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Abstract Background Osteoarthritis (OA) poses a significant risk to the mobility of patients. Carbonic anhydrase 12 (CA12) can boost apoptosis and inflammation in several cancers, but its role in OA is unknown. Methods Differentially expressed genes in OA were analyzed using the GEO database (GSE169077). RT-qPCR and western blot estimated relative mRNA and protein levels of CA12. Cell viability and apoptosis were estimated by cell counting and flow cytometry assays. Oxidative stress (OxS) was determined by detecting with ROS and MDA levels, as well as CAT and SOD activities. Cytokine levels of IL-6 and TNF-α were detected by ELISA. Parameters associated with apoptosis and extracellular matrix (ECM) were detected by western blot. The m6A modification profile was determined by methylated RNA immunoprecipitation assays. Results Relative CA12 and wilms’ tumor 1-associating protein (WTAP) mRNA and protein levels were overexpressed in OA articular cartilages and IL-1β-challenged chondrocytes CHON-001. CA12 silencing impaired IL-1β-induced cell apoptosis, inflammation, OxS, and ECM degradation in chondrocytes. Yet, CA12 overexpression exerted an opposing function. WTAP reinforced the stability of CA12 mRNA depending on the m6A modification. Furthermore, WTAP knockdown weakened cell apoptosis, inflammation, OxS, and ECM degradation in chondrocytes induced by IL-1β, but these changes were impaired after CA12 overexpression. In addition, WTAP knockdown mitigates cartilage degeneration in DMM-induced mouse models. Conclusion IL-1β-induced WTAP enhances CA12 mRNA stability depending on m6A modification, thus promoting chondrocyte apoptosis, inflammatory response, OxS, and ECM degradation, providing evidence to support the possibility of WTAP and CA12 as potential targets for OA treatment.

Abstract Background Subarachnoid hemorrhage (SAH) is a life -threatening cerebrovascular disease, where early brain injury (EBI) stands as a primary contributor to mortality and unfavorable patient outcomes. Neuronal ferroptosis emerges as a key pathological mechanism underlying EBI in SAH. Targeting ferroptosis for therapeutic intervention in SAH holds significant promise as a treatment strategy. Methods SAH model was induced via intravascular puncture and quantitatively assessed the presence of neuronal ferroptosis in the early phase of SAH using FJC staining, Prussian blue staining, as well as malondialdehyde (MDA) and glutathione (GSH) measurements. Hyaluronic acid-coated ursolic acid nanoparticles (HA-PEG-UA NPs) were prepared using the solvent evaporation method. We investigated the in vivo distribution of HA-PEG-UA NPs in SAH model through IVIS and fluorescence observation, and examined their impact on short-term neurological function and cortical neurological injury. Finally, we assessed the effect of UA on the Nrf-2/SLC7A11/GPX4 axis via Western Blot analysis. Results We successfully developed self-assembled UA NPs with hyaluronic acid to target the increased CD44 expression in the SAH-afflicted brain. The resulting HA-PEG-UA NPs facilitated delivery and enrichment of UA within the SAH-affected region. The targeted delivery of UA to the SAH region can effectively inhibit neuronal ferroptosis, improve neurological deficits, and prognosis in mice. Its mechanism of action is associated with the activation of the Nrf-2/SLC7A11/GPX4 signaling pathway. Conclusions Brain-targeted HA-PEG-UA NPs was successfully developed and hold the potential to enhance SAH prognosis by limiting neuronal ferroptosis via modulation of the Nrf-2/SLC7A11/GPX4 signal. Graphical Abstract

Abstract Drought poses a significant ecological threat that limits the production of crops worldwide. The objective of this study to examine the impact of soil applied biochar (BC) and peatmoss (PM) on the morpho-biochemical and quality traits of tobacco plants under drought conditions. In the present experiment work, a pot trial was conducted with two levels of drought severity (~ well-watered 75 ± 5% field capacity) and severe drought stress (~ 35 ± 5% field capacity), two levels of peatmoss (PM) @ 5% [PM+ (with peatmoss) and PM- (without peatmoss)] and three levels of rice straw biochar (BC0 = no biochar; BC1 = 150 mg kg− 1; and BC2 = 300 mg kg− 1 of soil) in tobacco plants. The results indicate that drought conditions significantly impacted the performance of tobacco plants. However, the combined approach of BC and PM significantly improved the growth, biomass, and total chlorophyll content (27.94%) and carotenoids (32.00%) of tobacco. This study further revealed that the drought conditions decreased the production of lipid peroxidation and proline accumulation. But the synergistic approach of BC and PM application increased soluble sugars (17.63 and 12.20%), soluble protein (31.16 and 15.88%), decreased the proline accumulation (13.92 and 9.03%), and MDA content (16.40 and 8.62%) under control and drought stressed conditions, respectively. Furthermore, the combined approach of BC and PM also improved the leaf potassium content (19.02%) by limiting the chloride ions (33.33%) under drought stressed conditions. Altogether, the balanced application of PM and BC has significant potential as an effective approach and sustainable method to increase the tolerance of tobacco plants subjected to drought conditions. This research uniquely highlights the combined potential of PM and BC as an eco-friendly strategy to enhance plant resilience under drought conditions, offering new insights into sustainable agricultural practices.

