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Background: Alarmin cytokine IL-25 promotes type 2 inflammatory responses in disorders such as asthma and chronic rhinosinusitis with nasal polyps (CRSwNP) and known targets include ILC2 and Th2 cells. However, other cellular targets for IL-25 remain poorly defined., Objective: To investigate induction and expression of IL-25 receptor (IL-17RB) by B cells and evaluate responsiveness of IL-17RB-expressing B cells to IL-25 in vitro., Methods: IL-17RB expression, regulation and function on B cells were evaluated in peripheral blood-derived B cells by flow cytometry and RT-PCR, including in response to IgE-inducing stimuli (anti-CD40 mAb and IL-4). Single-cell RNA sequencing was used to compare IL-17RB+ and IL-17RB-activated peripheral blood-derived B cells. To evaluate B cell IL-17RB expression within type 2 inflamed tissue, B cells were compared from nasal polyps, control turbinate tissue and matched peripheral blood., Results: Activation of B cells with anti-CD40 and IL-4 increased IL-17RB expression at both protein and mRNA level, which was further upregulated by IL-25. B cells induced to express IL-17RB responded to IL-25 with enhanced antibody production. Single-cell RNA-sequencing showed that IL17RB+ activated B cells expressed higher levels of IGHE, CCL17 and CCL22 compared to IL17RB- B cells. B cells from nasal polyp tissue expressed higher levels of surface IL-17RB compared with control tissue, correlating with patient-reported CRSwNP severity (SNOT-22)., Conclusion: Peripheral blood B cells activated under IgE-inducing conditions express surface IL-17RB, and tissue IL-17RB+ B cells are increased in type 2 inflammation. IL-17RB+ cells have a distinct transcriptional profile and respond to IL-25 with enhanced antibody production, highlighting the IL-25/IL-17RB pathway as a potential therapeutic target for CRSwNP and other type 2 inflammatory disorders., (© 2025 The Author(s). Allergy published by European Academy of Allergy and Clinical Immunology and John Wiley & Sons Ltd.)

Chronic obstructive pulmonary disease (COPD) is a chronic respiratory condition characterized by persistent inflammation and oxidative stress, which ultimately leads to progressive restriction of airflow. Extensive research findings have cogently suggested that the dysregulation of essential transition metal ions, notably iron, copper, and zinc, stands as a critical nexus in the perpetuation of inflammatory processes and oxidative damage within the lungs of COPD patients. Unraveling the intricate interplay between metal homeostasis, oxidative stress, and inflammatory signaling is of paramount importance in unraveling the intricacies of COPD pathogenesis. This comprehensive review aims to examine the current literature on the sources, regulation, and mechanisms by which metal dyshomeostasis contributes to COPD progression. We specifically focus on iron, copper, and zinc, given their well-characterized roles in orchestrating cytokine production, immune cell function, antioxidant depletion, and matrix remodeling. Despite the limited number of clinical trials investigating metal modulation in COPD, the advent of emerging methodologies tailored to monitor metal fluxes and gauge responses to chelation and supplementation hold great promise in unlocking the potential of metal-based interventions. We conclude that targeted restoration of metal homeostasis represents a promising frontier for ameliorating pathological processes driving COPD progression.

While the formation of an inorganic-rich solid electrolyte interphase (SEI) plays a crucial role, the persistent challenge lies in the formation of an organic-rich SEI due to the high solvent ratio in low-concentration electrolytes (LCEs), which hinders the achievement of high-performance lithium metal batteries. Herein, by incorporating di-fluoroethylene carbonate (DFEC) as a non-solvating cosolvent, a solvation structure dominated by anions is introduced in the innovative LCE, leading to the creation of a durable and stable inorganic-rich SEI. Leveraging this electrolyte design, the Li||NCM83 cell demonstrates exceptional cycling stability, maintaining 82.85% of its capacity over 500 cycles at 1 C. Additionally, Li||NCM83 cell with a low N/P ratio (≈2.57) and reduced electrolyte volume (30 µL) retain 87.58% of its capacity after 150 cycles at 0.5 C. Direct molecular information is utilized to reveal a strong correlation between solvation structures and reduction sequences, proving the anion-dominate solvation structure can impedes the preferential reduction of solvents and constructs an inorganic-rich SEI. These findings shed light on the pivotal role of solvation structures in dictating SEI composition and battery performance, offering valuable insights for the design of advanced electrolytes for next-generation lithium metal batteries., (© 2024 Wiley‐VCH GmbH.)

Accurate climate projections are critical for various applications and impact assessments in environmental science and management. This study presents HiCPC (High-resolution CMIP6 downscaled daily Climate Projections over China), a novel dataset tailored to China's specific needs. HiCPC leverages outputs from 22 global climate models (GCMs) from the Coupled Model Intercomparison Project Phase 6 (CMIP6). To address inherent biases in daily GCM simulations, an advanced Bias Correction and Spatial Disaggregation (BCSD) method is employed, using the China Meteorological Forcing Dataset (CMFD) as a reference. HiCPC offers detailed daily precipitation and temperature data across China at an enhanced spatial resolution of 0.1° × 0.1°. It covers both the historical period (1979-2014) and future projections (2015-2100) based on four CMIP6 Shared Socioeconomic Pathways (SSPs) scenarios (SSP1-2.6, SSP2-4.5, SSP3-7.0, and SSP5-8.5). Upon validation, HiCPC demonstrates good performance, surpassing CMIP6 GCMs across the historical period. This reinforces its significance for essential research in climate change evaluation and its associated implications within China., (© 2024. The Author(s).)

Objective: In a previous study we have shown that, in the presence of interleukin (IL)-33, repeated, per-nasal challenge of murine airways with Streptococcus pneumoniae ( S. pneumoniae ) organisms induces human asthma-like airways inflammation. It is not clear, however, whether this effect is unique or manifest in response to other common respiratory pathogens. Methods: To explore this, airways of BALB/c mice were repeatedly challenged per-nasally with formaldehyde-inactivated bacterial bodies in the presence or absence of murine recombinant IL-33. Serum concentrations of S.pneumoniae , Moraxella catarrhalis ( M.catarrhalis ) and Haemophilus influenzae ( H.influenzae ) lysates-specific IgE were measured in patients with asthma and control subjects. Results: We showed that in the presence of IL-33, repeated, per-nasal airways exposure to the bodies of these bacteria induced airways hyperresponsiveness (AHR) in the experimental mice. This was accompanied by cellular infiltration into bronchoalveolar lavage fluid (BALF), eosinophilic infiltration and mucous hypertrophy of the lung tissue, with elevated local expression of some type 2 cytokines and elevated, specific IgG and IgE in the serum. The precise characteristics of the inflammation evoked by exposure to each bacterial species were distinguishable. Conclusions: These results suggest that in the certain circumstances, inhaled or commensal bacterial body antigens of both Gram-positive ( S. pneumoniae ) and Gram-negative ( M. catarrhalis and H. influenzae ) respiratory tract bacteria may initiate type 2 inflammation typical of asthma in the airways. In addition, we demonstrated that human asthmatic patients manifest elevated serum concentrations of M.catarrhalis- and H.influenzae- specific IgE.

