Inhibition of PKM2 suppresses osteoclastogenesis and alleviates bone loss in mouse periodontitis.

BACKGROUND: Chronic periodontitis triggers an increase in osteoclastogenesis, with glycolysis playing a crucial role in this process. Pyruvate kinase M2 (PKM2) is a critical enzyme involved in glycolysis and pyruvate metabolism. Yet, the precise function of PKM2 in osteoclasts and their formation remains unclear and requires further investigation. METHODS: Bioinformatics was used to investigate critical biological processes in osteoclastogenesis. In vitro, osteoclastogenesis was analyzed using tartrate-resistant acid phosphatase (TRAP) staining, phalloidin staining, quantitative real­time PCR (RT-qPCR), and Western blotting. Small interfering RNA (siRNA) of PKM2 and Shikonin, a specific inhibitor of PKM2, were used to verify the role of PKM2 in osteoclastogenesis. The mouse model of periodontitis was used to assess the effect of shikonin on bone loss. Analyses included micro computed tomography, immunohistochemistry, flow cytometry, TRAP staining and HE staining. RESULTS: Bioinformatic analysis revealed a significant impact of glycolysis and pyruvate metabolism on osteoclastogenesis. Inhibition of PKM2 leads to a significant reduction in osteoclastogenesis. In vitro, co-culture of the heat-killed Porphyromonas gingivalis significantly promoted osteoclastogenesis, concomitant with an increased PKM2 expression in osteoclasts. Shikonin weakened the promoting effect of porphyromonas gingivalis on osteoclastogenesis. In vivo experiments demonstrated that inhibition of PKM2 by shikonin alleviated bone loss induced by periodontitis, suppressed excessive osteoclastogenesis in alveolar bone, and reduced tissue inflammation to some extent. CONCLUSION: PKM2 inhibition by shikonin, a specific inhibitor of this enzyme, attenuated osteoclastogenesis and bone resorption in periodontitis. Shikonin appears to be a promising therapeutic agent for treating periodontitis.

Research on the purification efficiency and mechanism for road runoff pollutants in pervious concrete with recycled aggregates.

The use of recycled aggregate (RA) in pervious concrete (PC) is a green approach that can effectively mitigate urban waterlogging, excessive RA, and runoff pollution, thereby enhancing the urban ecological environment. This article focuses on the long-term purification efficiency of runoff pollutants by PC at different porosities and RA dosages. Moreover, the purification mechanism of pollutants by recycled aggregate pervious concrete (RAPC) was revealed utilizing particle size analysis, microstructure, and elemental analysis. Finally, the recovery effects of different maintenance approaches on the purification capacity of RAPC were explored. The results indicate that an increase in the RA dosage reduced the effective porosity of PC, thereby decreasing the permeability of RAPC. In addition, PC with a lower porosity demonstrated a slightly greater purification effectiveness for pollutants. However, the utilization of RA significantly enhanced the purification capacity of PC for various pollutants, primarily by leveraging advantages in terms of pore structure, micromorphology, and surface chemical composition. Additionally, RAPC exhibited nearly 100 % retention effectiveness for particles larger than 68.95 µm but relatively lower purification efficiency for particles ranging from 1.541 to 17.11 µm. In particular, it displayed the poorest purification performance for particles with a diameter of 6.396 µm. The surface of RAPC's pore channels exhibited a loose state with high porosity and appeared rough and uneven with numerous pits and grooves. RAPC had a larger surface area and contained more components, such as SiO2, CaCO3, and Al2O3, than regular PC. Therefore, RAPC possessed a higher purification capacity. High-pressure flushing (HPF) and sodium citrate flushing (SCF) under different maintenance frequencies significantly contributed to the recovery of the purification efficiency of RAPC. However, overall, a lower maintenance frequency led to a less favorable recovery effect. Furthermore, SCF had a better recovery effect than HPF.

Preparation of Auricularia auricula polysaccharides and their protective effect on acute oxidative stress injury of Caenorhabditis elegans.

