Search

Abstract Rheumatoid arthritis (RA) involves chronic inflammation, oxidative stress, and complex immune cell interactions, leading to joint destruction. Traditional treatments are often limited by off-target effects and systemic toxicity. This study introduces a novel therapeutic approach using hyaluronic acid (HA)-conjugated, redox-responsive polyamino acid nanogels (HA-NG) to deliver tacrolimus (TAC) specifically to inflamed joints. The nanogels’ disulfide bonds enable controlled TAC release in response to high intracellular glutathione (GSH) levels in activated macrophages, prevalent in RA-affected tissues. In vitro results demonstrated that HA-NG/TAC significantly reduced TAC toxicity to normal macrophages and showed high biocompatibility. In vivo, HA-NG/TAC accumulated more in inflamed joints compared to non-targeted NG/TAC, enhancing therapeutic efficacy and minimizing side effects. Therapeutic evaluation in collagen-induced arthritis (CIA) mice revealed HA-NG/TAC substantially reduced paw swelling, arthritis scores, synovial inflammation, and bone erosion while suppressing pro-inflammatory cytokine levels. These findings suggest that HA-NG/TAC represents a promising targeted drug delivery system for RA, offering potential for more effective and safer clinical applications.

Abstract Au@Pt nanozyme, a bimetallic core–shell structure Au and Pt nanoparticle, has attracted significant attention due to its excellent catalytic activity and stability. Here, we propose that Au@Pt improves glucose tolerance and reduces TG after four weeks administration. The transcriptomic analysis of mouse liver tissues treated with Au@Pt nanozyme showed changes in genes related to glucose and lipid metabolism signaling pathways, including glycolysis/gluconeogenesis, pyruvate metabolism, PPAR signaling, and insulin signaling. Moreover, analysis of fecal samples from mice treated with Au@Pt nanozyme showed significant changes in the abundance of beneficial gut microbiota such as Dubosiella, Parvibacter, Enterorhabdus, Monoglobus, Lachnospiraceae_UCG-008, Lachnospiraceae_UCG-006, Lachnospiraceae_UCG-001, and Christensenellaceae_R-7_group. Combined multi-omics correlation analyses revealed that the modulation of glucose and lipid metabolism by Au@Pt was strongly correlated with changes in hepatic gene expression profiles as well as changes in gut microbial profiles. Overall, our integrated multi-omics analysis demonstrated that Au@Pt nanozyme could modulate glucose and lipid metabolism by regulating the expression of key genes in the liver and altering the composition of gut microbiota, providing new insights into the potential applications of Au@Pt nanozyme in the treatment of metabolic disorder. Graphical Abstract

Introduction: Chronic ankle instability (CAI) carries a high risk of progression to talar osteochondral lesions and post-traumatic osteoarthritis. It has been clinically hypothesized the progression is associated with abnormal joint motion and ligament elongation, but there is a lack of scientific evidence.Methods: A total of 12 patients with CAI were assessed during level walking with the use of dynamic biplane radiography (DBR) which can reproduce the in vivo positions of each bone. We evaluated the uninjured and CAI side of the tibiotalar and subtalar joint for three-dimensional kinematics differences. Elongation of the anterior talofibular ligament (ATFL), calcaneofibular ligament (CFL), and posterior talofibular ligament (PTFL) were also calculated bilaterally.Results: For patients with CAI, the dorsiflexion of the tibiotalar joint had reduced (21.73° ± 3.90° to 17.21° ± 4.35°), displacement of the talus increased (2.54 ± 0.64 mm to 3.12 ± 0.55 mm), and the inversion of subtalar joint increased (8.09° ± 2.21° to 11.80° ± 3.41°). Mean ATFL elongation was inversely related to mean dorsiflexion angle (CAI: rho = −0.82, P < 0.001; Control: rho = −0.92, P < 0.001), mean ATFL elongation was related to mean anterior translation (CAI: rho = 0.82, P < 0.001; Control: rho = 0.92, P < 0.001), mean CFL elongation was related to mean dorsiflexion angle (CAI: rho = 0.84, P < 0.001; Control: rho = 0.70, P < 0.001), and mean CFL elongation was inversely related to mean anterior translation (CAI: rho = −0.83, P < 0.001; Control: rho = −0.71, P < 0.001). Furthermore, ATFL elongation was significantly (CAI: rho = −0.82, P < 0.001; Control: rho = −0.78, P < 0.001) inversely correlated with CFL elongation.Discussion: Patients with CAI have significant changes in joint kinematics relative to the contralateral side. Throughout the stance phase of walking, ATFL increases in length during plantarflexion and talar anterior translation whereas the elongation trend of CFL was the opposite. This understanding can inform the development of targeted therapeutic exercises aimed at balancing ligament tension during different phases of gait. The interrelationship between two ligaments is that when one ligament shortens, the other lengthens. The occurrence of CAI didn’t change this trend. Surgeons might consider positioning the ankle in a neutral sagittal plane to ensure optimal outcomes during ATFL and CFL repair.

Stimulator of Interferon Genes (STING) is essential for the inflammatory response to cytosolic DNA. Despite that aberrant activation of STING is linked to an increasing number of inflammatory diseases, the development of inhibitors has been challenging, with no compounds in the pipeline beyond the preclinical stage. We previously identified endogenous nitrated fatty acids as novel reversible STING inhibitors. With the aim of improving the specificity and efficacy of these compounds, we developed and tested a library of nitroalkene-based compounds for in vitro and in vivo STING inhibition. The structure-activity relationship study revealed a robustly improved electrophilicity and reduced degrees of freedom of nitroalkenes by conjugation with an aromatic moiety. The lead compounds CP-36 and CP-45, featuring a β-nitrostyrene moiety, potently inhibited STING activity in vitro and relieved STING-dependent inflammation in vivo. This validates the potential for nitroalkene compounds as drug candidates for STING modulation to treat STING-driven inflammatory diseases, providing new robust leads for preclinical development.

