid was observed and the content and proportion of each element of the hybrid were found. CNT/NiCoAl-LDH showed excellent catalytic oxygen reduction reaction (ORR) ability by cyclic voltammetry (CV) and linear voltammetry (LSV) due to its abundant electrochemical active sites and excellent conductivity. The maximum output voltage of CNT/NiCoAl-LDH catalyst as MFC cathode was 450 mV, the maximum power density was 433.5 ± 14.8 mW/m2, and the maximum voltage stabilization time was 7–8 d. The results indicated that the CNT/NiCoAl-LDH hybrid was full potential as a high-performance, low-cost MFC cathode catalyst in future.

In recent years, a large number of studies have reported that neuroinflammation aggravates the occurrence of secondary injury after spinal cord injury. Gramine (GM), a natural indole alkaloid, possesses various pharmacological properties; however, the anti‐inflammation property remains unclear. In our study, Gramine was investigated in vitro and in vivo to explore the neuroprotection effects. In vitro experiment, our results suggest that Gramine treatment can inhibit release of pro‐inflammatory mediators. Moreover, Gramine prevented apoptosis of PC12 cells which was caused by activated HAPI microglia, and the inflammatory secretion ability of microglia was inhibited by Gramine through NF‐κB pathway. The in vivo experiment is that 80 mg/kg Gramine was injected orthotopically to rats after spinal cord injury (SCI). Behavioural and histological analyses demonstrated that Gramine treatment may alleviate microglia activation and then boost recovery of motor function after SCI. Overall, our research has demonstrated that Gramine exerts suppressed microglia activation and promotes motor functional recovery after SCI through NF‐κB pathway, which may put forward the prospect of clinical treatment of inflammation‐related central nervous diseases.

T follicular helper (Tfh) cells, one of the subsets of CD4

Combating the accumulated senescent cells and the healing of osteoporotic bone fractures in the older remains a significant challenge. Nicotinamide mononucleotide (NMN), a precursor of NAD+, is an excellent candidate for mitigating aging-related disorders. However, it is unknown if NMN can alleviate senescent cell induction and enhance osteoporotic bone fracture healing. Here we show that NMN treatment partially reverses the effects of tumor necrosis factor-alpha (TNF-α) on human primary osteoblasts (HOBs): senescent cell induction, diminished osteogenic differentiation ability, and intracellular NAD+ and NADH levels. Mechanistically, NMN restores the mitochondrial dysfunction in HOBs induced by TNF-α evidenced by increased mitochondrial membrane potential and reduced reactive oxidative species and mitochondrial mass. NMN also increases mitophagy activity by down-regulating P62 expression and up-regulating light chain 3B-II protein expression. In addition, the cell senescence protective effects of NMN on HOBs are mitigated by a mitophagy inhibitor (Bafilomycin A1). In vivo, NMN supplementation attenuates senescent cell induction in growth plates, partially prevents osteoporosis in an ovariectomized mouse model, and accelerates bone healing in osteoporotic mice. We conclude that NMN can be a novel and promising therapeutic candidate to enhance bone fracture healing capacity in the older.

To enhance the oxygen reduction reaction (ORR) activity and power generation capacity of a microbial fuel cell (MFC), MIL-53(Fe) (Fe-based Materials of Institute Lavoisier) as the electrochemical catalyst was synthesized using the hydrothermal method. The catalytic structure and morphology of all materials were comprehensively characterized by Fourier Transform infrared spectrometer (FTIR), X-ray diffraction (XRD), scanning electron microscope (SEM) and energy dispersive spectrometer (EDS). The results show that there were many nanopores on MIL-53(Fe), which improved the electrocatalytic activity. The MIL-53(Fe)-modified air cathode MFC had a voltage output of approximately 0.37 V and maintained that output for one week. The maximum power density was 397 ± 6.3 mW/m2. MIL-53(Fe) was an excellent electrochemical catalyst, significantly enhancing the catalytic oxygen reduction ability and promoting the power output of the MFC. This study provides a method to apply MIL-53(Fe) materials in microbial fuel cells.

