Circadian Biology and the Neurovascular Unit.

Mammalian physiology and cellular function are subject to significant oscillations over the course of every 24-hour day. It is likely that these daily rhythms will affect function as well as mechanisms of disease in the central nervous system. In this review, we attempt to survey and synthesize emerging studies that investigate how circadian biology may influence the neurovascular unit. We examine how circadian clocks may operate in neural, glial, and vascular compartments, review how circadian mechanisms regulate cell-cell signaling, assess interactions with aging and vascular comorbidities, and finally ask whether and how circadian effects and disruptions in rhythms may influence the risk and progression of pathophysiology in cerebrovascular disease. Overcoming identified challenges and leveraging opportunities for future research might support the development of novel circadian-based treatments for stroke.

Face-Specific Activity in the Ventral Stream Visual Cortex Linked to Conscious Face Perception.

When presented with visual stimuli of face images, the ventral stream visual cortex of the human brain exhibits face-specific activity that is modulated by the physical properties of the input images. However, it is still unclear whether this activity relates to conscious face perception. We explored this issue by using the human intracranial electroencephalography technique. Our results showed that face-specific activity in the ventral stream visual cortex was significantly higher when the subjects subjectively saw faces than when they did not, even when face stimuli were presented in both conditions. In addition, the face-specific neural activity exhibited a more reliable neural response and increased posterior-anterior direction information transfer in the "seen" condition than the "unseen" condition. Furthermore, the face-specific neural activity was significantly correlated with performance. These findings support the view that face-specific activity in the ventral stream visual cortex is linked to conscious face perception.

Common Sequential Organization of Face Processing in the Human Brain and Convolutional Neural Networks.

Face processing includes two crucial processing levels - face detection and face recognition. However, it remains unclear how human brains organize the two processing levels sequentially. While some studies found that faces are recognized as fast as they are detected, others have reported that faces are detected first, followed by recognition. We discriminated the two processing levels on a fine time scale by combining human intracranial EEG (two females, three males, and three subjects without reported sex information) and representation similarity analysis. Our results demonstrate that the human brain exhibits a "detection-first, recognition-later" pattern during face processing. In addition, we used convolutional neural networks to test the hypothesis that the sequential organization of the two face processing levels in the brain reflects computational optimization. Our findings showed that the networks trained on face recognition also exhibited the "detection-first, recognition-later" pattern. Moreover, this sequential organization mechanism developed gradually during the training of the networks and was observed only for correctly predicted images. These findings collectively support the computational account as to why the brain organizes them in this way.

Defensive and Ion Conductive Surface Layer Enables High Rate and Durable O3-type NaNi1/3 Fe1/3 Mn1/3 O2 Sodium-Ion Battery Cathode.

Na-based layered transition metal oxides with an O3-type structure are considered promising cathodes for sodium-ion batteries. However, rapid capacity fading, and poor rate performance caused by serious structural changes and interfacial degradation hamper their use. In this study, a NaPO3 surface modified O3-type layered NaNi1/3 Fe1/3 Mn1/3 O2 cathode is synthesized, with improved high-voltage stability through protecting layer against acid attack, which is achieved by a solid-gas reaction between the cathode particles and gaseous P2 O5 . The NaPO3 nanolayer on the surface effectively stabilizes the crystal structure by inhibiting surface parasitic reactions and increasing the observed average voltage. Superior cyclic stability is exhibited by the surface-modified cathode (80.1% vs 63.6%) after 150 cycles at 1 C in the wide voltage range of 2.0 V-4.2 V (vs Na+ /Na). Moreover, benefiting from the inherent ionic conduction of NaPO3 , the surface-modified cathode presents excellent rate capability (103 mAh g-1  vs 60 mAh g-1 ) at 10 C. The outcome of this study demonstrates a practically relevant approach to develop high rate and durable sodium-ion battery technology.

Pain sensitivity and quality of life of patients with burning mouth syndrome: a preliminary study in a Chinese population.

