Insights into drought stress response mechanism of tobacco during seed germination by integrated analysis of transcriptome and metabolome.

Drought stress inhibits seed germination, plant growth and development of tobacco, and seriously affects the yield and quality of tobacco leaves. However, the molecular mechanism underlying tobacco drought stress response remains largely unknown. In this study, integrated analysis of transcriptome and metabolome was performed on the germinated seeds of a cultivated variety K326 and its EMS mutagenic mutant M28 with great drought tolerance. The result showed that drought stress inhibited seed germination of the both varieties, while the germination rate of M28 was faster than that of K326 under drought stress. Besides, the levels of phytohormone ABA, GA19, and zeatin were increased by drought stress in M28. Five vital pathways were identified through integrated transcriptomic and metabolomic analysis, including zeatin biosynthesis, aspartate and glutamate synthesis, phenylamine metabolism, glutathione metabolism, and phenylpropanoid synthesis. Furthermore, 20 key metabolites in the above pathways were selected for further analysis of gene modular-trait relationship, and then four highly correlated modules were found. Then analysis of gene expression network was carried out of Top30 hub gene of these four modules, and 9 key candidate genes were identified, including HSP70s, XTH16s, APX, PHI-1, 14-3-3, SCP, PPO. In conclusion, our study uncovered some key drought-responsive pathways and genes of tobacco during seeds germination, providing new insights into the regulatory mechanisms of tobacco drought stress response.

Interfacial Co-O-Cu bonds prompt electrochemical nitrate reduction to ammonia in neutral electrolyte.

In this work, the formed interfacial Co-O-Cu bonds in Co-doped Cu(OH)2 (Co2-Cu(OH)2) sufficiently expose active sites and improve the reaction kinetics. As a result, the optimal Co2-Cu(OH)2 provides an amazing faradaic efficiency (91.6%), high selectivity (93.2%) and robust stability toward the NO3RR.

Timp1 Deletion Induces Anxiety-like Behavior in Mice.

The hippocampus is essential for learning and memory, but it also plays an important role in regulating emotional behavior, as hippocampal excitability and plasticity affect anxiety and fear. Brain synaptic plasticity may be regulated by tissue inhibitor of matrix metalloproteinase 1 (TIMP1), a known protein inhibitor of extracellular matrix (ECM), and the expression of TIMP1 in the hippocampus can be induced by neuronal excitation and various stimuli. However, the involvement of Timp1 in fear learning, anxiety, and hippocampal synaptic function remains to be established. Our study of Timp1 function in vivo revealed that Timp1 knockout mice exhibit anxiety-like behavior but normal fear learning. Electrophysiological results suggested that Timp1 knockout mice showed hyperactivity in the ventral CA1 region, but the basic synaptic transmission and plasticity were normal in the Schaffer collateral pathway. Taken together, our results suggest that deletion of Timp1 in vivo leads to the occurrence of anxiety behaviors, but that Timp1 is not crucial for fear learning.

The heat shock protein 20 gene family in large yellow croaker (Larimichthys crocea): Identification, phylogenetic relationships, expression analyses.

Large yellow croaker (Larimichthys crocea) is an economically important fish in China, but its aquaculture industry has been threatened by both biotic and abiotic stressors such as hypoxia and pathogens. In the current study, hsp20 genes were identified and analyzed systematically for the first time from the genome of large yellow croaker, and their roles in hypoxia response and Aeromonas hydrophila, Pseudomonas plecoglossicida infection were investigated. Herein, 11 hsp20 genes were identified and annotated, phylogenetic analysis and selection pressure analysis showed that the hsp20 genes were evolutionarily-constrained and their function was conserved among fishes. Besides, we observed the expression patterns of the hsp20 genes under hypoxia and two pathogens' stress. In brief, seven, four, seven genes responded to hypoxia stress, A. hydrophila infection and P. plecoglossicida challenge, respectively, which indicated that they were involved in hypoxia and disease responses. Furthermore, pathogen- and time-specific pattern was observed after A. hydrophila and P. plecoglossicida infection whereas tissue-specific pattern was observed after hypoxia exposure, revealing that hsp20 genes showed differential functions in response to hypoxia and immune stress. Taken together, these results provided preliminary information for future analysis of the roles of hsp20 genes in both biotic and abiotic stress response in fish.

