Genetic mechanisms underlying gray matter volume changes in patients with drug-naive first-episode schizophrenia.

Brain structural damage is a typical feature of schizophrenia. Investigating such disease phenotype in patients with drug-naive first-episode schizophrenia (DFSZ) may exclude the confounds of antipsychotics and illness chronicity. However, small sample sizes and marked clinical heterogeneity have precluded definitive identification of gray matter volume (GMV) changes in DFSZ as well as their underlying genetic mechanisms. Here, GMV changes in DFSZ were assessed using a neuroimaging meta-analysis of 19 original studies, including 605 patients and 637 controls. Gene expression data were derived from the Allen Human Brain Atlas and processed with a newly proposed standardized pipeline. Then, we used transcriptome-neuroimaging spatial correlations to identify genes associated with GMV changes in DFSZ, followed by a set of gene functional feature analyses. Meta-analysis revealed consistent GMV reduction in the right superior temporal gyrus, right insula and left inferior temporal gyrus in DFSZ. Moreover, we found that these GMV changes were spatially correlated with expression levels of 1,201 genes, which exhibited a wide range of functional features. Our findings may provide important insights into the genetic mechanisms underlying brain morphological abnormality in schizophrenia.

Unprecedented phytoplankton blooms in autumn/winter in the southern Bohai Sea (China) due to high Yellow River discharge: Implications of extreme rainfall events.

The occurrence of abnormal phytoplankton blooms is one of the significant changes in coastal ecosystems due to climate change. However, the underlying mechanism of such blooms remains poorly understood due to the complexity of the system. In this study, the data from numerous observations was used to elucidate the unprecedented phytoplankton blooms in the autumn and winter of 2021 in Laizhou Bay, a typical aquaculture bay in the southern Bohai Sea of China. The abundance of phytoplankton cells increased by more than tenfold in the southern waters compared to that in the same period from 2019 to 2020. The phytoplankton bloom was first observed in winter in the Bohai Sea, with the cell abundance in the southern bay exceeding 108 cells L-1 in December 2021. The diversity and evenness of phytoplankton communities decreased in the southern area. Cerataulina pelagica was the dominant algae, comprising 69 % of the total phytoplankton in October and 99 % in December. In autumn 2021, the largest flood of the Yellow River in recent decades occurred. This was attributed to extreme rainfall events within the river basin. The input of substantial riverine nutrients played a significant role in promoting phytoplankton blooms. Correlation analysis indicated the important cumulative impact of the Yellow River on phytoplankton blooms rather than a direct short-term effect. Numerical modeling results indicated that exceptionally high Yellow River discharge in autumn could significantly affect the entire bay from autumn to the following spring. This study may contribute to understanding the abnormal phytoplankton blooms in coastal waters and provide valuable insights for environmental management in river basins and coastal waters.

Brain function mediates the association between low vitamin D and neurocognitive status in female patients with major depressive disorder.

BACKGROUND: Vitamin D is engaged in various neural processes, with low vitamin D linked to depression and cognitive dysfunction. There are gender differences in depression and vitamin D level. However, the relationship between depression, gender, vitamin D, cognition, and brain function has yet to be determined. METHODS: One hundred and twenty-two patients with major depressive disorder (MDD) and 119 healthy controls underwent resting-state functional MRI and fractional amplitude of low-frequency fluctuations (fALFF) was calculated to assess brain function. Serum concentration of vitamin D (SCVD) and cognition (i.e. prospective memory and sustained attention) were also measured. RESULTS: We found a significant group-by-gender interaction effect on SCVD whereby MDD patients showed a reduction in SCVD relative to controls in females but not males. Concurrently, there was a female-specific association of SCVD with cognition and MDD-related fALFF alterations in widespread brain regions. Remarkably, MDD- and SCVD-related fALFF changes mediated the relation between SCVD and cognition in females. CONCLUSION: Apart from providing insights into the neural mechanisms by which low vitamin D contributes to cognitive impairment in MDD in a gender-dependent manner, these findings might have clinical implications for assignment of female patients with MDD and cognitive dysfunction to adjuvant vitamin D supplementation therapy, which may ultimately advance a precision approach to personalized antidepressant choice.

Role of brain-derived neurotrophic factor in endotoxaemia-induced acute lung injury.