Burdock (Arctium lappa L.), which belongs to the Compositae family, possesses significant medicinal and nutritional value. Burdock root is abundant in proteins and essential amino acids. In this study, proteins from burdock roots were extracted by alkali extraction and acid sedimentation, and the optimal conditions for protein extraction from burdock roots and protein characteristics were explored. The optimal conditions for protein extraction with a yield rate of 9.03% were 40°C extraction temperature, pH 8.0, a solid–liquid ratio of 1:25 (g/mL), and extraction time of 65 minutes. Single-factor and orthogonal tests on the extraction rate of burdock proteins under different conditions found that the extraction rate of protein from burdock roots were influenced by pH, extraction temperature, solid–liquid ratio, and extraction time, in this order. The solubility and gelation properties of burdock protein were similar to those of soybean protein; however, burdock protein demonstrated 1.2 times the oil absorption capacity and half the water retention capacity of soybean protein. Analysis of the amino acid contents of burdock protein revealed that burdock root contains significant essential and non-essential amino acids, with valine content being the highest.

Abstract Rheumatoid arthritis (RA) is a systemic autoimmune disease characterized by chronic inflammation of the joints and bone destruction. Because of systemic administration and poor targeting, traditional anti‐rheumatic drugs have unsatisfactory treatment efficacy and strong side effects, including myelosuppression, liver or kidney function damage, and malignant tumors. Consequently, mesenchymal stem cells (MSCs)‐involved therapy is proposed for RA therapy as a benefit of their immunosuppressive and tissue‐repairing effects. This review summarizes the progress of MSCs‐involved RA therapy through suppressing inflammation and promoting tissue regeneration and predicts their potential clinical application.

Type 2 diabetes mellitus (T2DM) as a chronic metabolic disease has become a global public health problem. Insulin resistance (IR) is the main pathogenesis of T2DM. Oxidative stress refers to an imbalance between free radical production and the antioxidant system, causing insulin resistance and contributing to the development of T2DM via several molecular mechanisms. Besides, the reduction in hepatic glycogen synthesis also leads to a decrease in peripheral insulin sensitivity. Thus, reducing oxidative stress and promoting glycogen synthesis are both targets for improving insulin resistance and treating T2DM. The current study aims to investigate the pharmacological effects of single-atom Ce-N-C nanozyme (SACe-N-C) on the improvement of insulin resistance and to elucidate its underlying mechanisms using HFD/STZ-induced C57BL/6J mice and insulin-resistant HepG2 cells. The results indicate that SACe-N-C significantly improves hepatic glycogen synthesis and reduces oxidative stress, as well as pancreatic and liver injury. Specifically, compared to the T2DM model group, fasting blood glucose decreased by 29%, hepatic glycogen synthesis increased by 17.13%, and insulin secretion increased by 18.87%. The sod and GPx in the liver increased by 17.80% and 25.28%, respectively. In terms of mechanism, SACe-N-C modulated glycogen synthesis through the PI3K/AKT/GSK3β signaling pathway and activated the Keap1/Nrf2 pathway to alleviate oxidative stress. Collectively, this study suggests that SACe-N-C has the potential to treat T2DM.

Objective The efficacy of sirolimus in treating severe or refractory systemic lupus erythematosus (SLE) has been confirmed by small-scale clinical trials. However, few studies focused on mild or moderate SLE. Therefore, in this study we elucidated clinical efficacy of add-on sirolimus in patients with mild or moderate SLE.Methods Data of 17 consecutive patients with SLE were retrospectively collected. SLE Disease Activity Index-2000 (SLEDAI-2K), clinical manifestation, laboratory data and peripheral T lymphocyte subsets with cytokines were collected before and 6 months after sirolimus add-on treatment. T cell subsets were detected by flow cytometry and cytokines were determined by multiplex bead-based flow fluorescent immunoassay simultaneously. Twenty healthy controls matched with age and sex were also included in our study.Results (1) The numbers of peripheral blood lymphocytes, T cells, T helper (Th) cells, regulatory T (Treg) cells, Th1 cells, Th2 cells and Treg/Th17 ratios in patients with SLE were significantly lower, while the numbers of Th17 cells were evidently higher than those of healthy control (p

Abstract Anti‐HER2 (human epidermal growth factor receptor 2) therapies significantly increase the overall survival of patients with HER2‐positive breast cancer. Unfortunately, a large fraction of patients may develop primary or acquired resistance. Further, a multidrug combination used to prevent this in the clinic places a significant burden on patients. To address this issue, this work develops a nanotherapeutic platform that incorporates bimetallic gold‐silver hollow nanoshells (AuAg HNSs) with exceptional near‐infrared (NIR) absorption capability, the small‐molecule tyrosine kinase inhibitor pyrotinib (PYR), and Herceptin (HCT). This platform realizes targeted delivery of multiple therapeutic effects, including chemo‐and photothermal activities, oxidative stress, and immune response. In vitro assays reveal that the HCT‐modified nanoparticles exhibit specific recognition ability and effective internalization by cells. The released PYR inhibit cell proliferation by downregulating HER2 and its associated pathways. NIR laser application induces a photothermal effect and tumor cell apoptosis, whereas an intracellular reactive oxygen species burst amplifies oxidative stress and triggers cancer cell ferroptosis. Importantly, this multimodal therapy also promotes the upregulation of genes related to TNF and NF‐κB signaling pathways, enhancing immune activation and immunogenic cell death. In vivo studies confirm a significant reduction in tumor volume after treatment, substantiating the potential effectiveness of these nanocarriers.

Abstract Oxyfluoride transparent glass‐ceramics (GC) are widely used as the matrix for rare‐earth (RE) ions due to their unique properties such as low phonon energy, high transmittance, and high solubility for RE ions. Tb3+ doped oxyfluoride glasses exhibit a large absorption cross section for ultraviolet (UV) excitation, high stability, high photoluminescence quantum efficiency, and sensitive spectral conversion characteristics, making them promising candidate materials for use as the spectral converter in UV photodetectors. Herein, a Tb3+ doped oxyfluoride GC is developed by using the melt‐quenching method, and the microstructure and optical properties of the GC sample are carefully investigated. By combining with a Si‐based photo‐resistor,a solar‐blind UV detector is fabricated, which exhibits a significant photoelectric response with a broad detection range from 188 to 400 nm. The results indicate that the designed UV photodetector is of great significance for the development of solar‐blind UV detectors.