Fibrous plants with higher biomass, particularly industrial hemp, have ability to withstand and accumulate significant quantities of heavy metals from contaminated environments. The present study aimed to evaluate the dynamics of different levels (ratios) of macronutrients nitrogen, phosphorus and potassium (NPK) viz., NPK1NPK (1:1:1); NPK2NPK (2:1:1); NPK3NPK (3:1:2); NPK4NPK (4:1:2) on hemp growth and Cu contents under various levels of Cu stress (100, 400 and 800 mg kg−1 on dry soil basis using CuSO4·5H2O). Results revealed that by increasing the Cu stress, growth and biomass decreased linearly and lipid per oxidation and enzymatic antioxidants increased. Balanced application of NPK improved the biomass and decreased the membrane damage by the modulation of malonaldehyde contents. Maximum concentration of Cu in roots (377.47 ± 4.90 mg kg−1), shoots (137.45 ± 5.60 mg kg−1) and (150.07 ± 3.57 mg kg−1) was recorded at Cu3NPK2 treatment as compared to control. Maximum translocation factor (TF) and bioaccumulation coefficients (BAC) in the shoots and leaves of hemp plant were noticed where Cu stress was applied at the rate of 100 mg kg−1. However, BAC and TF were below 1. The NPK2 treatment enhanced biomass and increase Cu content both in leaves and stems, rather than the roots. Our study suggests that balanced application of NPK is a practicable approach to alleviate Cu stress and improve biomass production of industrial hemp plant. These findings indicate that optimum nutrient supply, under Cu stress, can maximize the growth potential and overall health of industrial hemp, making it a viable option for phytoremediation and sustainable agriculture on contaminated soils.

Subarachnoid hemorrhage (SAH) is a life-threatening acute hemorrhagic cerebrovascular disease, with early brain injury (EBI) being the main cause of high mortality and severe neurological dysfunction. Oxidative stress plays a crucial role in the pathogenesis of EBI. In this study, we synthesized antioxidant stress nanoparticles based on self-assembled oleanolic acid (OA) using the solvent volatilization method. X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and transmission electron microscopy (TEM) techniques were employed to analyze and understand the self-assembly mechanism of oleic acid nanoparticles (OA NPs). The TUNEL assay, Nissl staining, and brain water content measurements were conducted to investigate the impact of OA NPs on cortical neuronal injury. Additionally, Western blot analysis was performed to investigate the antioxidant stress mechanism of OA NPs. The result showed that OA NPs exhibited a spherical structure with an average diameter of 168 nm. The application of OA NPs in SAH has been found to contribute to the reduction of keap1 protein levels and an increase in the nuclear level of Nrf2. As a result, the transcription of antioxidant stress proteins, including HO1 and NQO1, is triggered. The activation of the antioxidant stress pathway by OA NPs ultimately leads to a decrease in neuron damage and an improvement in neurological dysfunction. In conclusion, we successfully designed and synthesized OA NPs that can efficiently target the site of SAH. These nanoparticles have demonstrated their potential as antioxidants for the treatment of SAH, offering significant clinical applications.

Summary: Cytochrome b5 (CYB5) is a hemoprotein crucial for electron transfer to oxygenases. Although microsomal CYB5A is required for sterol C4-demethylation in vitro, cholesterol biosynthesis remains intact in Cyb5a knockout mice. Here, we show that knockout of mitochondrial CYB5B, rather than CYB5A, blocks cholesterol biosynthesis at the sterol-C4 oxidation step in HeLa cells, causing an accumulation of testis meiosis-activating sterol (T-MAS) and dihydro-T-MAS. Surprisingly, liver-specific Cyb5b knockout (L-Cyb5b−/−) mice exhibit normal cholesterol metabolism. Further knockdown of Cyb5a in L-Cyb5b−/− (L-Cyb5b−/−/short hairpin [sh]Cyb5a) mice leads to a marked accumulation of T-MAS and dihydro-T-MAS, indicating that either CYB5A or CYB5B is required for sterol C4-demethylation. The L-Cyb5b−/−/shCyb5a mice are largely normal, with lower sterol regulatory element-binding protein (SREBP)-target gene expression during refeeding and higher liver triglyceride levels while fasting, as T-MAS and dihydro-T-MAS inhibit the SREBP pathway and activate the PPARγ pathway. In summary, CYB5A and CYB5B compensate for sterol C4-demethylation, and T-MAS and dihydro-T-MAS can modulate the SREBP and PPARγ pathways.

