Knocking Down PIAS3 Reduces H2O2-induced Oxidative Stress Injury in HT22 Cells.

Oxidative stress is involved in the pathological processes of many neurodegenerative diseases. Protein modification by small ubiquitin-like modifiers (SUMOs) has been implicated in oxidative stress injury. By conjugating SUMOs to their selective protein substrates, SUMO ligases play critical roles in regulating functions of proteins involved in oxidative stress injury. In this study, we screened siRNAs to knockdown the SUMO ligase PIAS3 to assess its role in H2O2-induced injury in HT22 cells. H2O2 stimulation increased total protein SUMOylation, facilitated intracellular reactive oxygen species (ROS) release, increased cleaved caspase-3 levels, promoted p38 and JNK activation (phosphorylation), upregulated apoptosis, and decreased cell viability. The siRNA against PIAS3 329-347 (siPIAS3-329) markedly downregulated the protein expression of PIAS3 and reversed these effects, whereas siNC (negative control) had no effect. Our findings demonstrate that PIAS3-mediated SUMOylation facilitates oxidative stress injury and p38/JNK-mediated cell apoptosis and that PIAS3 is a potential target to protect against oxidative stress injury.

Wearable sampling of proteins from human exhaled aerosols for nano-liquid chromatography-tandem mass spectrometry analysis: A pilot study.

RATIONALE: Human exhaled breath usually contains unique proteins that may provide clues to characterize individual physiological activities and many diseases. However, the concentration of exhaled proteins in exhaled breath is extremely low and usually does not reach the detection limits of all online breath mass spectrometry instruments. Therefore, developing a new breath sampler for collecting and characterizing exhaled proteins is important. METHODS: In this study, a new mask-based wearable sampler was developed by fixing metal materials into the inner surface of the KN95 mask. Human exhaled proteins could be directly adsorbed onto the metal material while wearing the mask. After sampling, the collected proteins were eluted, digested, and identified using nano-liquid chromatography-tandem mass spectrometry (nano-LC-MS/MS). RESULTS: The adsorption of exhaled proteins was evaluated, showing that modified gold foil is an effective material for collecting exhaled proteins. Various endogenous proteins were successfully identified from exhaled breath, many of which can be potential biomarkers for disease diagnosis. CONCLUSIONS: By coupling the newly developed mask sampler with nano-LC-MS/MS, human exhaled proteins were successfully collected and identified. Our results show that the mask sampler is wearable, simple, and convenient, and the method is noninvasive for investigating disease diagnosis and human health.

Disrupting PIAS3-mediated SUMOylation of MLK3 ameliorates poststroke neuronal damage and deficits in cognitive and sensorimotor behaviors.

Activated small ubiquitin-like modifiers (SUMOs) have been implicated in neuropathological processes following ischemic stroke. However, the target proteins of SUMOylation and their contribution to neuronal injury remain to be elucidated. MLK3 (mixed-lineage kinase 3), a member of the mitogen-activated protein kinase kinase kinase (MAPKKK) family, is a critical regulator of neuronal lesions following cerebral ischemia. Here, we found that SUMOylation of MLK3 increases in both global and focal ischemic rodent models and primary neuronal models of oxygen and glucose deprivation (OGD). SUMO1 conjugation at the Lys401 site of MLK3 promoted its activation, stimulated its downstream p38/c-Jun N-terminal kinase (JNK) cascades, and led to cell apoptosis. The interaction of MLK3 with PIAS3, a SUMO ligase, was elevated following ischemia and reperfusion. The PINIT domain of PIAS3 was involved in direct interactions with MLK3. Overexpression of the PINIT domain of PIAS3 disrupted the MLK3-PIAS3 interaction, inhibited SUMOylation of MLK3, suppressed downstream signaling, and reduced cell apoptosis and neurite damage. In rodent ischemic models, the overexpression of the PINIT domain reduced brain lesions and alleviated deficits in learning, memory, and sensorimotor functions. Our findings demonstrate that brain ischemia-induced MLK3 SUMOylation by PIAS3 is a potential target against poststroke neuronal lesions and behavioral impairments.

Effects of two celery fibers on the structural properties and digestibility of glutinous rice starch: A comparative study.

The present study focused on the extraction of water-soluble dietary fiber (CSDF) and water-insoluble dietary fiber (CIDF) from celery. It investigated their effects on glutinous rice starch's (GRS) physicochemical, structural, and digestive properties. The results showed that as the addition of the two dietary fibers increased, they compounded with GRS to varying degrees, with the complexing index reaching 69.41 % and 60.81 %, respectively. The rheological results indicated that the two dietary fibers reduced the viscosity of GRS during pasting and inhibited the short-term regrowth of starch. The FTIR and XRD results revealed that the two fibers interacted with GRS through hydrogen bonding, effectively inhibiting starch retrogradation. Furthermore, both fibers increased the pasting temperature of GRS, thus delaying its pasting and exhibiting better thermal stability. Regarding digestibility, the starch gels containing dietary fibers exhibited significantly reduced digestibility, with RS significantly increased by 8.15 % and 8.95 %, respectively. This study provides insights into the interaction between two dietary fibers and GRS during processing. It enriches the theoretical model of dietary fiber-starch interaction and provides a reference for the application development of starch-based functional foods.