This research enhanced the extraction procedure for Auricularia auricula crude polysaccharides by utilizing a modified Fenton reagent as a solvent, and obtained A. auricula polysaccharides (AAPs-VH) via alcohol precipitation and deproteinization. The HPLC profile revealed that the purified AAPs-VH using Sepharose 6FF was mainly a heteropolysaccharide, consisting primarily of mannose, glucuronic acid, glucose, and xylose. The Mw and Mn of the purified AAPs-VH were 87.646 kDa and 48.854 kDa, respectively. The FT-IR and NMR spectra revealed that the purified AAPs-VH belonged to pyranose and were mainly formed by (1 â†’ 3)-linked-ß-D glucan formation. In vivo experiments conducted with Caenorhabditis elegans, AAPs-VH was found to notably influence the lifespan, improve the antioxidant system, and decrease the level of cell apoptosis. This might be achieved by up-regulating the expression of genes in the IIS and TOR pathways. The study concludes that the modified Fenton reagent can increase Auricularia auricula polysaccharide solubleness and active sites, which may be an essential prompt for future studies.

Engineering a Cl- -Modulated Light-Driven Na+ Pump.

Microbial Na+ -pumping rhodopsin (NaR) is a promising optogenetic tool due to its unique ability to transport Na+ . Like most rhodopsin-based tools, NaR is limited to light-based control. In this study, our objective was to develop a novel mode of modulation for NaR beyond light control. By introducing a potential Cl- binding site near the putative Na+ release cavity, we engineered Nonlabens dokdonensis rhodopsin 2 (NdR2) to be modulated by Cl- , an essential chemical in organisms. The engineered NdR2 demonstrated an approximately two-fold increase in Na+ pump activity in the presence of 100 mM Cl- compared to Cl- -free solution. Increasing Cl- concentration decreased the lifetimes of the M and O intermediates accordingly. The analysis of competitive ion uptake suggested the bound Cl- may increase the Na+ affinity and selectivity. This chemical modulation allows for more diverse and precise control over cellular processes, advancing the development of next-generation optogenetic tools. Notably, our Cl- -modulated NdR2 establishes an innovative mechanism for linking Cl- to Na+ -related processes, with potential applications in optogenetic therapies for related diseases.

Functionalized Fluorescent Organic Nanoparticles Based AIE Enabling Effectively Targeting Cancer Cell Imaging.

We report a fluorescent dye TM by incorporating the tetraphenylethylene (TPE) and cholesterol components into perylene bisimides (PBI) derivative. Fluorescence emission spectrum shows that the dye has stable red emission and aggregation-induced emission (AIE) characteristics. The incorporation of cholesterol components triggers TM to show induced chirality through supramolecular self-assembly. The cRGD-functionalized nanoparticles were prepared by encapsulating fluorescent dyes with amphiphilic polymer matrix. The functionalized fluorescent organic nanoparticles exhibit excellent biocompatibility, large Stokes' shift and good photostability, which make them effective fluorescent probes for targeting cancer cells with high fluorescence contrast.

Insulin Resistance Triggers Atherosclerosis: Caveolin 1 Cooperates with PKCzeta to Block Insulin Signaling in Vascular Endothelial Cells.

OBJECTIVE: To date, therapies for endothelial dysfunction have primarily focused on ameliorating identified atherosclerosis (AS) risk factors rather than explicitly addressing endothelium-based mechanism. An in-depth exploration of the pathological mechanisms of endothelial injury was performed herein. METHODS: Aortic caveolin 1 (Cav1) knockdown was achieved in mice using lentivirus, and AS was induced using a high-fat diet. Mouse body weight, blood glucose, insulin, lipid parameters, aortic plaque, endothelial injury, vascular nitric oxide synthase (eNOS), injury marker, and oxidative stress were examined. The effect of Cav1 knockdown on the content of PKCzeta and PI3K/Akt/eNOS pathway-related protein levels, as well as PKCzeta binding to Akt, was studied. ZIP, a PKCzeta inhibitor, was utilized to treat HUVECs in vitro, and the effect of ZIP on cell viability, inflammatory response, oxidative stress, and Akt activation was evaluated. RESULTS: Cav1 knockdown had no significant effect on body weight or blood glucose in mice over an 8-week period, whereas drastically reduced insulin, lipid parameters, endothelial damage, E-selectin, and oxidative stress and elevated eNOS levels. Moreover, Cav1 knockdown triggered decreased PKCzeta enrichment and the activation of the PI3K/Akt/eNOS pathway. PKCzeta has a positive effect on cells without being coupled by Cav1, and ZIP had no marked influence on PKCzeta-Akt binding following Cav1/PKCzeta coupling. CONCLUSION: Cav1/PKCzeta coupling antagonizes the activation of PI3K on Akt, leading to eNOS dysfunction, insulin resistance, and endothelial cell damage.