Nickel-rich layered oxide cathode material LiNi x Co y Mn z O 2 (NCM) has emerged as a promising candidate for next-generation lithium-ion batteries (LIBs). These cathode materials possess high theoretical specific capacity, fast electron/ion transfer rate, and high output voltage. However, their potential is impeded by interface instability, irreversible phase transition, and the resultant significant capacity loss, limiting their practical application in LIBs. In this work, a simple and scalable approach is proposed to prepare gradient cathode material (M-NCM) with excellent structural stability and rate performance. Taking advantage of the strong coordination of Ni 2+ with ammonia and the reduction reaction of KMnO 4 , the elemental compositions of the Ni-rich cathode are reasonably adjusted. The resulted gradient compositional design plays a crucial role in stabilizing the crystal structure, which effectively mitigates Li/Ni mixing and suppresses unwanted surficial parasitic reactions. As a result, the M-NCM cathode maintains 98.6% capacity after 200 cycles, and a rapid charging ability of 107.5 mAh g -1 at 15 C. Furthermore, a 1.2 Ah pouch cell configurated with graphite anode demonstrates a lifespan of over 500 cycles with only 8% capacity loss. This work provides a simple and scalable approach for the in situ construction of gradient cathode materials via cooperative coordination and deposition reactions., (© 2024 Wiley‐VCH GmbH.)

With increasing energy storage demands across various applications, reliable batteries capable of performing in harsh environments, such as extreme temperatures, are crucial. However, current lithium-ion batteries (LIBs) exhibit limitations in both low and high-temperature performance, restricting their use in critical fields like defense, military, and aerospace. These challenges stem from the narrow operational temperature range and safety concerns of existing electrolyte systems. To enable LIBs to function effectively under extreme temperatures, the optimization and design of novel electrolytes are essential. Given the urgency for LIBs operating in extreme temperatures and the notable progress in this research field, a comprehensive and timely review is imperative. This article presents an overview of challenges associated with extreme temperature applications and strategies used to design electrolytes with enhanced performance. Additionally, the significance of understanding underlying electrolyte behavior mechanisms and the role of different electrolyte components in determining battery performance are emphasized. Last, future research directions and perspectives on electrolyte design for LIBs under extreme temperatures are discussed. Overall, this article offers valuable insights into the development of electrolytes for LIBs capable of reliable operation in extreme conditions., (© 2023 The Authors. Advanced Materials published by Wiley‐VCH GmbH.)

Mass spectrometry (MS)-based glycoproteomics research requires additional sample pretreatment to improve the effective identification of low-abundance glycopeptides without interference from non-glycoproteins. Herein, an attractive strategy using resorcinol-formaldehyde (RF) resin and zirconium-based coordination polymer (Zr-BCP) was established to prepare one-dimensional porous coordination polymer composites for glycopeptide enrichment before MS analysis. The obtained Fe 3 O 4 @RF@Zr-BCP nanochains feature excellent magnetic response (42.26 emu/g), high hydrophilicity (16.0°), and large specific surface area (140.84 m 2 /g), which provides abundant affinity sites for specific capture of glycopeptides. The materials exhibit outstanding performance in the enrichment of glycopeptides in terms of sensitivity (15 fmol/μL IgG), selectivity (1:200, molar ratio of IgG/BSA), loading capacity (200 mg/g) and recovery (106.4 ± 3.5%). In addition, the developed method based on Fe 3 O 4 @RF@Zr-BCP has been successfully applied to capture glycopeptides in tryptic digest of mouse teratoma cell extracts. It is worth emphasizing that compared with dispersed nanoparticles, the one-dimensional chain structure brings extraordinary reusability to Fe 3 O 4 @RF@Zr-BCP nanochains, which is conducive to the rapid cyclic enrichment of glycopeptides. This present work provides a potential enrichment platform for comprehensive glycoprotein analysis, and opens a new avenue for the application of oriented-assembly nanochains., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Background: Asthma is a common chronic respiratory disease characterized by airways inflammation, hyperresponsiveness and remodeling. IL-37, an anti-inflammatory cytokine, consists of five splice isoforms, that is, a-e. Although it has been previously shown that recombinant human IL-37b is able to inhibit airway inflammation and hyperresponsiveness in animal models of asthma, the effects and difference of other IL-37 isoforms, such as IL-37a on features of asthma are unknown., Methods: Animal models of chronic asthma were established using IL-37a and IL-37b transgenic mice with C57BL/6J background and wild-type (WT) mice sensitized and nasally challenged with ovalbumin (OVA). Airway hyperresponsiveness was measured using FlexiVent apparatus, while histological and immunohistological stainings were employed to measure airways inflammation and remodeling indexes, including goblet cell metaplasia, mucus production, deposition of collagen, hypertrophy of airway smooth muscles and pulmonary angiogenesis., Results: Compared to WT mice, both IL-37a and IL-37b transgenic mice had significant reduced airway hyperresponsiveness and the declined total numbers of inflammatory cells, predominant eosinophils into airways and lung tissues. Furthermore, all features of airways remodeling, including degrees of mucus expression, collagen deposition, hypertrophy of smooth muscles, thickness of airways and neovascularization markedly decreased in IL-37 transgenic mice compared with OVA-treated WT mice., Conclusion: Our data suggest that both IL-37a and IL-37b isoforms are able to not only ameliorate airways inflammation and airways hyperresponsiveness, but also greatly reduce airways structural changes of animal models of chronic asthma., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Ratiometric fluorescence carbon dots (CDs) that serve as probes have attracted more attention on account of their unique optical properties, low toxicity, anti-interference, and internal reference. However, the facile fabrication of CDs with the aim of detecting multiple targets through mutually independent response channels is always a challenge. Herein, multifunctional label-free N-doped ratiometric fluorescence CDs (N-CDs) are developed from tea leaves extract and o -phenylenediamine by a mild solvothermal method. The prepared N-CDs are tailored with nitrogen- and oxygen-containing functional groups on the surface and contribute to splendid hydrophilia. Two completely independent ratiometric fluorescence channels of N-CDs, respectively, respond to Hg 2+ and H 2 O in a mutually independent manner. Based on the interactions of N-Hg and O-Hg, N-CDs achieve an excellently sensitive and selective detection for Hg 2+ in the channel of I 387 nm / I 351 nm , giving a linear relationship in the range of 0-50 μM. Also, a wide range of Hg 2+ concentration (0-100 μM) is linear to A 374 nm through UV-vis assay. Otherwise, the linear determination of H 2 O content (0-30%) is realized in another channel ( I green / I blue ). The good performance in the independent testing of Hg 2+ and H 2 O, demonstrate that the proposed N-CDs have potential in multifunctional detection.