Influence of the structure and physicochemical properties of OSA modified highland barley starch based on ball milling assisted treatment.

This study aimed to investigate the influence of ball milling assisted treatment on the degree of substitution of octenyl succinic anhydride (OSA) modified highland barley starch (HBS) and on the physicochemical properties and structure of HBS. Scanning electron microscopy (SEM) findings showed that with the increasing of ball milling time, the surface morphology of OSA modified HBS became rougher and rougher and the particle morphology and crystal structure were damaged. When the pretreatment time of ball milling was 40 min, the degree of substitution of OSA modified HBS was 1.32 times higher than that of the conventional modification method. In addition, the longer the ball milling assistant, the longer the short-range ordering of the OSA modified HBS significantly decreased, and the relative crystallinity decreased (from 16.68 % to 7.93 %), leading to a decrease in thermal stability too. However, it greatly enhanced the aging resistance and flowability. In terms of emulsification properties, the emulsification properties of OSA modified HBS increased from 60.67 % to 75.67 %. Therefore, the HBS with better freeze-thaw stability and higher degree of substitution can be prepared by ball milling pretreatment and OSA modification, which provides technical support for further development of starch resources.

Effects of starch multiscale structure on the physicochemical properties and digestibility of Radix Cynanchi bungei starch.

Radix Cynanchi bungei (RCb) contains 40-70 % starch, yet little is known about the structure and properties of RCb starch. In this study, the multiscale structure of two cultivars of RCb starch (YW201501 and BW201001) were characterized, and the effects of starch structure on its physicochemical properties were investigated. The differences in physicochemical properties of RCb starch were influenced by its multiscale structure. The starch granules were round and irregular polygon, with sizes ranging between 2 and 14 µm. YW201501 had a higher amylose (21.81 %) and lipid (0.96 %) content, molecular weight (59.5 × 106 g/mol), and A chain proportion (27.5 %), and a lower average granule size (6.14 µm), amylopectin average chain length (19.7), and B3 chain proportion (10.3 %). Both starches were B-type crystalline, with higher crystallinity (26.3 %) and R1047/1022 (0.74) for YW201501, resulting in large gelatinization enthalpy. In addition, the higher peak viscosity and larger retrogradation degree of YW201501 were correlated to its higher amylose content. In vitro digestibility revealed that the low rapidly digestible starch and high resistant starch of BW201001 were related to the fine structure of starch. YW201501 and BW201001 had a medium glycemic index (62.6-66.0) with potential for processing into healthy starchy foods.

MLK3 localizes mainly to the cytoplasm and promotes oxidative stress injury via a positive feedback loop.

Activation of mixed lineage kinase 3 (MLK3) by phosphorylation at Thr277/Ser281 stimulates downstream apoptotic pathways and ultimately leads to cell injury. MLK3 is reported to localize to both the cytoplasm and nucleus in human ovarian cancer cells and immortalized ovarian epithelial cells (T80 and T90 cells), and phosphorylation at Thr477 is required for the cytoplasmic retention of MLK3 in T80 cells. However, the subcellular distribution of MLK3 in other cell types has rarely been reported, and whether phosphorylation of MLK3 at Thr277/Ser281 affects its subcellular distribution is unknown. Here, our bioinformatics analysis predicted that MLK3 was mainly distributed in the cytoplasm and nucleus. In the human HEK293T embryonic kidney cell line and murine HT22 hippocampal neuronal cell line, endogenous MLK3 was more abundant in the cytoplasm and less abundant in the nucleus. In addition, overexpressed Myc-tagged MLK3 and EGFP-tagged MLK3 were also observed to localize mainly to the cytoplasm. MLK3 that was activated by phosphorylation at Thr277/Ser281 was mainly distributed in the cytoplasm, and phosphorylation deficient (T277A/S281A) and mimic (T277E/S281E) mutants both showed distributions similar to that of wild type (wt) MLK3, further proving that phosphorylation at Thr277/Ser281 was not involved in regulating MLK3 subcellular localization. In HEK293T cells, H2O2 stimulation accelerated MLK3 phosphorylation (activation), and this phosphorylation was reduced by the antioxidant N-acetylcysteine in a dose-dependent manner. Overexpressing wt MLK3 promoted the production of intracellular reactive oxygen species and increased cell apoptosis, both of which were enhanced by the phosphorylation-mimic (T277E/S281E) MLK3 variant but not by the phosphorylation-deficient (T277A/S281A) MLK3 variant. These findings provided additional evidence for the cytoplasmic and nuclear distribution of MLK3 in HEK293T cells or HT22 cells and revealed the pivotal role of MLK3 in the positive feedback loop of oxidative stress injury.

Pediatric tuina for the treatment of fever in children: A protocol for systematic review and meta-analysis.

