Dietary Fiber Intake Influences Changes in Ankylosing Spondylitis Disease Status.

The objectives of this study were to characterize dietary fiber (DF) intake in patients with ankylosing spondylitis (AS), to assess whether DF intake affects disease activity in AS, and to investigate the effect of DF intake on disease activity in AS in the context of functional bowel disease (FBD) symptoms. We recruited 165 patients with AS and divided them into two groups according to whether they had a high DF intake > 25 g/d to investigate the characteristics of people with high DF intake. Some 72 of the 165 AS patients (43%) met the criteria for high DF intake, which was more common in patients with negative FBD symptoms (68%). Data analysis revealed that DF intake was negatively associated with AS disease activity and did not differ statistically significantly from FBD symptoms. Multivariate adjusted models were used to explore the effect of DF intake on AS disease activity. ASDAS-CRP and BASDAI were stable and negatively correlated across models in both groups with and without FBD symptoms. Thus, DF intake positively affected disease activity in patients with AS. ASDAS-CRP and BASDAI were negatively correlated with DF intake.

Precise Design of Alginate Hydrogels Crosslinked with Microgels for Diabetic Wound Healing.

Alginate hydrogel has received great attention in diabetic wound healing. However, the limited tunability of the ionic crosslinking method prevents the delicate management of physical properties in response to diverse wound conditions. We addressed this issue by using a microgel particle (fabricated by zinc ions and coordinated through the complex of carboxymethyl chitosan and aldehyde hyaluronic acid) as a novel crosslinker. Then the cation was introduced as a second crosslinker to create a double crosslinked network. The method leads to the precise regulation of the hydrogel characters, including the biodegradation rate and the controlled release rate of the drug. As a result, the optimized hydrogels facilitated the live-cell infiltration in vitro and boosted the tissue regeneration of diabetic wounds in vivo. The results indicated that the addition of the microgel as a new crosslinker created flexibility during the construction of the alginate hydrogel, adapting for diverse applications during diabetic-induced wound therapy.

Heparin Protects Severe Acute Pancreatitis by Inhibiting HMGB-1 Active Secretion from Macrophages.

Heparin has shown benefits in severe acute pancreatitis (SAP) therapy, but the underlying mechanisms were unknown. Extracellular high-mobility group protein-1 (HMGB-1) has been regarded as a central mediator contributing to inflammation exacerbation and disease aggravation. We hypothesized heparin attenuated the disease by targeting HMGB-1-related pathways. In the present study, the possible therapeutic roles of heparin and its non-anticoagulant derivatives, 6-O-desulfulted heparin and N-acylated-heparin, were determined on mouse models induced by "Two-Hit" of L-arginine. The compounds exhibited potent efficiency by substantially decreasing the pancreatic necrosis, macrophage infiltration, and serum inflammatory cytokine (IL-6 and TNF-α) concentration. Moreover, they greatly reduced the rapidly increasing extracellular HMGB-1 levels in the L-arginine injured pancreases. As a result, multiple organ failure and mortality of the mice were inhibited. Furthermore, the drugs were incubated with the RAW264.7 cells activated with damaged pancreatic tissue of SAP mice in vitro. They were found to inhibit HMGB-1 transfer from the nucleus to the plasma, a critical step during HMGB-1 active secretion from macrophages. The results were carefully re-examined with a caerulein and LPS induced mouse model, and similar results were found. The paper demonstrated heparin alleviated SAP independent of the anti-coagulant functions. Therefore, non-anticoagulant heparin derivatives might become promising approaches to treat patients suffering from SAP.

A Severe Acute Pancreatitis Mouse Model Transited from Mild Symptoms Induced by a "Two-Hit" Strategy with L-Arginine.