Objectives Osteochondral lesions of the talus (OLTs) are common injuries in the general population. Abnormal mechanical conditions applied to defected cartilage are believed to be the culprits to deteriorating OLTs. This study aims to investigate the biomechanical effects of defect size of talar cartilage on OLTs during ankle movements. Methods A finite element model of the ankle joint was created based on the computed tomography images of a healthy male volunteer. Different defect sizes (S = 0.25, 0.5, 0.75, 1.0, 1.25, 1.5, 1.75, and 2.0 cm2) of talar cartilage were modeled to simulate the progression of OLTs. Mechanical moments were applied to the model to generate different ankle movements, including dorsiflexion, plantarflexion, inversion, and eversion. The effects of varying defect sizes on peak stress and its location were evaluated. Results The maximum stress on the talar cartilage increased as the area of the defect enlarged. Additionally, as the defect size of OLTs increased, the areas with peak stress on talar cartilage tended to move closer to where the injury was located. High stresses were present in the medial and lateral areas of the talus at the neutral position of the ankle joint. The concentrated stresses were mainly located in the anterior and posterior defect areas. The peak stress in the medial region was higher than on the lateral side. The order of peak stress from highest to lowest was dorsiflexion, internal rotation, inversion, external rotation, plantar flexion, and eversion. Conclusions Osteochondral defect size and ankle joint movements significantly modulate the biomechanical features of the articular cartilage in osteochondral lesions of the talus. The progression of osteochondral lesions in a talus deteriorates the biomechanical well‐being of the bone tissues of the talus.

Mechanical stimuli regulate the chondrogenic differentiation of mesenchymal stem cells and the homeostasis of chondrocytes, thus affecting implant success in cartilage tissue engineering. The mechanical microenvironment plays fundamental roles in the maturation and maintenance of natural articular cartilage, and the progression of osteoarthritis Hence, cartilage tissue engineering attempts to mimic this environment in vivo to obtain implants that enable a superior regeneration process. However, the specific type of mechanical loading, its optimal regime, and the underlying molecular mechanisms are still under investigation. First, this review delineates the composition and structure of articular cartilage, indicating that the morphology of chondrocytes and components of the extracellular matrix differ from each other to resist forces in three top-to-bottom overlapping zones. Moreover, results from research experiments and clinical trials focusing on the effect of compression, fluid shear stress, hydrostatic pressure, and osmotic pressure are presented and critically evaluated. As a key direction, the latest advances in mechanisms involved in the transduction of external mechanical signals into biological signals are discussed. These mechanical signals are sensed by receptors in the cell membrane, such as primary cilia, integrins, and ion channels, which next activate downstream pathways. Finally, biomaterials with various modifications to mimic the mechanical properties of natural cartilage and the self-designed bioreactors for experiment in vitro are outlined. An improved understanding of biomechanically driven cartilage tissue engineering and the underlying mechanisms is expected to lead to efficient articular cartilage repair for cartilage degeneration and disease.

Objectives We conducted a systematic review and meta-analysis to determine the efficacy of non-conventional synthetic disease-modifying antirheumatic drug (ncs-DMARD) strategies on patients with rheumatoid arthritis (RA)-associated interstitial lung disease (ILD).Methods PubMed, EMBASE, the Cochrane Library and Web of Science were searched for relevant articles from inception to 1 June 2022. The results obtained from the analysis were expressed as mean difference (MD), effect size and 95% CI.Results A total of 17 studies, including 1315 patients with RA-ILD, were eligible. The ncs-DMARDs included abatacept, rituximab, tocilizumab, tumour necrosis factor and Janus kinase inhibitors. Compared with the baseline, there were no significant changes in forced vital capacity (FVC), forced expiratory volume in the first second (FEV1) and diffusion lung capacity for carbon monoxide (DLCO) values in the pooled data after ncs-DMARD treatment (alone or combined with conventional therapy) (p=0.36 for FVC; p=0.96 for FEV1 and p=0.46 for DLCO). Of note, FVC was obviously increased in rituximab subgroup (MD=−4.62, 95% CI −8.90 to −0.33, p=0.03). Also, high-resolution CT non-progression rate and fatality rate due to ILD progression in patients with RA-ILD were 0.792 (95% CI 0.746 to 0.834, p=0.015) and 0.049 (95% CI 0.035 to 0.065, p=0.000), respectively.Conclusion ncs-DMARDs alone or combined with conventional therapy might be an optimal and promising treatment for stabilising or improving ILD in patients with RA-ILD.PROSPERO registration number CRD42022356816.

Nitro fatty acids (NO2-FAs) are endogenously generated lipid signaling mediators from metabolic and inflammatory reactions between conjugated diene fatty acids and nitric oxide or nitrite-derived reactive species. NO2-FAs undergo reversible Michael addition with hyperreactive protein cysteine thiolates to induce posttranslational protein modifications that can impact protein function. Herein, we report a novel mechanism of action of natural and non-natural nitroalkenes structurally similar to (E) 10-nitro-octadec-9-enoic acid (CP-6), recently de-risked by preclinical Investigational New Drug-enabling studies and Phase 1 and Phase 2 clinical trials and found to induce DNA damage in a TNBC xenograft by inhibiting homologous-recombination (HR)-mediated repair of DNA double-strand breaks (DSB). CP-6 specifically targets Cys319, essential in RAD51-controlled HR-mediated DNA DSB repair in cells. A nitroalkene library screen identified two structurally different nitroalkenes, a non-natural fatty acid [(E) 8-nitro-nonadec-7-enoic acid (CP-8)] and a dicarboxylate ester [dimethyl (E)nitro-oct-4-enedioate (CP-23)] superior to CP-6 in TNBC cells killing, synergism with three different inhibitors of the poly ADP-ribose polymerase (PARP) and γ-IR. CP-8 and CP-23 effectively inhibited γ-IR-induced RAD51 foci formation and HR in a GFP-reported assay but did not affect benign human epithelial cells or cell cycle phases. In vivo, CP-8 and CP-23's efficacies diverged as only CP-8 showed promising anticancer activities alone and combined with the PARP inhibitor talazoparib in an HR-proficient TNBC mouse model. As preliminary preclinical toxicology analysis also suggests CP-8 as safe, our data endorse CP-8 as a novel anticancer molecule for treating cancers sensitive to homologous recombination-mediated DNA repair inhibitors.