Regulating charge distribution of Cu3PdN nanocrystals for nitrate electroreduction to ammonia.

As-synthesized Cu3PdN nanocrystals displayed high faradaic efficiency and selectivity for nitrate-to-ammonia conversion. The excellent performances can be attributed to the charge redistribution in Cu3PdN as a result of modulations of the electronic structures of Pd and Cu atoms, which altered the adsorption activation energy of the intermediates during the nitrate reduction reaction process.

Predictive genomic biomarkers of therapeutic effects in renal cell carcinoma.

BACKGROUND: In recent years, there have been great improvements in the therapy of renal cell carcinoma. Nevertheless, the therapeutic effect varies significantly from person to person. To discern the effective treatment for different populations, predictive molecular biomarkers in response to target, immunological, and combined therapies are widely studied. CONCLUSION: This review summarized those studies from three perspectives (SNPs, mutation, and expression level) and listed the relationship between biomarkers and therapeutic effect, highlighting the great potential of predictive molecular biomarkers in metastatic RCC therapy. However, due to a series of reasons, most of these findings require further validation.

Monoexponential, biexponential, stretched-exponential and kurtosis models of diffusion-weighted imaging in kidney assessment: comparison between patients with primary aldosteronism and healthy controls.

PURPOSE: This study used various diffusion-weighted imaging (DWI) models (including monoexponential, biexponential, stretched-exponential and kurtosis models) in renal magnetic resonance imaging (MRI) to compare whether there were differences in each diffusion parameter between patients with primary aldosteronism (PA) and healthy volunteers. MATERIALS AND METHODS: Twenty-two (female:male, 14:8; age, 48 ± 10 years) patients with PA and 22 age- and sex-matched healthy controls (HCs) underwent MRI examinations of the kidneys. The independent-sample t test or the Mann‒Whitney U test was used to detect differences in the diffusion metrics of the kidneys between the two groups. Univariable and multivariable linear regression were applied to analyze the correlations between diffusion parameters and the clinical indicators. RESULTS: The mean diffusivity (MD, p < 0.001) and radial diffusivity (Dr, p < 0.001) values in the medulla were lower in the PA group than in the HC group. The medullary fractional anisotropy (FA, p < 0.001) was higher than that of HCs. The FA (p < 0.001) and axial diffusivity (Da, p < 0.001) values in the cortex were lower in the PA group. The cortical α (anomalous exponent term, p = 0.016) was higher in the PA patients than in the HCs. Linear regression analysis showed that log(plasma aldosterone concentration) and the estimated glomerular filtration rate (eGFR) were correlated with medullary FA. CONCLUSION: The stretched-exponential model (cortical α) and the kurtosis model (FA, MD and Dr in the medulla and FA and Da in the cortex) showed significant differences between PA patients and healthy volunteers and may have potential for noninvasive renal assessment in PA patients.

Risk factors for in-stent restenosis in maintenance hemodialysis patients with central venous occlusive disease and biomechanical assessment of stents.