BackgroundPrimary Sjogren’s syndrome (pSS) is a slowly progressive, inflammatory autoimmune disease characterized by lymphocytic infiltration into salivary and lacrimal glands. It becomes more recognized that morphology alterations of epithelial mitochondria are involved in altered cellular bioenergetics in pSS patients. The integrated analysis of the mitochondrial role in the pathogenesis and aberrant immune microenvironment in pSS remains unknown.MethodsThe mitochondria-related genes and gene expression data were downloaded from the MitoMiner, MitoCarta, and NCBI GEO databases. We performed novel transcriptomic analysis and constructed a network between the mitochondrial function and immune microenvironment in pSS-salivary glands by computer-aided algorithms. Subsequently, real-time PCR was performed in clinical samples in order to validate the bioinformatics results. Histological staining and transmission electron microscopy (TEM) were further studied on labial salivary gland samples of non-pSS and pSS patients characterized for mitochondria-related phenotypic observation in the different stages of the disease.ResultsThe bioinformatic analysis revealed that the expression of several mitochondria-related genes was altered in pSS. Quantitative real-time PCR showed that four hub genes, CD38, CMPK2, TBC1D9, and PYCR1, were differentially expressed in the pSS clinical samples. These hub genes were associated with the degree of immune cell infiltration in salivary glands, the mitochondrial respiratory chain complexes, mitochondrial metabolic pathway in gluconeogenesis, TCA cycle, and pyruvate/ketone/lipid/amino acid metabolism in pSS. Clinical data revealed that the gene expression of fission (Fis1, DRP1, and MFF) and fusion (MFN1, MFN2, and OPA1) was downregulated in pSS samples, consistent with the results from the public validation database. As the disease progressed, cytochrome c and Bcl-2 proteins were regionally distributed in salivary glands from pSS patients. TEM revealed cytoplasmic lipid droplets and progressively swollen mitochondria in salivary epithelial cells.ConclusionOur study revealed cross talk between mitochondrial dysfunction and the immune microenvironment in salivary glands of pSS patients, which may provide important insights into SS clinical management based on modulation of mitochondrial function.

BackgroundPrimary Sjögren’s syndrome (pSS) is a chronic systemic autoimmune disease of the exocrine glands characterized by specific pathological features. Previous studies have pointed out that salivary glands from pSS patients express a unique profile of cytokines, adhesion molecules, and chemokines compared to those from healthy controls. However, there is limited evidence supporting the utility of individual markers for different stages of pSS. This study aimed to explore potential biomarkers associated with pSS disease progression and analyze the associations between key genes and immune cells.MethodsWe combined our own RNA sequencing data with pSS datasets from the NCBI Gene Expression Omnibus (GEO) database to identify differentially expressed genes (DEGs)viabioinformatics analysis. Salivary gland biopsies were collected from 14 pSS patients, 6 non-pSS patients, and 6 controls. Histochemical staining and transmission electron micrographs (TEM) were performed to macroscopically and microscopically characterize morphological features of labial salivary glands in different disease stages. Then, we performed quantitative PCR to validate hub genes. Finally, we analyzed correlations between selected hub genes and immune cells using the CIBERSORT algorithm.ResultsWe identified twenty-eight DEGs that were upregulated in pSS patients compared to healthy controls. These were mainly involved in immune-related pathways and infection-related pathways. According to the morphological features of minor salivary glands, severe interlobular and periductal lymphocytic infiltrates, acinar atrophy and collagen in the interstitium, nuclear shrinkage, and microscopic organelle swelling were observed with pSS disease progression. Hub genes based on above twenty-eight DEGs, including MS4A1, CD19, TCL1A, CCL19, CXCL9, CD3G, and CD3D, were selected as potential biomarkers and verified by RT-PCR. Expression of these genes was correlated with T follicular helper cells, memory B cells and M1 macrophages.ConclusionUsing transcriptome sequencing and bioinformatics analysis combined with our clinical data, we identified seven key genes that have potential value for evaluating pSS severity.