Effect of the lateral bone cut end on pattern of lingual split during bilateral sagittal split osteotomy in patients with skeletal class III malocclusion.

This study examines the effect of the lateral bone cut end (LBCE) on the pattern of lingual split during bilateral sagittal split osteotomy (BSSO) in patients with skeletal class III malocclusion. A case-control study according to the pattern of the sagittal split osteotomy (SSO) lingual split line was conducted in patients who underwent BSSO. The primary predictor variable was the ratio of the LBCE. The primary outcome variable was the type of lingual fracture line classified according to the lingual split scale (LSS). Other variables included patients' weight, sex, age, left and right sides of the mandible, and experience of the surgeon. Logistic regression analysis or the chi-squared test was performed to determine the effect of these variables on various types of lingual fracture line. The significance level was 95% (p < 0.05). There were 271 patients enrolled in this study. The SSO lingual split lines were divided into LSS1 (329/542), LSS2 (82/542), LSS3 (93/542), and LSS4 (38/542) splits. Logistic regression analysis showed that the LSS3 split was more likely to appear when the LBCE was closer to the lingual side (p = 0.0017). The age of patients significantly affected the possibilities of LSS2 (p = 0.0008) and LSS3 (p = 0.0023) splits. A LBCE close to the lingual side was an inducer for the formation of a LSS3 split in patients with skeletal class III malocclusion during BSSO. The age of the patient also affected the possibility of LSS2 and LSS3 splits.

Establishment of a neutralization assay for Nipah virus using a high-titer pseudovirus system.

OBJECTIVE: To construct a high-titer Nipah pseudovirus packaging system using the HIV lentivirus backbone vector and establish a safe neutralization assay for Nipah pseudovirus in biosafety level 2 facilities. METHODS: Nipah virus (NiV) fusion protein (F) and glycoprotein (G) recombinant expression plasmids, psPAX2, and pLenti CMV Puro LUC (w168-1) were transiently transfected into 293T cells for 72 h for the generation of a NiV pseudovirus. The neutralization ability of Nipah virus F and G protein antibodies was assessed using the pseudovirus. RESULTS: A NiV pseudovirus was constructed using 293T cells. The ideal mass ratio of plasmid psPAX2: w168-1: F: G for transfection was determined to be 4:4:1:1. The specificity of recombinant F and G protein expression was indicated by indirect immunofluorescence and western blotting. The pseudovirus particles showed obvious spikes under a transmission electron microscope. The NiV pseudovirus titer was 4.73 × 105 median tissue culture infective dose per mL, and the pseudovirus could be effectively neutralized by polyclonal antibodies specifically targeting the F and G proteins respectively. CONCLUSIONS: A NiV pseudovirus was successfully generated using HIV vector systems, and was used as a platform for a safe and reliable pseudovirus-based neutralizing assay that can be performed in biosafety level 2 facilities.

Lignin detaching from the oxidative delignified softwood during enzymatic hydrolysis and its effect on carbohydrate saccharification.