To develop a severe acute pancreatitis (SAP) model transited from mild symptoms, we investigated a "two-hit" strategy with L-arginine in mice. The mice were intraperitoneally injected with ice-cold L-arginine (4 g/kg) twice at an interval of 1 h on the first day and subjected to the repeated operation 72 h afterwards. The results showed the "two-hit" strategy resulted in the destructive damage and extensive necrosis of acinar cells in the pancreas compared with the "one-hit" model. Meanwhile, excessive levels of pro-inflammatory mediators, namely IL-6 and TNF-α, were released in the serum. Remarkably, additional deleterious effects on multiple organs were observed, including high intestinal permeability, kidney injury, and severe acute lung injury. Therefore, we confirmed that the SAP animal model triggered by a "two-hit" strategy with L-arginine was successfully established, providing a solid foundation for a deeper understanding of SAP initiation and therapy research to prevent worsening of the disease.

Structure comparison and anti-coagulant effects of the glycosaminoglycans from porcine duodenum and jejunum.

To make full use of the porcine sources and develop better choice of novel GAGs as anti-coagulants, two fractions of GAGs from the porcine jejunum (A) and duodenum (C) have been separated & purified. The products were further sulfated to give B and D in order to test the influence of sulfate pattern on the bioactivity. The results showed that the relative molecular weight range of A was 3000-50,000 (Mw, g/mol), whereas C had an average molecular weight of 75,885 (Mw, g/mol). A was identified as a novel heparan sulfate through enzymatic hydrolysis analysis. C was a chondroitin like polysaccharide mainly composed of ß-d-GlcA-(1 â†’ 4) and ß-d-GalNAc-(1 â†’ 3). A possessed controllable anti-coagulant activity (7 IU/mg) in vitro. The activity of D almost achieved the same magnitude of A. This study demonstrated the anticoagulant potential of the polysaccharides, providing solid foundation for development of anti-coagulants from porcine intestine.

Zinc ions coordinated carboxymethyl chitosan-hyaluronic acid microgel for pulmonary drug delivery.

Microgel affords a porous and swollen microstructure for the establishment of pulmonary delivery system with sustained released properties. Here, we report a microgel (with the diameter around 4 µm) prepared with a precipitation method, synthesized by coordinating Zn2+ to the Schiff base cross-linked carboxymethyl chitosan and glycol split hyaluronate. The microgel has shown well swollen and pH sensitive behaviors, high safety and biocompatibility in vitro. Besides, the biomaterial could escape from macrophage phagocytosis, a key factor contribute to quick drug clearance in the lung after co-incubated with RAW 264.7 cells. In consist with this, the bovine serum albumin loaded in the microgel showed sustained release behavior in 24 h in vitro; meanwhile, the drug had a retention time up to 36 h in the lung and followed by clearance in ICR mice through pulmonary administration. Thus, our microgel platform provides a promising candidate for pulmonary drug delivery systems with controlled release rate.

Multifunctionalized Brush-Like Glycopolymers with High Affinity to P-Selectin and Antitumor Metastasis Activity.

Glycopolymers that can mimic natural glycosaminoglycan, such as heparin, have shown great potentials in inhibition of cancer metastasis. In the current work, a novel series of brush-like glycopolymers (BGPs) with simultaneous functionalization of various monosaccharide or disaccharide compositions have been synthesized through a new grafting-polymerization strategy, in order to mimic the activities of both heparin and P-selectin ligand PSGL-1. In the subsequent in vitro assays of antiadhesion, platelets activation, heparanase inhibition, and so on, BGP-SFH, as one of the BGPs with the composition of the combined three sugar units, sialic acids, fucoses, and heparin disaccharides, showed the highest antimetastasis ability, similar to its prototype heparin. Moreover, in a mouse metastatic melanoma model, the BGP-SFH also inhibited B16 cell metastasis effectively. Thus, the current work not only demonstrated a type of promising antimetastasis glycopolymer BGPs, but also illustrated an easy synthetic approach to multifunctionalized glycopolymers, leading to potential applications for broader biomedical research.

