Pericardial application as a new route for implanting stem-cell cardiospheres to treat myocardial infarction.

KEY POINTS: Cardiospheres (CSps) are a promising new form of cardiac stem cells with advantage over other stem cells for myocardial regeneration, but direct implantation of CSps by conventional routes has been limited due to potential embolism. We have implanted CSps into the pericardial cavity and systematically demonstrated its efficacy regarding myocardial infarction. Stem cell potency and cell viability can be optimized in vitro prior to implantation by pre-conditioning CSps with pericardial fluid and hydrogel packing. Transplantation of optimized CSps into the pericardial cavity improved cardiac function and alleviated myocardial fibrosis, increased myocardial cell survival and promoted angiogenesis. Mechanistically, CSps are able to directly differentiate into cardiomyocytes in vivo and promote regeneration of myocardial cells and blood vessels through a paracrine effect with released growth factors as potential paracrine mediators. These findings establish a new strategy for therapeutic myocardial regeneration to treat myocardial infarction. ABSTRACT: Cardiospheres (CSps) are a new form of cardiac stem cells with an advantage over other stem cells for myocardial regeneration. However, direct implantation of CSps by conventional routes to treat myocardial infarction has been limited due to potential embolism. We have implanted CSps into the pericardial cavity and systematically assessed its efficacy on myocardial infarction. Preconditioning with pericardial fluid enhanced the activity of CSps and matrix hydrogel prolonged their viability. This shows that pretransplant optimization of stem cell potency and maintenance of cell viability can be achieved with CSps. Transplantation of optimized CSps into the pericardial cavity improved cardiac function and alleviated myocardial fibrosis in the non-infarcted area, and increased myocardial cell survival and promoted angiogenesis in the infarcted area. Mechanistically, CSps were able to directly differentiate into cardiomyocytes in vivo and promoted regeneration of myocardial cells and blood vessels in the infarcted area through a paracrine effect with released growth factors in pericardial cavity serving as possible paracrine mediators. This is the first demonstration of direct pericardial administration of pre-optimized CSps, and its effectiveness on myocardial infarction by functional and morphological outcomes with distinct mechanisms. These findings establish a new strategy for therapeutic myocardial regeneration to treat myocardial infarction.

Soluble dietary fiber and cellulose from Saccharina japonica by-product ameliorate Loperamide-induced constipation via modulating enteric neurotransmitters, short-chain fatty acids and gut microbiota.

The effects of soluble dietary fiber (SDF) and cellulose (IDF) from Saccharina japonica by-product and their differences in improving constipation were further clarified in the present study. We demonstrated that SDF was mainly made up of d-mannuronic acid and d-mannose while IDF consisted of d-glucose , which is different from other reported dietary fibers of terrestrial plants. In this research, both SDF and IDF improved fecal-related indicators, gastrointestinal transit rate and histological morphology in Lop-induced mice. Moreover, they could increase the level of antioxidant enzymes (SOD and GSH-Px), restore the expression of enteric neurotransmitters, and maintain the function of ZO-1, JAM-1 as well as Occludin. Interestingly, SDF and IDF had a significant up-regulated effect on the proportion of Muribaculacea, Prevotellaceaen and Lachnospiraceae, which are critical to preserving intestinal immune homeostasis. Besides, they promoted the biosynthesis of short-chain fatty acids (SCFAs). The overall index showed that SDF is more effective for constipation due to its better water retention capacity. Thus, they can be used as a safe dietary supplement for the treatment of chronic or occasional constipation in humans.

Growth of grasses and forbs, nutrient concentration, and microbial activity in soil treated with microbeads.