Acute lung injury (ALI) or acute respiratory distress syndrome (ARDS), which is a pulmonary manifestation of a systemic reactive inflammatory syndrome, is a serious disease with high mortality, and sepsis is an important risk factor in the development of ALI. Brain-derived neurotrophic factor (BDNF) is a member of the nerve growth factor family. It plays an essential role in the regulation of the modification of synaptic efficacy and brain metabolic activity and enhances neuronal survival. However, the role and underlying mechanism of BDNF in sepsis-induced ALI remain unclear. Here, we sought to observe the expression of BDNF in the lung tissues of mice. C57BL/6J mice were divided randomly into two groups: saline (n = 4) and lipopolysaccharide (LPS) (n = 4). We found that BDNF expression was elevated in the lung tissues of septic mice. Furthermore, we found that BDNF colocalized with aquaporin 5, a marker for type I alveolar epithelial cells, by immunofluorescence staining. In addition, we also found that tropomyosin-related kinase B, the specific receptor of BDNF, colocalized with surfactant protein C, a marker for type II alveolar epithelial cells, by immunofluorescence staining. Finally, the present study indicated that BDNF may alleviate excessive LPS-induced autophagy in alveolar epithelial cells. Overall, we hypothesize that BDNF expression increases in the lung tissues of septic mice as a compensatory mechanism to ameliorate sepsis-induced ALI by inhibiting excessive alveolar epithelial cell autophagy.

Genetic Architecture Underlying Differential Resting-state Functional Connectivity of Subregions Within the Human Visual Cortex.

The human visual cortex is a heterogeneous entity that has multiple subregions showing substantial variability in their functions and connections. We aimed to identify genes associated with resting-state functional connectivity (rsFC) of visual subregions using transcriptome-neuroimaging spatial correlations in discovery and validation datasets. Results showed that rsFC of eight visual subregions were associated with expression measures of eight gene sets, which were specifically expressed in brain tissue and showed the strongest correlations with visual behavioral processes. Moreover, there was a significant divergence in these gene sets and their functional features between medial and lateral visual subregions. Relative to those associated with lateral subregions, more genes associated with medial subregions were found to be enriched for neuropsychiatric diseases and more diverse biological functions and pathways, and to be specifically expressed in multiple types of neurons and immune cells and during the middle and late stages of cortical development. In addition to shared behavioral processes, lateral subregion associated genes were uniquely correlated with high-order cognition. These findings of commonalities and differences in the identified rsFC-related genes and their functional features across visual subregions may improve our understanding of the functional heterogeneity of the visual cortex from the perspective of underlying genetic architecture.

PKD2/polycystin-2 inhibits LPS-induced acute lung injury in vitro and in vivo by activating autophagy.

Polycystin-2 (PC2), which is a transmembrane protein encoded by the PKD2 gene, plays an important role in kidney disease, but its role in lipopolysaccharide (LPS)-induced acute lung injury (ALI) is unclear. We overexpressed PKD2 in lung epithelial cells in vitro and in vivo and examined the role of PKD2 in the inflammatory response induced by LPS in vitro and in vivo. Overexpression of PKD2 significantly decreased production of the inflammatory factors TNF-α, IL-1ß, and IL-6 in LPS-treated lung epithelial cells. Moreover, pretreatment with 3-methyladenine (3-MA), an autophagy inhibitor, reversed the inhibitory effect of PKD2 overexpression on the secretion of inflammatory factors in LPS-treated lung epithelial cells. We further demonstrated that overexpression of PKD2 could inhibit LPS-induced downregulation of the LC3BII protein levels and upregulation of SQSTM1/P62 protein levels in lung epithelial cells. Moreover, we found that LPS-induced changes in the lung wet/dry (W/D) weight ratio and levels of the inflammatory cytokines TNF-α, IL-6 and IL-1ß in the lung tissue were significantly decreased in mice whose alveolar epithelial cells overexpressed PKD2. However, the protective effects of PKD2 overexpression against LPS-induced ALI were reversed by 3-MA pretreatment. Our study suggests that overexpression of PKD2 in the epithelium may alleviate LPS-induced ALI by activating autophagy.

Short-term changes in algal blooms and phytoplankton community after the passage of Super Typhoon Lekima in a temperate and inner sea (Bohai Sea) in China.