Abstract The multi-scale variability of global sea surface temperature (GSST), which is often dominated by secular trends, significantly impacts global and regional climate change. Previous studies were mainly carried out under linear assumptions. Even if the nonlinear evolution patterns have been discussed based on annual-mean data, the conclusions are still insufficient due to several factors. Here, based on the Ensemble Empirical Mode Decomposition (EEMD) method, the robustness of GSST trends tied to the sampling frequency and time interval selection is further explored. The main features derived from the annual-mean data are maintained. However, monthly and seasonal-mean data both mute the cooling in the equatorial central Pacific and the Southern Ocean in the Pacific sector, meanwhile intensify and expand the warming over the North Pacific. The results also highlight that early data cause a minimal effect on secular trends except for the portion near the start point of the interval due to the local temporal nature of EEMD. Overall, the long-term GSST trends extracted by EEMD have good robustness. Our research also clarifies that quadratic fitting cannot reveal all the meaningful evolution patterns, even as a nonlinear solution.

This paper aims to investigate passengers’ perception of the acoustic environment in airport terminals. Airport terminals typically consist of several interconnected large spaces and have a noisy environment in China. A questionnaire survey was conducted among 221 passengers at the T2 terminal of Tianjin Binhai International Airport in China to explore their perception and attitude towards the acoustic environment. On-site measurements were also taken to obtain the noise levels in different areas. The results show that the sound pressure levels vary periodically, ranging from 60.7 dB(A) to 79.1 dB(A) across different areas. The acoustic environment lacks eventfulness and pleasantness, with nearly two-thirds of respondents expressing dissatisfaction. Public announcement is generally perceived as the loudest but most preferred sound, while other audible sources are given varying degrees of negative attitudes. The perceived loudness of aircraft and footstep sounds is not high, but their absence significantly improves overall evaluations. Furthermore, age influences the perceived vitality of the acoustic environment. This study is important for the improvement in the indoor environment of the terminal building and the satisfaction of passengers.

The accumulation of reactive oxygen species (ROS) in the oxidative stress microenvironment of diabetic wounds often results in delayed wound healing. In this study, a multifunctional hydrogel with ROS-scavenging property was developed by cross-linking epigallocatechin-3-gallate (EGCG), 2-(hydroxyethyl) methacrylamide (HEMAA), acrylamide (AM), and borax. The resulting hydrogel showed adequate hemostatic properties, self-healing capability, tissue adhesiveness, and an appropriate degradation rate. Moreover, the green tea derivative-based hydrogel scavenged the accumulated ROS and protected skin-related cells (such as keratinocytes, fibroblasts, and endothelial cells) from ROS-mediated death and proliferation inhibition. In vivo, this hydrogel accelerated diabetic wound healing by decreasing the ROS level, inducing macrophage polarization to the M2 phenotype, alleviating excessive inflammation, and promoting proliferation, epithelialization, collagen deposition, and neovascularization. By day 16, the hydrogel dressing had almost completely healed the diabetic wounds, whereas the residual wound areas of the blank control and the commercial TegadermTM dressing were 55.57 ± 5.67 % and 35.15 ± 3.45 %, respectively. Overall, this advanced hydrogel provides an effective strategy for the treatment of diabetic wounds and may be applied for the treatment of tissue injury in other ROS-accumulated microenvironments.

Background: The current study investigated the application of three-dimensional (3D) printing technology in the treatment of talar avascular necrosis (TAN). Custom-made Vitallium talar prostheses were designed and generated via 3D printing. We hypothesized that these talar prostheses would facilitate more stable positioning, better ergonomically fit the ankle joint surfaces, and promote favorable long-term prognoses.Material and Methods: Computed tomography scans of both ankle joints were acquired from three patients diagnosed with TAN. The talar on the unaffected side was used as the design blueprint. Hence, with the aid of 3D printing technology a customized talar prosthesis made from a novel Vitallium alloy could be manufactured for each individual patient.Results: In all three cases there were no signs of prosthesis loosening or substantial degenerative change in the surrounding area of the joint, but small osteophytes were observed on the tibial side and navicular side. No chronic infection or other prosthesis-related complications were observed in any of the patients. All three were able to walk without pain at the most recent follow-up.Conclusion: With the aid of 3D printing and a novel Vitallium alloy, total talar replacement achieved encouraging results in 3/3 patients. All patients were satisfied with their joint function, and were able to return to their daily activities without limitations. Although more cases and longer-term follow-up periods are required, the success rate reported herein is encouraging.

Abstract Introduction Laccases are green biocatalysts that possess attractive for the treatment of resistant environmental pollutants and dye effluents. Purpose To exploit the laccase of Anoxybacillus ayderensis SK3-4 that possesses dye decolorization ability at room and higher temperature, we characterized the enzyme in considerable detail and investigated its ability to decolorize different dyes. Results A bacterial laccase gene designed as LacAn from Anoxybacillus ayderensis SK3-4 of hot springs was cloned and expressed in Escherichia coli. LacAn is a monomeric protein with a molecular weight of 29.8 kDa. The optimum pH and temperature for syringaldazine oxidation were 7.0 and 75 °C, respectively. LacAn was stable at pH values ranging from 6.5 to 8.5 above 65 °C. The enzyme activity was significantly enhanced by Cu2+ and Mg2+ but inhibited by Zn2+ and Fe2+. Furthermore, LacAn showed high decolorization capability toward five dyes (direct blue 6, acid black 1, direct green 6, direct black 19, and acid blue 93) in the absence of redox mediators. It also demonstrated a wide temperature range, and it can retain its high decolorization ability even at high temperatures. Conclusions These properties including better enzymatic properties and efficiency to decolorize dyes demonstrate that the bacterial laccase LacAn has potentials for further industrial applications.

The therapy of burns is a challenging clinical issue. Burns are long-term injuries, and numerous patients suffer from chronic pain. Burn treatment includes management, infection control, wound debridement and escharotomy, dressing coverage, skin transplantation, and the use of skin substitutes. The future of advanced care of burn wounds lies in the development of “active dressings”. Hydrogel dressings have been employed universally to accelerate wound healing based on their unique properties to overcome the limitations of existing treatment methods. This review briefly introduces the advantages of hydrogel dressings and discusses the development of new hydrogel dressings for wound healing along with skin regeneration. Further, the treatment strategies for burns, ranging from external to clinical, are reviewed, and the functional classifications of hydrogel dressings along with their clinical value for burns are discussed.