The Ciprofloxacin (CIP) toxicity and salinity stress in agricultural soils cause risk to environmental and food safety. Consequently, it is essential to devise or use more effective techniques for mitigating salinity and ciprofloxacin-induced stress in soil. This study includes the nZVI-loaded biochar synthesis, integrating the unique characteristics of raw biochar with nZVI. The present study examined the impact of raw and nZVI-loaded biochar on soil quality and the mitigation of salinity stress and Ciprofloxacin toxicity in wheat plants. The results showed that the application of nZVI-loaded biochar treatments led to substantial enhancement in shoot biomass, root biomass, grain biomass, and spike biomass by 152.1, 54.3 %, 59.8 %, and 151 %, respectively compared to control treatment. The treatment with nZVI-loaded biochar significantly increased the rates of photosynthesis and transpiration, as well as the conductance of stomata. It also resulted in higher levels of intercellular CO2, photosynthetic pigments, and water use efficiency with increases of 49 %, 59 %, 57 %, 37 %, 40 %, and 95 %, respectively. The nZVI-loaded biochar significantly decreased electrolyte leakage, malondialdehyde (MDA), and hydrogen peroxide levels compared to the NaCl treatment alone. It also enhanced the activities of enzymatic antioxidants such as peroxidase (POD), superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (GPX), and ascorbate peroxidase (APX). The non-enzymatic antioxidants including total soluble sugars (TSS), flavonoids, total soluble proteins (TSP), phenolics, ascorbic acid, anthocyanin, proline, and glycine betaine significantly influenced by the nZVI-loaded biochar. The nZVI-loaded biochar effectively alleviates the stress of soils that are contaminated with hazardous amounts of Ciprofloxacin while improving the soil and plant health.

Objective To investigate the impact of human leukocyte antigen (HLA) functional epitope mismatch (EM) on the efficacy of platelet transfusion in patients with hematological diseases. Methods HLA genotyping was performed on platelet donors and patients with hematological diseases who applied for platelet serological cross-matching and HLA antigen matching from June 2021 to June 2023 by PCR-SBT method. HLA platelet matching was based on the principle of CREG to select donors for patients. HLA Matchmaker 4.0 software was used to analyze donor-recipient HLA EM information. The expression level and gene distribution of related HLA functional epitope (Eplet) were searched from the international HLA Epitope registry website (www.Epregistry.com.br). Retrospective analysis was conducted on clinical platelet transfusion data to evaluate the impact of HLA EM on platelet transfusion effectiveness. Results Platelet transfusion efficacy showed no correlation with gender and age, but it was associated with platelet matching strategy. When the total number of HLA EMs was less than 20, a lower total number of donor-recipient HLA EMs resulted in higher platelet transfusion efficiency (χ2=19.311, P=0.001) and higher average value of 24 h corrected count increment (CCI)(F=7.737, P

Background Stroke is a leading cause of death worldwide, with a lack of effective treatments for improving the prognosis. The aim of the present study was to identify novel therapeutic targets for functional outcome after ischemic stroke . Methods and Results Cis‐expression quantitative trait loci data for druggable genes were used as instrumental variables. The primary outcome was the modified Rankin Scale score at 3 months after ischemic stroke, evaluated as a dichotomous variable (3–6 versus 0–2) and also as an ordinal variable. Drug target Mendelian randomization, Steiger filtering analysis, and colocalization analysis were performed. Additionally, phenome‐wide Mendelian randomization analysis was performed to identify the safety of the drug target genes at the genetic level. Among >2600 druggable genes, genetically predicted expression of 16 genes (ABCC2, ATRAID, BLK, CD93, CHST13, NR1H3, NRBP1, PI3, RIPK4, SEMG1, SLC22A4, SLC22A5, SLCO3A1, TEK, TLR4, and WNT10B) demonstrated the causal associations with ordinal modified Rankin Scale (P

Liver ischemia-reperfusion injury (LIRI) commonly occurs in liver resection, liver transplantation, shock, and other hemorrhagic conditions, resulting in profound local and systemic effects via associated inflammatory responses and hepatic cell death. Hepatocyte death is a significant component of LIRI and its mechanism was previously thought to be limited to apoptosis and necrosis. With the discovery of novel types of programmed cell death (PCD), necroptosis, ferroptosis, pyroptosis, autophagy, NETosis, and parthanatos have been shown to be involved in LIRI. Understanding the mechanisms underlying cell death following LIRI is indispensable to mitigating the widespread effects of LIRI. Here, we review the roles of different PCD and discuss potential therapy in LIRI.