Extensive multi-species harmful algal blooms (HABs) were triggered by Super Typhoon Lekima in Laizhou Bay (Bohai Sea) in August 2019. After conducting two field cruises before and after the typhoon passage, we employed both high-performance liquid chromatography (HPLC)-pigment and microscopic methods to study the changes in the phytoplankton community and biomass. Following the passage of Lekima, the average surface salinity decreased, while dissolved inorganic nitrogen and dissolved silicate concentrations increased in the study area. The phytoplankton abundance and Chl a significantly increased after the typhoon event. Post-typhoon, the highest abundance values of Pseudo-nitzschia spp., Noctiluca scintillans, and Coscinodiscus spp. reached 106 cells/L and those of Bacillaria paxillifera, Ceratium spp., and Gymnodinium catenatum were in the order of 105 cells/L. HPLC-pigment CHEMTAX analysis showed that the biomass (Chl a) of dinoflagellates, diatoms, cryptophytes, chlorophytes, and haptophytes increased significantly after the typhoon. The increase in Chl a concentration was mainly attributable to large-sized phytoplankton, which are mostly diatoms and dinoflagellates. This study highlights that typhoons may cause HABs by introducing large amounts of freshwater and nutrients and change the phytoplankton community in a temperate and inner bay.

Numerical Study on Heat-Transfer Characteristics of Convection Melting in Metal Foam under Sinusoidal Temperature Boundary Conditions.

Convection melting in metal foam under sinusoidal temperature boundary conditions is numerically studied in the present study. A multiple-relaxation-time lattice Boltzmann method, in conjunction with the enthalpy approach, is constructed to model the melting process without iteration steps. The effects of the porosity, phase deviation, and periodicity parameter on the heat-transfer characteristics are investigated. For the cases considered in this work, it is found that the effects of the phase deviation and periodicity parameter on the melting rate are weak, but the melting front can be significantly affected by the sinusoidal temperature boundary conditions.

Microglial TLR4-induced TAK1 phosphorylation and NLRP3 activation mediates neuroinflammation and contributes to chronic morphine-induced antinociceptive tolerance.

BACKGROUND AND PURPOSE: The aim of this work was to investigate the role and signal transduction of toll-like receptor 4 (TLR4), TGF-ß-activated kinase 1 (TAK1) and nod-like receptor protein 3 (NLRP3) in microglial in the development of morphine-induced antinociceptive tolerance. METHODS: TLR4 and NLRP3 knockout mice and 5Z-7-oxozeaeno (a selective inhibitor against TAK1 activity) were used to observe their effect on the development of morphine tolerance. Intrathecal injections of morphine (0.75 mg/kg once daily for 7 days) were used to establish anti-nociceptive tolerance, which was measured by the tail-flick test. Spinal TLR4, TAK1, and NLRP3 expression levels and phosphorylation of TAK1 were evaluated by Western blotting and immunofluorescence. RESULTS: Repeated treatment with morphine increased total expression of spinal TLR4, TAK1, and NLRP3 and phosphorylation of TAK1 in wild-type mice. TLR4 knockout attenuated morphine-induced tolerance and inhibited the chronic morphine-induced increase in NLRP3 and phosphorylation of TAK1. Compared with controls, mice that received 5Z-7-oxozeaenol showed decreased development of morphine tolerance and inhibition on repeated morphine-induced increase of NLRP3 but not TLR4. NLRP3 knockout mice showed resistance to morphine-induced analgesic tolerance with no effect on chronic morphine-induced expression of TLR4 and TAK1. TLR4, TAK1, and NLRP3 were collectively co-localized together and with the microglia marker Iba1. CONCLUSIONS: Microglial TLR4 regulates TAK1 expression and phosphorylation and results in NLRP3 activation contributes to the development of morphine tolerance through regulating neuroinflammation. Targeting TLR4-TAK1-NLRP3 signaling to regulate neuro-inflammation will be alternative therapeutics and strategies for chronic morphine-induced antinociceptive tolerance.

Transferable Active Centers of Strongly Coupled MoS2@Sulfur and Molybdenum Co-doped g-C3N4 Heterostructure Electrocatalysts for Boosting Hydrogen Evolution Reaction in Both Acidic and Alkaline Media.