A series of binary composites Bi4O5Br2/BiPO4 (PBX) was fabricated through a simple mechanical ball milling protocol. Relevant microstructural, morphological, and optical properties were thoroughly analyzed via various techniques. The integration of both components was confirmed to produce heterojunction domains at the phase boundaries. Upon exposure to visible light irradiation, the as-achieved PBX series possessed the reinforced photocatalytic NOx removal efficiencies and the weakened generation of toxic intermediate NO2 in comparison to both bare components, chiefly attributed to the efficient transport and separation of carriers and boosted production of superoxide radicals (·O2−) through the combination of a wide-bandgap ornament BiPO4 as an electron acceptor. In particular, the composite PB5 with the optimal phase composition exhibited the highest NOx removal of 40% with the lowest NO2 formation of 40 ppb among all tested candidates. According to the band structures’ estimation and reactive species’ detection, a reasonable mechanism was ultimately proposed to describe the migration of charge carriers and the enhancement of photocatalytic performance.

Plantar fascia plays an important role in human foot biomechanics; however, the morphology and mechanical properties of plantar fascia in patients with flexible flatfoot are unknown. In this study, 15 flexible flatfeet were studied, each plantar fascia was divided into 12 positions, and the morphologies and mechanical properties in the 12 positions were measured in vivo with B-mode ultrasound and shear wave elastography (SWE). Peak pressures under the first to fifth metatarsal heads (MH) were measured with FreeStep. Statistical analysis included 95% confidence interval, intragroup correlation coefficient (ICC1,1), one-way analysis of variance (one-way ANOVA), and least significant difference. The results showed that thickness and Young’s modulus of plantar fascia were the largest at the proximal fascia (PF) and decreased gradually from the proximal end to the distal end. Among the five distal branches (DB) of the fascia, the thickness and Young’s modulus of the second and third DB were larger. The peak pressures were also higher under the second and third MH. This study found a gradient distribution in that the thickness and Young’s modulus gradient decreased from the proximal end to the distal end of plantar fascia in the longitudinal arch of flexible flatfeet. In the transverse arch, the thickness and Young’s modulus under the second and third DB were larger than those under the other three DB in flexible flatfoot, and the peak pressures under the second and third MH were also larger than those under the other three MH in patients with flexible flatfoot. These findings deepen our understanding of the changes of biomechanical properties and may be meaningful for the study of pathological mechanisms and therapy for flexible flatfoot.

Abstract Background Dishevelled-associated activator of morphogenesis 1 (DAAM1) is a member of microfilament-related formins and mediates cell motility in breast cancer (BrCa). However, the genetic mutation status of DAAM1 mRNA and its correlation with pathological characteristics are still unclearly. Methods A patient cohort and BrCa cells were recruited to demonstrate the role of functional SNP in microRNA-208a-5p binding site of DAAM1 3′-UTR and underlying mechanism in BrCa metastasis. Results The expression and activation of DAAM1 increased markedly in lymphnode metastatic tissues. A genetic variant (rs79036859 A/G) was validated in the miR-208a-5p binding site of DAAM1 3′-UTR. The G genotype (AG/GG) was a risk genotype for the metastasis of BrCa by reducing binding affinity of miR-208a-5p for the DAAM1 3′-UTR. Furthermore, the miR-208a-5p expression level was significantly suppressed in lymphnode metastatic tissues compared with that in non-lymphnode metastatic tissues. Overexpression of miR-208a-5p inhibited DAAM1/RhoA signaling pathway, thereby leading to the decrease of the migratory ability. Conclusion Overall, the rs79036859 G variant of DAAM1 3′-UTR was identified as a relevant role in BrCa metastasis via the diversity of miR-208a-5p binding affinity.

Three-dimensional (3D) ZnO hierarchical microspheres composed of well-ordered nanorods were synthesized by a facile one-step hydrothermal method without any surfactants. The effects of reaction conditions including reaction temperature, reaction time on the microstructure of hierarchical microspheres were investigated. The results demonstrated that the reaction conditions played an important role in the morphology of 3D ZnO hierarchical microspheres. A three-stage reaction mechanism of 3D ZnO hierarchical microspheres was proposed. Firstly, Zn2+ reacts with OH− and NH4+ to produce nanoparticles. Then the nanoparticles are self-assembled into nucleis. Finally is the formation and growth of the hierarchical structure on the nuclear surface. When the as-synthesized ZnO were used as the anode of the dye sensitization solar cells (DSSCs), the 3D ZnO hierarchical microspheres composed of well-ordered nanorods exhibits the best photoelectric properties, which was attributed to the increased surface area and advantageous electronic transmission path. Keywords: Hierarchical microspheres, Conditions, Mechanism, Photoelectric properties

The wide diffusion of information in social networks can be exploited to solve searching-for-a-target (SFT) problems including those of missing individuals. Incentive mechanisms that promote active individual participation can be designed to favor a clear propagation direction to help efficiently find a target. However, the existing incentive research rarely focuses on a clear propagation direction based on a specific goal. Thus, we propose an effective contribution-driven information propagation incentive mechanism (IPIM) that exploits ego networks to solve the SFT problem. First, we use an all-pay auction-inspired model to determine the propagation of alters in each ego network. We then propose a novel algorithm, the node propagation utility, based on effective contributions, to focus the propagation toward the target rather than searching indiscriminately and inefficiently. The theoretical analyses and simulation results indicate that IPIM guarantees the truthfulness, individual rationality, and budget feasibility. The simulations are conducted based on real and public social datasets. The IPIM shows increased efficiencies of 951.18 % of success rate, of 215.65 % in propagation hops, and of 514.41 % in participation scale, compared with a typical incentive mechanism. In conclusion, the IPIM shows significant value in the potential application in SFT.

Background The role of tumor necrosis factor (TNF)-α small interfering (si)RNA alveolar epithelial cell (AEC)-targeting nanoparticles in lung injury is unclear. Methods Sixty C57BL/6J mice with sepsis were divided into normal, control, sham, 25 mg/kg, 50 mg/kg, and 100 mg/kg siRNA AEC-targeting nanoparticles groups (n = 10 per group). The wet:dry lung weight ratio, and hematoxylin and eosin staining, western blotting, and enzyme-linked immunosorbent assays for inflammatory factors were conducted to compare differences among groups. Results The wet:dry ratio was significantly lower in control and sham groups than other groups. TNF-α siRNA AEC-targeting nanoparticles significantly reduced the number of eosinophils, with significantly lower numbers in the 50 mg/kg group than in 25 mg/kg and 100 mg/kg groups. The nanoparticles also significantly reduced the expression of TNF-α, B-cell lymphoma-2, caspase 3, interleukin (IL)-1β, and IL-6, with TNF-α expression being significantly lower in the 50 mg/kg group than in 25 mg/kg and 100 mg/kg groups. Conclusion TNF-α siRNA AEC-targeting nanoparticles appear to be effective at improving lung injury-related sepsis, and 50 mg/kg may be a preferred dose option for administration.