Caffeic acid (CA) is a phenolic compound, which has a multifaceted role in the response of plants to various abiotic stresses. Plants must adapt to external stimuli, prompting extensive research into phenolic and flavonoid compounds as potential mitigators of environmental stress impacts. Phenylpropanoids are emphasized for their importance in maintaining plant integrity under stress, while the significance of CA, a cinnamic acid, is interpreted as a key component involved in lignin synthesis and regulating physiological processes. CA deals with different kinds of stress, like salinity-induced stress, chilling stress, and especially drought-induced stress in plants. CA helps plants make certain chemicals called phenolics, which protect them from damage caused by oxidative stress. Researchers are investigating the possible ways that CA functions, including its roles in regulating signals, defending against antioxidants, and preserving the integrity of cells. There is a need to understand the response of CA to abiotic stress factors, which can provide information about its adaptive mechanisms and applications in agriculture. This review proposes genetic advances to increase crop tolerance and provides an overview of the complexities surrounding the role of CA in plant responses to various abiotic conditions. This highlights the need for further research to identify specific mechanisms and interactions with other compounds, as well as the importance of comparative analysis of CA responses in different plant species. It emphasized the importance of meeting the challenges of stress response research and encouraged collaborative efforts for sustainable agriculture and human well-being. This demonstrates CA's importance in enhancing stress resistance and provides a blueprint for future research directions, including the study of specific mechanisms, interactions between components, and applications in the development of stress-resistant products.

IntroductionUnderstanding the pattern of species diversity and underlying ecological determinants driving a forest ecosystem is fundamental to conservation biology and forest management. Boreal forests play an irreplaceable role in providing ecosystem services and maintaining the carbon cycle globally, yet research attention remains disproportionately limited and lacking throughout time.MethodsBased on field measurement data from a large (25 ha) fully-mapped coniferous forest plot, the present study quantified patterns of species diversity and their determinants in Kanas of Xinjiang, northwest China. We applied linear regression analysis to test the effects of biotic and soil factors on alpha-diversity and local contribution of beta diversity (LCBD), and then we adopted path analysis to test the determinants that affected the species diversity index.Results and discussionOur results revealed that alpha-diversity indices did not vary greatly across different subplots, and richness value (between 2 and 6) was low in Kanas. Noteworthy is the discerned negative association between the average diameter at breast height (DBH) and species richness, suggesting that areas with smaller DBH values tend to harbor greater species richness. For beta-diversity, a higher value was observed in the substory layer (0.221) compared to both the canopy layer (0.161) and the understory layer (0.158). We also found that the species abundance distance matrix of biological and soil environmental factors were significantly correlated with species geographic distance matrices. More importantly, our results showed that average DBH and soil pH would affect the alpha diversity indices, and average DBH, soil Ph, average height and soil total Phosphorous would affect the beta diversity indices. Soil pH also indirectly affected the LCBDunder, LCBDsub, and LCBDcan (p ≤ 0.001), upon mediation of alpha diversity indices. Overall, our results provide crucial revelations about species diversity patterns in boreal forests, and insights that can support the protection of forest biodiversity in China.

Stem cells offer new therapeutic avenues for the repair and replacement of damaged tissues and organs owing to their self-renewal and multipotent differentiation capabilities. In this paper, we conduct a systematic review of the characteristics of various types of stem cells and offer insights into their potential applications in both cellular and cell-free therapies. In addition, we provide a comprehensive summary of the technical routes of stem cell therapy and discuss in detail current challenges, including safety issues and differentiation control. Although some issues remain, stem cell therapy demonstrates excellent potential in the field of regenerative medicine and provides novel tactics and methodologies for managing a wider spectrum of illnesses and traumas.

ObjectiveThis study aimed to explore the bidirectional causal relationship between periodontal disease-related phenotype (PDRP) and knee osteoarthritis (KOA) in a European population using a two-sample Mendelian Randomization (MR) approach.MethodsWe leveraged publicly available GWAS summary statistics for PDRP (n = 975) and KOA (n = 403,124), assessing their roles as both exposures and outcomes. Our comprehensive MR analysis employed various methods, including inverse variance weighting (IVW), weighted median, Egger regression, simple mode, and weighted mode, to enhance the robustness of our findings. To ensure the reliability of our instrumental variables, we implemented a rigorous screening process based on p-values and F-values, utilized Phenoscanner to investigate potential confounders, and conducted sensitivity analyses.ResultsOur analysis identified five SNPs associated with PDRP and three SNPs with KOA, all surpassing the genome-wide significance threshold, as instrumental variables. The IVW method demonstrated a significant causal relationship from PDRP to KOA (beta = 0.013, SE = 0.007, P = 0.035), without evidence of directional pleiotropy (MR-Egger regression intercept = 0.021, P = 0.706). No support was found for reverse causality from KOA to PDRP, as further MR analyses yielded non-significant P-values. Additionally, funnel plots and Cochran's Q test detected no significant heterogeneity or directional pleiotropy, confirming the robustness of our results. In multivariate analysis, when considering smoking, alcohol consumption, BMI collectively no direct causal relationship between KOA and PDRP. Conversely, smoking and higher BMI were independently associated with an increased risk of KOA.ConclusionIn conclusion, our analysis revealed no direct causal relationship from KOA to PDRP. However, a causal relationship from PDRP to KOA was observed. Notably, when adjusting for potential confounders like smoking, alcohol intake, and BMI, both the causal connection from PDRP to KOA and the inverse relationship were not substantiated.