Most studies about the influence of lignin on enzymatic digestibility of lignocellulose have focused on the content and properties, but less on the detaching behavior of lignin. The samples were prepared from Pinus massoniana wood chips by kraft cooking followed by delignification using oxygen/alkali (KP-O) and chlorine dioxide (KP-D), respectively. Two oxidative delignified samples with a similar lignin content were subject to enzymatic hydrolysis at both pH of 5.0 and 5.5 to investigate the effects of lignin detached rate (LDR) on substrate enzymatic digestibility (SED). The LDRs and the SEDs from both samples increased with the enzymatic hydrolysis time, and the situations of KP-D were much higher than those of KP-O under the same enzymatic hydrolysis time. The results of enzymatic hydrolysis at an elevated pH of 5.5 and the changes in concentration of free cellulase of the two samples indicated that the lignin detaching increased the free cellulase concentration, and thus promoted the enzymatic digestibility. Moreover, lignin distribution analysis by X-ray photoelectric spectroscopy and confocal laser scanning microscopy indicated surface lignin being preferentially detached. This work provided a reference for rationally designing pretreatment strategies, which can improve the efficiency of enzyme hydrolysis of lignocellulosic biomass.

Human metapneumovirus in hospitalized children with acute respiratory tract infections in Beijing, China.

BACKGROUND: This study aims to described the epidemiology and genotypic diversity of Human metapneumovirus (HMPV) and the impact of SARS-CoV-2 on the prevalence of HMPV in hospitalized children with Acute respiratory tract infections (ARTIs) in Beijing, China. METHODS: From April 2018 to March 2019 and from September 2020 to August 2021, nasopharyngeal aspirates (NPAs) from hospitalized children with ARTIs in Beijing were collected and subjected to real-time polymerase chain reaction tests for HMPV. Then genotyping, detection of 15 common respiratory viruses and clinical characteristics were analyzed on HMPV positive samples. RESULTS: 7.9% (124/1572) enrolled pediatric patients were identified as having HMPV infection, and the majority of children under the age of 5 (78.2%, 92/124), From April 2018 to March 2019. The detection rate of HMPV in spring and winter is significantly higher than that in summer and autumn. The co-infection rate were 37.1% (46/124), the most common co-infected virus were parainfluenza virus type 3 (HPIV-3). The main diagnosis of HMPV infection was pneumonia (92.7%,115/124), most patient have cough and fever. Of 78 HMPV-positive specimens, A2b (82.1%,64/78) were the main epidemic subtypes. Hospitalized children with HMPV genotype A infection had a higher viral load compared to genotype B. During the COVID-19 outbreak, Among 232 samples, only 4 cases were HMPV-positive. After statistical test, the detection rate of HMPV during the COVID-19 pandemic has decreased significantly compared with that before the epidemic (p = 0.001). CONCLUSIONS: HMPV is an important cause of ARTIs in children under 5 years old. The epidemic peak is generally in winter and spring, and the A2b subtype is the most common. However, under the prevention and control of the COVID-19 pandemic, the HMPV infection of hospitalized children with ARTIs has decreased significantly.

Comprehensive virome analysis of the viral spectrum in paediatric patients diagnosed with Mycoplasma pneumoniae pneumonia.

BACKGROUND: Among hospitalized children suffering from community-acquired pneumonia, Mycoplasma pneumoniae (MP) is one of the most common pathogens. MP often exists as a co-infection with bacteria or viruses, which can exacerbate the clinical symptoms. We investigated the pathogen spectrum in MP-positive and MP-negative samples from hospitalized children with respiratory tract infections in Beijing, China. METHOD: This study included 1038 samples of nasopharyngeal aspirates obtained between April, 2017 and March, 2018 from hospitalized children under 6 years of age with respiratory tract infections. To explore the impact of MP infection on the composition of the pathogen spectrum, 185 nasopharyngeal aspirates (83 MP-positive/102 MP-negative) were randomly selected for next-generation sequencing and comprehensive metagenomics analysis. Real-time PCR was used to detect and verify common respiratory viruses. RESULTS: Of the 1038 samples, 454 (43.7%) were infected with MP. In children < 6 years of age, the MP infection rate gradually increased with age, with the highest rate of 74.2% in 5-6-year-olds. The results of metagenomics analysis revealed 11 human, animal and plant virus families, and bacteriophages, including common respiratory viruses, enteroviruses and anelloviruses. The virus family with the highest number of reads in both MP-positive and MP-negative samples was the Pneumoviridae, and the number of reads for human respiratory syncytial virus (HRSV) in MP-positive samples was higher than that in MP-negative samples. Among the 83 MP-positive samples, 47 (56.63%) were co-infected with viruses, the most common of which was influenza virus (IFV). The durations of hospitalization and fever were higher in patients with MP co-infection than MP single infection, but the difference was not statistically significant. CONCLUSION: The viral family with the highest number of reads in both groups was Pneumoviridae, and the number of reads matched to HRSV in MP-positive samples was much higher than MP-negative samples. Co-infection of MP and IFV infection were the most cases.