In this study, a simple two-step hydrothermal method was used to prepare the cathode catalyst of the microbial fuel cell (MFC). NiCoAl- layered double hydroxide (LDH) nanosheets were grown vertically on multi-wall carbon nanotubes (MWCNTs) in situ; Ni-catecholate-based metal organic framework (Ni-CAT MOF) were modified on the surface of the nanosheets. The maximum output voltage of Ni-CAT/NiCoAl-LDH/MWCNTs was 475 mV, the maximum stabilization time was 8 d, the maximum output power was 448.5 ± 12.0 mW/m2, which was 1.03 times that of NiCoAl-LDH/MWCNT-MFC (433.5 ± 14.8 mW/m2) and 1.35 times of NiCoAl-LDH- MFC (329.9 ± 2.9 mW/m2). The layer structure of LDH, conductivity of Ni-CAT and MWCNT improved the flow efficiency of ions between layers and effectively reduced transmission resistance, and these have effectively enhanced the cycle stability and power generation efficiency of the electrode.

Our previous work indicated that fibrinogen-related protein (AjFREP) from sea cucumber (Apostichopus japonicus) plays important roles in innate immunity. To understand AjFREP expression regulation in A. japonicus, we cloned the AjFREP promoter and characterized the putative transcription-factor binding motifs. The AjFREP promoter region spans 1365bp, containing several transcription-factor binding sites. The full-length sequence and all truncated fragments exhibited high promoter activity in HEK-293 cells. Luciferase activity significantly increased for P1(-1365/+16) after LPS exposure, suggesting that the promoter responded to LPS. We also found that two potential NF-κB binding sites were involved in the promoter region, and co-transfection assay demonstrated that the first binding site was necessary for AjFREP transcription. The expression of AjNF-κB/Rel after AjFREP knock-down followed by LPS injection in vivo was further investigated. We found that AjNF-κB/Rel transcript significantly decreased after silencing AjFREP with LPS challenge compared with the control at 4 and 8h, suggesting that activated AjFREP in turn affected the NF-κB pathway under immune responses. These results provided novel insights into the activation mechanism of AjFREP, i.e., that selectively changing AjFREP expression may prevent pathogen infection in A. japonicus.

Bone fractures and critical-sized bone defects present significant health threats for the elderly who have limited capacity for regeneration due to the presence of functionally compromised senescent cells. A wide range of synthetic materials has been developed to promote the regeneration of critical-sized bone defects, but it is largely unknown if a synthetic biomaterial (scaffold) can modulate cellular senescence and improve bone regeneration in aged scenarios. The current study investigates the interaction of Baghdadite (Ca3ZrSi2O9) ceramic scaffolds with senescent human primary osteoblast-like cells (HOBs) and its bone regeneration capacity in aged rats. A senescent HOB model was established by repeatedly passaging HOBs till passage 7 (P7). Compared to the clinically used hydroxyapatite/tricalcium phosphate (HA/TCP), Baghdadite prevented senescence induction in P7 HOBs and markedly negated the paracrine effect of P7 HOB secretomes that mediated the up-regulations of cellular senescence-associated gene expression levels in P2 HOBs. We further demonstrated that conditioned media extracted from Baghdadite corrected the dysfunctional mitochondria in P7 HOBs. In vivo, the bone regeneration capacity was enhanced when 3D printed Baghdadite scaffolds were implanted in a calvaria critical-sized bone defect model in both young and aged rats compared to HA/TCP scaffolds, but a better effect was observed in aged rats than in young rats. This study suggests that Baghdadite ceramic represents a novel and promising biomaterial approach to promote bone regeneration capacity in the elderly by providing an anti-senescent microenvironment.

With the well improvement of motor and power electronic technology, the technology of Electro-mechanical actuator (EMA)has come into a new stage. A high-reliability driving and controlling technology with a double redundancy feedback loop is described in this paper. In this method, the position of electromechanical actuator and the resolver position of motor can be back up for each other by some compensate algorithms. This method can improve the reliability of the control system without extra hardware and increasing cost.