BACKGROUND: Infantile fever is a common symptom of the pediatric diseases, which is often caused by cold, food accumulation, or other pathogenic factors. Pediatric tuina is regarded as an acceptable non-pharmaceutical therapy for children with optimal effects, which has been widely used for infantile fever around China. But there is still a lack of systematic evaluation and research on its safety and effectiveness during the treatment of infantile fever. Thus the protocol is to collect clinical evidence and demonstrate the efficacy and safety of antipyretic manipulation by pediatric tuina. METHODS: The systematic electronic search will be executed in Cochrane Library (1991-2020.6), EMBASE (1980-2020.6), PubMed (1996-2020.6), WHOICTRP (2004-2020.6), Web of Science (1900-2020.6), CNKI (1994-2020.6), CBM (1994-2020.6), WANFANG (1980-2020.6), and VIP (2000-2020.6) Database. The Review Manager (V.5.3) will be use to assess the risk of bias and data analyses. The methodological quality will be assessed by using the online GRADEpro tool. If the quality of numeric data is favorable, a meta-analysis will be carried out. RESULTS: A high-quality evidence of pediatric tuina for the treatment of infantile fever. CONCLUSION: The systematic review will provide a reliable basis for judging whether pediatric tuina is safe and effective in the treatment of pediatric fever. INPLASY REGISTRATION NUMBER: INPLASY202060032.

[Coupling between ecological environment vulnerability and multidimensional poverty: An empirical analysis from 1586 poverty-stricken villages in Hechi City, Guangxi, Southwest China]. / ç”Ÿæ€çŽ¯å¢ƒè„†å¼±æ€§ä¸Žå¤šç»´è´«å›°çš„è€¦åˆå ³ç³»­­åŸºäºŽå¹¿è¥¿æ²³æ± 市1586个贫困村的实证分析.

Examining the interaction mechanism between ecological environment and regional poverty at fine scale is an important prerequisite and guarantee for sustainable development of resources, environment and social economy from the combined perspectives of precise poverty alleviation and rural revitalization. We established the evaluation models of ecological vulnerability, multidimensional poverty, and the coupling coordination degree model, beased on which we systematically and comprehensively revealed the interaction between ecological environment and multidimensional poverty. The test results from 1586 administrative villages showed that the higher-vulnerability area of ecological environment was mainly distributed in the southeast region. Multidimensional poverty was increasing from northwest to southeast, with a spatial distribution pattern of "overall dispersion, local aggregation". In the coordinated development zone, ecological environment vulnerability and multidimensional poverty was positively correlated. In the disordered declining zone, the degree of coupling between ecological environment vulnerability and multidimensional poverty was lower, wth significantly negative correlation, indicating that the improvement of ecological environment quality and the development of multidimensional poverty reduction were asynchronous. There was a close and complex relationship between eco-environmental vulnerability and multidimensional poverty. Therefore, during poverty alleviation and development, we should pay more attention to environmental protection to prevent the poverty-stricken areas falling into the trap of ecological poverty.

Multidimensional poverty measure and analysis: a case study from Hechi City, China.

Aiming at the anti-poverty outline of China and the human-environment sustainable development, we propose a multidimensional poverty measure and analysis methodology for measuring the poverty-stricken counties and their contributing factors. We build a set of multidimensional poverty indicators with Chinese characteristics, integrating A-F double cutoffs, dimensional aggregation and decomposition approach, and GIS spatial analysis to evaluate the poor's multidimensional poverty characteristics under different geographic and socioeconomic conditions. The case study from 11 counties of Hechi City shows that, firstly, each county existed at least four respects of poverty, and overall the poverty level showed the spatial pattern of surrounding higher versus middle lower. Secondly, three main poverty contributing factors were unsafe housing, family health and adults' illiteracy, while the secondary factors include fuel type and children enrollment rate, etc., generally demonstrating strong autocorrelation; in terms of poverty degree, the western of the research area shows a significant aggregation effect, whereas the central and the eastern represent significant spatial heterogeneous distribution. Thirdly, under three kinds of socioeconomic classifications, the intra-classification diversities of H, A, and MPI are greater than their inter-classification ones, while each of the three indexes has a positive correlation with both the rocky desertification degree and topographic fragmentation degree, respectively. This study could help policymakers better understand the local poverty by identifying the poor, locating them and describing their characteristics, so as to take differentiated poverty alleviation measures according to specific conditions of each county.

[Studies on light catalysis oxidation degradation of Malachite Green by Photo-Fenton reagent].

In the present paper, light catalysis oxidation degradation of Malachite Green by Photo-Fenton reagent was researched. The influences of various parameters such as the wave length-absorbency curve of Malachite Green, the concentration-absorbency curve of Malachite Green, initial pH, optimum dosage of Fe2+, optimum dosage of H2O2, different light sources and cation-exchange resin, on dye degradation were researched. Through numerous experiments, the optimum condition for Malachite Green degradation was given. Under the optimal conditions, the sun light can promote this reaction apparently and the reaction time can greatly be shortened too. After the cation-exchange resin was introduced into Fenton system, the activation of Fenton reagent for degradation reaction was enhanced to a great extent, and the degradation effect of Malachite Green was better.