Surface Modification by Amino Group Inducing for Highly Efficient Catalytic Oxidation of Toluene over a Pd/KIT-6 Catalyst.

Toluene is one of the typical volatile organic compounds in industry, particularly in energy and fuels production processes, which is required to be eliminated effectively to protect the environment. Catalytic oxidation of toluene is widely studied for its high efficiency, and rational design and synthesis of metal catalysts are keys for toluene oxidation. In this study, an efficient catalyst was designed and synthesized by introducing -NH2 groups on the ordered mesoporous silica (KIT-6) surface to anchor and disperse Pd species, leading to Pd nanoparticles being highly dispersed with uniform particle size distribution. Meanwhile, it was found that the introduction of -NH2 made Pd centers present an electron-rich state, and the active Pd centers could activate O2 molecules to generate more reactive oxygen species and promote the conversion of toluene, which was verified by in situ XPS and O2-TPD characterization. Compared with the catalysts prepared by an impregnation method, the catalytic performance of the Pd/NH2-KIT-6 (0.5 wt %) catalyst was significantly improved. A conversion of 90% for toluene (2400 ppm, 24,000 mL·g-1·h-1) was achieved at 171 °C, and the toluene conversion was maintained above 90% for 900 min, displaying the excellent activity and stability of the Pd/NH2-KIT-6 catalyst.

Porphyromonas gingivalis Induces Bisphosphonate-Related Osteonecrosis of the Femur in Mice.

One of the most prominent characteristics of bisphosphonate-related osteonecrosis of the jaw(BRONJ) is its site-specificity. Osteonecrosis tends to occur specifically in maxillofacial bones, in spite of a systemic administration of the medicine. Previous studies suggested rich blood supply and fast bone turnover might be reasons for BRONJ. Yet, a sound scientific basis explaining its occurrence is still lacking. The present study aimed to explore the role of Porphyromonas gingivalis (P. gingivalis), an important oral pathogen, on the site-specificity of bisphosphonate-induced osteonecrosis and to elucidate its underlying mechanism. Mice were intraperitoneally injected with zoledronic acid (ZA) or saline for 3 weeks. In the third week, the right mandibular first molars were extracted and circular bone defects with a diameter of 1 mm were created in right femurs. After the operation, drug administration was continued, and P. gingivalis suspension was applied to the oral cavities and femur defects. The mice were killed after four or eight weeks postoperatively. The right mandibles and femurs were harvested for micro-CT and histological analyses. A poor healing of bone defects of both jaws and femurs was noted in mice injected with both ZA and P. gingivalis. Micro-CT analysis showed a decreased bone volume, and histological staining showed an increased number of empty osteocyte lacunae, a decreased collagen regeneration, an increased inflammatory infiltration and a decreased number of osteoclasts. In addition, the left femurs were collected for isolation of osteoclast precursors (OCPs). The osteoclastogenesis potential of OCPs was analyzed in vitro. OCPs extracted from mice of ZA-treated groups were shown to have a lower osteoclast differentiation potential and the expression level of related genes and proteins was declined. In conclusion, we established a mouse model of bisphosphonate-related osteonecrosis of both the jaw and femur. P. gingivalis could inhibit the healing of femur defects under the administration of ZA. These findings suggest that P. gingivalis in the oral cavity might be one of the steering compounds for BRONJ to occur.

Effect of Enzyme-Assisted Extraction on the Chemical Properties and Antioxidant Activities of Polysaccharides Obtained from the Wood Ear Mushroom, Auricularia auricula (Agaricomycetes).