OBJECTIVES: To investigate the risk factors and biomechanical mechanisms of in-stent restenosis (ISR) in central venous occlusive disease (CVOD). PATIENTS AND METHODS: This retrospective study consecutively included 77 maintenance hemodialysis (MHD) patients with CVOD who received the first percutaneous transluminal angioplasty with stenting (PTS) due to symptomatic CVOD in a tertiary hospital. The mean age was 59.7 ± 14.0 years, and 51.9% of patients were male. The clinical characteristics, occurrence of ISR and patency rates were recorded. Finite element method was applied to assess the biomechanical properties of stents. RESULTS: Among 77 patients with a mean CVS score of 8.0 ± 2.8, 20.8%, 62.3%, and 16.9% of patients had the main vein of CVOD in the subclavian vein, brachiocephalic vein, and superior vena cava, respectively. A total of 72 (93.5%) patients received successful PTS treatment, for which the stents implanted were mainly Fluency covered stent (48.6%) and SMART bare stent (31.9%). During 15 (10-24)-months of follow-up, ISR occurred in 36.1% of the 72 patients. The primary and assisted primary patency rates at 6, 12, and 18 months were 78%, 56%, 42% and 95%, 90%, 87%, respectively. A prolonged dialysis vintage was an independent risk factor for ISR, yet the stent type or the main vein location was not correlated with ISR. Among three laser-engraving stents, the SMART stent was the best in terms of flexibility, stress, and strain on stents but worst in stress or strain on vessels. The Luminexx stent was the best in radial force and worst in stress or strain on stents. The Vici stent was the best in stress and strain on vessels and worst in radial force and flexibility. CONCLUSIONS: An unsatisfactory comprehensive biomechanical performance from configurations rooted in existing stents may account for the high incidence of ISR in CVOD.

Molecular Mechanism Based on Histopathology, Antioxidant System and Transcriptomic Profiles in Heat Stress Response in the Gills of Japanese Flounder.

As an economically important flatfish in Asia, Japanese flounder is threatened by continuously rising temperatures due to global warming. To understand the molecular responses of this species to temperature stress, adult Japanese flounder individuals were treated with two kinds of heat stress-a gradual temperature rise (GTR) and an abrupt temperature rise (ATR)-in aquaria under experimental conditions. Changes in histopathology, programmed cell death levels and the oxidative stress status of gills were investigated. Histopathology showed that the damage caused by ATR stress was more serious. TUNEL signals confirmed this result, showing more programmed cell death in the ATR group. In addition, reactive oxygen species (ROS) levels and the 8-O-hDG contents of both the GTR and ATR groups increased significantly, and the total superoxide dismutase (T-SOD) activities and total antioxidant capacity (T-AOC) levels decreased in the two stressed groups, which showed damage to antioxidant systems. Meanwhile, RNA-seq was utilized to illustrate the molecular mechanisms underyling gill damage. Compared to the control group of 18 °C, 507 differentially expressed genes (DEGs) were screened in the GTR group; 341 were up-regulated and 166 were down-regulated, and pathway enrichment analysis indicated that they were involved in regulation and adaptation, including chaperone and folding catalyst pathways, the mitogen-activated protein kinase signaling (MAPK) pathway and DNA replication protein pathways. After ATR stress, 1070 DEGs were identified, 627 were up-regulated and 423 were down-regulated, and most DEGs were involved in chaperone and folding catalyst and DNA-related pathways, such as DNA replication proteins and nucleotide excision repair. The annotation of DEGs showed the great importance of heat shock proteins (HSPs) in protecting Japanese flounder from heat stress injury; 12 hsp genes were found after GTR, while 5 hsp genes were found after ATR. In summary, our study records gill dysfunction after heat stress, with different response patterns observed in the two experimental designs; chaperones were activated to defend heat stress after GTR, while replication was almost abandoned due to the severe damage consequent on ATR stress.

Transcriptome dataset of sago palm in peat soil.