Cartilage injuries are typically caused by trauma, chronic overload, and autoimmune diseases. Owing to the avascular structure and low metabolic activities of chondrocytes, cartilage generally does not self-repair following an injury. Currently, clinical interventions for cartilage injuries include chondrocyte implantation, microfracture, and osteochondral transplantation. However, rather than restoring cartilage integrity, these methods only postpone further cartilage deterioration. Stem cell therapies, especially mesenchymal stem cell (MSCs) therapies, were found to be a feasible strategy in the treatment of cartilage injuries. MSCs can easily be isolated from mesenchymal tissue and be differentiated into chondrocytes with the support of chondrogenic factors or scaffolds to repair damaged cartilage tissue. In this review, we highlighted the full success of cartilage repair using MSCs, or MSCs in combination with chondrogenic factors and scaffolds, and predicted their pros and cons for prospective translation to clinical practice.

Micro-electro-mechanical system (MEMS) blazed gratings are the critical optical elements in high resolution micro-spectrometer systems. The MEMS blazed gratings fabricated by conventional methods are not ideal saw-tooth profiles. After the photolithographic and anisotropic etching process, a tiny platform of silicon will be left on the top of the grating groove because of the silica mask, which will lead to amounts of stray light and reduce the diffraction efficiency of gratings. This paper presents a novel design and fabrication method for fabricating MEMS blazed grating, which utilizes off-cut silicon (111) wafers. The blazed angle of gratings is revised from 7.54° to 8.0° according to the tiny platform width on the top of grating grooves. In the procedure of grating fabrication, the oxidation sharpening method is used to cut down the width of the tiny platform. The experimental result shows that the platform width of the grating groove has sharpened from 1000 nm to 540 nm, and the fabricated blazed grating has high diffraction efficiency. The result indicates that the process to fabricate blazed gratings through the anisotropic etching and oxidation sharpening technology is quite effective.

Invertases catalyze the hydrolysis of sucrose into fructose and glucose and can be employed as an alternative in producing high fructose syrup. In this study, we reported the heterologous expression of an invertase gene (GspInv) of Gongronella sp. w5 in Komagataella pastoris. GspInv activity reached 147.6 ± 0.4 U/mL after 5 days of methanol induction. GspInv is invertase with a high specific activity of 2,776.1 ± 124.2 U/mg toward sucrose. GspInv showed high tolerance to sucrose (IC50 = 1.2 M), glucose (IC50 > 2 M), fructose (IC50 = 1.5 M), and a variety of metal ions that make it an ideal candidate for high fructose syrup production. A carbohydrate-binding module was sequence-optimized and fused to the N-terminus of GspInv. The fusion protein had the highest immobilization efficiency at room temperature within 1 h adsorption, with 1 g of cellulose absorption up to 8,000 U protein. The cellulose-immobilized fusion protein retained the unique properties of GspInv. When applied in high fructose syrup preparation by using 1 M sucrose as the substrate, the sucrose conversion efficiency of the fused protein remained at approximately 95% after 50 h of continuous hydrolysis on a packed bed reactor. The fused protein can also hydrolyze completely the sucrose in sugarcane molasses. Our results suggest that GspInv is an unusual invertase and a promising candidate for high fructose syrup preparation.

Abstract Background β-Glucosidase has attracted substantial attention in the scientific community because of its pivotal role in cellulose degradation, glycoside transformation and many other industrial processes. However, the tedious and costly expression and purification procedures have severely thwarted the industrial applications of β-glucosidase. Thus development of new strategies to express β-glucosidases with cost-effective and simple procedure to meet the increasing demands on enzymes for biocatalysis is of paramount importance. Results Light activated cassette YF1/FixJ and the SRRz lysis system were successfully constructed to produce Bgl1A(A24S/F297Y), a mutant β-glucosidase tolerant to both glucose and ethanol. By optimizing the parameters for light induction, Bgl1A(A24S/F297Y) activity reached 33.22 ± 2.0 U/mL and 249.92 ± 12.25 U/mL in 250-mL flask and 3-L fermentation tank, respectively, comparable to the controls of 34.02 ± 1.96 U/mL and 322.21 ± 10.16 U/mL under similar culture conditions with IPTG induction. To further simplify the production of our target protein, the SRRz lysis gene cassette from bacteriophage Lambda was introduced to trigger cell autolysis. As high as 84.53 ± 6.79% and 77.21 ± 4.79% of the total β-glucosidase were released into the lysate after cell autolysis in 250 mL flasks and 3-L scale fermentation with lactose as inducer of SRRz. In order to reduce the cost of protein purification, a cellulose-binding module (CBM) from Clostridium thermocellum was fused into the C-terminal of Bgl1A(A24S/F297Y) and cellulose was used as an economic material to adsorb the fusion enzyme from the lysate. The yield of the fusion protein could reach 92.20 ± 2.27% after one-hour adsorption at 25 °C. Conclusions We have developed an efficient and inexpensive way to produce β-glucosidase for potential industrial applications by using the combination of light induction, cell autolysis, and CBM purification strategy.

Four kinds of Al-TiO2 solid samples with different Ti/Al ratios of 1 : 0.1, 1 : 0.09, 1 : 0.07, and 1 : 0.05 were synthesized via a solid-phase synthesis method and characterized by XRD, SEM, EDS, BET, and other techniques. The prepared solids were used for separation of the bioderived product danshensu, the content of which was determined by UV spectrophotometry. Moreover, the effects of extract concentration, PH value, adsorption time, and ethanol elution volume were investigated. The results showed that these Al-TiO2 samples had good adsorption and desorption ability. Especially, the solid Al-TiO2 with a Ti/Al ratio of 1 : 0.05 is more suitable for the separation of danshensu, exhibiting a higher adsorption (77.70%) under 2 h adsorption time and pH = 3; meanwhile, the high desorption rate (70.29%）was received under 80% ethanol and the sample concentration of 3.0 mg/mL.