An effective method of polysaccharide extraction from Auricularia auricula (AAPs) by mannanase was developed and optimized by response surface methodology in which the ABTS+ [diammonium 2,2'-azino-bis(3-ethylben-zothiazoline-6-sulfonate radical] scavenging rate was the response. AAPs were graded by stepwise ethanol precipitation with concentrations of 5, 10, 15, and 20% ethanol successively. The fractions with a strong radical scavenging rate were obtained, and then their antioxidant stress effect was studied using Caenorhabditis elegans as a model organism. The ABTS+ scavenging rate of AAPs could reach 37.95 ± 0.53% at a temperature of 55°C, a time of 4 h, a liquid-to-material ratio of 58 mL/1 g, and an enzyme-to-substrate ratio of 2.97%. AAP-20 obtained by 20% ethanol with a strong radical scavenging rate was a heteropolysaccharide composed of mannose, glucose, galactose, xylose, and glucuronic acid. AAP-20 could significantly prolong the lifespan of C. elegans under oxidative stress conditions induced by methyl viologen or hydrogen peroxide, and it could also enhance the activity of antioxidant enzymes including catalase, glutathione reductase, and superoxide dismutase at 0.50 mg/mL (P < 0.05). These studies showed that AAPs prepared with mannanase had a significant protective effect against damage induced by intracellular radical generating agents.

P2RY8 variants in lupus patients uncover a role for the receptor in immunological tolerance.

B cell self-tolerance is maintained through multiple checkpoints, including restraints on intracellular signaling and cell trafficking. P2RY8 is a receptor with established roles in germinal center (GC) B cell migration inhibition and growth regulation. Somatic P2RY8 variants are common in GC-derived B cell lymphomas. Here, we identify germline novel or rare P2RY8 missense variants in lupus kindreds or the related antiphospholipid syndrome, including a "de novo" variant in a child with severe nephritis. All variants decreased protein expression, F-actin abundance, and GPCR-RhoA signaling, and those with stronger effects increased AKT and ERK activity and cell migration. Remarkably, P2RY8 was reduced in B cell subsets from some SLE patients lacking P2RY8 gene variants. Low P2RY8 correlated with lupus nephritis and increased age-associated B cells and plasma cells. By contrast, P2RY8 overexpression in cells and mice restrained plasma cell development and reinforced negative selection of DNA-reactive developing B cells. These findings uncover a role of P2RY8 in immunological tolerance and lupus pathogenesis.

Neuroprotective effects of arbutin against oxygen and glucose deprivation-induced oxidative stress and neuroinflammation in rat cortical neurons.

In this study, the neuroprotective potential of arbutin (100 µmol L-1) pre-treatment and post-treatment against oxygen/ glucose deprivation (OGD) and reoxygenation (R) induced ischemic injury in cultured rat cortical neurons was explored. The OGD (60 min) and reoxygenation (24 h) treatment significantly (p < 0.001) compromised the antioxidant defence in cultured neurons. Subsequently, an increase (p < 0.001) in lipid peroxidation and inflammatory cytokines (tumour necrosis factor-α and nuclear factor kappa-B) declined neuron survival. In pre- and post-condition experiments, treatment with arbutin enhanced both survival (p < 0.01) and integrity (p < 0.05) of cultured neurons. Results showed that arbutin protects (p < 0.05) against peroxidative changes, inflammation, and enhanced the antioxidant activity (e.g., glutathione, superoxide dismutase and catalase) in cultured neurons subjected to OGD/R. It can be inferred that arbutin could protect against ischemic injuries and stroke. The anti-ischemic activity of arbutin can arrest post-stroke damage to the brain.

Preparation of the Enzymatic Hydrolysates from Chlorella vulgaris Protein and Assessment of Their Antioxidant Potential Using Caenorhabditis elegans.