ABSTRACT: In the current era, the most crucial matter is to increase crop yields sustainably. Biological agents’ usage is an effective and environmentally friendly way to boost plant growth by activating the antioxidative defense system and improving soil health. The present investigation aimed at evaluating the bacterial (Bacillus subtilis, Bacillus amyloliquefaciens) and fungal (Trichoderma harzianum) bioagents in the sole and synergistic forms to improve tobacco (cultivar: Yunyan 87) growth by the mediation of antioxidant defense system and soil health. Bioagents were applied as root application at the rate of 100 ml suspension per plant after that same concentrations were applied 25 days of planting with four times taking 15 days intervals. Results revealed that soil application of these bioagents significantly enhanced growth, photosynthetic attributes, enzymatic antioxidants, and soil enzymatic activities. Combined application of bacterial and fungal bioagents improved the leaf area (121.50%), total chlorophyll contents (131.48%), polyphenol oxidase (PPO) (77.57%) and phenylalanine ammonia-lyase (PAL) activities (43.52%) as compared with control. Fungal bioagents marked better performance as compared to bacterial bioagents and control. The co-application of bioagents significantly enhanced the leaf quality attributes by decreasing the accumulation of proline, malondialdehyde content, and accumulation of chloride ions in tobacco leaves. The higher soil enzyme activities and essential nutrients were observed in the treatment of bacterial and fungal bioagents (application of Bacillus and Trichoderma spp.). In crux, the findings confirmed that the combined use of Bacillus and Trichoderma spp could contribute as a new cultivation practice for sustainable growth, productivity and quality of tobacco crop and can be recommended as safe and eco-friendly alternatives to preserve plant, soil, and human health for achieving the sustaibalility.

Accurate intracellular cholesterol traffic plays crucial roles. Niemann Pick type C (NPC) proteins NPC1 and NPC2, are two lysosomal cholesterol transporters that mediate the cholesterol exit from lysosomes. However, other proteins involved in this process remain poorly defined. Here, we find that the previously unannotated protein TMEM241 is required for cholesterol egressing from lysosomes through amphotericin B-based genome-wide CRISPR-Cas9 KO screening. Ablation of TMEM241 caused impaired sorting of NPC2, a protein utilizes the mannose-6-phosphate (M6P) modification for lysosomal targeting, resulting in cholesterol accumulation in the lysosomes. TMEM241 is a member of solute transporters 35 nucleotide sugar transporters family and localizes on the cis-Golgi network. Our data indicate that TMEM241 transports UDP-N-acetylglucosamine (UDP-GlcNAc) into Golgi lumen and UDP-GlcNAc is used for the M6P modification of proteins including NPC2. Furthermore, Tmem241-deficient mice display cholesterol accumulation in pulmonary cells and behave pulmonary injury and hypokinesia. Taken together, we demonstrate that TMEM241 is a Golgi-localized UDP-GlcNAc transporter and loss of TMEM241 causes cholesterol accumulation in lysosomes because of the impaired M6P-dependent lysosomal targeting of NPC2.

AbstractBackground: Cardiovascular disease (CVD) is widely observed in modern society. CVDs are responsible for the majority of fatalities, with heart attacks and strokes accounting for approximately 80% of these cases. Furthermore, a significant proportion of these deaths, precisely one-third, occurs in individuals under 70. Metabolic syndrome encompasses a range of diseases characterized by various physiological dysfunctions. These include increased inflammation in adipose tissue, enhanced cholesterol synthesis in the liver, impaired insulin secretion, insulin resistance, compromised vascular tone and integrity, endothelial dysfunction, and atheroma formation. These factors contribute to the development of metabolic disorders and significantly increase the likelihood of experiencing cardiovascular complications.Method: We selected studies that proposed hypotheses regarding metabolic disease syndrome and cardiovascular disease (CVD) and the role of Nrf2/HO-1 and factor regulation in CVD research investigations based on our searches of Medline and PubMed.Results: A total of 118 articles were included in the review, 16 of which exclusively addressed hypotheses about the role of Nrf2 on Glucose regulation, while 16 involved Cholesterol regulation. Likewise, 14 references were used to prove the importance of mitochondria on Nrf2. Multiple studies have provided evidence suggesting the involvement of Nrf2/HO-1 in various physiological processes, including metabolism and immune response. A total of 48 research articles and reviews have been used to highlight the role of metabolic syndrome and CVD.Conclusion: This review provides an overview of the literature on Nrf2/HO-1 and its role in metabolic disease syndrome and CVD.