Microplastics have emerged as an important threat to terrestrial ecosystems. To date, little research has been conducted on investigating the effects of microplastics on ecosystem functions and multifunctionality. In this study, we conducted the pot experiments containing five plant communities consisting of Phragmites australis, Cynanchum chinense, Setaria viridis, Glycine soja, Artemisia capillaris, Suaeda glauca, and Limonium sinense and added polyethylene (PE) and polystyrene (PS) microbeads to the soil (contained a mixture of 1.5 kg loam and 3 kg sand) at two concentrations of 0.15 g/kg (lower concentration, hereinafter referred to as PE-L and PS-L) and 0.5 g/kg (higher concentration, hereinafter referred to as PE-H and PS-H) to explore the effects of microplastics on total plant biomass, microbial activity, nutrient supply, and multifunctionality. The results showed that PS-L significantly decreased the total plant biomass (p = 0.034), primarily by inhibiting the growth of the roots. ß-glucosaminidase decreased with PS-L, PS-H, and PE-L (p < 0.001) while the phosphatase was noticeably augmented (p < 0.001). The observation suggests that the microplastics diminished the nitrogen requirements and increased the phosphorus requirements of the microbes. The decrease in ß-glucosaminidase diminished ammonium content (p < 0.001). Moreover, PS-L, PS-H, and PE-H reduced the soil total nitrogen content (p < 0.001), and only PS-H considerably reduced the soil total phosphorus content (p < 0.001), affecting the ratio of N/P markedly (p = 0.024). Of interest, the impacts of microplastics on total plant biomass, ß-glucosaminidase, phosphatase, and ammonium content did not become larger at the higher concentration, and it is observable that microplastics conspicuously depressed the ecosystem multifunctionality, as microplastics depreciated single functions such as total plant biomass, ß-glucosaminidase, and nutrient supply. In perspective, measures to counteract this new pollutant and eliminate its impact on ecosystem functions and multifunctionality are necessary.

Lignin particles as green pore-forming agents for the fabrication of microporous polysulfone membranes.

Templating polymeric membranes with micro-nano-scaled solid materials is an effective method to simultaneously improve the water flux and retention ratio. However, the fabrication of a green, recyclable, and size-controlled template material remains a challenge. Here, a new green pore-forming agent, lignin particles (LP), was developed to prepare porous polysulfone (PSF) membranes via the phase inversion technique. A series of LP have uniform sizes from ~200 nm to ~1800 nm. The performances of the templated PSF membranes cast at different sizes and contents of LP were examined for their surface and crosssection morphologies. The LP-templated PSF membranes displayed a remarkable enhancement in flux, porosity, and moisture content. Particularly, the PSF membranes cast with LP from ~200 to 1800 nm broke the traditional trade-off to a certain degree, which possessed stable retentions of bovine serum albumin (> 85 %) and significantly improved water flux (174.275 to 254.775 L m-2 h-1). In addition, the LP pore-forming agent is low-cost and environmentally friendly as it was prepared from industrial by-products and can be easily recycled. Overall, this study shows that lignin particles are green pore-forming agents that can be used for the fabrication of porous polymeric membranes with improved performance for water treatment.

Inhibition of lysosome-tethered Ragulator-Rag-3D complex restricts the replication of Enterovirus 71 and Coxsackie A16.

Enterovirus 71 (EV71) and Coxsackie A16 (CVA16) are two major causative agents of hand, foot, and mouth disease (HFMD) in young children. However, the mechanisms regulating the replication and pathogenesis of EV71/CVA16 remain incompletely understood. We performed a genome-wide CRISPR-Cas9 knockout screen and identified Ragulator as a mediator of EV71-induced apoptosis and pyroptosis. The Ragulator-Rag complex is required for EV71 and CVA16 replication. Upon infection, the Ragulator-Rag complex recruits viral 3D protein to the lysosomal surface through the interaction between 3D and RagB. Disruption of the lysosome-tethered Ragulator-Rag-3D complex significantly impairs the replication of EV71/CVA16. We discovered a novel EV71 inhibitor, ZHSI-1, which interacts with 3D and significantly reduces the lysosomal tethering of 3D. ZHSI-1 treatment significantly represses replication of EV71/CVA16 as well as virus-induced pyroptosis associated with viral pathogenesis. Importantly, ZHSI-1 treatment effectively protects against EV71 infection in neonatal and young mice. Thus, our study indicates that targeting lysosome-tethered Ragulator-Rag-3D may be an effective therapeutic strategy for HFMD.

Adsorption behavior and mechanism of five pesticides on microplastics from agricultural polyethylene films.

Polyethylene (PE) agricultural soil films are easily embrittled and decomposed to microplastics (MPs) in environment. As widely used pesticides in vegetable farmland, carbendazim, dipterex, diflubenzuron, malathion, difenoconazole have potential environmental and human safety risks. They are often coexisting with MPs in the environment, and may cause consequential pollution to the ecosystem. Studying the adsorption behavior between pesticides and PE agricultural soil films MPs would be helpful for the risk assessment of co-exposure of pesticides and MPs. Herein, a systematic study on batch adsorption experiments was performed to determine the adsorption process of pesticides on MPs, the environmental factors on adsorption capacity were evaluated, and the adsorption mechanisms were discussed. Results suggested that all these five pesticides can adsorb on MPs, especially for diflubenzuron and difenoconazole. The adsorption kinetics and isotherm fitted to the Pseudo-second-order and Freundlich model, respectively, indicating that besides the adsorption onto surface sites, mass transfer and intraparticle diffusion were involved in the adsorption process, and the adsorption process was mostly controlled by physical and chemical interactions. The adsorption amounts of 5 pesticides on PE MPs follow the order of DIF > DIFE > MAL > CAR > DIP with KF correlated positively with octanol-water partition coefficients (LogKow). The thermodynamic study indicates the adsorption of all pesticides as spontaneous and exothermic processes. The results of this study illustrated that PE MPs can be a good carrier of pesticides in agricultural field.