Aiming at the carbon fiber reinforced plastics (CFRP) of poor electrical conductivity which will lead to thermal stress damage due to intense Joule effect under lightning strikes, electro-thermal-structure coupling models of CFRP lightning strikes were established. By analyzing the thermal strain, thermal stress and status of the modified and unmodified CFRP, their thermal expansion characteristics were compared. Compared with the unmodified CFRP, the thermal strain peak value of the modified CFRP is respectively increased by 36.33%, 53.92%, 66.62%, and the thermal stress peak value of them is respectively reduced by 22.63%, 26.53%, and 9.45% in the absence of dielectric breakdown. But in the breakdown state, both specimens appear complete delamination damage, the protective effect of matrix modification becomes invalid. The thermal stress damage is found having little effect on the damage morphology of the specimens after lightning strikes by comparing their CT images.

A new copper-based supramolecular (β-cyclodextrins, β-CD) catalyst was synthesized and used for transesterification of Xanthium sibiricum Patr oil to biodiesel. This catalyst exhibited high activity (88.63% FAME yield) in transesterification under the ratio of methanol-oil: 40 : 1; catalyst dosage: 8 wt.%; reaction temperature: 120°C; and reaction time: 9 h. The XRD, SEM, TEM, XPS, and BET characterization results showed that Cu-β-CD catalyst was amorphous and had clear mesoporous structure (17.2 nm) as compared with the native β-CD. This phenomenon is attributed to the coordination of Cu and β-CD.

Mesoporous solid acids Ta3W7 and Nb3W7 were prepared from TaCl5 and NbCl5 with WCl6 in the presence of stearic acid (SA) via a sol-gel method, respectively. For comparison, mesoporous Ta3W7-P123 mixed oxides and mesoporous Nb3W7-P123 mixed oxides were synthesized in the same way. The catalysts were characterized through TGA, XRD, SEM, TEM, BET, and NH3-TPD. Experimental results showed that Ta3W7-SA and Nb3W7-SA exhibited several advantages such as higher activity, shorter preparation period, lower cost, stronger acid sites, and higher surface area, which had potential to be used as mesoporous heterogeneous catalysts in biodiesel production.

In recent decades, the biomedical applications of mesenchymal stem cells (MSCs) have attracted increasing attention. MSCs are easily extracted from the bone marrow, fat, and synovium, and differentiate into various cell lineages according to the requirements of specific biomedical applications. As MSCs do not express significant histocompatibility complexes and immune stimulating molecules, they are not detected by immune surveillance and do not lead to graft rejection after transplantation. These properties make them competent biomedical candidates, especially in tissue engineering. We present a brief overview of MSC extraction methods and subsequent potential for differentiation, and a comprehensive overview of their preclinical and clinical applications in regenerative medicine, and discuss future challenges.

This paper presents a dispersive near-infrared spectrometer with features of miniaturization, portability and low cost. The application of a resonantly-driven scanning grating mirror (SGM) as a dispersive element in a crossed Czerny–Turner configuration enables the design of a miniaturized spectrometer that can detect the full spectra using only one single InGaAs diode. In addition, a high accuracy recalculation is realized, which can convert time-dependent measurements to spectrum information by utilizing the deflection position detector integrated on SGM and its associated closed-loop control circuit. Finally, the spectrometer prototype is subjected to a series of tests to characterize the instrument’s performance fully. The results of the experiment show that the spectrometer works in a spectral range of 800 nm–1800 nm with a resolution of less than 10 nm, a size of 9 × 7 × 7 cm3, a wavelength stability better than ±1 nm and a measuring time of less than 1 ms. Furthermore, the power consumption of the instrument is 3 W at 5 V DC, and the signal-to-noise ratio is 3267 at full scale. Therefore, this spectrometer could be a potential alternative to classical spectrometers in process control applications or could be used as a portable or airborne spectroscopic sensor.

The persistent inflammation of rheumatoid arthritis (RA) always leads to partial synovial hyperplasia and the destruction of articular cartilage. Bone marrow mesenchymal stem cells (BMMSCs) have been proven to possess immunosuppressive effects, and widely explored in the treatment of autoimmune diseases. However, poor inhibitory effect on local inflammatory state and limited capacity of preventing destruction of articular cartilage by systemic BMMSCs transplantation were observed. Herein, toward the classical type II collagen-induced arthritis in rats, the combination treatment of microfracture and in situ transplantation of thermogel-encapsulated BMMSCs was verified to obviously down-regulate the ratio of CD4+ to CD8+ T lymphocytes in peripheral blood. In addition, it resulted in the decreased levels of inflammatory cytokines, such as interleukin-1β, tumor necrosis factor-α and anti-collagen type II antibody, in the serum. Simultaneously, the combination therapy also could inhibit the proliferation of antigen specific lymphocytes and local joint inflammatory condition, and prevent the articular cartilage damage. The results indicated that the treatment programs could effectively stimulate the endogenous and exogenous BMMSCs to exhibit the immunosuppression and cartilage protection capability. This study provided a new therapeutic strategy for autoimmune inflammatory diseases, such as RA.

Bacterial laccases are ideal alternatives of fungal laccases for specific industrial applications due to specific characteristics such as alkalescence dependence and high chloride tolerance. However, some bacterial laccases presented as inclusion bodies when expressing in Escherichia coli and showed thermal instability. In this study, rational design was employed to enhance the solubility and the thermostablity of the bacterial laccase Lac15-His6 based on the crystal structure obtained previously. After deletion of His-tag and residues323-332, the obtained Lac15D was completely expressed in soluble form even at a higher temperature of 28°C, compared to only 50% of Lac15-His6 expressed solubly at 16°C. It showed a two-time higher activity at temperatures lower than 35°C and a half-life increasing from 72 min to 150 min at 45°C. When used in chromogenic reactions, Lac15D showed constant activity toward dye precursors and their combinations under alkaline conditions, demonstrating its application potential in hair coloring biotechnology.

In an on-road environment, motor-engines severely disturb the wireless link of a sensor node, leading to high package loss rate, high delivery delay, and poor radio communication quality. The existing data delivery mechanisms, such as the ACK-based retransmission mechanism and window-based link quality estimation mechanism, could not handle these challenges well. To solve this challenge, we propose a Target-Prediction-based Link quality Estimation scheme (TPLE) to realize high quality data delivery in an on-road environment. To perform on-road link quality estimation, TPLE dynamically calculates the track of a nearby vehicle target and estimates target impact on wireless link. Based on the local estimation of link quality, TPLE schedules radio communication tasks effectively. Simulations indicate that our proposed TPLE scheme produces a 94% data delivery rate, its average retransmission number is around 0.8. Our conducted on-road data delivery experiments also indicated a similar result as the computer simulation.