COVID-19 is a disease caused by a virus named SARS-CoV-2. SARS-CoV-2 is a single-stranded positive-sense RNA virus. Reverse transcription quantitative PCR (RT–qPCR) assays are the gold standard molecular test for detection of RNA viruses. The aim of this study was to construct an RNA-positive control based on MS2 phage-like particles (MS2 VLPs) to detect SARS-CoV-2 RNA. pCDFDuet-1 was used as a one-plasmid double-expression system to construct MS2 VLPs containing ssRNA of SARS-CoV-2. The sequence encoding one copy of maturase, His-tag and coat protein dimer was cloned and inserted into MCS1 of the plasmid; the fragment encoding protein N and ORF1ab from SARS-CoV-2 was cloned and inserted into MCS2. The prepared plasmid was transformed into Escherichia coli strain BL2 (DE3), and expression of the construct was induced by 1 mM isopropyl-L-thio-D-galactopyranoside (IPTG) at 30°C for 12 hours. MS2 VLPs were purified and collected with Ni-NTA affinity chromatography columns. The size and shape of the MS2 VLPs were verified by transmission electron microscopy, and the stability of MS2 VLP packaged RNA was evaluated by treatment with RNase A. Effects of storage temperature and buffer on MS2 VLP stability were also investigated. The results showed that SARS-CoV-2 MS2 VLPs could be successfully produced by this one-plasmid double-expression system. MS2 VLPs showed high stability and may be used as a positive control in molecular diagnosis of COVID-19.

Abstract Neuromyelitis optica spectrum disorder (NMOSD) is an inflammatory demyelinating disorder of the central nervous system (CNS) triggered by autoimmune mechanisms. Microglia are activated and play a pivotal role in response to tissue injury. Triggering receptor expressed on myeloid cells 2 (TREM2) is expressed by microglia and promotes microglial activation, survival and phagocytosis. Here, we identify a critical role for TREM2 in microglial activation and function during AQP4-IgG and complement-induced demyelination. TREM2-deficient mice had more severe tissue damage and neurological impairment, as well as fewer oligodendrocytes with suppressed proliferation and maturation. The number of microglia clustering in NMOSD lesions and their proliferation were reduced in TREM2-deficient mice. Moreover, morphology analysis and expression of classic markers showed compromised activation of microglia in TREM2-deficient mice, which was accompanied by suppressed phagocytosis and degradation of myelin debris by microglia. These results overall indicate that TREM2 is a key regulator of microglial activation and exert neuroprotective effects in NMOSD demyelination. Graphical Abstract

Accurate stroke assessment and consequent favorable clinical outcomes rely on the early identification and quantification of aneurysmal subarachnoid hemorrhage (aSAH) in non-contrast computed tomography (NCCT) images. However, hemorrhagic lesions can be complex and difficult to distinguish manually. To solve these problems, here we propose a novel Hybrid 2D/3D UNet deep-learning framework for automatic aSAH identification and quantification in NCCT images. We evaluated 1824 consecutive patients admitted with aSAH to four hospitals in China between June 2018 and May 2022. Accuracy and precision, Dice scores and intersection over union (IoU), and interclass correlation coefficients (ICC) were calculated to assess model performance, segmentation performance, and correlations between automatic and manual segmentation, respectively. A total of 1355 patients with aSAH were enrolled: 931, 101, 179, and 144 in four datasets, of whom 326 were scanned with Siemens, 640 with Philips, and 389 with GE Medical Systems scanners. Our proposed deep-learning method accurately identified (accuracies 0.993–0.999) and segmented (Dice scores 0.550–0.897) hemorrhage in both the internal and external datasets, even combinations of hemorrhage subtypes. We further developed a convenient AI-assisted platform based on our algorithm to assist clinical workflows, whose performance was comparable to manual measurements by experienced neurosurgeons (ICCs 0.815–0.957) but with greater efficiency and reduced cost. While this tool has not yet been prospectively tested in clinical practice, our innovative hybrid network algorithm and platform can accurately identify and quantify aSAH, paving the way for fast and cheap NCCT interpretation and a reliable AI-based approach to expedite clinical decision-making for aSAH patients.

Background Storage of potato tubers is an essential stage of the supply chain, from farm to consumer, to efficiently match supply and demand. However, the quality and yield of potatoes are influenced by physiological changes during storage. Methods This study tested the physiological and biochemical indices in three potato varieties (YunSu 108, YunSu 304 and YunSu 306) during their dormancy periods. Results Three potato varieties with different dormancy periods were used to follow changes in starch, protein and several enzymes during storage. The starch and sugar content of the long-dormant variety (YunSu 108, LDV) were stable, whereas those of the short-dormant variety (YunSu 306, SDV) were variable. Starch synthase activity in the three varieties was initially high, then decreased; the starch content of LDV was relatively stable, that of the medium-dormant variety (YunSu 304, MDV) increased with storage time and peaked at sprouting, and that of SDV was low but variable. The sucrose synthase activity of LDV was significantly higher (p

The genus Metarhizium is one of the most significant entomopathogenic fungi with diverse morphological characteristics and host species. Species of Metarhizium have been widely used for pest control as an environmentally safe alternative to chemical pesticides. This study reports a new species of Metarhizium and a new record of M. bibionidarum from China. The taxonomic positions of the two species within Metarhizium were assessed by morphological and multi-gene phylogenetic data. This assessment confirmed that the new species M. dianzhongense on white grubs (Coleoptera) is a sister to M. ellipsoideum on adult leafhoppers (Hemiptera) and represents a distinctive fungus according to the morphological and phylogenetic evidence. The two species, M. dianzhongense and M. bibionidarum, were described and illustrated. Pathogenicity tests by M. bibionidarum and M. dianzhongense were performed on early instar larvae of the significant agricultural pest Spodoptera frugipera (Lepidoptera). The results demonstrated that both M. bibionidarum and M. dianzhongense exhibit significant insecticidal activity against larvae of S. frugipera, providing new fungal resources for the development of an eco-friendly biocontrol agent against this pest.