Hydrophilic Astaxanthin: PEGylated Astaxanthin Fights Diabetes by Enhancing the Solubility and Oral Absorbability.

To develop hydrophilic astaxanthin with significantly enhanced solubility and stability, astaxanthin polyethylene glycol succinate (APGS) was synthesized by esterification of an astaxanthin succinate diester with polyethylene glycol 1000. The chemical structure of the hydrophilic derivative was confirmed by 1H nuclear magnetic resonance and mass spectra. APGS showed better solubility than free astaxanthin in water and enhanced bioavailability compared to that of free astaxanthin. Additionally, testing the effects on diabetes and inflammation in a high-fat- and high-sucrose-diet-induced insulin-resistant mouse model demonstrated its benefits, suggesting that APGS maintains the health-promoting properties of astaxanthin. These results suggest that APGS could be a better source of hydrophilic astaxanthin.

Impact of lignin content on alkaline-sulfite pretreatment of Hybrid Pennisetum.

This work focuses to investigate the impact of lignin content on chemical compositions, crystallinity, surface characterizations, cellulase adsorption profiles and hydrolysability of Hybrid Pennisetum (HP) after alkaline sulfite pretreatment (ASP). For the HP with lower lignin content, the increase of the cellulose content by ASP was more obvious than raw HP. ASP decreased total lignin content and surface lignin content of HP substrates. HP with lower lignin content (e.g., ∼15%) is suitable for ASP, because a pretty perfect glucose yield (91%) was obtained using a low dosage of enzyme loadings (5 FPU of cellulases/g dry matter). The study provides a potential strategy to efficiently produce platform sugars from HP with reduced lignin content, indicating the importance of reduction HP lignin content properly by breeding or transgenesis programs. The work could also help elucidate the mechanism of alkaline sulfite pretreatment for efficient production of fermentable sugars from lignocelluloses.

Exploring surface characterization and electrostatic property of Hybrid Pennisetum during alkaline sulfite pretreatment for enhanced enzymatic hydrolysability.

The surface characterization and electrostatic property of Hybrid Pennisetum (HP) after alkaline sulfite pretreatment were explored for enhanced enzymatic hydrolysability. The O/C ratio in HP increased from 0.34 to 0.60, and C1 concentration decreased from 62.5% to 31.6%, indicating that alkaline sulfite pretreatment caused poorer lignin but richer carbohydrate on HP surface. Zeta potential and sulfur element analysis indicated that more enzymes would preferably adsorb on the carbohydrate surface of alkaline sulfite pretreated HP because the lignin was sulfonated, which facilitated the decrease of non-productive adsorption. Glucose yield of alkaline sulfite pretreated HP reached to 100% by synergistic action of cellulase and xylanase in the hydrolysis, which was significantly higher than that of NaOH pretreated, and the concentration of glucose released was 1.52times higher. The results suggested that alkaline sulfite pretreatment had potential for improving the HP hydrolysability, and the surface characterization and electrostatic property facilitated the enzymatic digestibility.

Structural properties and hydrolysabilities of Chinese Pennisetum and Hybrid Pennisetum: Effect of aqueous ammonia pretreatment.

The effects of aqueous ammonia pretreatment on structural properties and hydrolysabilities of Chinese Pennisetum and Hybrid Pennisetum were investigated. Aqueous ammonia pretreatment increased cellulose crystallinities and hydrolysabilities of Chinese Pennisetum and Hybrid Pennisetum. Compared with Chinese Pennisetum, Hybrid Pennisetum showed better enzymatic digestibility. Xylanase supplementation was more effective than the increase of cellulase loadings in the hydrolysis of aqueous ammonia pretreated Chinese Pennisetum and Hybrid Pennisetum. After supplementation of 2mg of xylanase/g dry matter to 5 FPU of cellulases/g dry matter, the hydrolysis yields of cellulose of aqueous ammonia pretreated Chinese Pennisetum and Hybrid Pennisetum were 92.3-95.4%, and the hydrolysis yields of xylan were 86.9-94.2%. High hydrolysability and low dosage of enzyme loadings together with the advantages of high yield and widely distribution demonstrated the potential of Chinese Pennisetum and Hybrid Pennisetum for the production of platform sugars.

