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This study was performed to evaluate the role of TSC2 (tuberous sclerosis complex 2) in the regulation of growth inhibition of fish exposed to chronic ammonia nitrogen. Firstly, the full-length cDNA of TSC2 in juvenile blunt snout bream, Megalobrama amblycephala was cloned using RACE technology (5’ and 3’ rapid amplification of cDNA ends). Then, TSC2 was analyzed by bioinformatics methods containing Seqman, BLAST, DNAMAN 8.0, EXPASY, and MEGA 6.0 program. Finally, the growth and gene expression level of AMKP-mTOR signaling pathway of blunt snout bream under ammonia nitrogen were investigated. The results showed that the full length of TSC2 was 6145 bp containing 5’- untranslated region of 179 bp, 3’-UTR of 992 bp and an open reading frame of 4974 bp encoding a peptide of 1657 amino acids. The estimated theoretical PI and MW (molecular weight) of TSC2 protein were 7.13 and 183016 Da, respectively. Meanwhile, TSC2 protein had three structural domains: DUF3384 domain (residues 5–303), Tuberin domain (residues 386–735) and Rap-GAP domain (residues 1409–1597). TSC2 from Megalobrama amblycephala gathered Pimephales promelas and Danio rerio clade belonging to Cyprinidae, and showed further distance to Ictalurus punctatus and Tachysurus fulvidraco. Final body weight and weight gain of fish under ammonia nitrogen stress significantly decreased (p < 0.05). Meanwhile, ammonia nitrogen stress significantly decreased Rheb, mTOR and S6K1 mRNA expression levels of blunt snout bream (p < 0.05). However, ammonia nitrogen stress significantly up-regulated the AMPKα1, AMPKα2, TSC2, 4E-BP2 mRNA expression levels (p < 0.05). Taken together, ammonia nitrogen stress inhibited protein synthesis of muscle via activating the AMKP-mTOR signaling pathway mediated by TSC2, which led to decreased body weight of blunt snout bream. The results obtained here will provide the theoretical foundation for completing the research of TSC2 functions involving protein synthesis in fish exposed to chronic ammonia nitrogen.

Abstract Background Accumulating evidence indicated that necroptosis plays an essential role in the pathogenesis of inflammatory bowel disease (IBD). The O-linked β-N-acetylglucosaminylation (O-GlcNAcylation) of necroptotic signal molecule receptor-interacting serine-threonine kinase 3 (RIPK3) was reported to exert a protective effect in gut inflammation. Our recent study suggested traditional Chinese herbal formula Wu-Mei-Wan (WMW) as an effective prescription in mouse colitis. However, the potential mechanisms are not fully understood. Considering the crucial role of necroptosis in the pathogenesis of IBD, therefore, this study was designed to explain whether the anti-colitis effect of WMW is mediated by modulating necroptosis and its related mechanisms. Methods The protective effects of WMW on colitis have been determined by detecting colitis mice body weight, disease activity index (DAI), survival rate and colon length. Colonic inflammation was examined by inflammatory cells infiltration and local cytokines levels. After then, we measured the levels of necroptosis and O-GlcNAcylation. C O-immunoprecipitation experiments were used to address whether elevated O-GlcNAcylation can inhibit necroptotic signal transduction in the treatment of WMW. Finally, the key enzymes in O-GlcNAcylation: O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) were examined and molecular docking analysis was used to determine effective natural compounds in the regulation on OGT and OGA activities. Results Our results showed that WMW significantly improved mice body weight, survival rate and colon length, decreased DAI in TNBS-induced colitis. WMW obviously alleviated colonic inflammatory responses with reduced macrophages, neutrophils infiltration and local IL-1β, IL-6, TNF-α and IFN-γ levels. It was found that WMW increased colonic O-GlcNAcylation level and inhibited the activation of RIPK1, RIPK3 and MLKL. Then, further experiments revealed that WMW enhanced OGT activity and suppressed OGA activity, thereby increasing RIPK3 O-GlcNAcylation and inhibiting the binding of RIPK3 and MLKL, which led to the inhibition of necroptosis. Additionally, docking analysis demonstrated that hesperidin, coptisine and ginsenoside Rb1 may exert a major role in the regulation on OGT and OGA activities by WMW. Conclusion Our work demonstrated that WMW can alleviate TNBS-induced colitis in mice by inhibiting necroptosis through increasing RIPK3 O-GlcNAcylation.