Background: It is unclear whether serum calcium on admission is associated with clinical outcomes in dilated cardiomyopathy (DCM). In this study, we conducted a retrospective study spanning a decade to investigate the prognostic value of baseline calcium in elderly patients with DCM. Methods: A total of 1,089 consecutive elderly patients (age ≥60 years) diagnosed with DCM were retrospectively enrolled from January 2010 to December 2019. Univariate and multivariate analyses were performed to investigate the association of serum calcium with their clinical outcomes. Results: In this study, the average age of the subjects was 68.36 ± 6.31 years. Receiver operating characteristic (ROC) curve analysis showed that serum calcium level had a great sensitivity and specificity for predicting in-hospital death, with an AUC of 0.732. Kaplan–Meier survival analysis showed that patients with a serum calcium >8.62 mg/dL had a better prognosis than those with a serum calcium ≤8.62 mg/dL (log-rank χ2 40.84, p < 0.001). After adjusting for several common risk factors, a serum calcium ≤8.62 mg/dL was related to a higher risk of long-term mortality (HR: 1.449; 95% CI: 1.115~1.882; p = 0.005). Conclusions: Serum calcium level could be served as a simple and affordable tool to evaluate patients’ prognosis in DCM.

A novel mode-locking method based on nonlinear multimode interference (NLMI) using a distributed large-core (105 μm) graded-index multimode fiber (GIMF)-based saturable absorber (SA) capable of generating four pulse modes is proposed. The distributed SA geometry consists of two GIMFs located at different positions in the resonant cavity. The coupling and joint operation not only facilitate resistance to pulse fragmentation but also provide a sophisticated and widely tunable transmission with saturable and reverse saturable absorption phenomena. Based on this, dissipative soliton (DS), dissipative soliton resonance (DSR), wedge-shaped, and staircase pulses are achieved without additional filters. The DS has accessible output power, pulse energy, bandwidth, and duration of up to 15.33 mW, 2.02 nJ, 22.63 nm, and ~1.68 ps. The DSR has an achievable pulse duration and energy of ~32.39 ns, 30.3 nJ. The dispersion range that allows DS operation is studied, and the dynamics of the evolution from DS to DSR are observed. The versatility, flexibility, and simplicity of the SA device, combined with the possibility of scaling the pulse energy, make it highly attractive for ultrafast optics and nonlinear dynamics.

Stroke is the leading cause of death and disability. Currently, there is no effective pharmacological treatment for this disease, which can be partially attributed to the inability to efficiently deliver therapeutics to the brain. Here we report the development of natural compound-derived nanoparticles (NPs), which function both as a potent therapeutic agent for stroke treatment and as an efficient carrier for drug delivery to the ischemic brain. First, we screened a collection of natural nanomaterials and identified betulinic acid (BA) as one of the most potent antioxidants for stroke treatment. Next, we engineered BA NPs for preferential drug release in acidic ischemic tissue through chemically converting BA to betulinic amine (BAM) and for targeted drug delivery through surface conjugation of AMD3100, a CXCR4 antagonist. The resulting AMD3100-conjugated BAM NPs, or A-BAM NPs, were then assessed as a therapeutic agent for stroke treatment and as a carrier for delivery of NA1, a neuroprotective peptide. We show that intravenous administration of A-BAM NPs effectively improved recovery from stroke and its efficacy was further enhanced when NA1 was encapsulated. Due to their multifunctionality and significant efficacy, we anticipate that A-BAM NPs have the potential to be translated both as a therapeutic agent and as a drug carrier to improve the treatment of stroke.