Designing an excellent acidic and alkaline general-purpose hydrogen evolution electrocatalyst plays an important role in promoting the development of the energy field. Here, a feasible strategy is reported to use the strongly coupled MoS2@sulfur and molybdenum co-doped g-C3N4 (MoS2@Mo-S-C3N4) heterostructure with transferable active centers for catalytic reactions in acidic and alkaline media. Research studies have shown that the unsaturated S site at the edge of MoS2 and the active N atom on the Mo-S-C3N4 substrate are, respectively, the active centers of acidic and alkaline hydrogen evolution reaction. Specifically, Mo-S-C3N4 is regarded as a synergistic catalyst for the active species MoS2 in acidic hydrogen evolution, while MoS2 acts as a co-catalyst when the alkaline active species are transferred to Mo-S-C3N4. The coordination of the electrons between the interfaces achieves a synergistic balance, which provides the optimal sites for the adsorption of the reactants. Such an electrocatalyst exhibits overpotentials of 193 and 290 mV at 10 mA cm-2 in 0.5 M H2SO4 and 1 M KOH, respectively, which was better than numerous previous reports. This research provides an outstanding avenue to realize multifunctional electrocatalysts.

Ursodeoxycholic acid protects against lung injury induced by fat embolism syndrome.

Acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) is a life-threatening disease with a high mortality rate, which was a common complication of fat embolism syndrome (FES). Ursodeoxycholic acid (UDCA) has been reported to exert potent anti-inflammatory effects under various conditions. In vivo, perinephric fat was injected via tail vein to establish a rat FES model, the anti-inflammatory effects of UDCA on FES-induced lung injury were investigated through histological examination, ELISA, qRT-PCR, Western blot and immunofluorescence. In vitro, human lung microvascular endothelial cells (HPMECs) were employed to understand the protective effects of UDCA. The extent of ALI/ARDS was evaluated and validated by reduced PaO2 /FiO2 ratios, increased lung wet/dry (W/D) ratios and impaired alveolar-capillary barrier, up-regulation of ALI-related proteins in lung tissues (including myeloperoxidase [MPO], vascular cell adhesion molecule 1 [VCAM-1], intercellular cell adhesion molecule-1 [ICAM-1]), elevated protein concentration and increased proinflammatory cytokines levels (TNF-α and IL-1ß) in bronchoalveolar lavage fluid (BALF). Pre-treatment with UDCA remarkably alleviated these pathologic and biochemical changes of FES-induced ALI/ARDS; our data demonstrated that pre-treatment with UDCA attenuated the pathologic and biochemical changes of FES-induced ARDS, which provided a possible preventive therapy for lung injury caused by FES.

Comparative Study on Soil Microbial Diversity and Structure Under Wastewater and Groundwater Irrigation Conditions.

Wastewater (WW) irrigation to agricultural soils is one of the most economical and effective water-saving strategies. The effects of WW irrigation on soil microbial communities have gained increasing focus as these effects are not well understood. In this study, the effects of WW and groundwater (GW) irrigation on microbial diversity and structure were compared using the high-throughput sequencing analysis of 16S rDNA amplicons. Soil samples irrigated by WW for several decades and maize soil (loamy) samples irrigated by GW were collected from Luancheng Town, Shijiazhuang City, China. Compared to the GW groups, WW groups exhibited non-significant soil bacterial community abundance at the 0-20 and 20-40 cm depths. WW irrigation significantly altered the bacterial community composition and structures compared to GW irrigation. The relative abundance of Proteobacteria and Firmicutes increased in WW irrigated soil, while Actinobacteria decreased. Moreover, 14 significantly abundant biomarkers from Proteobacteria and Firmicutes that corresponded with WW irrigation were identified. Additionally, WW irrigation enriched some KEGG pathways that corresponded with metabolism and human diseases. The physical and chemical properties of WW irrigated soil may shape the compositions and structures of soil bacterial communities. The findings of this study illuminated the effects of wastewater irrigation on microbial characteristics, which is important for estimating the effects of long-term wastewater irrigation on soil environmental health.

Vertical distribution and affecting factors of Escherichia coli over a 0-400 cm soil profile irrigated with sewage effluents in northern China.