Sago palm (Metroxylon sagu Rottb.) is an important agricultural starch-producing palm that contributes to Malaysia's economics, especially in the State of Sarawak, Malaysian Borneo. In this palm tree, the central part of the plant storage-starch. Under normal condition, sago palm develop its trunk after 4-5 years being planted. However, sago palms planted on deep-peat soil failed to develop their trunk even after 17 years of being planted. This phenomenon is known as 'non-trunking', which eliminates the economic value of the palms. Numerous research has been done to address the phenomenon, but the molecular mechanisms of sago palm responding toward the responsible stresses are still lacking. Therefore, in this study, leaf samples were collected from trunking (normal) and non-trunking sago palms planted on peat soil for total RNA extraction, followed by next-generation sequencing using the BGISEQ-500 platform. The raw reads were cleaned, and de novo assembled using TRINITY software package. A total of 40.11 Gb bases were sequenced from the sago palm leaf samples. The assembled sequence produced 102,447 unigenes, with N50 score 1809 bp and GC ratio of 44.34%. The alignment of unigenes with seven functional databases (NR, NT, GO, KOG, KEGG, SwissProt and InterPro) resulted in the annotation of 65,523 (63.96%) unigenes. Functional annotation results in the detection of 46,335 coding DNA sequences by Transdecoder. A total of 30,039 simple-sequence repeats distributed on 21,676 unigenes were detected using Primer3 software, and 2355 transcription factor coding unigenes were predicted using getorf and hmmseach software. This work is registered under NCBI BioProject PRJNA781491. The raw RNA sequencing data are available in Sequence Read Archive (SRA) database with accession numbers SRX13165895, SRX13165896, SRX13165897, SRX13165898, SRX13165899, and SRX13165900. Gene expression and annotation information are accessible in public functional genomics data repository Gene Expression Omnibus (GEO) with accession number GSE189085.

Prognostic value of ground glass opacity on computed tomography in pathological stage I pulmonary adenocarcinoma: A meta-analysis.

BACKGROUND: The clinical role of ground glass opacity (GGO) on computed tomography (CT) in stage I pulmonary adenocarcinoma patients currently remains unclear. AIM: To explore the prognostic value of GGO on CT in lung adenocarcinoma patients who were pathologically diagnosed with tumor-node-metastasis stage I. METHODS: A comprehensive and systematic search was conducted through the PubMed, EMBASE and Web of Science databases up to April 3, 2021. The hazard ratio (HR) and corresponding 95% confidence interval (CI) were combined to assess the association between the presence of GGO and prognosis, representing overall survival and disease-free survival. Subgroup analysis based on the ratio of GGO was also conducted. STATA 12.0 software was used for statistical analysis. RESULTS: A total of 12 studies involving 4467 patients were included. The pooled results indicated that the GGO predicted favorable overall survival (HR = 0.44, 95%CI: 0.34-0.59, P < 0.001) and disease-free survival (HR = 0.35, 95%CI: 0.18-0.70, P = 0.003). Subgroup analysis based on the ratio of GGO further demonstrated that the proportion of GGO was a good prognostic indicator in pathological stage I pulmonary adenocarcinoma patients, and patients with a higher ratio of GGO showed better prognosis than patients with a lower GGO ratio did. CONCLUSION: This meta-analysis manifested that the presence of GGO on CT predicted favorable prognosis in tumor-node-metastasis stage I lung adenocarcinoma. Patients with a higher GGO ratio were more likely to have a better prognosis than patients with a lower GGO ratio.

Identification, evolution and expression analyses of mapk gene family in Japanese flounder (Paralichthys olivaceus) provide insight into its divergent functions on biotic and abiotic stresses response.

Mitogen-activated protein kinase (MAPK) are a series of serine/threonine protein kinases showing evolutionary conservation, which can be activated by many stimulus signals and then transfer them from cell membrane to nucleus. MAPKs regulate a variety of biological processes, such as apoptosis, hormone signaling and immune response. In this study, 14 putative mapk genes in Japanese flounder were identified, and their basic physical and chemical properties were characterized. Phylogenetic analysis showed that mapk genes were divided into three main subfamilies, including ERK, JNK and the p38 MAPK. Selection pressure analysis revealed they were evolutionarily-constrained and undergone strong purifying selection. Gene structure and conserved protein motif comparison suggested high levels of conservation in members of mapk gene family. The expression patterns were further investigated in each embryonic and larval development stages and different tissues. In addition, RNA-seq analyses after bacteria and temperature stresses suggested mapk genes had different expression patterns. Three mapk genes showed significant differences in response to E. tarda challenge and five were induced significantly after temperature stress, indicating their potential functions. This systematic analysis provided valuable information for further understanding of the regulation mechanism of mapk gene family under different stresses in Japanese flounder.