Abstract Background Extending service scope of primary care facilities (PCFs) has been widely concerned in China. However, no current data about association between service scope of PCFs with patient outcomes are available. This study aims to investigate association between service scope of PCFs and patient outcomes. Methods A multistage, stratified clustered sampling method was used to collect information about service scope of PCFs from rural Guizhou, China. Claim data of 299,633 inpatient cases covered by 64 PCFs were derived from local information system of New Rural Cooperation Medical Scheme. Service scope of PCFs was collected with self-administrated questionnaires. Primary outcomes were (1) level of inpatient institutions, (2) length of stay, (3) per capita total health cost, (4) per capita out-of-pocket cost, (5) reimbursement ratio, (6) 30-day readmission. A total of 64 PCFs were categorized into five groups per facility-level service scope scores. Generalized linear regression models, logistic regression model, and ordinal regression model were conducted to identify association between service scope of PCFs and patient outcomes. Results On average, the median service scope score of PCFs was 20, with wide variation across PCFs. After controlling for demographic and clinical characteristics, patients living in communities with PCFs of greatest service scope (Quintile V vs. I) tended to have smaller rates of admission by county-level hospitals (-6.2 % [-6.5 %, -5.9 %], city-level hospitals (-1.9 % [-2.0 %, -1.8 %]), and provincial hospitals (-2.1 % [-2.2 %, -2.0 %]), smaller rate of 30-day readmission (-0.5 % [-0.7 %, -0.2 %]), less total health cost (-201.8 [-257.9, -145.8]) and out-of-pocket cost (-210.2 [-237.2, -183.2]), and greater reimbursement ratio (2.3 % [1.9 %, 2.8 %]) than their counterparts from communities with PCFs of least service scope. Conclusions Service scope of PCFs varied a lot in rural Guizhou, China. Greater service scope was associated with a reduction in secondary and tertiary hospital admission, reduced total cost and out-of-pocket cost, and 30-day readmission and increased reimbursement ratio. These results raised concerns about access to care for patients discharged from hospitals, which suggests potential opportunities for cost savings and improvement of quality of care. However, further evidence is warranted to investigate whether extending service scope of PCFs is cost-effective and sustainable.

Phototherapy and multimodal synergistic phototherapy (including synergistic photothermal and photodynamic therapy as well as combined phototherapy and other therapies) are promising to achieve accurate diagnosis and efficient treatment for tumor, providing a novel opportunity to overcome cancer. Notably, various nanomaterials have made significant contributions to phototherapy through both improving therapeutic efficiency and reducing side effects. The most key factor affecting the performance of phototherapeutic nanomaterials is their microstructure which in principle determines their physicochemical properties and the resulting phototherapeutic efficiency. Vacancy defects ubiquitously existing in phototherapeutic nanomaterials have a great influence on their microstructure, and constructing and regulating vacancy defect in phototherapeutic nanomaterials is an essential and effective strategy for modulating their microstructure and improving their phototherapeutic efficacy. Thus, this inspires growing research interest in vacancy engineering strategies and vacancy-engineered nanomaterials for phototherapy. In this review, we summarize the understanding, construction, and application of vacancy defects in phototherapeutic nanomaterials. Starting from the perspective of defect chemistry and engineering, we also review the types, structural features, and properties of vacancy defects in phototherapeutic nanomaterials. Finally, we focus on the representative vacancy defective nanomaterials recently developed through vacancy engineering for phototherapy, and discuss the significant influence and role of vacancy defects on phototherapy and multimodal synergistic phototherapy. Therefore, we sincerely hope that this review can provide a profound understanding and inspiration for the design of advanced phototherapeutic nanomaterials, and significantly promote the development of the efficient therapies against tumor.