Analysis of the external acoustic meatus for ergonomic design: part I - measurement of the external acoustic meatus using casting, scanning and rapid estimation approaches.

Numerous ear-related wearables require precise measurements of the external acoustic meatus (EAM) to optimise function and comfort. The purpose of this study is to describe a novel methodology for measuring the EAM. A total of 23 measurement variables (18 novel) of the EAM from the entrance to the second bend were collected on 700 Chinese subjects (age: 15-83) using casting and 3D scanning over seven age spans: 10, 20, 30, 40, 50, 60 s and 70+. The ear horizontal plane was identified as a new reference plane for measurements and the medial concha was selected as the reference point for positioning the entrance. A detailed approach to characterising the EAM was developed as was an approach for the rapid estimation of circumference and area using regression equations making it ideal for use in early design conceptualizations. Practitioner summary: This study provides a scalable measurement methodology for determining anthropometric measurements of the external acoustic meatus. The measurement methodology and its application to the design and fitting of ear-related wearables are important to optimising their function and comfort.

Estimations of the effective Young's modulus of specimens prepared by fused filament fabrication.

The elastic response of homogeneous isotropic materials is most commonly represented by their Young's modulus (E), but geometric variability associated with additive manufacturing results in materials that are neither homogeneous nor isotropic. Here we investigated methods to estimate the effective elastic modulus (Eeff) of samples fabricated by fused filament fabrication. We conducted finite element analysis (FEA) on printed samples based on material properties and CT-scanned geometries. The analysis revealed how the layer structure of a specimen altered the internal stress distribution and the resulting Eeff. We also investigated different empirical methods to estimate Eeff as guides. We envision the findings from our study can provide guidelines for modulus estimation of as-printed specimens, with the potential of applying to other extrusion-based additive manufacturing technologies.

Interfacial Diffusion of Hydrated Ion on Graphene Surface: A Molecular Simulation Study.

Hydration plays an important role in the diffusion and sieving of ions within nanochannels. However, it is hard to quantitatively analyze the contribution of hydration to the diffusion rates due to the complex hydrogen-bond and charge interactions between atoms. Here, we quantitatively investigated the interfacial diffusion rates of a single hydrated ion with different number of water molecules on graphene surface through molecular dynamics simulation. The simulation results show the ballistic diffusion mode by analyzing the mean-square displacement, and the diffusion rates change nonmonotonically with the hydration number. The potential energy profiles with the changing position of the hydrated ion on graphene surface were further analyzed, which shows the dominant factor for interfacial diffusion changing from ion-graphene interaction to water-graphene interaction as the number of water molecules increases. Besides, it was found that the surface hydrophilicity weakened the influence of hydration number on the diffusion rates of hydrated ion. Finally, the diffusion properties of different hydrated ions on graphene surface were investigated, and the hydrated Li+, Na+, and K+ containing three, four, and five water molecules, respectively, show the fastest diffusion rate. This work demonstrates the interfacial diffusion behavior and mechanism of hydrated ions at the molecular level, which can provide valuable guidance in nanosensors, seawater desalination, and other hydrated ion-related fields.

Effects of Environmental Temperature and Humidity on the Geometry and Strength of Polycarbonate Specimens Prepared by Fused Filament Fabrication.

It is widely known that the printing quality of fused filament fabrication (FFF) is heavily affected by environmental temperature and humidity, taking the form of warping and porosity. However, there is little understanding about the quantitative relations between environmental conditions, geometry, and the mechanical properties of printed parts. In this study, we systematically investigated those relations using bisphenol A polycarbonate as a model material system. For the environmental temperature, an in-situ infrared imaging analysis revealed the presence of an up to 5.4 °C/mm thermal gradient when printing using an open-chamber printer and a heated build plate. For the environmental humidity, an analysis of X-ray micro-computed tomography (micro-CT) scans showed an up to 11.7% porosity that was brought by polymer water content absorbed from environmental moisture. Meanwhile, tensile tests showed a mechanical performance loss associated with those defects, but, surprisingly, the transverse direction ductility had the potential to increase at a higher porosity. Furthermore, the experimental results were combined with analytical and parametrical studies to elucidate quantitative relations between environmental conditions and printing quality. Based on the results, quantitative guidelines for the estimation of printing quality based on environmental conditions are provided that would also help users to obtain desired printing results with a better understanding of the effects of environmental conditions.