BACKGROUND: Burning mouth syndrome (BMS) is an oral-facial pain disorder involving the central and peripheral nervous systems, but the evidence for altered pain sensitivity remains inconclusive. The aim of this study was to investigate pain sensitivity and oral health-related quality of life (OHRQoL) in patients with BMS and to assess the relationship between them. METHODS: Fifty Chinese patients with BMS (57.82 ± 11.2 years) and fifty age- and gender-matched healthy subjects (55.64 ± 10.1 years) participated in the study. The Pain Sensitivity Questionnaire (PSQ) was used to assess participants' pain sensitivity. The Oral Health Impact Profile (OHIP-14) was used to evaluate participants' OHRQoL. RESULTS: The PSQ total score (p = 0.009), the PSQ minor score (p = 0.003) and the OHIP-14 score (p<0.05) of patients with BMS were significantly higher than those of the healthy subjects. Simple linear regression showed that the PSQ minor score was significantly associated with the OHIP-14 score in patients with BMS (ß = 0.338, p = 0.016). CONCLUSION: Patients with BMS have higher pain sensitivity than healthy subjects. Reducing pain sensitivity might help to improve the quality of life of patients with BMS.

Research on boundary optimization of adjacent mining areas in open pit coal mine based on calculation of sectional stripping ratio.

To address the problem of excessive local secondary stripping between adjacent mining areas in open pit mines caused by internal row raising, a multivariate function was fitted to the model of the main mining seam of Zhundong open pit coal mine in Xinjiang, and the different locations of the end gang of the second mining area were divided into multiple sections at certain step sizes and calculated by integration, resulting in stripping ratios for each section, which were fitted to a stripping ratio curve. The optimal location of the mining area boundary was found to be 55 m westward offset from the mining area boundary in the inner row of the raised section, and numerical simulations based on the strength reduction method were applied to analyse the slope stability of the end gang at this location. The results of the study show that the analysed slope meets the stability requirements and the optimised new boundary avoids the stripping of approximately 65,837,376 m3 of economically unreasonable section.

Enhancing Wheat Gluten Content and Processing Quality: An Analysis of Drip Irrigation Nitrogen Frequency.

Drip irrigation is a water-saving and fertilizer-saving application technology used in recent years, with which the frequency of drip irrigation nitrogen application has not yet been determined. In order to investigate the effects of different drip irrigation nitrogen application frequencies on the processing quality of medium-gluten wheat (Jimai22) and strong-gluten wheat (Jimai20 and Shiluan02-1), a two-year field experiment was carried out. Two frequencies of water and N application were set under the same conditions of total N application (210 kg·ha-1) and total irrigation (120 mm): DIF4, consisting of four equal applications of water and N (each of 30 kg·ha-1 of N application and 30 mm of irrigation) and DIF2, consisting of two equal applications of water and N (each of 60 kg·ha-1 of N application and 60 mm of irrigation). The results showed that IF4 significantly increased protein content by 2-8.6%, wet gluten content by 4.5-22.1%, and hardness value (p > 0.05), and PC2 was considered as a protein factor; the sedimentation value was highly significantly correlated with most of the parameters of the flour stretch (p < 0.01). DIF4 improved the stretching quality, and the flour quality of Jima22 was decreased, the flour quality of strong-gluten wheats Jimai20 and Shiluan02-1 was improved, and PC1 was considered to be the dough factor. In conclusion, although the frequency of nitrogen application by drip irrigation increased the protein factor and improved the tensile quality, the flour quality was not necessarily enhanced.

O-GlcNAcylation is essential for therapeutic mitochondrial transplantation.