Respiratory tract infection early in life plays a significant role in the pathogenesis of asthma. In the present study we examine, using a murine surrogate, the effects of early life respiratory infection with Streptococcus pneumoniae (SP) on adult asthma induced by sensitisation and exposure to house dust mite (HDM) allergen. Mice (one week old) were infected with SP, then 3 weeks later sensitised to HDM emulsified with Al (OH)3 intraperitoneally and challenged intranasally with same allergen for up to a further 5 weeks to establish the asthma surrogate. Outcome measures were quantified using the FlexiVent apparatus, histology and immunohistology, ELISA and flow cytometry. The murine surrogates of asthma infected with SP early in life exhibited significantly more severe disease compared with the controls of mice without SP infection, as shown by airways responsiveness, inflammatory cellular infiltration of the airways, expression of markers of airways remodelling, serum concentrations of HDM-specific IgE and the concentrations of Th2-type cytokines and the numbers of activated Th2 and ILC2 cells in the lung tissues. These data are compatible with the hypothesis that early-life infection of the airways with SP exacerbates, at least in some individuals, subsequent HDM-induced allergic airways inflammation and associated asthma in adulthood in this murine surrogate., (Copyright © 2022. Published by Elsevier Inc.)

While environmental aeroallergens and epithelial alarmins such as IL-33 are firmly implicated in asthma, the possible role of Streptococcus pneumoniae (S. pneumoniae) antigens is less clear. To explore this, wild-type BALB/c mice were repeatedly challenged per-nasally with IL-33 and inactivated S. pneumoniae, either agent alone or diluent control. Some animals were rested then later re-challenged with inactivated S. pneumoniae alone. Serum concentrations of S. pneumoniae lysates-specific IgE were measured in patients with asthma and control subjects. Interestingly, in the presence of IL-33, repeated exposure to inactivated S. pneumoniae induced asthma-like pathological changes accompanied by a systemic adaptive immune response. Subsequent re-exposure of the sensitized animals to inactivated S. pneumoniae alone was able to induce such changes. The concentration of S. pneumoniae lysates-specific IgE was significantly elevated in the asthma patients. These data suggest that antigens derived from infectious microorganisms may participate in generating the mucosal inflammation which characterizes asthma., (Copyright © 2021 Elsevier Inc. All rights reserved.)

To reduce the possibility of bacterial infection and implant-related complications, surface modification on polyurethane (PU) film is an ideal solution to endow hydrophobic PU with antibacterial and antifouling properties. In this work, a variety of polyhexamethylene guanidine/ hyaluronic acid (PHMG/HA) multilayer films were self-assembled layer-by-layer on PU films using polyanions, carboxyl-activated HA, and polycations PHMG by controlling the concentration of these polyelectrolytes as well as the number of layers self-assembled. Attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) spectra, water contact angle (WCA), and A Atomic force microscope (AFM) of PU and modified PU films were studied. Protein adsorption and bacterial adhesion as well as the cytotoxicity against L929 of the film on selected PU-(PHMG/HA) 5 /5-5 were estimated. The results showed that PU-(PHMG/HA) 5 /5-5 had the best hydrophilicity among all the prepared films, possessing the lowest level of protein adsorption. Meanwhile, this film showed efficient broad-spectrum antibacterial performance as well as significant resistance of bacterial adhesion of more than a 99.9% drop for the selected bacteria. Moreover, almost no influence on cell viability of L929 enhanced the biocompatibility of film. Therefore, the modified PU films with admirable protein absorption resistance, antimicrobial performance, and biocompatibility would have promising applications in biomedical aspect.

Glycosylation and phosphorylation are two of the most common and important post-translational modifications (PTMs) of proteins, which play critical roles in regulating a variety of complex biological processes and involvement in many diseases. Due to the low abundance of phosphopeptides and glycopeptides, highly selective enrichment methods are crucial to the identification of protein phosphorylation and glycosylation by mass spectrometry (MS). Here, monodisperse uniform Al 3+ -doping-TiO 2 mixed oxide microspheres were easily synthesized. The morphology was controlled by a sol-gel method, during the hydrothermal treatment. The obtained microspheres with uniform particle size distribution (about 1-2 μm)，high surface area and improved pore structures, were characterized by SEM, TEM, XRD and N 2 adsorption-desorption isotherms. Al 3+ -doping-TiO 2 was applied in enriching glycopeptides and phosphopeptides respectively or simultaneously by using different enrichment conditions, achieving selective enrichment of glycopeptides and phosphopeptides. 20 glycopeptides and 25 phosphopeptides enriched from the tryptic digest mixtures of human serum immunoglobulin G (IgG) and α-casein (molar ratio of 1:1) were obviously observed with greatly improved signal-to-noise (S/N) ratio. Meanwhile, the enrichment results of non-fat milk and human serum also show the enrichment selectivity from complex biological samples. This study will provide a novel insight for selective enrichment of glycopeptides and phosphopeptides in post-translational modification proteomics research., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Asthma is an inflammatory disease of the airways and numerous cytokines contribute to this pathogenesis. It is shown that challenge of airways with IL-33 induces asthma-like pathological changes in mice, but the possible downstream cytokines in this process remain to be characterised. To explore this, we compared changes in the airways of wildtype (WT) and IL-9 deficient mice challenged with IL-33. In line with previous report, per-nasal challenge of WT mice with IL-33 significantly increased the responsiveness of the airways along with infiltration of inflammatory cells, goblet cell hyperplasia, collagen deposition and smooth muscle hypertrophy, and the expression of cytokines compared with control group. Surprisingly, all of these pathological changes were significantly attenuated in IL-9 deficient mice following identical IL-33 challenge. These data suggest that IL-9 is one downstream cytokine relevant to the effects of IL-33 in asthmatic airways and consequently a potential therapeutic target for the treatment of asthma., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Post-treatment was performed for poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) films screen-printed on fluorine-doped tin oxide (FTO) substrates, to improve their charge transfer efficiency. Different H 2 SO 4 solutions, including concentrated H 2 SO 4 and H 2 SO 4 diluted with H 2 O or dimethyl sulfoxide (DMSO), were adopted during the post-treatment. The adhesion of the as-treated films was evaluated by adhesive tape peeling tests, the surface morphology and vertical charge transfer from the films to the substrates were investigated by current-sensing atomic force microscopy, and the catalytic activities toward I 3 - reduction of PEDOT:PSS films were characterized by electrochemical measurements. It is discovered that selecting proper H 2 SO 4 solutions is crucial to improve the charge transfer efficiency and catalytic performance while maintaining reliable adhesion of the film on the substrates, with H 2 SO 4 /DMSO performing best as the solution for post-treatment. A mechanistic explanationis proposed based on different interactions among solution, PEDOT:PSS, and the substrate for various post-treatment solutions., (© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Wnt5a signalling plays pathological roles in synovial inflammation and bone destruction. In the present study, we designed four human Wnt5a-based DNA recombinants and detected their effects on immunogenicity and anti-rheumatism in a collagen-induced arthritis (CIA) model. Histomorphometry and micro-CT scanning showed that the phWnt5a-NL was superior to other recombinants because it resulted in decreased severity of arthritis, histopathological scores of synovial inflammation and bone erosion in CIA mice. In addition, ELISA and TRAP staining showed that the phWnt5a-NL-immunized CIA mice had reductions in the serum concentrations of the rheumatoid-associated cytokines IL-1β and RANKL and in osteoclastogenesis. Furthermore, flow cytometry showed that the phWnt5a-NL treatment increased the percentage of Treg cells. Finally, western blotting analysis showed that the phWnt5a-NL-immunization interrupted β-catenin and JNK expression in osteoclast precursors derived from the CIA mice. The results suggest that depleting the carboxy-terminus in hWnt5a-based DNA recombinants may be beneficial for the treatment of chronic inflammatory disorders involving bone resorption., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Objective: Histone deacetylases (HDACs) play an important role in dysregulation of histone acetylation/deacetylation, which is the main driving force of the progression of pulmonary fibrosis. Here we investigated the changes in histone acetylation/deacetylation, and the contribution of specific class I and class II HDACs in the progression of pulmonary fibrosis., Methods: Male C57BL/6J mice received a single dose of tracheal administration of bleomycin to establish the pulmonary fibrosis model. The changes in acetylation rate of histone 3 (H3) and histone 4 (H4), and the activity of HDAC2 and HDAC4 in the lung tissue during the progression from alveolitis to pulmonary fibrosis were measured., Results: The acetylation rate of H3/H4 significantly decreased during alveolitis and the early and middle stages of fibrosis, but restored in the late stage of fibrosis. Correlation analysis showed that H4 deacetylation affected both alveolitis and pulmonary fibrosis. H3 deacetylation only affected alveolitis. HDAC2 activity significantly increased in the middle and late stages of pulmonary fibrosis. There was no significant difference in HDAC4 activity between bleomycin and saline groups. However, HDAC4 activity changed significantly with the progression of the disease in bleomycin group. The changes in HDAC2 and HDAC4 activity were different. HDAC2 had long-lasting effects, while HDAC4 had transient effects. Correlation analysis showed that HDAC2 and HDAC4 activity was positively correlated with alveolitis score and fibrosis score., Conclusions: The changes in histone acetylation may directly regulate the gene expression of inflammatory cytokines/fibronectin and thus affect the progression of pulmonary fibrosis. The injury-induced histone deacetylation switched into acetylation at the late stage of pulmonary fibrosis, which may be involved in the repair process. HDAC2 is mainly involved in the chronic progression of pulmonary fibrosis, and HDAC4 is mainly involved in early stress response to pulmonary fibrosis., (Copyright © 2017. Published by Elsevier B.V.)