The improvement of cathode performance has always been the bottleneck and research hot spot for microbial fuel cells (MFCs). An Fe

This work studied bacterial community structure and gene function prediction in long-term running of dual graphene modified bioelectrode bioelectrochemical systems (LT D-GM-BE BES, 2 year). The maximum power density of LT D-GM-BE BES was 99.03 ± 3.64 mW/m2, which was 3.66 times of dual control BES (D-C-BE BES), and the transfer resistance of LT GM-BE was just approximately 1/4 of control bioelectrode (C-BE). Proteobacteria and Firmicutes were dominant bacteria in long-term modified bioanode (LT GM-BA, 30.03% and 45.64%), and in long-term modified biocathode (LT GM-BC) was Armatimonadetes (47.14%) in phylum level. The dominant bacteria in LT GM-BA was Clostridium (30.56%), in GM-BC was Chthonomonas (47.14%) in genus level. Gene function related with substrate, energy metabolism and environmental adaptation were enriched. LT GM-BE was tended to enrich dominant bacteria and enrich gene to adapt to micro-environmental changes. This study would provide metagenomics information for long-term running of BES in future.

Angiogenesis mediated by vascular endothelial growth factor (VEGF) is known to play an important role in regulating cartilage remodelling and endochondral ossification. However, the details of how VEGF signalling mechanisms affect condyle remodelling in response to alterations in functional loading remains unclear. To explore this, eighty 16‐day‐old male SD rats were divided into two equal groups which were fed either a soft/powdery diet or a hard diet for 4 weeks; the stiffness of the diet results in alteration of mastication force and hence temporomandibular joint (TMJ) development. We performed a proteomic analysis of rat condylar cartilage using isobaric tags for relative and absolute quantification (iTRAQ) labelling, followed by 2D nano‐high performance liquid chromatography and MALDI‐TOF/time‐of‐flight technology. After protein identification, we used biological information analysis to identify the differentially expressed proteins associated with the VEGF signalling pathway. Among the identified differentially expressed proteins, we found VEGF signalling mainly via the p44/42 MAPK and p38 mitogen‐activated protein kinase (MAPK) pathways in condylar cartilage, including VEGFD, VGFR2, KPCB, KPCT, KPCZ, ARAF, RASN, PLCG2, PLCG1, JUN and M3K12. Furthermore, four representative protein candidates, VEGF, p38 MAPK and p44/42 MAPK/phospho‐p44/42 MAPK, were confirmed by immunohistochemical staining and western blot. Our data suggest that VEGF might play an important role in TMJ development and remodelling in response to alterations in functional loading through the p44/42 MAPK and p38 MAPK signalling pathway. This study provides new clues to the understanding of the signalling mechanism responsible for VEGF production in response to different masticatory functions at the protein level.

The improvement of cathode performance has always been the bottleneck and research hot spot for microbial fuel cells (MFCs). An Fe3O4@NiFe-LDH composite with a nanoscale core–shell structure containing an Fe3O4 magnetic core and a layered double hydroxide (LDH) shell was prepared by the hydrothermal method. The Fe3O4@NiFe-LDH was characterized by FT-IR, XRD, SEM and EDS. The characterization results showed that the composite had a unique cauliflower-like nanoflake structure and special pore size distribution, which greatly improved the ORR performance. Moreover, the use of the synthesized Fe3O4@NiFe-LDH core–shell structure as an electrode in an MFC was characterized by CV and LSV, which showed that the Fe3O4@NiFe-LDH exhibited excellent ORR catalytic properties. The voltage output of the Fe3O4@NiFe-LDH MFC was maintained at approximately 0.39 V, with insignificant variations over 110 h. The maximum power density was 211.40 ± 2.27 mW/m2, which was 34 times that of the blank control group MFC and was caused by the many electroactive sites, good rate capability and remarkable cycling stability of LDH. This study provides the possibility for using Fe3O4@NiFe-LDH in cathodes to operate continuously and at low cost in fuel cells.

Aberrant promoter methylation of multiple genes is associated with various diseases, including Alzheimer's disease (AD). The goal of the present study was to determine whether dopamine receptor D4 (DRD4) promoter methylation is associated with AD. In the current study, the methylation levels of the DRD4 promoter were measured in 46 AD patients and 61 controls using bisulfite pyrosequencing technology. The results of the present study demonstrated that DRD4 promoter methylation was significantly higher in AD patients than in controls. A further breakdown analysis by gender revealed that there was a significant association of DRD4 promoter methylation with AD in males (23 patients and 45 controls). In conclusion, the results of the present study demonstrated that elevated DRD4 promoter methylation was associated with AD risk in males.

