The role of carbonic anhydrase III and autophagy in type 2 diabetes with cardio-cerebrovascular disease.

Type 2 diabetes mellitus (T2DM) is one of the most common chronic diseases among the elderly people. The T2DM increases the risk of cardio-cerebrovascular disease (CCD), and the main pathological change of the CCD is atherosclerosis (AS). Meanwhile, the carbonic anhydrases (CAs) are involved in the formation and progression of plaques in AS. However, the exact physiological mechanism of carbonic anhydrase III (CAIII) has not been clear yet, and there are also no correlation study between CAIII protein and T2DM with CCD. The 8-week old diabetic mice (db/db-/- mice) and wild-type mice (wt mice) were feed by a normal diet till 32 weeks, and detected the carotid artery vascular opening angle using the method of biomechanics; The changes of cerebral cortex and myocardium were watched by the ultrastructure, and the autophagy were observed by electron microscope; The tissue structure, inflammation and cell injury were observed by Hematoxylin and eosin (HE) staining; The apoptosis of cells were observed by TUNEL staining; The protein levels of CAIII, IL-17, p53 were detected by immunohistochemical and Western Blot, and the Beclin-1, LC3, NF-κB were detected by Western Blot. All statistical analysis is performed using PRISM software. Compared with wt mice, db/db-/- mice' carotid artery open angle increased significantly. Electron microscope results indicated that autophagy in db/db-/- mice cerebral cortex and heart tissue decreased and intracellular organelle ultrastructure were damaged. HE staining indicated that, db/db-/- mice' cerebral cortex and heart tissue stained lighter, inflammatory cells infiltration, cell edema were obvious, myocardial fibers were disorder, and myocardial cells showed different degrees of degeneration. Compared with wt mice, TUNEL staining showed that there was obviously increase in db/db-/- mice cortex and heart tissue cell apoptosis. The results of immunohistochemistry and Western Blot indicated that CAIII, Beclin-1 and LC3II/I expression levels conspicuously decreased in cortex and heart tissue of db/db-/- mice, and the expression level of IL-17, NF-κB and p53 obviously increased. The carotid artery' vascular stiffness was increased and which was probably related with formation of AS in diabetic mice. And the autophagy participated in the occurrence and development of diabetic CCD. CAIII protein might somehow be involved in the regulation of autophagy probably through affecting cell apoptosis and inflammation, but the underlying mechanism remains to be further studied.

Effects of substrate stiffness on mast cell migration.

Mast cells (MCs) play important roles in multiple pathologies, including fibrosis; however, their behaviors in different extracellular matrix (ECM) environments have not been fully elucidated. Accordingly, in this study, the migration of MCs on substrates with different stiffnesses was investigated using time-lapse video microscopy. Our results showed that MCs could appear in round, spindle, and star-like shapes; spindle-shaped cells accounted for 80-90 % of the total observed cells. The migration speed of round cells was significantly lower than that of cells with other shapes. Interestingly, spindle-shaped MCs migrated in a jiggling and wiggling motion between protrusions. The persistence index of MC migration was slightly higher on stiffer substrates. Moreover, we found that there was an intermediate optimal stiffness at which the migration efficiency was the highest. These findings may help to improve our understanding of MC-induced pathologies and the roles of MC migration in the immune system.

Platelet-Derived Extracellular Vesicles Increase Col8a1 Secretion and Vascular Stiffness in Intimal Injury.