This study aimed to assess the antioxidant potential of Chlorella vulgaris protein-derived enzymatic hydrolysate using Caenorhabditis elegans. Protein extraction was performed using an alkali solution after complete C. vulgaris swelling and hydrolysis using four commercial proteases (alcalase, neutrase, protamex, and flavourzyme). The results showed that the flavourzyme hydrolysates exhibited the strongest antioxidant activity both in vitro and in vivo. Under the optimum conditions of the enzymatic hydrolysis, the half-maximal effective concentration of the hydrolysates for superoxide and hydroxyl radicals was 0.323 mg/mL and 0.139 mg/mL, respectively. The hydrolysates could significantly extend the lifespan, improve the resistance to methyl viologen-induced oxidative stress, reduce the levels of reactive oxygen species, and enhance the activity of catalase and superoxide dismutase in C. elegans.

The lipid lowering and antioxidative stress potential of polysaccharide from Auricularia auricula prepared by enzymatic method.

An efficient extraction method of Auricularia auricula polysaccharides (AAPs) by neutral protease was developed and optimized by response surface methodology. AAPs were graded by stepwise ethanol precipitation, the fraction with high recovery rate and strong radical scavenging rate were obtained, then its antioxidant and lipid lowering effect were studied using Caenorhabditis elegans as model organism. The extract yield and ABTS+ scavenging rates of AAPs could reach 14.90% and 86.0% at 50 °C, 75 mL/g of liquid-to-material ratio and pH 9.0. AAP3 obtained by 15% ethanol was a heteropolysaccharide comprised of mannose, glucose, glucuronic acid, xylose, galactose and glucosamine. AAP3 could significantly prolong the lifespan of C. elegans and enhance the activity of antioxidant enzymes including superoxide dismutase (SOD), catalases (CAT) at 0.25 mg/mL (p < 0.05). The qRT-PCR results showed that AAP3 could up regulate mRNA expression levels of daf-16 and skn-1 (>1.6 fold) at 0.25 mg/mL. Besides, AAP3 could significantly reduce the level of body fat and triglyceride in C. elegans (p < 0.05). These studies demonstrated that A. auricula polysaccharides prepared by neutral protease had a prominent protective effect to the damage induced by the intracellular free radical generating agents.

The antioxidant capacity evaluation of polysaccharide hydrolyzates from pumpkin using Caenorhabditis elegans model.

Our previous study has optimized the acid hydrolysis process of pumpkin polysaccharides (PPe) with scavenging ability based on central composite design. The aim of this study was to explore the in vivo-antioxidant ability of PPe and pumpkin polysaccharides acid-hydrolysis (PPe-S) using Caenorhabditis elegans. In composition analysis, the constituents of total sugar, protein, uronic acid, and sulfur groups in PPe-S were 87.03 ± 1.21%, 1.25 ± 0.78%, 37.61 ± 0.97%, and 0.14 ± 0.04%, respectively. Besides, results of antioxidant ability showed that PPe and PPe-S could reduce the oxidative stress (OS) induced by methyl viologen, extend lifespan of worms, and reduce reactive oxygen species (ROS) level under oxidative conditions significantly (p < .05). Furthermore, PPe and PPe-S could enhance the stress-resistance related antioxidant enzymes including catalase (CAT) and superoxide dismutase (SOD) significantly (p < .05). Moreover, the antioxidant effect of PPe-S was superior to PPe at the concentration of 4.0 mg/ml. In summary, this study demonstrated that the derived hydrolyzates from PPe had protective effects on the damage induced by the generation of intracellular free radical agents. PRACTICAL APPLICATIONS: OS plays an important role in the pathogenesis of metabolic diseases, including type 2 diabetes. It is widely acknowledged that diabetes and its complications pose a threat to human's health, and the number of people with diabetes will expand to 640 million in the 2040 year. Current studies have shown that all diabetes drugs have a kind of side effects. Fortunately, researchers have found and confirmed that plant-derived polysaccharide had a notable hypoglycemic effect via reducing the OS level in cell and tissue, and could decrease the diabetes symptoms as well. In this study, we proved that the polysaccharide derived from pumpkin could effectively ameliorate the OS level in C. elegans, including decreasing the damage of biofilm and ROS level. Therefore, our study shows that there is a high potential for pumpkin-derived polysaccharide and its hydrolyzates to be a bioactive component to prevent diabetes. In other words, this research can be applied to diabetes prevention and other diseases induced by OS.