BACKGROUND: Transplantation of mitochondria is increasingly explored as a novel therapy in central nervous system (CNS) injury and disease. However, there are limitations in safety and efficacy because mitochondria are vulnerable in extracellular environments and damaged mitochondria can induce unfavorable danger signals. METHODS: Mitochondrial O-GlcNAc-modification was amplified by recombinant O-GlcNAc transferase (OGT) and UDP-GlcNAc. O-GlcNAcylated mitochondrial proteins were identified by mass spectrometry and the antiglycation ability of O-GlcNAcylated DJ1 was determined by loss-of-function via mutagenesis. Therapeutic efficacy of O-GlcNAcylated mitochondria was assessed in a mouse model of transient focal cerebral ischemia-reperfusion. To explore translational potential, we evaluated O-GlcNAcylated DJ1 in CSF collected from patients with subarachnoid hemorrhagic stroke (SAH). RESULTS: We show that isolated mitochondria are susceptible to advanced glycation end product (AGE) modification, and these glycated mitochondria induce the receptor for advanced glycation end product (RAGE)-mediated autophagy and oxidative stress when transferred into neurons. However, modifying mitochondria with O-GlcNAcylation counteracts glycation, diminishes RAGE-mediated effects, and improves viability of mitochondria recipient neurons. In a mouse model of stroke, treatment with extracellular mitochondria modified by O-GlcNAcylation reduces neuronal injury and improves neurologic deficits. In cerebrospinal fluid (CSF) samples from SAH patients, levels of O-GlcNAcylation in extracellular mitochondria correlate with better clinical outcomes. CONCLUSIONS: These findings suggest that AGE-modification in extracellular mitochondria may induce danger signals, but O-GlcNAcylation can prevent glycation and improve the therapeutic efficacy of transplanted mitochondria in the CNS.

Mitochondria are the part of a cell that generate most of its energy to perform its functions. In injury or disease, mitochondrial function can become disrupted. Transplantation of healthy mitochondria is being explored as a potential therapy to replace damaged mitochondria and restore normal cellular function. However, this approach is difficult to perform because mitochondria are not able to maintain their healthy state outside of cells. Here, we show that one of the reasons for this is due to a molecular process called advanced glycation end product modification. We show that simple modification of mitochondria with a sugar prevents this process and helps to improve the success of therapeutic mitochondrial transplantation in cells and in a mouse model of stroke. Our findings may help to guide future efforts to develop therapies based on mitochondrial transplantation.

Number of positive lymph nodes is superior to neck stage of the 8th AJCC in predicting prognosis in parotid mucoepidermoid carcinoma.

BACKGROUND: To clarify the impact of the number of positive lymph nodes (LNs) on the prognosis of parotid mucoepidermoid carcinoma (MEC). METHODS: Patients who underwent neck dissection for parotid MEC were retrospectively enrolled. The primary outcome variable was overall survival (OS). Associations between OS and LN factors, including the AJCC N stage, intraparotid LN metastasis, number of positive LNs, LN size, and extranodal extension (ENE), were evaluated using Cox proportional hazard regression analyses. RESULTS: A total of 720 patients were included with a mean age of 56 ± 16 years. There was no additional survival compromise until two positive LNs were presented. After adjusting for the number of positive LNs, intraparotid LN metastasis, ENE, and LN size were not related to prognosis. Our proposed N stage based on the number of metastatic LNs (0/1 vs. 2-4 vs. 5+) showed a superior C-index to the AJCC N stage in OS prediction. CONCLUSION: Quantitative LN burden was an important determinant of prognosis, and the proposed N stage provided better OS stratification than the AJCC N stage.

Single cell profiling of female breast fibroadenoma reveals distinct epithelial cell compositions and therapeutic targets.

Fibroadenomas (FAs) are the most common breast tumors in women. No pharmacological agents are currently approved for FA intervention owing to its unclear mechanisms and a shortage of reproducible human models. Here, using single-cell RNA sequencing of human FAs and normal breast tissues, we observe distinct cellular composition and epithelial structural changes in FAs. Interestingly, epithelial cells exhibit hormone-responsive functional signatures and synchronous activation of estrogen-sensitive and hormone-resistant mechanisms (ERBB2, BCL2 and CCND1 pathways). We develop a human expandable FA organoid system and observe that most organoids seem to be resistant to tamoxifen. Individualized combinations of tamoxifen with ERBB2, BCL2 or CCND1 inhibitors could significantly suppress the viability of tamoxifen-resistant organoids. Thus, our study presents an overview of human FA at single-cell resolution that outlines the structural and functional differences between FA and normal breast epithelium and, in particular, provides a potential therapeutic strategy for breast FAs.

A Minimally-Invasive Method for Serial Cerebrospinal Fluid Collection and Injection in Rodents with High Survival Rates.