To improve the activity of oxygen reduction reaction (ORR) of microbial fuel cell (MFC), the cathode was modified by using NiFe layered double hydroxide (LDH)@Co3O4 as catalyst to increase the power generation of MFC. The unique NiFe-LDH@Co3O4 core-shell structure was prepared by using NiFe-LDH as metal source, Co3O4 as support matrix and urea-doped nitrogen in LDH. XRD, FTIR, SEM and EDS showed that LDH was successfully grown on the surface of Co3O4, and had typical LDH nanostructure. The voltage output of NiFe-LDH@Co3O4-MFC was maintained at about 0.43 V, with insignificant variations over 8 d, which indicated that NiFe-LDH@Co3O4 catalyst has remarkable stability and durability. Moreover, the maximum power density was 467.35 ± 8.27 mW/m2, which was produced by NiFe-LDH@Co3O4. The results implied that the improvement of ORR activity of NiFe-LDH@Co3O4 was due to its high conductivity and abundant active sites, and therefore, the power generation capacity of MFC was improved. The electrode prepared by NiFe-LDH@Co3O4 modified air cathode was a promising material for fuel cell.

Periodontitis is a chronic, destructive inflammatory disease that injures tooth- supporting tissues, eventually leading to tooth loss. Complete eradication of periodontal pathogenic microorganisms is fundamental to allow periodontal healing and commonly precedes periodontal tissue regeneration. To address this challenge, we report a strategy for developing an enzyme-mediated periodontal membrane for targeted antibiotic delivery into infectious periodontal pockets; the unique components of the membrane will also benefit periodontal alveolar bone repair. In this approach, a chitosan membrane containing polyphosphoester and minocycline hydrochloride (PPEM) was prepared. Physical, morphological, and ultrastructural analyses were carried out in order to assess cellular compatibility, drug release and antibacterial activity in vitro. Additionally, the functionality of the PPEM membrane was evaluated in vivo with a periodontal defect model in rats. The results confirm that the PPEM membrane exhibits good physical properties with excellent antibacterial activity and successfully promotes periodontal tissue repair, making it promising for periodontal treatment.

Interleukin 1 receptor-associated kinase 4 (IRAK4) is a key factor in TLR-mediated host immune function. In this study, an IRAK4 homologue molecule was identified from Apostichopus japonicus (designated as AjIRAK4) by RACE approach. The full-length cDNA of AjIRAK4 was of 2024 bp containing an open reading frame of 1311 bp encoding a 436-amino-acid (aa) residue polyprotein with the typical death domain (10-113aa) and the kinase domain (160-426aa). The mRNA transcripts of AjIRAK4 displayed constitutively expressed in all examined tissues with highest expression in the muscles (7.20-fold increase compared to the coelomocytes). The pathogen Vibrio splendidus challenge and LPS exposure could both significantly up-regulate the mRNA expression of AjIRAK4. Silencing AjIRAK4 in vitro and in vivo could inhibit the expression of TLR members at mRNA and protein levels, suggesting AjIRAK4 was an important component of TLR cascade in sea cucumber. More importantly, knockdown of AjIRAK4 by specific siRNA resulted in the significant promotion of coelomocyte apoptosis by 1.82-fold increase in vitro and 1.95-fold in vivo. Taken together, all our results suggested that AjIRAK4 might be served as coelomocyte apoptosis regulator via TLR cascade.

Fibrinogen-related proteins (FREPs) play important roles in innate immunity by recognizing pathogen associated molecular patterns on pathogenic bacteria surfaces via conserved fibrinogen-like domain (FBG). In this paper, the full-length cDNA of Apostichopus japonicus FREP (designated as AjFREP) was cloned combined with rapid amplification of cDNA ends (RACE) and transcriptome sequencing. The full-length cDNA of AjFREP was of 2110 bp with an open reading frame (ORF) of 1659 bp. SMART analysis revealed that the AjFREP contained a typical signal peptide of 19 amino acid residues, a FBG and two unusual epidermal growth factor-like domains (EGFs). Multiple sequence alignments suggested that FBG domain shared a remarkably high structural conservation in polypeptide binding site and Ca2+ binding site. Tissue distribution analysis revealed that AjFREP was constitutively expressed in all examined tissues with the largest magnitude in coelomocytes, indicating AjFREP might play an important role in immune defense. The mRNA level of AjFREP in coelomocytes was sharply up-regulated by Vibrio splendidus challenge, and reached its peak expression at 48 h. Knock-down AjFREP by specific siRNA could significantly repress the coelomocyte phagocytosis rate. Meantime, the survival number of V. splendidus in the coelomic fluid was promoted. All these current results indicated that AjFREP might be involved in pathogen clearance through mediating coelomocytes phagocytosis activity.