Topographic Anatomy of the Zygomatico-Orbital Artery: Implications for Improving the Safety of Temporal Augmentation.

BACKGROUND: Anatomical knowledge of the zygomatico-orbital artery and its most relevant clinical applications is essential for ensuring the safety of filler injection into the temporal region. The purpose of this study was to provide the precise position, detailed course, and relationship with surrounding structures of the zygomatico-orbital artery. METHODS: Fifty-eight patients who underwent head contrast-enhanced three-dimensional computed tomography and 10 fresh frozen cadavers were investigated. RESULTS: The zygomatico-orbital artery was identified in 93 percent of the samples in this work. Ninety-four percent of the zygomatico-orbital arteries derived directly from the superficial temporal artery, and the remaining arteries started from the frontal branch of the superficial temporal artery. According to the origin of the zygomatico-orbital artery, it was classified into type I and type II. Type I arteries were then classified into three subtypes. The trunk of the zygomatico-orbital artery was located between the deep temporal fascia and the superficial temporal fascia. Deep branches of the zygomatico-orbital artery pierced the superficial layer of the deep temporal fascia. The zygomatico-orbital artery originated from 11.3 mm in front of the midpoint of the apex of the tragus, and most of its trunks were located less than 20.0 mm above the zygomatic arch. The mean diameter of the zygomatico-orbital artery was 1.2 ± 0.2 mm. There were extensive anastomoses between the zygomatico-orbital artery and various periorbital arteries at the lateral orbital rim. CONCLUSION: The precise anatomical knowledge of the zygomatico-orbital artery described in this study could be helpful for cosmetic physicians for improving the safety of temporal augmentation.

Decreased neuronal synaptosome associated protein 29 contributes to poststroke cognitive impairment by disrupting presynaptic maintenance.

Background: Poststroke cognitive impairments are common in stroke survivors, and pose a high risk of incident dementia. However, the cause of these cognitive impairments is obscure and required an investigation. Methods: Oxygen-glucose deprivation (OGD) model and middle cerebral artery occlusion (MCAO) model were used to imitate in vitro or in vivo acute cerebral ischemia, respectively. The differentially expressed synaptosome associated protein 29 (SNAP29)-interacting proteins upon ischemia and reperfusion were analyzed with bioinformatics analysis and the results indicated that the changes of SNAP29 after acute ischemia were mainly involved in the synaptic functions. The outcomes of SNAP29 reduction were assessed with SNAP29 knockdown, which mimicked the distribution of SNAP29 along neuronal processes after acute ischemia. Using the whole-cell patch clamp recording method and transmission electron microscope, the pre-synaptic function and readily releasable pool (RRP) were observed after SNAP29 knock down. Using photogenetic manipulations and behavioral tests, the neuronal projection and cognitive functions of mice with SNAP29 knock down in hippocampus CA1 region were evaluated. Results: It was found that SNAP29 protein levels decreased in both in vitro and in vivo ischemic models. Further, the SNAP29 reduction wasn't associated with impaired autophagy flux and neuronal survival. When SNAP29 was knocked down in primary cortical neurons, the frequency of AMPARs-mediated mEPSCs, but not the amplitude, significantly decreased. Meanwhile, the mice with SNAP29 knockdown at CA1 region of hippocampus developed an impairment in hippocampus-mPFC (middle prefrontal cortex) circuit and behavioral dysfunctions. Moreover, the size of RRP at presynaptic sites was diminished. Conclusion: Since SNAP29 protein levels didn't significantly influence the neuronal survival and its decrease was sufficient to disturb the neural circuit via a presynaptic manner, the SNAP29-associated strategies may be an efficient target against poststroke synaptic dysfunction and cognitive deficits.

[Acute cerebral ischemia-induced down-regulation of Sirt3 protein expression contributes to neuronal injury via damaging mitochondrial function].