The contributions of aryl hydrocarbon receptor (Ahr) to the pathogenesis of rheumatoid arthritis (RA), particularly bone loss, have not been clearly explored. The imbalance between osteoblasts and osteoclasts is a major reason for bone loss. The dysfunction of osteoblasts, which are derived from mesenchymal stem cells (MSCs), induced bone erosion occurs earlier and is characterized as more insidious. Here, we showed that the nuclear expression and translocation of Ahr were both significantly increased in MSCs from collagen-induced arthritis (CIA) mice. The enhanced Ahr suppressed the mRNA levels of osteoblastic markers including Alkaline phosphatase (Alp) and Runt-related transcription factor 2 (Runx2) in the differentiation of MSCs to osteoblasts in CIA. The 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD)-mediated activation of Ahr dose-dependently suppressed the expression of osteoblastic markers. In addition, the expression of β-catenin was reduced in CIA MSCs compared with control, and the TCDD-mediated activation of the Ahr significantly inhibited β-catenin expression. The Wnt3a-induced the activation of Wnt/β-catenin pathway partly rescued the osteogenesis decline induced by TCDD. Taken together, these results indicate that activated Ahr plays a negative role in CIA MSCs osteogenesis, possibly by suppressing the expression of β-catenin., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Peptide-based therapy, such as modified peptides, has attracted increased attention. IL-17 is a promising therapeutic target for autoimmune diseases, and levels of circulating bioactive IL-17 are associated with rheumatoid arthritis severity. In this study, a modified truncated IL-17RC is generated to ameliorate inflammation and bone destruction in arthritis. The truncated IL-17RC binds to both IL-17A and IL-17F with higher binding capacity compared to nonmodified IL-17RC. In addition, the truncated IL-17RC reduces the secretion of inflammatory and osteoclastogenic factors induced by IL-17A/F in vitro. Moreover, the administration of truncated IL-17RC dramatically improves symptoms of inflammation and inhibited bone destruction in collagen-induced arthritis mice. Collectively, these data demonstrate that modified truncated IL-17RC peptide may be a more effective treatment strategy in the simultaneous inhibition of both IL-17A and IL-17F signaling, whereas the existing agents neutralize IL-17A or IL-17F alone. These suggest that the truncated IL-17RC may be a potential candidate in the treatment of inflammatory associated bone diseases., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Context: Paclitaxel (PTX) is widely used in chemotherapy for cancer treatment; however, it has some serious side effects. Andrographolide (Andro) is a potential cancer therapeutic agent isolated from Andrographis paniculata (Burm. f.) Nees (Acanthaceae)., Objective: The objective of this study is to evaluate the effects of PTX combined with Andro against A549 cells., Materials and Methods: The effects of 24-48 h treatment with 0.48-60.75 nM PTX and 5.10-328.0 μM Andro on cellular proliferation, apoptosis, cell cycle and intracellular reactive oxygen species (ROS) were determined by sulphorhodamine B assay, Annexin V-FITC/PI apoptosis detection, PI staining and ROS assay, respectively. Synergy was determined using combination index. The antitumour efficacy of 20 mg/kg PTX with 100 mg/kg Andro was studied in a xenograft murine model., Results: IC 50 value of the PTX combined with Andro against A549 cells was 0.5-7.4 nM, which was significantly lower than that of PTX (15.9 nM). PTX with 10 μM Andro caused (1.22-1.27)-fold apoptosis and 1.7-fold ROS accumulation compared with PTX alone. N-Acetylcysteine, a ROS scavenger, blocked this synergy in vitro. In contrast, G2/M phase cell cycle arrest resulting from PTX was not potentiated by Andro. Moreover, PTX in combination with Andro inhibited the growth of A549 transplanted tumours by 98%., Discussion and Conclusion: The results indicate that the combination of PTX and Andro exert significant synergistic anticancer effect on A549 cells in vitro and in vivo. The synergy may be the result of the accumulation of ROS. The combination of Andro and PTX represents a potential strategy for the treatment of A549 cells.

In this paper, the doxorubicin (DOX)-loaded micelles were prepared based on a novel folic acid conjugated pH-dependent thermo-sensitive copolymer poly(D,L-lactic acid)-b-poly(N-isopropyl methacrylamide-co-N-isopropylmaelic acid-co-10-undecenoic acid) (PLA-PNNUA-FA) constructed to provide an active targeting drug delivery and triggered drug release system. The micelles were able to target tumors through the interaction between folic acid and its receptors which are overexpressed on the tumor cell membrane, and achieved pH-dependent thermo induced drug release in the intracellular mild acidic media such as endosomes and lysosomes after the micelles enter the cells. The results of cell assays and animal experiments showed that the micelles exhibited obvious tumor penetration efficiency in vivo, also improved DOX cell uptake and cytotoxicity in vitro. It was suggested that copolymer PLA-PNNUA-FA might be a potential targeted drug carrier to deliver chemotherapeutic drugs achieving better efficacy of chemotherapy.