Hyperhomocysteinemia (HHcy) is an established and potent independent risk factor for degenerative diseases, including cardiovascular disease (CVD), Alzheimer disease, type II diabetes mellitus, and chronic kidney disease. HHcy has been shown to inhibit proliferation and promote inflammatory responses in endothelial cells (EC), and impair endothelial function, a hallmark for vascular injury. However, metabolic processes and molecular mechanisms mediating HHcy-induced endothelial injury remains to be elucidated. This study examined the effects of HHcy on the expression of microRNA (miRNA) and mRNA in human aortic EC treated with a pathophysiologically relevant concentration of homocysteine (Hcy 500 μM). We performed a set of extensive bioinformatics analyses to identify HHcy-altered metabolic and molecular processes. The global functional implications and molecular network were determined by Gene Set Enrichment Analysis (GSEA) followed by Cytoscape analysis. We identified 244 significantly differentially expressed (SDE) mRNA, their relevant functional pathways, and 45 SDE miRNA. HHcy-altered SDE inversely correlated miRNA-mRNA pairs (45 induced/14 reduced mRNA) were discovered and applied to network construction using an experimentally verified database. We established a hypothetical model to describe the biochemical and molecular network with these specified miRNA/mRNA axes, finding: 1) HHcy causes metabolic reprogramming by increasing glucose uptake and oxidation, by glycogen debranching and NAD+/CoA synthesis, and by stimulating mitochondrial reactive oxygen species production via NNT/IDH2 suppression-induced NAD+/NADP-NADPH/NADP+ metabolism disruption; 2) HHcy activates inflammatory responses by activating inflammasome-pyroptosis mainly through ↓miR193b→↑CASP-9 signaling and by inducing IL-1β and adhesion molecules through the ↓miR29c→↑NEDD9 and the ↓miR1256→↑ICAM-1 axes, as well as GPCR and interferon α/β signaling; 3) HHcy promotes cell degradation by the activation of lysosome autophagy and ubiquitin proteasome systems; 4) HHcy causes cell cycle arrest at G1/S and S/G2 transitions, suppresses spindle checkpoint complex and cytokinetic abscission, and suppresses proliferation through ↓miRNA335/↑VASH1 and other axes. These findings are in accordance with our previous studies and add a wealth of heretofore-unexplored molecular and metabolic mechanisms underlying HHcy-induced endothelial injury. This is the first study to consider the effects of HHcy on both global mRNA and miRNA expression changes for mechanism identification. Molecular axes and biochemical processes identified in this study are useful not only for the understanding of mechanisms underlying HHcy-induced endothelial injury, but also for discovering therapeutic targets for CVD in general.

Frequent itemsets mining with differential privacy refers to the problem of mining all frequent itemsets whose supports are above a given threshold in a given transactional dataset, with the constraint that the mined results should not break the privacy of any single transaction. Current solutions for this problem cannot well balance efficiency, privacy, and data utility over large-scale data. Toward this end, we propose an efficient, differential private frequent itemsets mining algorithm over large-scale data. Based on the ideas of sampling and transaction truncation using length constraints, our algorithm reduces the computation intensity, reduces mining sensitivity, and thus improves data utility given a fixed privacy budget. Experimental results show that our algorithm achieves better performance than prior approaches on multiple datasets.

Species-area relationships (SARs), also known as species-area curves, are fundamental scaling tools for biodiversity research. Sampling design and taxonomic groups affect the widely cited forms of species-area curves. However, the influence of sampling design and related environmental heterogeneity on SAR curves is rarely considered. Here, we investigated the SAR among different plant life forms (herbaceous plants, shrubs, and trees) in a 25.2-ha ForestGEO plot, the Wanglang Plot, in Sichuan, southwestern China, using a non-contiguous quadrat sampling method and power-law model. We compared the estimated parameters (the intercept c and the slope z) of the power-law models among different plant life forms, tested whether the SAR curve forms varied with sampling starting location, and assessed the effect of environmental heterogeneity accumulating with sampling area on curve variation. We found a wider range of variations in the SARs. The estimated c, z-values of power SAR were higher for the herbaceous plants than for the woody plants. A wider variation of SARs for the herbaceous plants than those for the woody plants. The selection of sampling starting location affected the SAR curve forms because of the roles of soil and topographic heterogeneity. We concluded that environmental heterogeneity regulates SAR curves sampled from different starting locations through spatial distribution of plant life forms. Thus, we recommend considering the design of sampling starting location when constructing SAR curves, especially in a heterogeneous habitat with unrandom distribution patterns of species. [ABSTRACT FROM AUTHOR]