This study was aimed to determine the effect of acute cerebral ischemia on the protein expression level of silent mating type information regulator 2 homolog 3 (Sirt3) in the neurons and clarify the pathological role of Sirt3 in acute cerebral ischemia. The mice with middle cerebral artery occlusion (MCAO) and primary cultured rat hippocampal neurons with oxygen glucose deprivation (OGD) were used as acute cerebral ischemia models in vivo and in vitro, respectively. Sirt3 overexpression was induced in rat hippocampal neurons by lentivirus transfection. Western blot was utilized to measure the changes in Sirt3 protein expression level. CCK8 assay was used to detect cell viability. Immunofluorescent staining was used to detect mitochondrial function. Transmission electron microscope was used to detect mitochondrial autophagy. The results showed that, compared with the normoxia group, hippocampal neurons from OGD1 h/reoxygenation 2 h (R2 h) and OGD1 h/R12 h groups exhibited down-regulated Sirt3 protein expression levels. Compared with contralateral normal brain tissue, the ipsilateral penumbra region from MCAO1 h/reperfusion 24 h (R24 h) and MCAO1 h/R72 h groups exhibited down-regulated Sirt3 protein expression levels, while there was no significant difference between the Sirt3 protein levels on both sides of sham group. OGD1 h/R12 h treatment damaged mitochondrial function, activated mitochondrial autophagy and reduced cell viability in hippocampal neurons, whereas Sirt3 over-expression attenuated the above damage effects of OGD1 h/R12 h treatment. These results suggest that acute cerebral ischemia results in a decrease in Sirt3 protein level. Sirt3 overexpression can alleviate acute cerebral ischemia-induced neural injuries by improving the mitochondrial function. The current study sheds light on a novel strategy against neural injuries caused by acute cerebral ischemia.

Ischemia-induced upregulation of autophagy preludes dysfunctional lysosomal storage and associated synaptic impairments in neurons.

Macroautophagy/autophagy is vital for neuronal homeostasis and functions. Accumulating evidence suggest that autophagy is impaired during cerebral ischemia, contributing to neuronal dysfunction and neurodegeneration. However, the outcomes after transient modification in autophagy machinery are not fully understood. This study investigated the effects of ischemic stress on autophagy and synaptic structures using a rat model of oxygen-glucose deprivation (OGD) in hippocampal neurons and a mouse model of middle cerebral artery occlusion (MCAO). Upon acute ischemia, an initial autophagy modification occurred in an upregulation manner. Following, the number of lysosomes increased, as well as lysosomal volume, indicating dysfunctional lysosomal storage. These changes were prevented by inhibiting autophagy via 3-methyladenine (3-MA) treatment or ATG7 (autophagy related 7) knockdown, or were mimicked by rapamycin (RAPA), a known activator of autophagy. This suggests that dysfunctional lysosomal storage is associated with the early burst of autophagy. Dysfunctional lysosomal storage contributed to autophagy dysfunction because the basal level of MTOR-dependent lysosomal biogenesis in the reperfusion was not sufficient to clear undegraded cargoes after transient autophagy upregulation. Further investigation revealed that impairment of synaptic ultra-structures, accompanied by dysfunctional lysosomal storage, may result from a failure in dynamic turnover of synaptic proteins. This indicates a vital role of autophagy-lysosomal machinery in the maintenance of synaptic structures. This study supports previous evidence that dysfunctional lysosomal storage may occur following the upregulation of autophagy in neurons. Appropriate autophagosome-lysosomal functioning is vital for maintenance of neuronal synaptic function and impacts more than the few known synaptic proteins.Abbreviations: 3-MA: 3-methyladenine; ACTB: actin beta; AD: Alzheimer disease; ALR: autophagic lysosome reformation; ATG7: autophagy related 7; CTSB: cathepsin B; CTSD: cathepsin D; DAPI: 4',6-diamidino-2-phenylindole; DEGs: differentially expressed genes; DMEM: Dulbecco's modified Eagle's medium; DMSO: dimethyl sulfoxide; GO: Gene Ontology; HBSS: Hanks' balanced salt solution; HPCA: hippocalcin; i.c.v: intracerebroventricular; KEGG: kyoto encyclopedia of genes and genomes; LAMP1: lysosomal-associated membrane protein 1; MAP1LC3B/LC3: microtubule-associated protein 1 light chain 3 beta; LSDs: lysosomal storage disorders; MAP2: microtubule-associated protein 2; MCAO: middle cerebral artery occlusion; mCTSB: mature CTSB; mCTSD: mature CTSD; MOI: multiplicity of infection; MTOR: mechanistic target of rapamycin kinase; OGD/R: oxygen-glucose deprivation/reoxygenation; PBS: phosphate-buffered saline; PRKAA/AMPKα: protein kinase AMP-activated catalytic subunit alpha; proCTSD: pro-cathepsin D; RAPA: rapamycin; RNA-seq: RNA sequencing; RPS6KB/p70S6K: ribosomal protein S6 kinase; SDS-PAGE: sodium dodecyl sulfate-polyacrylamide gel electrophoresis; SIM: Structured Illumination Microscopy; SNAP25: synaptosomal-associated protein 25; SQSTM1/p62: sequestosome 1; SYN1: synapsin I; SYT1: synaptotagmin I; TBST: tris-buffered saline Tween-20; TEM: transmission electron microscopy; TFEB: transcription factor EB; tMCAO: transient middle cerebral artery occlusion; TTC: 2,3,5-triphenyltetrazolium chloride; TUBB3: tubulin, beta 3 class III.