Dickkopf-1 (DKK1), a secretory inhibitor of canonical Wnt signaling, plays a critical role in certain bone loss diseases. Studies have shown that serum levels of DKK1 are significantly higher in rheumatoid arthritis (RA) patients and are correlated with the severity of the disease, which indicates the possibility that bone erosion in RA may be inhibited by neutralizing the biological activity of DKK1. In this study, we selected a panel of twelve peptides using the software DNASTAR 7.1 and screened high affinity and immunogenicity epitopes in vitro and in vivo assays. Furthermore, we optimized four B cell epitopes to design a novel DKK1 multiepitope DNA vaccine and evaluated its bone protective effects in collagen-induced arthritis (CIA), a mouse model of RA. High level expression of the designed vaccine was measured in supernatant of COS7 cells. In addition, intramuscular immunization of BALB/c mice with this vaccine was also highly expressed and sufficient to induce the production of long-term IgG, which neutralized natural DKK1 in vivo. Importantly, this vaccine significantly attenuated bone erosion in CIA mice compared with positive control mice. These results provide evidence for the development of a DNA vaccine targeted against DKK1 to attenuate bone erosion.

Functional polymeric micelles play an important role in the efficient delivery of therapeutic drugs into tumours. In this study, a functional drug delivery platform with ligands for targeting and fluorescent imaging was designed based on Pluronic F127 (PF127). Using folic acid (FA) and fluorescein isothiocyanate (FITC) to chemically conjugate with PF127, two functional polymers, Pluronic F127-FA (PF127-FA) and Pluronic F127-FITC (PF127-FITC), were synthesized. Solasodine-loaded micelles were then prepared via the thin-film hydration method. By employing A549 and HeLa cells, the results of in vitro cell assays performed using confocal laser scanning microscopy and flow cytometry suggested that the proposed micelles could provide the desired specific targeting and fluorescent imaging functions. In addition, the results of in vitro cytotoxicity experiments showed that the growth inhibition rates of A549 and HeLa cells treated with solasodine-loaded micelles were remarkably higher than those of cells treated with free solasodine. Solasodine-loaded micelles exhibited a more distinct inhibitory effect against HeLa cells than against A549 cells. Thus, an effective drug delivery system for targeting and imaging cancer cells was developed.

Murine models for rheumatoid arthritis (RA) research can provide important insights for understanding RA pathogenesis and evaluating the efficacy of novel treatments. However, simultaneously imaging both murine articular cartilage and subchondral bone using conventional techniques is challenging because of low spatial resolution and poor soft tissue contrast. X-ray phase-contrast imaging (XPCI) is a new technique that offers high spatial resolution for the visualisation of cartilage and skeletal tissues. The purpose of this study was to utilise XPCI to observe articular cartilage and subchondral bone in a collagen-induced arthritis (CIA) murine model and quantitatively assess changes in the joint microstructure. XPCI was performed on the two treatment groups (the control group and CIA group, n = 9 per group) to monitor the progression of damage to the femur from the knee joint in a longitudinal study (at 0, 4 and 8 weeks after primary injection). For quantitative assessment, morphologic parameters were measured in three-dimensional (3D) images using appropriate image analysis software. Our results showed that the average femoral cartilage volume, surface area and thickness were significantly decreased (P<0.05) in the CIA group compared to the control group. Meanwhile, these decreases were accompanied by obvious destruction of the surface of subchondral bone and a loss of trabecular bone in the CIA group. This study confirms that XPCI technology has the ability to qualitatively and quantitatively evaluate microstructural changes in mouse joints. This technique has the potential to become a routine analysis method for accurately monitoring joint damage and comprehensively assessing treatment efficacy.

Ahr activation is known to be associated with synovitis and exacerbated rheumatoid arthritis (RA), but its contributions to bone loss have not been completely elucidated. Osteoblast proliferation and differentiation are abnormal at the erosion site in RA. Here, we reported that the expression of Ahr was increased in the hind paws' bone upon collagen-induced arthritis (CIA) in mice, and the levels of Ahr were negatively correlated with bone mineral density (BMD). In addition, immunofluorescent staining showed that the high expression of Ahr was mainly localized in osteoblasts from the CIA mice compared to normal controls. Moreover, the luciferase intensity of Ahr in the nucleus increased by 12.5% in CIA osteoblasts compared to that in normal controls. In addition, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) activation of the Ahr inhibited pre-osteoblast MC3T3-E1 cellular proliferation and differentiation in a dose-dependent manner. Interestingly, the levels of alkaline phosphatase (ALP) mRNA expression in the osteoblasts of CIA mice were reduced compared to normal controls. In contrast, decreased ALP expression by activated Ahr was completely reversed after pretreatment with an Ahr inhibitor (CH-223191) in MC3T3-E1 cell lines and primary osteoblasts on day 5. Our data further showed that activation of Ahr promoted the phosphorylation of ERK after 5days. Moreover, Ahr-dependent activation of the ERK signaling pathway decreased the levels of proliferation cells and inhibited ALP activity in MC3T3-E1 cells. These results demonstrated that the high expression of Ahr may suppress osteoblast proliferation and differentiation through activation of the ERK signaling pathway, further enabling bone erosion in CIA mice., (Copyright © 2014 Elsevier Inc. All rights reserved.)

To neutralize the pathological activities of tumor necrosis factor-α (TNF-α) and receptor activator of NF-κB ligand (RANKL), we engineered and characterized a humanized 8G12 (h8G12) antibody that targeted TNF-α and RANKL. Standard molecular biological and complementarity determining region (CDR)-grafting techniques were used to engineer the h8G12 antibody, and enzyme-linked immunosorbent assays (ELISAs) and Western blotting were employed to determine its binding activation and specificity. TNF-α-mediated cytotoxicity and RANKL-induced osteoclastogenesis assays were used to evaluate the neutralizing effects of the antibody. The cDNA sequences were established by grafting the murine monoclonal antibody (mAb) 8G12 CDRs into the heavy and light chain (HC and LC) variable regions (VH and VL) of the human mAbs 3DGG&#95;B and 1I9R&#95;L, respectively. The recombinant plasmids were transfected into Chinese hamster ovary (CHO) cells to produce the h8G12 antibody, which could simultaneously recognize TNF-α and RANKL. In addition, the h8G12 antibody reduced the TNF-α-mediated apoptosis of L929 cells by 25.84%. Furthermore, the h8G12 antibody significantly inhibited leukocyte infiltration in a murine allergic contact inflammation model. Concurrent with the inhibition of apoptosis, the h8G12 antibody significantly reduced the number of osteoclast-like cells in a dose-dependent manner. These results demonstrated that the h8G12 antibody neutralized the activities of TNF-α and RANKL and that it might be a potential candidate for the treatment of inflammatory bone diseases, such as rheumatoid arthritis (RA)., (Copyright © 2014 Elsevier Inc. All rights reserved.)