Benefits from additives and xylanase during enzymatic hydrolysis of bamboo shoot and mature bamboo.

Effects of additives (BSA, PEG 6000, and Tween 80) on enzymatic hydrolysis of bamboo shoot and mature bamboo fractions (bamboo green, bamboo timber, bamboo yellow, bamboo node, and bamboo branches) by cellulases and/or xylanase were evaluated. The addition of additives was comparable to the increase of cellulase loadings in the conversion of cellulose and xylan in bamboo fractions. Supplementation of xylanase (1 mg/g DM) with cellulases (10 FPU/g DM) in the hydrolysis of bamboo fractions was more efficient than addition of additives in the production of glucose and xylose. Moreover, addition of additives could further increase the glucose release from different bamboo fractions by cellulases and xylanase. Bamboo green exhibited the lowest hydrolyzability. Almost all of the polysaccharides in pretreated bamboo shoot fractions were hydrolyzed by cellulases with the addition of additives or xylanase. Additives and xylanase showed great potential for reducing cellulase requirement in the hydrolysis of bamboo.

Microstructure, mechanical and bio-corrosion properties of Mn-doped Mg-Zn-Ca bulk metallic glass composites.

The effects of Mn substitution for Mg on the microstructure, mechanical properties, and corrosion behavior of Mg69-xZn27Ca4Mnx (x=0, 0.5 and 1at.%) alloys were investigated using X-ray diffraction, compressive tests, electrochemical treatments, and immersion tests, respectively. Microstructural observations showed that the Mg69Zn27Ca4 alloy was mainly amorphous. The addition of Mn decreases the glass-forming ability, which results in a decreased strength from 545 MPa to 364 MPa. However, this strength is still suitable for implant application. Polarization and immersion tests in the simulated body fluid at 37 °C revealed that the Mn-doped Mg-Zn-Ca alloys have significantly higher corrosion resistance than traditional ZK60 and pure Mg alloys. Cytotoxicity test showed that cell viabilities of osteoblasts cultured with Mn-doped Mg-Zn-Ca alloys extracts were higher than that of pure Mg. Mg68.5Zn27Ca4Mn0.5 exhibits the highest bio-corrosion resistance, biocompatibility and has desirable mechanical properties, which could suggest to be used as biomedical materials in the future.

[Characteristics of the extracellular polymeric substances of a heterotrophic nitrifying bacterium strain].

To study the variation of extracellular polymers substances (EPS) composition of the heterotrophic denitrifying bacteria (Acinetobacter sp. YY-5) at different growth stages, The EPS of YY-5 was extracted by thermal, and the composition of protein, polysaccharides, nucleic acid and free amino acids were analyzed at different growth stages. Subsequently, the proteins in extracts were hydrolyzed into amino acids, the amino acid's changes and effect on physical and chemical properties of proteins were investigated. The results showed the strain has a high EPS contents which mainly consist of proteins, and the EPS content achieved the highest value at stable stage. The amount of proteins increased from 14.599 mg x g(-1) to 28.489 mg x g(-1), then declined to 15.139 mg x g(-1). Polysaccharides content increased from 6.757 mg x g(-1) to 10.199 mg x g(-1), then declined to 7.857 mg x g(-1). The nucleic acid contents increased from 1.56 mg x g(-1) to 6.287 mg x g(-1) in the whole growth stages. The free amino acids contents increased from 3.713 mg x g(-1) to 4.374 mg x g(-1), then obviously declined to 1.299 mg x g(-1). After the proteins were hydrolyzed into amino acids, the amount of polar amino acids showed the trend that increased earlier and declined later, the contents of nonpolar amino acids increased at all growth stages. The amino acids with negative charges were more than that with the positive.

Immune potential of a novel multiple-epitope vaccine to FMDV type Asia 1 in guinea pigs and sheep.