A heparin derivatives library constructed by chemical modification and enzymatic depolymerization for exploitation of non-anticoagulant functions.

Non-anticoagulant biological functions of heparin-based drugs have drawn increasing attention. However, the exploration into the non-anticoagulant activities of various low molecular weight heparins was associated with bleeding risks in clinical practice and often led to controversial conclusions due to the structural differences. In this study, we aimed to establish a process to produce a library of heparin derivatives with structural diversity and reduced/abolished anticoagulant activity through the combination of chemical modifications and enzymatic cleavage of heparins. The depolymerization characteristics of various selectively modified heparin derivatives by three heparinases were comprehensively analyzed. The order of periodate treatment and heparinase-I depolymerization was proved to significantly change the structural characteristics of the oligosaccharide products. Finally, among several heparin derivatives that screened in the bleomycin-induced cell apoptosis model, the low molecular weight partially 6-O-/N-desulfated heparins showed the strongest anti-apoptotic activities. This study provided a useful approach for future development of novel heparin-derivative medications.

Pien Tze Huang accelerated wound healing by inhibition of abnormal fibroblast apoptosis in Streptozotocin induced diabetic mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Diabetic foot ulcer is one of the most serious complications of diabetes. Effective medical treatment regarding improvement of ulcer healing in patients is essential. Pien Tze Huang (PZH), a valuable Chinese traditional medicine, has been found significant efficacy on the curing of diabetic wound in clinic recently. AIM OF THE STUDY: This work was conducted to confirm the efficacy, and compare the therapeutic effect through the oral administration and local delivery route, providing a rationale for the new PZH form development; besides, the mechanisms through which PZH promoted the wound healing was also discussed. MATERIALS AND METHODS: First, the chemical composition of PZH was characterized by 1H-NMR and HPLC. The anti-apoptosis effects of PZH on high concentration glucose injured epidermal fibroblast (HFF-1) was investigated in a dose dependent way. Then, the effects of the systematical administration of PZH, and the topical used route on excisional wounds of Streptozotocin (STZ) induced diabetic mice were compared. RESULTS: The results illustrated that PZH decreased the reactive oxygen species (ROS) levels in cells, preventing cell damage/apoptosis through an ROS/Bcl-2/Bax/Caspase-3 pathway. The in vivo study proved that topical use of PZH exceeded the systematical route both in accelerating the wound closure and improving the healing quality. Meanwhile, PZH promoted wound closure through stimulating the secretion of Col-I, decreasing fibroblast apoptosis, and enhancing myo-fibroblast differentiation, in consistent with the mechanism study in vitro. CONCLUSIONS: Local used PZH improves wound healing by inhibiting the abnormal HFF-1 apoptosis and senescence. The study held a great promise for development of a topical dosage form of PZH for diabetic wound healing.

A Sponge-Like Double-Layer Wound Dressing with Chitosan and Decellularized Bovine Amniotic Membrane for Promoting Diabetic Wound Healing.