In the centrosymmetric tetranuclear title complex, [Ni4(C12H15NO3)2(CH3COO)2(N3)2(CH3OH)2]·2CH3OH, the asymmetric unit comprises half of a complex molecule and a methanol solvent molecule. The NiII ions display two different coordination environments: (i) two O atoms from the Schiff base ligand, two O atoms from symmetry-related methanol molecules and an O atom from an acetate group, one N atom from the azide group, and (ii) two O atoms and one N atom from the Schiff base, one O atom from methanol, one O atom from the acetate anion, and one N atom from the azide group. Four coplanar NiII ions are connected by two μ2-bridging O atoms from the two deprotonated Schiff bases, two μ3-O atoms from methanol molecules, two μ1,1-N atoms from two azide ions, and four O atoms from acetate groups. The shortest Ni...Ni distance in the tetranuclear unit is 2.962 (2) Å. O—H...O hydrogen bonds between the methanol solvent molecule and an acetate O atom feature in the crystal packing.

The digestion of lignin and lignin-related phenolic compounds from bamboo by giant pandas has puzzled scientists because of the lack of lignin-degrading genes in the genome of the bamboo-feeding animals. We constructed a 16S rRNA gene library from the microorganisms derived from the giant panda feces to identify the possibility for the presence of potential lignin-degrading bacteria. Phylogenetic analysis showed that the phylotypes of the intestinal bacteria were affiliated with the phyla Proteobacteria (53%) and Firmicutes (47%). Two phylotypes were affiliated with the known lignin-degrading bacterium Pseudomonas putida and the mangrove forest bacteria. To test the hypothesis that microbes in the giant panda gut help degrade lignin, a metagenomic library of the intestinal bacteria was constructed and screened for clones that contained genes encoding laccase, a lignin-degrading related enzyme. A multicopper oxidase gene, designated as lac51, was identified from a metagenomic clone. Sequence analysis and copper content determination indicated that Lac51 is a laccase rather than a metallo-oxidase and may work outside its original host cell because it has a TAT-type signal peptide and a transmembrane segment at its N-terminus. Lac51 oxidizes a variety of lignin-related phenolic compounds, including syringaldazine, 2,6-dimethoxyphenol, ferulic acid, veratryl alcohol, guaiacol, and sinapinic acid at conditions that simulate the physiologic environment in giant panda intestines. Furthermore, in the presence of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), syringic acid, or ferulic acid as mediators, the oxidative ability of Lac51 on lignin was promoted. The absorbance of lignin at 445 nm decreased to 36% for ABTS, 51% for syringic acid, and 51% for ferulic acid after incubation for 10 h. Our findings demonstrate that the intestinal bacteria of giant pandas may facilitate the oxidation of lignin moieties, thereby clarifying the digestion of bamboo lignin by the animal.

Follicle-stimulating hormone (FSH) is a pituitary glycoprotein that, together with luteinizing hormone, plays a crucial role in ovarian folliculogenesis and female fertility. We previously found that FSH beta is a major gene controlling high prolificacy of Chinese Erhualian pigs. To directly study the biological effects on reproductive function of porcine FSH (pFSH) for polyovulatory species, we generated a novel gain-of-function mouse model using a bacterial artificial chromosome (BAC) system to jointly introduce 92 kb and 165 kb genomic fragments comprising the pFSH α- and β-subunit genes. These directed the physiological expression of pFSH with the same temporal and spatial pattern as endogenous FSH in female transgenic (TG) mice. Serum levels of biologically active pFSH heterodimers in independent TG lines ranged from 6.36 to 19.83 IU/L. High basal pFSH activity led to a significant reduction of serum LH and testosterone levels in TG females compared to wild-type (WT) littermates, yet endogenous FSH and estradiol levels were significantly elevated. Interestingly, ovarian histology showed that the number of corpora lutea was significantly higher at 14 and 28 weeks of age in TG females and breeding curves revealed that mean litter sizes of TG females were obviously larger than for WT littermates before 52 weeks of age. These findings indicate that pituitary-specific overexpression of pFSH within physiological boundaries can increase ovulation rate and litter size, but it does not cause reproductive defects. Therefore, our TG mouse model provides exciting insights for investigating the actions of pFSH in vivo.

This study investigates the performance and cycling fading of a solid-state fluoride-ion battery (FIB) based on the CuF2 electrode. The cathode and solid electrolyte of CuF2 composite and La0.9Ba0.1F2.9 are prepared by ballmilling. Meanwhile, the anode materials are used as Sn and Pb. All FIB cells with sandwich structures are fabricated by compressing under a pressure of 4.5 tons/cm². Electrochemical measurements of discharge/charge are performed at 423 K and under 40 μA/cm². The resultant cycling stability of the cell with the Pb anode is higher than that of the cell with the Sn anode. Concerning the cell with Pb anode, the first and tenth discharge capacities of 150 and 90 mAh/g are obtained. X-ray photoelectron analysis demonstrates that the cycling fading of the cell with the Sn (or Pb) anode may be attributed to the irreversible formation of materials (e.g., SnF4 or PbF4) during the electrochemical reaction. [ABSTRACT FROM AUTHOR]

This study aims to investigate the CO2 uptake performance of mesostructured adsorbents, such as Mobil Composition of Matter No. 41 (MCM-41), Santa Barbara Amorphous-15 (SBA-15), and multi-walled carbon nanotubes (MWNTs), modified with polyethylenimine (PEI). Mesoporous materials are loaded with 50 wt.% PEI using a wet impregnation method. CO2 kinetic experiments of the PEI-modified adsorbents are conducted by a thermogravimetric method. The results reveal that the CO2 adsorption capacities of the PEI/MCM-41, PEI/SBA-15, and PEI/MWNTs composites are 2.02, 3.06, and 2.93 mmol/g, respectively, under 15% CO2 flow at 348 K. The lower CO2 adsorption capacity of PEI/MCM-41 (2.02 mmol/g) is attributed to its poor porosity. The PEI/MWNTs composite has the fastest CO2 adsorption and desorption kinetics at the same temperature, compared to other PEImodified adsorbents. These results suggest that MWNTs might play a significant "separator" role in effectively dispersing the PEI molecular chains on the mesostructured adsorbent. [ABSTRACT FROM AUTHOR]