Quantification and evaluation of the spatial distribution and the primary factors that affect Escherichia coli (E. coli) distribution in soils is important to assess soil pollution and potential contamination of groundwater. However, little information is available on distribution of E. coli in deep soil layers. To analyze the spatial distribution and factors affecting E. coli over a 0-400 cm soil profile, soil samples were collected from two land use type in the sewage irrigation fields. The primary factors dominating the spatial distribution of E. coli were quantified by the model of principal component analysis with multiple linear regression (PCA-MLR). The results indicated that the number of E. coli under cropland decreased greatly with soil depth. The average number of E. coli over the 0-400 cm profile under forestland was 49 × 104 colony-forming unit/g (cfu/g), which was significantly higher than that under cropland (20 × 104 cfu/g). For forestland and cropland, the average number of E. coli at depths of 300-400 cm decreased by 85% and 88%, respectively, compared to that at depths of 0-100 cm. The presence of E. coli at the depths of 300-400 cm was at high level (forestland: 3 × 104 cfu/g; cropland: 2 × 104 cfu/g) for the potential risks of shallow groundwater. The PCA-MLR model estimated that the factors of soil organism, soil salt and land type use contributed 28%, 29% and 43%, respectively, to the distribution of E. coli. According to the Monte Carlo simulation, the average number of E. coli over the 0-400 cm profile was 46 ± 17 × 104 cfu/g in the sewage irrigated area, and the interval distribution with a probability of 95% varied between 14 × 104 cfu/g and 78 × 104 cfu/g. The findings of this study are useful for understanding negative effects of sewage irrigation on pathogens in deep soil and are critical to assess the potential risks of groundwater pollution.

Ursodeoxycholic acid stimulates alveolar fluid clearance in LPS-induced pulmonary edema via ALX/cAMP/PI3K pathway.

This study aims to examine the impact of ursodeoxycholic acid (UDCA) on pulmonary edema and explore the underlying molecular mechanisms. The effects of UDCA on pulmonary edema were assessed through hematoxylin and eosin (H&E) staining, lung dry/wet (W/D) ratio, TNF-α/IL-1ß levels of bronchoalveolar lavage fluid (BALF), protein expression of epithelial sodium channel (ENaC), and Na+ /K+ -ATPase. Besides, the detailed mechanisms were explored in primary rat alveolar type (AT) II epithelial cells by determining the effects of BOC-2 (ALX [lipoxin A4 receptor] inhibitor), Rp-cAMP (cAMP inhibitor), LY294002 (PI3K inhibitor), and H89 (PKA inhibitor) on the therapeutic effects of UDCA against lipopolysaccharide (LPS)-induced changes. Histological examination suggested that LPS-induced lung injury was obviously attenuated by UDCA. BALF TNF-α/IL-1ß levels and lung W/D ratios were decreased by UDCA in LPS model rats. UDCA stimulated alveolar fluid clearance (AFC) though the upregulation of ENaC and Na+ /K+ -ATPase. BOC-2, Rp-cAMP, and LY294002 largely suppressed the therapeutic effects of UDCA. Significant attenuation of pulmonary edema and lung inflammation was revealed in LPS-challenged rats after the UDCA treatment. The therapeutic efficacy of UDCA against LPS was mainly achieved through the ALX/cAMP/PI3K pathway. Our results suggested that UDCA might be a potential drug for the treatment of pulmonary edema induced by LPS.

Post-operative delirium associated with metabolic alterations following hemi-arthroplasty in older patients.

BACKGROUND: post-operative delirium (POD) is a common complication in older patients, though a possible link between metabolic changes and POD development has yet to be investigated. METHODS: older patients with hip fracture who underwent hemi-arthroplasty were recruited, and delirious states were assessed for 3 days after surgery using the confusion assessment method-Chinese revision. Simultaneously, fasting blood samples were collected on the morning of surgery and on the first post-operative day. Ultimately, 244 older patients who met the inclusion and exclusion criteria were assessed. Blood samples from 60 patients with POD and 60 matched controls were analysed using metabolomics platforms. RESULTS: sixty patients (24.6%) developed POD. Principal component analysis scores plot and cross-validated scores plots from orthogonal partial least squares-discriminant analysis were implemented to visualise the differences in metabolites between the two groups before and after surgery (P < 0.05). Our data indicate that levels of ω3 and ω6 fatty acids were lower in the POD group than in the NPOD (non-POD) group both before and after surgery; tricarboxylic cycle intermediate levels were lower in the POD group than in the NPOD group, but glycolysis products were higher in the POD group than in the NPOD group after surgery. Furthermore, the branched-chain amino acid (BCAA)/aromatic amino acid ratio was lower in the POD group than in the NPOD group after surgery. CONCLUSIONS: metabolic abnormalities, including deficiencies in ω3 and ω6 fatty acids, perturbations in tricarboxylic cycle and oxidative stress and metabolic imbalances in BCAA and AAA might contribute to POD development.