The decreased bone mass and impaired osteogenesis capacities that occur with aging may influence the outcome of dental implants . Parathyroid hormone (PTH) (1–34) is an anabolic agent for the treatment of osteoporosis. However, little is known about its effects and mechanisms on vascularized bone regeneration and implant osseointegration in aging. In current study, we adopted both in vivo and in vitro approaches to explore the mechanisms of early actions of PTH (1–34) on the angiogenic and osteogenic microenvironment to enhance implant osseointegration in aged rats. Daily subcutaneous injections of 30 μg/kg PTH (1–34) were given to female rats aged 20 months beginning on next day of implantation and lasting for 5 weeks. Radiological and histological analysis confirmed that PTH (1–34) improved new bone formation, angiogenesis and implant osseointegration in aged rats in the early stage. The osteogenic potential of aged bone mesenchymal stem cells (BMSCs) was enhanced, while their adipogenesis capacity was attenuated. Furthermore, PTH (1–34) was shown to promote angiogenesis directly via endothelial cell migration and blood vessel formation in vitro. Meanwhile, PTH (1–34) stimulated more osteoclasts participation in bone remodeling by secreting angiogenic and osteogenic growth factors to induce early vascularization and stimulate the migration or differentiation of BMSCs indirectly. Together, these results demonstrate mechanistic insight into how PTH (1–34) regulates the angiogenic and osteogenic microenvironment to result in more active bone remodeling and new bone formation, making it an excellent potential therapeutic agent for rapid vascularized bone regeneration and implant osseointegration in the aged population.

Bis(propyl)-cognitin (B3C), derived from tacrine linked with three methylene (-CH2-) groups, is a dimerized molecule interacting multiple targets. During the past several years, it has been reported as a promising therapeutic drug for Alzheimer's disease (AD) and other neurodegenerative disorders. However, the therapeutic mechanism of B3C for AD needs further demonstration. Based on a combination of behavioral tests, electrophysiological technique, immunocytochemistry, and live cell imaging, we studied the effects and the underlying mechanism of B3C on the impairments of cognitive function, synapse formation, and synaptic plasticity induced by soluble amyloid-β protein (Aβ) oligomers. Our study showed that spatial learning and memory in a Morris water maze task and recognition memory in a novel object recognition task were significantly decreased in the AD model mice created by hippocampal injection of Aβ. Chronic administration of B3C for 21 days prevented the memory impairments of the AD model mice in a dose-dependent manner. Live cell imaging study showed that 2-h pretreatment of B3C prevented the decrease in the number of filopodia and synapses induced by Aβ (0.5 μM) in a dose-dependent manner. Besides, electrophysiological recording data showed that the inhibition of long-term potentiation (LTP) induced by Aβ1-42 oligomers in the dentate gyrus (DG) of hippocampus was prevented by B3C in a dose-dependent manner. Furthermore, we found that the neuroprotective effect of B3C against Aβ-oligomer-induced impairments of synaptic formation and plasticity could be partially blocked by a specific phosphatidylinositol 3-kinase (PI3-K) inhibitor LY294002 (50 μM). Therefore, these results indicate that B3C can prevent Aβ-oligomer-induced cognitive deficits, synaptic formation impairments, and synaptic plasticity impairments in a concentration-dependent manner. These effects of B3C are partially mediated via the PI3-K pathway. This study provides novel insights into the cellular mechanisms for the protective effects of B3C on AD.