The diabetic wounds do not heal easily in part because they are susceptible to infection due to environmental influences. Wound dressing is crucial to wound healing, as it can basically protect the wound from external damages and provide a suitable microenvironment for tissue regeneration. In this study, a double-layer membrane that consists of chitosan sponge and decellularized bovine amniotic membrane (dBAM) has been developed by freeze-casting method. The results showed that the porous structure of the sponge layer improved the performances of blood coagulation and swelling. The dense dBAM can optimize the mechanical property of wound dressing. In vitro studies revealed that the bilayer membrane had favorable biocompatible, especially for human foreskin fibroblast cells (HFF-1) cell adhesion and proliferation. Moreover, the full-thickness skin defects of diabetic model mice that treated with bilayer membrane showed over 80% closure in 8 days. Our findings imply that the double-layer dressing has great potentials to be used in diabetic patients.

Design and synthesis of a native heparin disaccharide grafted poly­2­aminoethyl methacrylate glycopolymer for inhibition of melanoma cell metastasis.

Numerous studies have proved that heparin, a sub-group of glycosaminoglycan, possesses great potential as anti-metastasis agent. However, the native strong anti-coagulant activity which causes serious side effects, such as bleeding, has limited its clinical applications for safety concern. To overcome this problem, we synthesized a panel of novel glycopolymers that mimic heparin structure with substantially reduced anti-coagulant activity by a simple grafted-on strategy. The influence of molecular weight & distribution, substituting degree and sulfonic density on cytotoxicity were determined by systematic MTT analysis to select the candidates with highly bio-compatibility. Among these glycopolymers, a sulfated poly­2­aminoethyl methacrylate grafted with heparin disaccharide (abbreviated as SGPHD) has shown potent efficacy in inhibition of heparanase activity and microvascular endothelial cell proliferation in vitro. Further experiments demonstrated that SGPHD inhibited B16 murine melanoma cell migration, invasion and adhesion to platelets or microvascular endothelial cells, thus, presented as a possible anti-metastatic agent by providing a whole course protection during tumor metastasis. The results will have significant impacts for the further rational design of glycopolymer medicine.

Structural characterization and in vitro antioxidant activities of chondroitin sulfate purified from Andrias davidianus cartilage.

Origin and manufacturing process are key factors affecting the biological activities of chondroitin sulfate (CS), which can be utilized as a nutraceutical in dietary supplements. Herein, we extracted and purified CS from the cartilage of artificially breeding Andrias davidianus (ADCS), i.e., Chinese giant salamander (CGS), one of the prospective functional food source materials in China. Low molecular weight CS (LMWADCS) was then prepared by free radical depolymerization of ADCS. High-performance gel permeation chromatography (HPGPC) analysis showed that the average molecular weight (Mw) of ADCS was 49.2 kDa, while the Mw of LMWADCS was 6.4 kDa. After the eliminative degradation of ADCS by chondroitinase ABC, strong anion-exchange high-performance liquid chromatography (SAX-HPLC) analysis showed that the disaccharide composition of ADCS was 14.6% ΔDi0S, 60.9% ΔDi6S and 24.5% ΔDi4S. Then, in vitro antioxidant assays were performed with ADCS, LMWADCS and CS from a commercial source. Our results showed that LMWADCS exerted the highest total antioxidant activity out of the total antioxidant capacity, including the capacity of scavenging DPPH radicals, hydroxyl radicals and superoxide anion radicals. From the results of this study, we can conclude that the Mw and composition of ADCS are different from those reported for bovine and shark CS, and LMWADCS can be utilized as a valuable and potential nutraceutical for the functional food industry.

Design of Fusion Proteins for Efficient and Soluble Production of Immunogenic Ebola Virus Glycoprotein in Escherichia coli.

The Ebola hemorrhagic fever caused by Ebola virus is an extremely dangerous disease, and effective therapeutic agents are still lacking. Platforms for the efficient production of vaccines are crucial to ensure quick response against an Ebola virus outbreak. Ebola virus glycoprotein (EbolaGP) on the virion surface is responsible for membrane binding and virus entry, thus becoming the key target for vaccine development. However, heterologous expression of this protein still faces engineering challenges such as low production levels and insoluble aggregation. Here, the authors design and compare various fusion strategies, attaching great importance to the solubility-enhancing effect, and tag removal process. It is found that a C-terminal intein-based tag greatly enhances the solubility of EbolaGP and allows one-step chromatographic purification of the untagged EbolaGP through thiol-catalyzed self-cleavage. The purified untagged EbolaGP alone or with Freund's adjuvant are highly immunogenic, as confirmed in a mouse model. Consequently, the present study puts forward a new strategy for the efficient and soluble expression of untagged immunogenic EbolaGP. The intein-based protein fusion approach may be of importance for the large-scale production of Ebola virus subunit vaccine.