The Na+/K+ ATPase Inhibitor Ouabain Attenuates Stemness and Chemoresistance of Osteosarcoma Cells.

BACKGROUND The purpose of this study was to explore the effects of the Na+/K+ ATPase inhibitor ouabain in regulating osteosarcoma (OS) cell stemness. MATERIAL AND METHODS Western blot, qPCR, sphere-forming analysis, DNA methylation analysis, and Ca²âº concentration detection were performed to evaluate the stem-like traits of cells and ouabain-induced effects and related mechanisms on OS cell stemness. Cell viability assessment was performed to evaluate the effect of ouabain on OS cell chemosensitivity. RESULTS Ouabain reduced the ALDH1 activity, the expression of critical stemness regulators, sphere size and number, and migration, invasion, and adhesion ability, but had little effects on cell viability. Additionally, the intracellular Ca²âº concentration and methylation level of the critical stemness regulators were higher in OS cells than in spheres formed by OS cells. Mechanistic studies revealed that ouabain leads to DNA methylation of stemness markers through increasing intracellular Ca²âº concentration. Notably, inhibition of Ca²âº channel or DNA methylation rescued the inhibition of ouabain on OS cell stemness. Additionally, ouabain enhances cisplatin sensitivity of OS cells, which is involved in Ca²âº channel and DNA methylation. CONCLUSIONS This work provides a potential compound for treating OS patients, especially OS patients with chemoresistance.

A2BAR activation attenuates acute lung injury by inhibiting alveolar epithelial cell apoptosis both in vivo and in vitro.

The epithelial barrier of the lung is destroyed during acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) due to the apoptosis of alveolar epithelial cells (AECs). Therefore, treatments that block AEC apoptosis might be a therapeutic strategy to ameliorate ALI. Based on recent evidence, A2B adenosine receptor (A2BAR) plays an important role in ALI in several different animal models, but its exact function in AECs has not been clarified. We investigated the role of A2BAR in AEC apoptosis in a mouse model of oleic acid (OA)-induced ALI and in hydrogen peroxide (H2O2)-induced AEC (A549 cells and MLE-12 cells) injury. Mice treated with BAY60-6583, a selective A2BAR agonist, showed lower AEC apoptosis rates than mice treated with OA. However, the role of BAY60-6583 in OA-induced ALI was attenuated by a specific blocker of A2BAR, PSB1115. A2BAR activation decreased H2O2-induced cell apoptosis in vitro, as characterized by the translocation of apoptotic proteins, the release of cytochrome c, and the activation of caspase-3 and poly (ADP ribose) polymerase 1 (PARP-1). In addition, apoptosis was required for the phosphorylation of ERK1/2, p38, and JNK. Importantly, compared with cells transfected with the A2BAR-siRNA, an ERK inhibitor or p38 inhibitor exhibited decreased apoptotic ratios and cleaved caspase-9 and cleaved PARP-1 levels, whereas the JNK inhibitor displayed increases in these parameters. In conclusion, A2BAR activation effectively attenuated OA-induced ALI by inhibiting AEC apoptosis and mitigated H2O2-induced AEC injury by suppressing the p38 and ERK1/2-mediated mitochondrial apoptosis pathway.

Rapamycin alleviated pulmonary injury induced by fat embolism syndrome in rats.