The arterial mechanical microenvironment, including stiffness, is a crucial pathophysiological feature of vascular remodeling, such as neointimal hyperplasia after carotid endarterectomy and balloon dilatation surgeries. In this study, we examined changes in neointimal stiffness in a Sprague-Dawley rat carotid artery intimal injury model and revealed that extracellular matrix (ECM) secretion and vascular stiffness were increased. Once the endothelial layer is damaged in vivo, activated platelets adhere to the intima and may secrete platelet-derived extracellular vesicles (pEVs) and communicate with vascular smooth muscle cells (VSMCs). In vitro, pEVs stimulated VSMCs to promote collagen secretion and cell adhesion. MRNA sequencing analysis of a carotid artery intimal injury model showed that ECM factors, including col8a1, col8a2, col12a1, and elastin, were upregulated. Subsequently, ingenuity pathway analysis (IPA) was used to examine the possible signaling pathways involved in the formation of ECM, of which the Akt pathway played a central role. In vitro, pEVs activated Akt signaling through the PIP3 pathway and induced the production of Col8a1. MicroRNA (miR) sequencing of pEVs released from activated platelets revealed that 14 of the top 30 miRs in pEVs targeted PTEN, which could promote the activation of the Akt pathway. Further research showed that the most abundant miR targeting PTEN was miR-92a-3p, which promoted Col8a1 expression. Interestingly, knockdown of Col8a1 expression in vivo abrogated the increase in carotid artery stiffness and simultaneously increased the degree of neointimal hyperplasia. Our results revealed that pEVs may deliver miR-92a-3p to VSMCs to induce the production and secretion of Col8a1 via the PTEN/PIP3/Akt pathway, subsequently increasing vascular stiffness. Therefore, pEVs and key molecules may be potential therapeutic targets for treating neointimal hyperplasia.

In situ generation of H2O2 using MWCNT-Al/O2 system and possible application for glyphosate degradation.

Hydrogen peroxide (H2O2), as a green oxidant, has been widely applied into advanced oxidation processes (AOPs) for the degradation of toxic organic pollutants. The in situ generation of H2O2 can not only improve the storage and transportation safety of H2O2 but also reduce the capital and operation costs. In the present work, a novel system, i.e., multi-walled carbon nanotube­aluminum (MWCNT-Al) composite was used to in situ generate H2O2 through micro-electrolysis. The MWCNT-Al composite was characterized and optimized. The accumulation concentration of H2O2 reached 947 mg/L at the initial pH of 9.0, the MWCNT-Al composite dosage of 8 g/L and oxygen gas flow rate of 400 mL/min after 60 min. The in situ generation of H2O2 was achieved by MWCNT-Al/O2 system, mainly owing to the direct contact between Al0 and MWCNT in MWCNT-Al composite, which accelerated the transfer of electrons from Al0 to O2, as well as the excellent electrocatalytic activity of MWCNT toward the two-electron reduction of oxygen. When H2O2 in situ generation technology was used in peroxone process (O3/H2O2 process) to degrade glyphosate in aqueous solution, the removal efficiency of TOC and total phosphorus was 68.35% and 73.27%, respectively. Finally, the possible mechanism of in situ generation of H2O2 in MWCNT-Al/O2 system was temporarily proposed.

An investigation of the distribution and location of mast cells affected by the stiffness of substrates as a mechanical niche.

The distribution and location of mast cells are closely related to their physiological and pathological functions, such as allergic responses, immunity, and fibrosis, and are used in acupuncture. In this study, the distribution of mast cells in vivo was observed, and mechanical clues for understanding their distribution based on mechanical niches were explored. By toluidine blue staining and immunohistochemical staining, we examined the distribution and location of mast cells in rat skin and found that mast cells are distributed in a spatially nonuniform manner, preferring to locate at regions in the tissue and extracellular matrix with stiffness changes. In vitro experiments for studying the distribution of rat basophilic leukemia (RBL-2H3) mast cell line on poly-di-methyl-siloxane (PDMS) substrates with stiffness variations were performed. It was found that RBL-2H3 cells migrate and tend to remain in the areas with stiffness variations. The present research suggests that changing the stiffness of local tissues may stimulate mast cell recruitment, which may be the method by which some traditional Chinese medicine treatments, such as acupuncture. On the basis of the origin of mast cells and our experimental results, we predict that mast cells exist in tissues that contain permeable capillaries and prefer regions with stiffness changes. We discussed this prediction using examples of specific tissues from some cases.

Zn0-CNTs-Fe3O4 catalytic in situ generation of H2O2 for heterogeneous Fenton degradation of 4-chlorophenol.