Effects of Enzymatically Depolymerized Low Molecular Weight Heparins on CCl4-Induced Liver Fibrosis.

With regard to identifying the effective components of LMWH drugs curing hepatic fibrosis disease, we carried out a comparative study on the efficacy of enzymatically depolymerized LMWHs on CCl4 induced mouse liver fibrosis. The results showed that the controlled enzymatic depolymerization conditions resulted in LMWHs with significantly different activities. The LMWH product depolymerized by Heparinase I (I-11) with a Mw of 7160, exhibited a significant advantage in reducing the liver inflammation by suppressing TNF-α and IL-1ß secretion, and minimizing hepatic fibrogenesis. The products prepared by only Heparinase II (II-11), and combined Heparinase III and II (III-II-5) showed limited positive effect on hepatic inflammation and fibrosis. On the contrary, the products by combined Heparinase III and I (III-I-9, III-I-5) showed no effect or stimulation effect on the hepatic fibrogenesis. Our results provided the basis for structure-activity relationship insight for inhibition of liver fibrosis activities of LMWHs, which might have significant implications for generic anti-fibrosis disease drug development.

Discovery of enzymatically depolymerized heparins capable of treating Bleomycin-induced pulmonary injury and fibrosis in mice.

Heparin has recently been shown to slow down idiopathic pulmonary fibrosis (IPF) process and improve survival of patients in some cases. To improve the anti-IPF function while minimizing their side effects, we developed heparin libraries with different structures depolymerized by single or combined heparinases, and systematically screened the efficacy of the different heparins for treatment of Bleomycin-induced pulmonary injury and fibrosis using mice model. Then we characterized the structural properties of the components capable of treating pulmonary injury and fibrosis by use of chip-based amide hydrophilic interaction chromatography (HILIC)-fourier transform (FT)-ESI-MS, polyacrylamide gel electrophoresis (PAGE), and high performance liquid chromatography (HPLC). Our results showed that the depolymerized heparins with relative higher molecular weight (I-2 and III-2) by the respective heparinase I and III protected mice from the induced pulmonary injury and fibrosis. In addition, the selected depolymerized heparins inhibited high-mobility group protein B1 (HMGB-1) expression, prevented E-cadhesin from downregulation, and reduced fibroblasts accumulation in the mouse lung tissue. Our study suggested that the depolymerized heparins of I-2 and III-2 with the most significant efficacy might target several pathways in alleviating the induced pulmonary fibrosis.

Non-anticoagulant effects of low molecular weight heparins in inflammatory disorders: A review.

Low molecular weight heparins (LMWHs) are produced by chemical or enzymatic depolymerization of unfractionated heparin (UFH). Besides their well-known anticoagulant effects, LMWHs have also been reported to exhibit numerous anti-inflammatory properties. Previous studies have, however, shown that different production processes result in unique structural characteristics of LMWHs. The structural variations may help explain the different therapeutic spectrums in disease treatment for non-anticoagulant effects. In the present review, we summarize major advances in understanding and exploiting the anti-inflammatory disorder activities of LMWHs, based on mechanistic studies, preclinical experiments and clinical trials. We highlight differences in these activities of commercially available LMWHs produced using different manufacturing processes. We stress the importance of structure-activity relationship (SAR) studies on the non-anticoagulant effects of LMWHs and discuss strategies for exploring new clinical indications.