This study aimed to generate a monoclonal antibody (mAb) targeting both tumor necrosis factor-α (TNF-α) and receptor activator of NF-κB ligand (RANKL) and to evaluate the therapeutic effects of this antibody on acute inflammation and osteoporosis. We used hybridoma techniques to generate potential mAbs and enzyme-linked immunosorbent assay (ELISA) to determine their specificity. Crystal violet staining was performed to measure the effective dose of the candidate mAbs. The neutralizing effect of the mAbs was evaluated by TNF-α-mediated cytotoxicity and RANKL-induced osteoclastogenesis assays. We further assessed the therapeutic effect of the mAbs in BALB/c mice with carrageenan-induced acute inflammation and ovariectomy-induced osteoporosis. We successfully generated an IgG1 isotype mAb that recognizes human TNF-α and RANKL, which we named 8G12. The 50% effective dose of 8G12 was approximately 1μg/mL. L929 cells treated with 8G12 exhibited decreased levels of apoptosis (20.04% compared to 63.28% in the positive controls). In addition, treatment with 8G12 inhibited osteoclastogenesis in a dose-dependent manner in vitro. Carrageenan-induced paw edema was significantly reduced in the 8G12-treated mice compared to the positive controls. Treatment with 8G12 also reduced the number of infiltrating leukocytes by more than 50%. The 8G12 treatment not only prevented bone loss but also increased the number, thickness and volume of trabeculae and reduced trabecular separation in ovariectomized mice. Our data suggest that the 8G12 effectively neutralizes the bioactivity of TNF-α and RANKL, ameliorating osteoporosis and inflammation. We therefore propose that 8G12 could be a candidate for generating therapeutic antibodies for treating inflammatory bone diseases., (Copyright © 2014 Elsevier B.V. All rights reserved.)

In the present work, we propose a new multifunctional micelle, co-self-assembled from different monofunctional copolymers, for tumor targeting and fluorescent and electron spin resonance (ESR) dual detection. Firstly, a poly(D,L-lactic acid)-b-poly(N-isopropyl methacrylamide-co-N-isopropylmaelic acid-co-10-undecenoic acid)-b-poly(N-acryloxysuccinimide) (PLA-PNNUA-PNAS) copolymer, with pH-dependent thermo-responsive properties, was synthesized. The copolymer was synthesized using reversible addition fragmentation chain transfer (RAFT) polymerization method, after which it was further used as a parent copolymer to combine with folic acid (FA), fluorescein isothiocyanate (FITC) and 4-amino-2,2,6,6-tetramethylpiperidin-1-oxyl (4-NH2-TEMPO), respectively, resulting into three new monofunctional copolymers. Finally, the multifunctional copolymer micelle was easily then fabricated, through co-self-assembly, using the monofunctional copolymers. The results from in vitro cell assays indicated that the proposed micelle was able to provide desired multifunctional properties, including tumor specific targeting and fluorescent and ESR dual detection. Additionally, the parent copolymer allowed conjugation with other ligands to prepare them for more functional copolymers attachment for future potential applications. More importantly, the multifunctional properties of the copolymer micelles could be rationally tailored, able for given purposes.

Objective: To study the relationship between the human secreted protein stabilin-1-interacting chitinase-like protein (SI-CLP) and rheumatoid arthritis (RA)., Methods: The expression of SI-CLP in peripheral blood mononuclear cells (PBMCs) and synovial fluid from patients with RA and the effects of cytokines on SI-CLP expression were examined by Western blotting. Fluorescence-activated cell sorting analysis was performed to investigate the binding between SI-CLP and cells. Bone marrow-derived macrophages were isolated from wild-type and SI-CLP(-/-) mice, and real-time quantitative polymerase chain reaction was performed to detect the levels of messenger RNA for cytokines or SI-CLP in SI-CLP- or cytokine-treated macrophages. Histologic studies were conducted to evaluate inflammation and the expression of interleukin-12 (IL-12), IL-13, and SI-CLP in lesions. Enzyme-linked immunosorbent assays were used to detect the cytokine levels in bone marrow-derived macrophages. Rats or mice with collagen-induced arthritis (CIA) and SI-CLP(-/-) mice were used to study the function of SI-CLP in RA., Results: SI-CLP expression was increased in PBMCs and detectable in synovial fluid from patients with RA. Administration of SI-CLP to rats with CIA aggravated arthritis-associated inflammation. SI-CLP was specifically attached to the surface protein of macrophages, which elevated the expression of IL-1β, IL-6, IL-12, and IL-13 in macrophages and mouse bone marrow-derived macrophages, up-regulating ERK phosphorylation. Moreover, SI-CLP was up-regulated by both IL-12 and IL-13 through JNK and JAK/STAT signaling, respectively. Knockout of SI-CLP resulted in a decrease in the expression of IL-1β, IL-6, IL-12, and IL-13 and lower susceptibility to CIA compared with wild-type mice. SI-CLP treatment also aggravated arthritis-related inflammation in wild-type and SI-CLP(-/-) mice., Conclusion: SI-CLP functions as a regulator of the inflammatory response by macrophages. The decrease in inflammation-associated cytokine levels resulting from SI-CLP knockout may explain the lower susceptibility to CIA in SI-CLP(-/-) mice., (Copyright © 2014 by the American College of Rheumatology.)

Novel macrocyclic molecules comprising sulfonyl and acyl moiety at the position N-12a of 16-membered azalides (6a-n) and azalactams (10a-r) scaffold were synthesized from cyclododecanone 1 as starting material via 5 steps and 4 steps, respectively. The antifungal activity of these compounds against Sclerotinia sclerotiorum, Pyricularia oryzae, Botrytis cinerea, Rhizoctonia solani and Phytophthora capsici were evaluated and found that compounds possessing α-exomethylene (6c, 6d, 6e and 6g) showed antifungal activity comparable to commercial fungicide Chlorothalonil against P. oryzae and compounds possessing p-chlorobenzoyl exhibited enhanced antifungal activity than those with other substituents against S. sclerotiorum, P. oryzae, and B. cinerea. These findings suggested that the α-exomethylene and p-chlorobenzoyl may be two potential pharmacological active groups with antifungal activities., (Copyright © 2013 Elsevier Masson SAS. All rights reserved.)

Highly biocompatible sub-50-nm monodisperse superparamagnetic Fe3O4@SiO2 core/shell nanoparticles with luminescent silica shells were synthesized by a w/o-microemulsion technique. And then these nanoparticles were coated with the covalently bonded biocompatible polymer poly(ethylene glycol) (PEG) and modified with the biological cancer targeting ligand folic acid (FA). After characterized by means of powder X-ray diffraction (XRD), transmission electron microscopy (TEM), dynamic light scattering (DLS), Fourier transformed infrared spectroscopy (FT-IR), Thermogravimetric analysis (TGA), vibrating sample magnetometer (VSM), UV-vis, fluorescence spectroscopy and confocal laser scanning microscopy (CLSM), we confirmed that Fe3O4@SiO2 (FITC)-PEG-FA nanocomposites (SMNPs-FA) could be efficiently taken up by HeLa cancer cells and KB cells which are of over-expression of folate receptors. The multifunctional nanomaterials exhibited superparamagnetic, monodisperse, highly biocompatible, intensively fluorescent and capable of recognizing and binding cells that overexpress folate receptors, which would be useful for targeting cell imaging and provide an excellent platform for further development of an efficient cancer therapy.