Fat embolism syndrome (FES) is a serious complication after trauma, surgery and fat emulsion input and can lead to serious pulmonary injury. Autophagy controls the cell survival and homeostasis by removing the mis-folded proteins and damaged organelles as well as intracellular pathogens through a lysosomal degradation pathway. Increasing research documented that autophagy was wildly involved in variety of human diseases and had huge therapeutic potential. However, the role and mechanism of autophagy in FES remains largely unknown. The rat model of FES was established by tail vein injection with fat and was assessed by Wet-to-Dry (W/D) ratio analysis, hematoxylin-eosin (HE) analysis, staining Oil red staining analysis and qPCR analysis. Western blots were employed to detect the expression of autophagy markers. The changes of pulmonary injury were observed after premedication of rapamycin (an autophagy activator). The alveolar structural damage, red free fat substances in the blood vessels of lung, increased the lung ratio, and the up-regulated MPO expression and activity were showed in the FES models. The expressions of autophagy markers were decreased and meanwhile, apoptosis markers were increased in the FES model. Rapamycin restored the expression of autophagy markers and inhibited the apoptosis and further, resulting in the improvement of the pulmonary injury. Thus, our study demonstrated that autophagy was inhibited and apoptosis was promoted in FES and further Rapamycin alleviated the pulmonary damage in FES via restoring the autophagy and inhibiting the apoptosis.

O-GlcNAcylation modulates PKA-CREB signaling in a manner specific to PKA catalytic subunit isoforms.

O-GlcNAcylation is a post-translational modification of proteins. Protein kinase A (PKA)-cAMP response element binding protein (CREB) signaling plays critical roles in multiple biological processes. Isoforms α and ß of PKA catalytic subunit (PKAc) and CREB are modified by O-GlcNAcylation. In the present study, we determined the role of O-GlcNAcylation in PKAc isoform-specific CREB signaling. We found that up-regulation of O-GlcNAcylation enhanced CREB phosphorylation, but suppressed CREB expression in exogenous PKAc isoform-unspecific manner. PKAc isoforms affected exogenous expression of OGT or OGA and protein O-GlcNAcylation differently. Up-regulation of O-GlcNAcylation did not significantly affect net PKAcα-CREB signaling, but enhanced PKAcß-CREB signaling. The role of O-GlcNAcylation in PKA-CREB signaling was desensitized by insulin treatment. This study suggests a role of O-GlcNAcylation in PKA-CREB signaling by affecting phosphorylation of CREB in a PKAc isoform-specific manner.

ITRAQ-Based Proteomics Analysis of Acute Lung Injury Induced by Oleic Acid in Mice.

BACKGROUND/AIMS: This study was conducted to investigate the relationship between differentially expressed proteins (DEPs) and the pathogenesis of oleic acid (OA)-induced acute lung injury (ALI) in mice. METHODS: Eight-week-old male C57BL/6 mice were injected with OA through the tail vein and sacrificed 6 hours after OA administration to identify protein expression levels in lung tissue using isobaric tags for relative and absolute quantification (iTRAQ) technology. Then, DEPs such as antithrombin III (AT III), 12-lipoxygenase (12-LO), dedicator of cytokinesis 2 (DOCK2), polycystin-2 and plasminogen were identified by western blotting. Subsequently, we focused on investigating the effect of AT III on endothelial integrity using siRNA interference technology. The levels of IL-6, IL-1ß, TNF-α and TGF-ß expression were detected using an enzyme-linked immunosorbent assay (ELISA). Alterations in the tight junction component ZO-1 and the phosphorylation of myosin light chain (pMLC) were determined by western blotting. The stress fiber F-actin were also detected by immunofluorescence staining. In addition, endothelial permeability was determined via a transwell permeability assay. RESULTS: A total of 5152 proteins were found to be expressed in lung tissues from the OA-treated and saline-treated mice. Among these proteins, 849 were differentially expressed between the two groups, including 545 upregulated and 304 downregulated proteins. After AT III knockdown, the levels of inflammatory factors and endothelial permeability were elevated, the expression of ZO-1 was decreased, and the expression of F-actin and pMLC was increased. All these results illustrated that AT III knockdown exaggerated the disruption of endothelial integrity mediated by OA. CONCLUSION: These findings using iTRAQ technology demonstrate, for the first time, differences in the lung tissue expression levels of proteins between OA-treated mice and saline-treated mice. This study reveals that 12-LO, DOCK2 and especially AT III may be candidate biomarkers for OA-induced acute lung injury.