Deposits of amyloid-β (Aβ) protein are one of the hallmarks of Alzheimer's disease (AD). Numerous studies report that the Aβ peptide, especially in the oligomeric form, causes memory decline and other cognitive deficits. However, there have been very few effective interventions for termination or even delay of AD progression. Brilliant Blue G (BBG), a safe triphenylmethane dye and P2X7 antagonist, has been reported to have protective effects on neuroinflammation, ischemia, spinal injury and neurodegenerative disorders. Here we report that systematic administration of BBG diminishes spatial memory impairment and cognitive deficits in a mouse AD model produced by injecting soluble Aβ peptide into the hippocampal CA1 region. In addition, we show that Aβ-induced loss of filopodia and spine density in cultured hippocampal neurons was prevented by administration of BBG. We conclude that BBG prevents the learning and memory impairment and cognitive deficits induced by the toxicity of soluble Aβ, and improves the development of dendritic spines in hippocampal neurons in an AD model mouse. Considering the safety and blood-brain-barrier (BBB)-permeability of BBG, our data suggest a potential for BBG as a new therapy for AD.

Aberrant DNA methylation has been observed in the patients with Alzheimer's disease (AD), a common neurodegenerative disorder in the elderly. OPRD1 encodes the delta opioid receptor, a member of the opioid family of G-protein-coupled receptors. In the current study, we compare the DNA methylation levels of OPRD1 promoter CpG sites (CpG1, CpG2, and CpG3) between 51 AD cases and 63 controls using the bisulfite pyrosequencing technology. Our results show that significantly higher CpG3 methylation is found in AD cases than controls. Significant associations are found between several biochemical parameters (including HDL-C and ALP) and CpG3 methylation. Subsequent luciferase reporter gene assay shows that DNA fragment containing the three OPRD1 promoter CpGs is able to regulate gene expression. In summary, our results suggest that OPRD1 promoter hypermethylation is associated with the risk of AD.

Objective To evaluate microarchitectural changes in condylar cartilage and associated subchondral bone after bisphosphonates treatment using an ovariectomized (OVX) osteoporosis rat model. Methods Thirty six-month-old female Sprague-Dawley rats were randomly divided into sham, OVX, and risedronate (RIS)-treated groups. Both OVX and RIS groups received bilateral ovariectomy. OVX group was treated subcutaneously with saline, whereas RIS group received risedronate treatment (2.4 μg/kg) subcutaneously for 3 months. At the end of 3 months, animals were sacrificed and the entire condyles were harvested for micro-CT and histological analyses. Immunohistochemistry (IHC) was performed to assess the expression of type I/II collagen protein by semiquantitative imaging analysis. Results Micro-CT analysis showed OVX group had significant condylar subchondral bone loss compared to sham as shown by significant decrease in bone volume fraction (P = 0.028), trabecular thickness (P = 0.041), and significant increase in trabecular spacing (P = 0.003). In RIS group, partial inhibition of OVX-induced bone loss was detected. HE staining showed proliferative layer of condylar cartilage reduced, while hypertrophic chondrocyte layer increased significantly in RIS group compared to sham and OVX groups. IHC showed reduced expression of Col I in both the OVX and RIS groups, whereas expression of Col II was reduced in the OVX group but increased in the RIS group. Conclusion Our findings suggest that systemic bisphosphonate treatment influences the structure and ossification of condylar cartilage and it has a dual action on condyle in a postmenopausal osteoporosis rat model which raises the concerns for the potential side effects of BPs on condyle to elder patients.

Background Recent studies have shown Growth differentiation factor–15 (GDF-15) that is a member of the transforming growth factor β (TGF-β) superfamily might be a potential predictive cytokine for the prognosis of Acute coronary syndrome (ACS). However, there are discrepancies in these studies. Methods Publication searches of the PubMed/Medline and EMBASE databases were performed without any time or ethnicity restrictions. The inclusion and exclusion criteria, when clear, were addressed. Random effects models were used for all analyses. Publication bias was tested using funnel plots and the Egger test. Results We identified eight eligible studies that provided mortality data. Five of these studies provided recurrent myocardial infarction (MI) data. The maximal duration of follow-up ranged from 6 months to 6 years. A significant association was found between the patients with the highest and lowest GDF-15 levels (overall analyses) in terms of mortality (p