A water-soluble polysaccharide (FAAP-02), composed of N-acetyl-D-glucosamine, glucose, mannose, galactose, rhamnose, arabinose, xylose and ribose, with an average molecular weight of 5169 Da, was isolated from Artemisia argyi. The antitumor and immunomodulatory activities of FAAP-02 were evaluated in Sarcoma 180 (S180) tumor-bearing mice by intraperitoneal administration. As a result, FAAP-02 significantly inhibited the growth of the S180 transplanted tumors and prolonged the survival time of the tumor-bearing mice. Moreover, FAAP-02 could obviously increase the thymus and spleen indices, the levels of serum Interleukin 2 (IL-2), Interleukin 6 (IL-6), Interleukin 12 (IL-12) and tumor necrosis factor-α (TNF-α), and the expression of CD4+ and CD8+ splenic T lymphocytes which were suppressed by the transplanted tumor or/and 5-fluorouracil (5-FU) in the mice. These results indicated that the antitumor activity of FAAP-02 might be associated with its immunostimulatory effects., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

In this study, thermosensitive and folate functionalized poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide)-ploy(N-isopropylacrylamide-co-hydroxyethyl methacrylate) (FA-Pluronic-PNH) copolymer was synthesized. The structure and molecular weight of the copolymer were confirmed by 1H NMR, FT-IR and GPC, respectively. The lower critical solution temperature (LCST) of the copolymer was 39.8 degrees C. By employing doxorubicin (DOX) as a model drug, folate receptor-targeted DOX-loaded micelles were further formed on the copolymer. The blank and DOX-loaded micelles both exhibited nearly spherical shapes and their average diameters were 35 nm and 50 nm, respectively. The in vitro release behaviors of the DOX-loaded micelles were temperature-dependent and the release rate of DOX at 42 degrees C (above LCST) was faster than that at 37 degrees C (below LCST). Furthermore, the cytotoxicity assays of free DOX and DOX-loaded micelles on human cervical cancer cell lines HeLa and human lung cancer cell lines A549 demonstrated that folate increased the cellular uptake of the micelles within targeted cells that vastly over-expressed folate receptors.

Chitosan (CS) grafted with β-cyclodextrin (CD-g-CS) nanoparticles as a new carrier for poorly water-soluble drugs has been developed. The CD-g-CS polymer is readily synthesized from chitosan and mono-6-deoxy-6-(p-toluenesulfonyl)-β-cyclodextrin. Three different degrees of substitution (DS) of β-cyclodextrin (β-CD) on CD-g-CS (9.6, 14.0 and 20.0%) are designed and evaluated by controlling the mole ratio of β-CD to chitosan. Then CD-g-CS nanoparticles are prepared by an ionic gelation method, with the controlled size of 202.0-589.0 nm. Stable colloidal dispersion of the nanoparticles has been formed with the zeta potential of +23.0 to +43.0 mV. In vitro stability test indicates that CD-g-CS nanoparticles are more stable in phosphate-buffered saline compared with CS nanoparticles. Finally, the poorly water-soluble drug, ketoprofen (KTP), is used as a model drug to evaluate the efficiency of the new drug delivery carrier. It is found that the encapsulation efficiency of KTP in the nanoparticles with 20% DS of CD is as high as 1.36-fold than that of CS nanoparticles. Moreover, notably KTP is released from the nanoparticles in a controlled-release manner and is pH-responsive on DS of CD. In summary, these results suggest that the CD-g-CS nanoparticles, as a general promising drug delivery system, can be used as a potential biodegradable nano-drug delivery system for controlled release of poorly water-soluble drugs with pH-responsive capability., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Targeting tumor necrosis factor-α (TNF-α) and activator of NF-κB ligand (RANKL) has been proved highly successful in rheumatoid arthritis (RA) models and patients. This raises a possibility whether a single agent simultaneously targeting TNF-α and RANKL provides a potential therapeutic opportunity. This study aimed to design a dual functional vaccine and evaluate its therapeutic effects in RA mice model. Standard molecular biological techniques were used to generate human RANKL-TNF-like core fusion protein (RTFP-2) vaccine. High titers of antibodies against human TNF-α and RANKL were elicited and the RTFP-2 antiserum decreased TNF-α mediated apoptosis of L929 cells to 41% compared with 90% in positive controls. In addition, the antiserum completely abrogated osteoclastogenesis in vitro. Immunization with RTFP-2 also reduced the mortality of TNF-α induced cachexia from 56% to 28%. The RANKL-mediated hypercalcemic effects were significantly attenuated in RTFP-2 vaccinated mice. Furthermore, RTFP-2 vaccine significantly mitigated the incidence and severity of CIA via inhibition of inflammation and bone resorption. Our results showed the RTFP-2 vaccine of dual targets ameliorated the symptoms of CIA mice, suggesting the potential possibility to treat inflammatory bone diseases such as RA., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Receptor activator of NF-κB ligand (RANKL), its signaling receptor RANK, and its decoy receptor osteoprotegerin (OPG) constitute a molecular triad that is critical in regulating bone remodeling, and also plays multiple roles in the immune system. OPG binds RANKL directly to block its interaction with RANK. In this article, we report the 2.7-Å crystal structure of human RANKL trimer in complex with the N-terminal fragment of human OPG containing four cysteine-rich TNFR homologous domains (OPG-CRD). The structure shows that RANKL trimer uses three equivalent grooves between two neighboring monomers to interact with three OPG-CRD monomers symmetrically. A loop from the CRD3 domain of OPG-CRD inserts into the shallow groove of RANKL, providing the major binding determinant that is further confirmed by affinity measurement and osteoclast differentiation assay. These results, together with a previously reported mouse RANKL/RANK complex structure, reveal that OPG exerts its decoy receptor function by directly blocking the accessibilities of important interacting residues of RANKL for RANK recognition. Structural comparison with TRAIL/death receptor 5 complex also reveals structural basis for the cross-reactivity of OPG to TRAIL.

In this study, the size-uniform (5-6 nm), nearly spherical, and well-dispersed aqueous Fe3o4 magnetic nanoparticles were prepared by an improved chemical coprecipitation method. The DDAT-terminated (S-1-Dodecyl-S'-(alpha,alpha'-dimethyl-alpha"-acetic acid) trithiocarbonate) polymethacrylic (PMA-DDAT) was chosen as the apt surfactant, and the terminal DDAT can be used as a high efficient RAFT chain-transfer agent for further functionalization. Then, the functionalized Fe3O4 reacted with 4-amino-2,2,6,6-tetramethyl-piperidine-oxyl (4-NH2-TEMPO) to give the spin labeling magnetic nanoparticles. Finally, the multifunctional MNPs was characterized by X-ray powder diffraction (XRD), transmission electron microscopy (TEM), electron paramagnetic resonance (EPR), Fourier transform infrared spectrometer (FT-IR), and vibrating-sample magnetometer (VSM). The obtained highly water-soluble, superparamagnetic, and multifunctional magnetic nanoparticles should find potential applications in biomedical research.