A novel Zn0-CNTs-Fe3O4 composite was synthesized by the chemical co-precipitation combined with high sintering process at nitrogen atmosphere. The as-prepared composite was characterized by SEM, EDS, XRD, XPS, VSM and N2 adsorption/desorption experiments. A novel heterogeneous Fenton-like system, composed of Zn0-CNTs-Fe3O4 composite and dissolved oxygen (O2) in solution, which can in situ generate H2O2 and OH, was used for the degradation of 4-chlorophenol (4-CP). The influences of various operational parameters, including the initial pH, dosage of Zn0-CNTs-Fe3O4 and initial concentration of 4-CP on the removal of 4-CP were investigated. The removal efficiencies of 4-CP and total organic carbon (TOC) were 99% and 57%, respectively, at the initial pH of 1.5, Zn0-CNTs-Fe3O4 dosage of 2 g/L, 4-CP initial concentration of 50 mg/L and oxygen flow rate of 400 mL/min. Based on the results of the radical scavenger effect study, the hydroxyl radical was considered as the main reactive oxidants in Zn0-CNTs-Fe3O4/O2 system and a possible degradation pathway of 4-CP was proposed.

Arterial wall remodeling under sustained axial twisting in rats.

Blood vessels often experience torsion along their axes and it is essential to understand their biological responses and wall remodeling under torsion. To this end, a rat model was developed to investigate the arterial wall remodeling under sustained axial twisting in vivo. Rat carotid arteries were twisted at 180° along the longitudinal axis through a surgical procedure and maintained for different durations up to 4weeks. The wall remodeling in these twisted arteries was examined using histology, immunohistochemistry and fluorescent microscopy. Our data showed that arteries remodeled under twisting in a time-dependent manner during the 4weeks post-surgery. Cell proliferation, MMP-2 and MMP-9 expressions, medial wall thickness and lumen diameter increased while collagen to elastin ratio decreased. The size and number of internal elastic lamina fenestrae increased with elongated shapes, while the endothelial cells elongated and aligned towards the blood flow direction gradually. These results demonstrated that sustained axial twisting results in artery remodeling in vivo. The rat carotid artery twisting model is an effective in vivo model for studying arterial wall remodeling under long-term torsion. These results enrich our understanding of vascular biology and arterial wall remodeling under mechanical stresses.

Pilot-scale bioelectrochemical system for efficient conversion of 4-chloronitrobenzene.

4-Chloronitrobenzene (4-CNB) is one of the highly toxic contaminants that may lead to acute, chronic or persistent physiological toxicity to ecology and environment. Conventional methods for removing 4-CNB from aquatic environment may be problematic due to inefficiency, high cost and low sustainability. This study develops a pilot-scale bioelectrochemical system (BES, effective volume of 18 L) and examines its performance of bioelectrochemical transformation of 4-CNB to 4-chloroaniline (4-CAN) under continuous operation. The results demonstrate that the initial 4-CNB concentration in the influent and hydraulic retention time (HRT) has a significant impact on 4-CNB reduction and 4-CAN formation. Compared with the conventional anaerobic process in the absence of external power supplied, the 4-CNB conversion efficiency can be enhanced with power supplied due to microbial-mediated electron transfer at the negative cathode potential. At a voltage of 0.4 V and HRT of 48 h, the 4-CNB reduction and 4-CAN formation efficiency reached 99% and 94.1%, respectively. Based on a small external voltage applied, the pilot-scale BES is effective in the conversion of 4-CNB to 4-CAN, an intermediate that is of less toxicity and higher bioavailability for subsequent treatment. This study provides a new strategy and methods for eliminating 4-CNB, making wastewater treatment more economical and more sustainable.

Improved interfacial oxygen reduction by ethylenediamine tetraacetic acid in the cathode of microbial fuel cell.

In this study, ethylenediamine tetraacetic acid (EDTA) was investigated as a new kind of non-polymeric catalyst binder to improve interfacial oxygen reduction reaction (ORR) for the cathode of microbial fuel cell (MFC). The electrochemical analysis and MFC tests show negative correlation between ORR activity and molar concentration of EDTA applied during electrode preparation. In particular, the 0.02mol/L-EDTA yields higher ORR activity than other binder materials like Nafion, water, 0.1mol/L-EDTA and 0.2mol/L-EDTA, as indicated by the strongest response of ORR current and the smallest charge-transfer resistance. Accordingly, the MFC with cathode of 0.02mol/L-EDTA produced a maximum power density of 722mW/m(2), accounting for a value approximately 42% higher than that of commercial Nafion binder (5wt%, 507mW/m(2)). The improved ORR activity should be attributed to the enhanced proton transfer from phosphate ions to EDTA-involved three-phase boundary as a result of dipole ion bonds on nitrogen atoms having unshared pair of electrons in EDTA molecule.