Cerebrospinal fluid (CSF) is an important sample source for diagnosing diseases in the central nervous system (CNS), but collecting and injecting CSF in small animals is technically challenging and often results in high mortality rates. Here, we present a cost-effective and efficient method for accessing the CSF in live rodents for fluid collection and infusion purposes. The key element of this protocol is a metal needle tool bent at a unique angle and length, allowing the successful access of the CSF through the foramen magnum. With this method, we can collect 5-10 µL of the CSF from mice and 70-100 µL from rats for downstream analyses, including mass spectrometry. Moreover, our minimally-invasive procedure enables iterative CSF collection from the same animal every few days, representing a significant improvement over prior protocols. Additionally, our method can be used to inject solutions into mice cisterna magna with high success rates and high postoperative recovery rates. In summary, we provide an efficient and minimally-invasive protocol for collecting and infusing reagents into the CSF in live rodents. We envision this protocol will facilitate biomarker discovery and drug development for diseases in the central nervous system.

Molecular engineering of fluorescence probe for real-time non-destructive visual screening of meat freshness.

Spoiled meat poses a great challenge to food security and human health, which should be addressed by the early monitoring and warning of the meat freshness. We herein exploited a molecular engineering strategy to construct a set of fluorescence probes (PTPY, PTAC, and PTCN) with phenothiazine as fluorophore and cyanovinyl as recognition site for the facile and efficient monitoring of meat freshness. These probes produce an obvious fluorescence color transition from dark red to bright cyan in response to cadaverine (Cad) through the nucleophilic addition/elimination reaction. The sensing performances were elaborately improved to achieve quick response (16 s), low detection limit (LOD = 3.9 nM), and high contrast fluorescence color change by enhancing the electron-withdrawing strength of cyanovinyl moiety. Furthermore, PTCN test strips were fabricated for portable and naked-eye detection of Cad vapor with fluorescence color change from crimson to cyan, and accurate determination of Cad vapor level with RGB color (red, green, blue) mode analysis. The test strips were employed to detect the freshness of real beef samples, and demonstrated a good capability of non-destructive, non-contact and visual screening meat freshness on site.

Clinical parameters predictors of malignant transformation of recurrent parotid pleomorphic adenoma.

Malignant transformation (MT) in recurrent parotid pleomorphic adenomas (PAs) is rare; therefore its occurrence lacks reliable predictive factors. Our goal was to clarify the predictors for MT of recurrent parotid PAs based on preoperative clinical parameters. Patients with a clinical diagnosis of recurrent parotid PA were retrospectively enrolled. The association between clinicopathologic variables and MT of PA was assessed using univariate and multivariate analyses. MT occurred in 11.8% of the 467 patients. In univariate analysis, three or more previous recurrences, newly developed facial nerve paralysis, difficulty in mouth opening, tumors with the largest tumor diameter ≥ 2.4 cm, and abnormal neck lymph node enlargement were associated with MT occurrence. Further, multivariate analysis showed that three or more previous recurrences, newly developed facial nerve paralysis, difficulty in mouth opening, and abnormal neck lymph node enlargement were independently related to MT. MT of recurrent PA was not uncommon. Clinical signs of malignancy included newly developed facial nerve paralysis, difficulty in mouth opening, three or more previous recurrences, and abnormal neck lymph node enlargement.

FUT8-AS1/miR-944/Fused in Sarcoma/Transcription Factor 4 Feedback Loop Participates in the Development of Oral Squamous Cell Carcinoma through Activation of Wnt/ß-Catenin Signaling Pathway.

As a common type of head and neck squamous cell carcinoma, oral squamous cell carcinoma (OSCC) is a lethal and deforming disease. Long noncoding RNAs have emerged as critical modulators in different malignancies. However, the role of fucosyltransferase 8 antisense RNA 1 (FUT8-AS1) in OSCC still remains elusive. In this study, quantitative RT-PCR and Western blot were used for the measurement of RNAs and proteins. Mechanism assays explored the putative correlation among genes. In vitro assays evaluated the changes in OSCC cell malignant phenotype, whereas in vivo assays highlighted the influence of FUT8-AS1 on tumor growth. FUT8-AS1, aberrantly up-regulated in OSCC tissues and cells, could exacerbate OSCC cell malignant behaviors. The cancerogenic property of FUT8-AS1 in OSCC was further confirmed via animal experiments. Furthermore, FUT8-AS1 enhanced the expression of transcription factor 4 (TCF4) via sponging miR-944 and recruiting fused in sarcoma (FUS), thus affecting OSCC cell biological behaviors via modulation on Wnt/ß-catenin signaling activity. In addition, TCF4 was validated as the transcriptional activator of FUT8-AS1. To conclude, TCF4-mediated FUT8-AS1 could exacerbate OSCC cell malignant behaviors and facilitate tumor growth via modulation on miR-944/FUS/TCF4.