Neurotoxicity was investigated in nano-SiO2-treated cultured PC12 cells, an in vitro neuronal cell model, in order to define a relatively safe dose range for its application. The following were observed in the present study: (1) A dose-dependent increase in the level of reactive oxygen species (ROS) with a corresponding decrease in the level of glutathione (R2=0.965) suggesting 20- and 50-nm SiO2-induced free radical generation and glutathione depletion. (2) A dose- and time-dependent decrease in cell viability that was associated with elevation of ROS level, especially after 24-h nano-SiO2 exposure (R2=0.965), suggesting the role of oxidative stress on nano-SiO2 induced cell death. (3) An increase in the level of thiobarbituric-acid reactive species that correlated reversely with cell viability of the PC12 cells treated with nano-SiO2 (R2=0.945) suggesting nano-SiO2-induced membrane damage caused by lipid peroxidation. (4) A dose-dependent increase in sub-G1 population in SiO2-exposed cells along with cell shrinkage and nuclear condensation from morphological examination suggesting nano-SiO2-induced cell apoptosis. Furthermore, nano-SiO2 exposure diminished the ability of neurite extension in response to nerve growth factor in treated PC12 cells. In summary, SiO2 nanoparticle exposure resulted in dose-dependent neurotoxicity in cultured PC12 cells that was probably associated with oxidative stress and induced apoptosis., (Crown Copyright © 2011. Published by Elsevier Ltd. All rights reserved.)

The fabrication of a new amphiphilic block copolymer composed of a poly DL-lactic acid (PLA) hydrophobic backbone and pH dependent thermo-sensitive poly (N-isopropyl) methacrylamide-co-N-isopropylmaleamic acid-co-10-undecenoic acid (PNIPAAm-co-NIPMMA-co-UA) entities as hydrophilic domains as well as carrying 2,2,6,6-tetramethylpiperidin-1-oxyl (TEMPO) residues for electron spin resonance (ESR) labeling is reported. The lower critical solution temperature (LCST) of the copolymer was determined by optical absorbance measurements. The LCST was pH dependent and varied in a narrow practical range, 35.4 °C at pH 5.0, 37.5 °C at pH 6.5 and 39.4 °C at pH 7.4, which was below, near and above nominal physiological temperature respectively. The assembly of the copolymer into micelles in aqueous solution at temperatures below the LCST was confirmed by FT-IR, (1)H NMR and fluorescence spectroscopy. It is demonstrated that the anticancer drug, 5-fluorouracil (5-FU) can be loaded effectively within the polymeric micelles and released in response to environmental stimuli- namely, pH and temperature., (Crown Copyright © 2011. Published by Elsevier Inc. All rights reserved.)

A series of spin-labeled melphalan and chlorambucil derivatives, coupling the alkylating agents with 2,2,6,6-tetramethyl-1-piperidinyloxy (TEMPO) radicals, were synthesized, characterized, and their biological properties in vitro were evaluated. These compounds showed much higher cytotoxic activity against human leukemia cell line K562 in vitro than their parent compounds.

The effects of the addition of three different organosiloxanes, 3-aminopropyltrimethoxysilane (APTMS), 3-aminopropyltriethoxysilane (APTES) and 3-mercaptopropyltrimethoxysilane (MPTMS), on the phase behavior and the pore size of the surfactant-extracted materials have been investigated using powder XRD, TEM, Nitrogen adsorption/desorption isotherms and elemental analysis. The organosiloxanes were added to a typical synthesis, which results in a KIT-6 type in the absence of organosiloxanes. Their addition resulted in mesophase transformation or not, depending on the nature of functional groups of organosiloxanes. Increasing the content of APTMS or APTES in the silica sources not only led to the transformation of mesostructures from cubic bicontinuous Ia-3d to mixed mesostructure then towards two-dimensional (2-D) hexagonal p6mm and eventually to disordered structure, but also caused an increase of the pore size. However, the d spacing and the pore size of the cubic bicontinuous Ia-3d mesoporous solids can be decreased by addition of MPTMS between 0 and 2.5 mol% in the silica sources without a distinct decrease in the long-range order of the material. These results are discussed in terms of differences in the ability of the organosiloxanes to interact with the surfactant P123. The potential applications of these mesoporous materials are supports for the immobilization of enzymes, heavy metal sorbents, base catalysts, etc.

In order to elucidate the nanoparticle-induced cytotoxicity and its mechanism, the effects of 20 and 50 nm silica nanoparticles on cultured human embryonic kidney (HEK293) cells were investigated. Cell viability, mitochondrial function, cell morphology, reactive oxygen species (ROS), glutathione (GSH), thiobarbituric acid reactive substance (TBARS), cell cycle and apoptosis were assessed under control and silica exposed conditions. Exposure to 20 or 50 nm SiO(2) nanoparticles at dosage levels between 20 and 100 microg/ml decreased cell viability in a dose-dependent manner. Median lethal dose (LD(50)) of 24h exposure was 80.2+/-6.4 and 140.3+/-8.6 microg/ml for 20 and 50 nm SiO(2) nanoparticles, respectively. Morphological examination revealed cell shrinkage and nuclear condensation after SiO(2) nanoparticle exposure. Increase in intracellular ROS level and reduction in GSH content were also observed in SiO(2) nanoparticle-exposed HEK293 cells. Increase in the amount of TBARS suggested an elevated level of lipid peroxidation. Flow cytometric analysis showed that SiO(2) nanoparticles can cause G2/M phase arrest and apoptotic sub-G1 population increase in a dose-dependent manner. In summary, exposure to SiO(2) nanoparticles resulted in a dose-dependent cytotoxicity in cultured HEK293 cells that was associated with increased oxidative stress.

The direct electron transfer of glucose oxidase (GOx) immobilized on a large-pore mesoporous silica KIT-6 matrix fixed to a screen-printed electrode (SPE) and its application as a disposable biosensor were studied. KIT-6 plays a vital role in this method through immobilizing GOx and facilitating the electron communication between the active centers of GOx and the surface of the SPE. GOx immobilized on KIT-6 maintains its bioactivity and structure and displays stable and well-defined redox peaks with a formal potential of -428 mV (vs. Saturated Calomel Electrode), with a heterogeneous electron transfer rate constant of 4.15 s(-1) in phosphate buffer (0.05 M, pH 6.0). Upon the addition of glucose into air-saturated phosphate buffer solution, the reduction peak current decreases, which can be used analytically for glucose detection. The biosensor exhibits a linear response to glucose concentration ranging from 0 to 2.83 mM and a sensitivity of 7.29 microA (mM)(-1) cm(-2) at an applied potential of -0.5 V. The present work provides a promising strategy for fabricating a novel and disposable mediator-free glucose biosensor, which could be potentially mass-produced through further development.