Plant debris, including woody debris and litter, is an essential but frequently overlooked component of carbon (C) storage in forest ecosystems. Here, we examined the C storage of plant debris and its contribution to total ecosystem C storage in an age sequence of six larch (Larix gmelinii) forest stands (15, 36, 45, 54, 65, and 138 years old) in northeastern China. The plant debris C storage increased from 6.0±0.5 Mg⋅C⋅ha-1 in the 15-year-old stand to a maximum of 9.3±1.8 Mg⋅C⋅ha-1 in the 138-year-old stand. The C storage of woody debris increased during stand development in a sigmoidal pattern, increasing from 0.7 - 0.2 Mg C ha 1 in the 15-year-old stand to 4.7± 1.3 Mg⋅C⋅ha-1 in the 138-year-old stand. However, the C storage of litter (4.6-5.4 Mg⋅C⋅ha-1) did not vary with stand age in this larch chronosequence. In addition, the ratio of woody debris to live tree biomass C storage was relatively stable across stands (approximately 3.3%). These results highlight the importance of considering successional development and stand characteristics in assessing changes of plant debris and total ecosystem C storage in the larch forest ecosystem. [ABSTRACT FROM AUTHOR]

Eukaryotic cell membrane dynamics change in curvature during physiological and pathological processes. In the past ten years, a novel protein family, Fes/CIP4 homology-Bin/Amphiphysin/Rvs (F-BAR) domain proteins, has been identified to be the most important coordinators in membrane curvature regulation. The F-BAR domain family is a member of the Bin/Amphiphysin/Rvs (BAR) domain superfamily that is associated with dynamic changes in cell membrane. However, the molecular basis in membrane structure regulation and the biological functions of F-BAR protein are unclear. The pathophysiological role of F-BAR protein is unknown. This review summarizes the current understanding of structure and function in the BAR domain superfamily, classifies F-BAR family proteins into nine subfamilies based on domain structure, and characterizes F-BAR protein structure, domain interaction, and functional relevance. In general, F-BAR protein binds to cell membrane via F-BAR domain association with membrane phospholipids and initiates membrane curvature and scission via Src homology-3 (SH3) domain interaction with its partner proteins. This process causes membrane dynamic changes and leads to seven important cellular biological functions, which include endocytosis, phagocytosis, filopodium, lamellipodium, cytokinesis, adhesion, and podosome formation, via distinct signaling pathways determined by specific domain-binding partners. These cellular functions play important roles in many physiological and pathophysiological processes. We further summarize F-BAR protein expression and mutation changes observed in various diseases and developmental disorders. Considering the structure feature and functional implication of F-BAR proteins, we anticipate that F-BAR proteins modulate physiological and pathophysiological processes via transferring extracellular materials, regulating cell trafficking and mobility, presenting antigens, mediating extracellular matrix degradation, and transmitting signaling for cell proliferation. [ABSTRACT FROM AUTHOR]

Deficient angiogenesismay contribute to worsen the prognosis of myocardial ischemia, peripheral arterial disease, ischemic stroke, etc. Dyslipidemic and inflammatory environments attenuate endothelial cell (EC) proliferation and angiogenesis, worsening the prognosis of ischemia. Under these dyslipidemic and inflammatory environments, EC-caspase-1 becomes activated and induces inflammatory cell death that is defined as pyroptosis. However, the underlying mechanism that correlates caspase-1 activation with angiogenic impairment and the prognosis of ischemia remains poorly defined. By using flow cytometric analysis, enzyme and receptor inhibitors, and hind limb ischemia model incaspase-1 knock-out (KO)mice, we examined our novel hypothesis, i.e. inhibition of caspase-1 in ECs under dyslipidemic and inflammatory environments attenuates EC pyroptosis, improves EC survival mediated by vascular endothelial growth factor receptor 2 (VEGFR-2), angiogenesis, and the prognosis of ischemia. We have made the following findings. Proatherogenic lipids induce higher caspase-1 activation in larger sizes of human aortic endothelial cells (HAECs) than in smaller sizes of HAECs. Proatherogenic lipids increase pyroptosis significantly more in smaller sizes of HAECs than in larger sizes of the cells. VEGFR-2 inhibition increases caspase-1 activation in HAECs induced by lysophos-phatidylcholine treatment.Caspase-1 activation inhibits VEGFR-2 expression. Caspase-1 inhibition improves the tube formation of lysophosphatidylcholine-treatedHAECs. Finally, caspase-1 depletion improves angiogenesis and blood flow in mouse hind limb ischemic tissues. Our results have demonstrated for the first time that inhibition of proatherogenic caspase-1 activation in ECs improves angiogenesis and the prognosis of ischemia. [ABSTRACT FROM AUTHOR]