Sirt1-ROS-TRAF6 Signaling-Induced Pyroptosis Contributes to Early Injury in Ischemic Mice.

Stroke is an acute cerebro-vascular disease with high incidence and poor prognosis, most commonly ischemic in nature. In recent years, increasing attention has been paid to inflammatory reactions as symptoms of a stroke. However, the role of inflammation in stroke and its underlying mechanisms require exploration. In this study, we evaluated the inflammatory reactions induced by acute ischemia and found that pyroptosis occurred after acute ischemia both in vivo and in vitro, as determined by interleukin-1ß, apoptosis-associated speck-like protein, and caspase-1. The early inflammation resulted in irreversible ischemic injury, indicating that it deserves thorough investigation. Meanwhile, acute ischemia decreased the Sirtuin 1 (Sirt1) protein levels, and increased the TRAF6 (TNF receptor associated factor 6) protein and reactive oxygen species (ROS) levels. In further exploration, both Sirt1 suppression and TRAF6 activation were found to contribute to this pyroptosis. Reduced Sirt1 levels were responsible for the production of ROS and increased TRAF6 protein levels after ischemic exposure. Moreover, N-acetyl-L-cysteine, an ROS scavenger, suppressed the TRAF6 accumulation induced by oxygen-glucose deprivation via suppression of ROS bursts. These phenomena indicate that Sirt1 is upstream of ROS, and ROS bursts result in increased TRAF6 levels. Further, the activation of Sirt1 during the period of ischemia reduced ischemia-induced injury after 72 h of reperfusion in mice with middle cerebral artery occlusion. In sum, these results indicate that pyroptosis-dependent machinery contributes to the neural injury during acute ischemia via the Sirt1-ROS-TRAF6 signaling pathway. We propose that inflammatory reactions occur soon after oxidative stress and are detrimental to neuronal survival; this provides a promising therapeutic target against ischemic injuries such as a stroke.

Roles of piwil1 gene in gonad development and gametogenesis in Japanese flounder, Paralichthys olivaceus.