Effects of microplastics on the water characteristic curve of soils with different textures.

Microplastic (MP) pollution in the soil severely damages the soil structure and affects the soil water-holding property, thereby affecting the soil water characteristic curve (SWCC). After polyethylene MP (PE-MP) addition at three concentrations (0.5%, 1%, and 2%) under three particle sizes (150 µm, 550 µm, and 950 µm) and two soil textures (sandy soil and loamy soil), SWCCs were measured and fitted with the van Genuchten model. The soil pore structure characteristics were obtained based on CT scanning combined with soil pore three-dimensional reconstruction to quantitatively analyze the relationships between MP properties and soil structure and the SWCC. Low concentrations (0.5%) of PE-MPs did not significantly affect the soil water content, while the accumulation of PE-MPs at a high concentration (2%) strongly affected the soil water-holding property, with small PE-MPs (150 µm) exerting significantly positive effects on the water-holding capacity of loamy soil and 950-µm MPs reducing the soil water content more strongly in sandy soil. The contributions of MP properties and soil textures to the SWCCs differed, and the impact of soil texture on the SWCCs was significantly higher than those of MP concentrations and particle sizes. Differences in MP occurrence characteristics and soil textures also led to variations in the fitted hydraulic parameters of the SWCCs. The addition of 2% 150-µm PE-MPs to loamy soil increased the soil porosity and surface area, while the addition of a higher concentration of large PE-MPs (2%, 950 µm) to sandy soil reduced soil porosity and circularity. This is related to the addition of a large number of small MPs, which may adsorb and bind many smaller soil particles to form larger, water-stable agglomerated structures, while the addition of high concentrations of large MPs in sandy soils may be related to the destruction of the original capillary pore structure of sandy soils and the weakening of soil capillarity. This study provides a theoretical basis for agroecological risk assessments.

Annexin A2 promotes angiogenesis after ischemic stroke via annexin A2 receptor - AKT/ERK pathways.

Promoting angiogenesis to restore circulation to the ischemic tissue is still an important therapeutic target in stroke. Our group and others previously reported that the Ca2+-regulated, phospholipid-and membrane-binding protein-Annexin A2 (ANXA2) functions in cerebrovascular integrity and retinal neoangiogenesis. Here, we hypothesized that ANXA2 may regulate angiogenesis after stroke, ultimately improve neurological outcomes. Compared with wild type (WT) mice, the density of microvessels in brain and the number of new vessels sprouting from aortic ring were significantly increased in Anxa2 knock-in (Anxa2 KI) mice. After focal cerebral ischemia, proliferation of brain endothelial cells in Anxa2 KI mice was significantly elevated at 7 days post-stroke, which further improved behavioral recovery. To assess the pro-angiogenic mechanisms of ANXA2, we used brain endothelial cells cultures to investigate its effects on cell tube-numbers and migration. Recombinant ANXA2 increased tube-numbers and migration of brain endothelial cells either under normal condition or after oxygen glucose deprivation (OGD) injury. Co-immunoprecipitation experiments demonstrated that these protective effects of recombinant ANXA2 were regulated by interaction with ANXA2 receptor (A2R), a protein found in cancer cells that can interact with ANXA2 to promote cell migration and proliferation, and the ability of ANXA2-A2R to activate AKT/ERK pathways. Inhibition of AKT/ERK diminished recombinant ANXA2-induced angiogenesis in vitro. Taken together, our study indicates that ANXA2 might be involved in angiogenesis after ischemic stroke. Further investigation of ANXA2-A2R will provide a new therapeutic target for stroke.

Endothelial cells regulate astrocyte to neural progenitor cell trans-differentiation in a mouse model of stroke.