To develop a safe and efficient recombinant subunit vaccine to foot-and-mouth disease virus (FMDV) type Asia 1 in sheep, a tandem repeated multiple-epitope gene consisting of residues 137-160 and 197-211 of the VP1 gene of FMDV was designed and artificially synthesized. The biologically functional molecule, the ovine IgG heavy constant region (oIgG) as a protein carrier was introduced for design of the multiple-epitope recombinant vaccine and recombinant expression plasmids pET-30a-RE and pET-30a-RE-oIgG were successfully constructed. The recombinant proteins, RE and RE-oIgG, were expressed as a formation of inclusion bodies in E. coli. The immune potential of this vaccine regime in guinea pigs and sheep was evaluated. The results showed that IgG could significantly enhance the immune potential of antigenic epitopes. The recombinant protein RE-oIgG could not only elicit the high levels of neutralizing antibodies and lymphocytes proliferation responses in the vaccinated guinea pigs, but confer complete protection in guinea pigs against virus challenge. Although the recombinant protein RE could not confer protection in the vaccinated animals, it could delay the appearance of the clinical signs and reduce the severity of disease. Inspiringly, the titers of anti-FMDV neutralizing antibodies elicited in sheep vaccinated with RE-oIgG was significantly higher than that for the RE vaccination. Therefore, we speculated that this vaccine formulation may be a promising strategy for designing a novel vaccine against FMDV in the future.

[Fusion expression of Asia I type FMDV neutralizing epitope with heavy chain constant region of sheep IgG and the assessment of its immunogenicity].

VP1 is a major antigenic protein of foot-and-mouth disease virus(FMDV), which induces the immune response against FMDV infection, and contains several epitopes of the virus. We designed and chemically synthesized a DNA fragment which encoding a tandem repeat protein of 136-160aa and 198-211aa of a strain of type Asia I FMDV, and cloned the gene of heavy chain constant region of sheep IgG. By using the BamH I, EcoR I and Xho I sites, both genes were cloned into pPROExHTb vector in turn to form a recombinant plasmid pPRO-FshIgG A chimeric protein, named FshIgG, was obtained after transforming the pPRO-FshIgG into Escherichia coli BL21 (DE3) host cell and induced by IPTG. Inoculation with 100 microg FsIgG induced strong neutralizing antibody response in guinea pigs, and FshIgG inoculated guinea pigs were also protected against 200 ID50 FMDV challenge. Our study indicated that the heavy chain constant region of sheep IgG can act as the carrier protein for FMDV peptide epitopes, and FshIgG is a potential multiepitope peptide vaccine candidate to prevent FMDV infection.

[Sludge characteristics in an aerobic granular sludge membrane bioreactor].

Granular sludge membrane bioreactor (GMBR), operated with alternating anaerobic-aerobic mode, showed good organics removal and simultaneous nitrification and denitrification (SND) performances during 120 days operation. Moreover, sludge size distribution examination showed that the increase of sludge concentration in GMBR was mostly due to the increase of small granular sludge with diameter between 0.18 - 0.45 mm and flocculent sludge smaller than 0.18 mm, and most of granular sludge larger than 0.45 mm was stably existed in GMBR, at the end of operation granular sludge with diameter larger than 0.18mm remained 60%-65% in GMBR. Sludge surface negative charge increased with the sludge size distribution variation, and surface charge remained -0.42 - -0.80 meq x g(-1) after 80 days operation. The increase of sludge surface negative charge was mostly caused by sludge smaller than 0.45 mm, thereinto sludge smaller than 0.18 mm had the greatest effect. Moreover, results revealed that the sludge surface negative charge decreased as the sludge size increased, and a linear correlation can be obtained between sludge surface negative charge and sludge size. Sludge in GMBR had good settleability, and SVI was about 60-90 mL/g. Furthermore results showed that sludge surface charge was correlated with SVI, sludge SVI increased with surface negative charge increased.

[Characteristics of nitrogen removal in aerobic granular sludge membrane bioreactor].

An aerobic granular sludge membrane bioreactor(GMBR) had been stably operated for 71 days, and it showed good organics removal and simultaneous nitrification and denitrification (SND) performances. TOC, ammonium and total nitrogen removal efficiencies of GMBR remained 84.7% - 91.9%, 85.4% - 99.7% and 41.7% - 78.4% respectively as influent TOC were 56.8 - 132.6 mg/L and ammonium nitrogen were 28.1 - 38.4 mg/L. Moreover, further batch tests of different size sludge revealed that floclike sludge nearly had not SND ability, the SND performances of GMBR was mostly come from granular sludge, and denitrification rate and total nitrogen removal efficiencies under aerobic condition increased with sludge size increased. Analyzing the morphology and section characteristics of granular sludge under scan electron microscope, the SND process was also described.