PIWI family member piwil1, which associates with Piwi-interacting RNA (piRNA), is responsible in regulation of germ cell differentiation and maintenance of reproductive stem cells. In this study, we analyzed the piwil1 gene in Paralichthys olivaceus. Bioinformatics analysis and structure prediction showed that piwil1 had the conserved domains: PAZ domain and PIWI domain. Expression analysis during embryonic development implied that piwil1 gene was maternally inherited. The tissue distribution showed a sexually dimorphic gene expression pattern, with higher expression level in testis than ovary. In situ hybridization results demonstrated that piwil1 was predominantly distributed in oogonia, oocytes, sertoli cells and spermatocytes. A CpG island was predicted in the 5'-flanking region of piwil1 gene, and its methylation levels showed significant disparity between males and females, indicating that the sexually dimorphic expression of piwil1 gene might be regulated by methylation. Furthermore, we explored the distinct roles of human chorionic gonadotropin and 17α-methyltestosterone in regulating the expression of piwil1, and found that piwil1 was interacting with the HPG axis hormones. These results indicated that piwil1 might play a crucial role in gonadal development and gametogenesis in Paralichthys olivaceus.

Nasopharyngeal carcinoma segmentation based on enhanced convolutional neural networks using multi-modal metric learning.

Multi-modality examinations have been extensively applied in current clinical cancer management. Leveraging multi-modality medical images can be highly beneficial for automated tumor segmentation as they provide complementary information that could make the segmentation of tumors more accurate. This paper investigates CNN-based methods for automated nasopharyngeal carcinoma (NPC) segmentation using computed tomography (CT) and magnetic resonance (MR) images. Specially, a multi-modality convolutional neural network (M-CNN) is designed to jointly learn a multi-modal similarity metric and segmentation of paired CT-MR images. By jointly optimizing the similarity learning error and the segmentation error, the feature learning processes of both modalities are mutually guided. In doing so, the segmentation sub-networks are able to take advantage of the other modality's information. Considering that each modality possesses certain distinctive characteristics, we combine the higher-layer features extracted by a single-modality CNN (S-CNN) and M-CNN to form a combined CNN (C-CNN) for each modality, which is able to further utilize the complementary information of different modalities and improve the segmentation performance. The proposed M-CNN and C-CNN were evaluated on 90 CT-MR images of NPC patients. Experimental results demonstrate that our methods achieve improved segmentation performance compared to their counterparts without multi-modal information fusion and the existing CNN-based multi-modality segmentation methods.

The dual role of KCNQ/M channels upon OGD or OGD/R insults in cultured cortical neurons of mice: Timing is crucial in targeting M-channels against ischemic injur ies.

KCNQ/M potassium channels play a vital role in neuronal excitability; however, it is required to explore their pharmacological modulation on N-Methyl- d-aspartic acid receptors (NMDARs)-mediated glutamatergic transmission of neurons upon ischemic insults. In the current study, both presynaptic glutamatergic release and activities of NMDARs were measured by NMDAR-induced miniature excitatory postsynaptic currents (mEPSCs) in cultured cortical neurons of C57 mice undergoing oxygen and glucose deprivation (OGD) or OGD/reperfusion (OGD/R). The KCNQ/M-channel opener, retigabine (RTG), suppressed the overactivation of postsynaptic NMDARs induced by OGD and then NO transient; RTG also decreased OGD-induced neuronal death measured with MTT assay, suggesting the beneficial role of KCNQ/M-channels for the neurons exposed to ischemic insults. However, when the neurons exposed to the subsequent reperfusion, KCNQ/M-channels played a differential role from its protective effect. OGD/R increased presynaptic glutamatergic release, which was further augmented by RTG or decreased by KCNQ/M-channel blocker, XE991. Reactive oxygen species (ROS) were produced partly in a NO-dependent manner. In addition, XE991 decreased neuronal injuries upon reperfusion measured with DCF and PI staining. Meanwhile, the addition of RTG upon OGD or XE991 upon reperfusion can reverse OGD or OGD/R-reduced mitochondrial membrane potential. Our present study indicates the dual role of KCNQ/M-channels in OGD and OGD/R, which will decide the fate of neurons. Provided that activation of KCNQ/M-channels has differential effects on neuronal injuries during OGD or OGD/R, we propose that therapy targeting KCNQ/M-channels may be effective for ischemic injuries but the proper timing is so crucial for the corresponding treatment.