A novel stainless steel mesh/cobalt oxide hybrid electrode for efficient catalysis of oxygen reduction in a microbial fuel cell.

To explore efficient and cost-effective cathode material for microbial fuel cells (MFCs), the present study fabricates a new type of binder-free gas diffusion electrode made of cobalt oxide (Co3O4) micro-particles directly grown on stainless steel mesh (SSM) by using an ammonia-evaporation-induced method. In various electrochemical analyses and evaluations in batch-fed dual-chamber MFCs, the SSM/Co3O4 hybrid electrode demonstrates improved performances in terms of electrocatalytic activity, selectivity, durability and economics toward oxygen reduction reaction (ORR) in pH-neutral solution, in comparison with conventional carbon supported platinum catalyst. This study suggests a new strategy to fabricate a more effective electrode for ORR in MFCs, making it more technically and economically viable to produce electrical energy from organic materials for practical applications.

Fabrication of stainless steel mesh gas diffusion electrode for power generation in microbial fuel cell.

This study reports the fabrication of a new membrane electrode assembly by using stainless steel mesh (SSM) as raw material and its effectiveness as gas diffusion electrode (GDE) for electrochemical oxygen reduction in microbial fuel cell (MFC). Based on feeding glucose (0.5 g L(-1)) substrate to a single-chambered MFC, power generation using SSM-based GDE was increased with the decrease of polytetrafluoroethylene (PTFE) content applied during fabrication, reaching the optimum power density of 951.6 mW m(-2) at 20% PTFE. Repeatable cell voltage of 0.51 V (external resistance of 400 Ω) and maximum power density of 951.6 mW m(-2) produced for the MFC with SSM-based GDE are comparable to that of 0.52 V and 972.6 mW m(-2), respectively obtained for the MFC containing typical carbon cloth (CC)-made GDE. Besides, Coulombic efficiency (CE) is found higher for GDE (SSM or CC) with membrane assembly than without, which results preliminarily from the mitigation of Coulombic loss being associated with oxygen diffusion and substrate crossover. This study demonstrates that with its good electrical conductivity and much lower cost, the SSM-made GDE suggests a promising alternative as efficient and more economically viable material to conventional typical carbon for power production from biomass in MFC.

Preliminary ex vivo feasibility study on targeted cell surgery by high intensity focused ultrasound (HIFU).

High intensity focused ultrasound (HIFU) has become a new noninvasive surgical modality in medicine. A portion of tissue seated inside a patient's body may experience coagulative necrosis after a few seconds of insonification by high intensity focused ultrasound (US) generated by an extracorporeal focusing US transducer. The region of tissue affected by coagulative necrosis (CN) usually has an ellipsoidal shape when the thermal effect due to US absorption plays the dominant role. Its long and short axes are parallel and perpendicular to the US propagation direction respectively. It was shown by numerical computations using a nonlinear Gaussian beams model to describe the sound field in a focal zone and ex vivo experiments that the dimension of the short and long axes of the tissue which experiences CN can be as small as 50µm and 250µm respectively after one second exposure of US pulse (the spatial and pulse average acoustic power is on the order of tens of Watts and the local acoustic spatial and temporal pulse averaged intensity is on the order of 3×10(4)W/cm(2)) generated by a 1.6MHz HIFU transducer of 12cm diameter and 11cm geometric focal length (f-number=0.92). The concept of thermal dose of cumulative equivalent minutes was used to describe the possible tissue coagulative necrosis generated by HIFU. The numbers of cells which suffered CN were estimated to be on the order of 40. This result suggests that HIFU is able to interact with tens of cells at/near its focal zone while keeping the neighboring cells minimally affected, and thus the targeted cell surgery may be achievable.