The concept of the neurovascular unit emphasizes the importance of cell-cell signaling between neural, glial, and vascular compartments. In neurogenesis, for example, brain endothelial cells play a key role by supplying trophic support to neural progenitors. Here, we describe a surprising phenomenon where brain endothelial cells may release trans-differentiation signals that convert astrocytes into neural progenitor cells in male mice after stroke. After oxygen-glucose deprivation, brain endothelial cells release microvesicles containing pro-neural factor Ascl1 that enter into astrocytes to induce their trans-differentiation into neural progenitors. In mouse models of focal cerebral ischemia, Ascl1 is upregulated in endothelium prior to astrocytic conversion into neural progenitor cells. Injecting brain endothelial-derived microvesicles amplifies the process of astrocyte trans-differentiation. Endothelial-specific overexpression of Ascl1 increases the local conversion of astrocytes into neural progenitors and improves behavioral recovery. Our findings describe an unexpected vascular-regulated mechanism of neuroplasticity that may open up therapeutic opportunities for improving outcomes after stroke.

Diurnal Differences in Immune Response in Brain, Blood and Spleen After Focal Cerebral Ischemia in Mice.

BACKGROUND: The immune response to acute cerebral ischemia is a major factor in stroke pathobiology. Circadian biology modulates some aspects of immune response. The goal of this study is to compare key parameters of immune response during the active/awake phase versus inactive/sleep phase in a mouse model of transient focal cerebral ischemia. METHODS: Mice were housed in normal or reversed light cycle rooms for 3 weeks, and then they were blindly subjected to transient focal cerebral ischemia. Flow cytometry was used to examine immune responses in blood, spleen, and brain at 3 days after ischemic onset. RESULTS: In blood, there were higher levels of circulating T cells in mice subjected to focal ischemia during zeitgeber time (ZT)1-3 (inactive or sleep phase) versus ZT13-15 mice (active or awake phase). In the spleen, organ weight and immune cell numbers were lower in ZT1-3 versus ZT13-15 mice. Consistent with these results, there was an increased infiltration of activated T cells into brain at ZT1-3 compared with ZT13-15. CONCLUSIONS: This proof-of-concept study indicates that there are significant diurnal effects on the immune response after focal cerebral ischemia in mice. Hence, therapeutic strategies focused on immune targets should be reassessed to account for the effects of diurnal rhythms and circadian biology in nocturnal rodent models of stroke.

A novel cuproptosis-related LncRNA signature: Prognostic and therapeutic value for acute myeloid leukemia.

Background: Cuproptosis is a type of programmed cell death that is involved in multiple physiological and pathological processes, including cancer. We constructed a prognostic cuproptosis-related long non-coding RNA (lncRNA) signature for acute myeloid leukemia (AML). Methods: RNA-seq and clinical data for AML patients were acquired from The Cancer Genome Atlas (TCGA) database. The cuproptosis-related prognostic lncRNAs were identified by co-expression and univariate Cox regression analysis. The least absolute shrinkage and selection operator (LASSO) was performed to construct a cuproptosis-related lncRNA signature, after which the AML patients were classified into two risk groups based on the risk model. Kaplan-Meier, ROC, univariate and multivariate Cox regression, nomogram, and calibration curves analyses were used to evaluate the prognostic value of the model. Then, expression levels of the lncRNAs in the signature were investigated in AML samples by quantitative polymerase chain reaction (qPCR). KEGG functional analysis, single-sample GSEA (ssGSEA), and the ESTIMATE algorithm were used to analyze the mechanisms and immune status between the different risk groups. The sensitivities for potential therapeutic drugs for AML were also investigated. Results: Five hundred and three lncRNAs related to 19 CRGs in AML samples from the TCGA database were obtained, and 21 differentially expressed lncRNAs were identified based on the 2-year overall survival (OS) outcomes of AML patients. A 4-cuproptosis-related lncRNA signature for survival was constructed by LASSO Cox regression. High-risk AML patients exhibited worse outcomes. Univariate and multivariate Cox regression analyses demonstrated the independent prognostic value of the model. ROC, nomogram, and calibration curves analyses revealed the predictive power of the signature. KEGG pathway and ssGSEA analyses showed that the high-risk group had higher immune activities. Lastly, AML patients from different risk groups showed differential responses to various agents. Conclusion: A cuproptosis-related lncRNA signature was established to predict the prognosis and inform on potential therapeutic strategies for AML patients.