Abstract Background Karst-adapted plant, Lysionotus pauciflours accumulates special secondary metabolites with a wide range of pharmacological effects for surviving in drought and high salty areas, while researchers focused more on their environmental adaptations and evolutions. Nevadensin (5,7-dihydroxy-6,8,4'-trimethoxyflavone), the main active component in L. pauciflours, has unique bioactivity of such as anti-inflammatory, anti-tubercular, and anti-tumor or cancer. Complex decoration of nevadensin, such as hydroxylation and glycosylation of the flavone skeleton determines its diversity and biological activities. The lack of omics data limits the exploration of accumulation mode and biosynthetic pathway. Herein, we integrated transcriptomics, metabolomics, and microbial recombinant protein system to reveal hydroxylation and glycosylation involving nevadensin biosynthesis in L. pauciflours. Results Up to 275 flavonoids were found to exist in L. pauciflorus by UPLC-MS/MS based on widely targeted metabolome analysis. The special flavone nevadensin (5,7-dihydroxy-6,8,4'-trimethoxyflavone) is enriched in different tissues, as are its related glycosides. The flavonoid biosynthesis pathway was drawn based on differential transcripts analysis, including 9 PAL, 5 C4H, 8 4CL, 6 CHS, 3 CHI, 1 FNSII, and over 20 OMTs. Total 310 LpCYP450s were classified into 9 clans, 36 families, and 35 subfamilies, with 56% being A-type CYP450s by phylogenetic evolutionary analysis. According to the phylogenetic tree with AtUGTs, 187 LpUGTs clustered into 14 evolutionary groups (A-N), with 74% being E, A, D, G, and K groups. Two LpCYP82D members and LpUGT95 were functionally identified in Saccharomyces cerevisiae and Escherichia coli, respectively. CYP82D-8 and CYP82D-1 specially hydroxylate the 6- or 8-position of A ring in vivo and in vitro, dislike the function of F6H or F8H discovered in basil which functioned depending on A-ring substituted methoxy. These results refreshed the starting mode that apigenin can be firstly hydroxylated on A ring in nevadensin biosynthesis. Furthermore, LpUGT95 clustered into the 7-OGT family was verified to catalyze 7-O glucosylation of nevadensin accompanied with weak nevadensin 5-O glucosylation function, firstly revealed glycosylation modification of flavones with completely substituted A-ring. Conclusions Metabolomic and full-length transcriptomic association analysis unveiled the accumulation mode and biosynthetic pathway of the secondary metabolites in the karst-adapted plant L. pauciflorus. Moreover, functional identification of two LpCYP82D members and one LpUGT in microbe reconstructed the pathway of nevadensin biosynthesis.

Neuromyelitis optica spectrum disorders (NMOSD) are demyelinating diseases of the central nervous system, have drawn the attention of many researchers due to the relapsing courses and cumulative disability. A first bibliometric analysis of NMOSD was conducted to identify the research hotspots and emerging trends. Articles relevant to NMOSD published in the core collection of Web of Science were retrieved and analyzed through visualized analysis using CiteSpace and VOSviewer, focusing on annual publication trends, countries, institutions, authors, journals, and keywords. The analysis showed that over the past 30 years, publications related to NMOSD had shown steady growth with slight fluctuations. The United States played an important part in this field, with the highest outputs and the greatest number of citations. Research hotspots of NMOSD had gradually shifted from the definition, biomarkers, and diagnostic criteria to diagnosis and treatment, particularly immunotherapy. This bibliometric analysis provides researchers with a theoretical basis for studying NMOSD and offers guidance for future research directions.

Background2021 World Health Organization (WHO) Central Nervous System (CNS) tumor classification increasingly emphasizes the important role of molecular markers in glioma diagnoses. Preoperatively non-invasive “integrated diagnosis” will bring great benefits to the treatment and prognosis of these patients with special tumor locations that cannot receive craniotomy or needle biopsy. Magnetic resonance imaging (MRI) radiomics and liquid biopsy (LB) have great potential for non-invasive diagnosis of molecular markers and grading since they are both easy to perform. This study aims to build a novel multi-task deep learning (DL) radiomic model to achieve preoperative non-invasive “integrated diagnosis” of glioma based on the 2021 WHO-CNS classification and explore whether the DL model with LB parameters can improve the performance of glioma diagnosis.MethodsThis is a double-center, ambispective, diagnostical observational study. One public database named the 2019 Brain Tumor Segmentation challenge dataset (BraTS) and two original datasets, including the Second Affiliated Hospital of Nanchang University, and Renmin Hospital of Wuhan University, will be used to develop the multi-task DL radiomic model. As one of the LB techniques, circulating tumor cell (CTC) parameters will be additionally applied in the DL radiomic model for assisting the “integrated diagnosis” of glioma. The segmentation model will be evaluated with the Dice index, and the performance of the DL model for WHO grading and all molecular subtype will be evaluated with the indicators of accuracy, precision, and recall.DiscussionSimply relying on radiomics features to find the correlation with the molecular subtypes of gliomas can no longer meet the need for “precisely integrated prediction.” CTC features are a promising biomarker that may provide new directions in the exploration of “precision integrated prediction” based on the radiomics, and this is the first original study that combination of radiomics and LB technology for glioma diagnosis. We firmly believe that this innovative work will surely lay a good foundation for the “precisely integrated prediction” of glioma and point out further directions for future research.Clinical trail registrationThis study was registered on ClinicalTrails.gov on 09/10/2022 with Identifier NCT05536024.

Objective: To investigate the effects of IMR, iDose4 and FBP reconstruction algorithms on the image quality of multi-row spiral CT in the chest before costal cartilage rhinoplasty. Methods: A total of 60 patients who met the inclusion criteria and planned to receive autogenous costal cartilage rhinoplasty were collected. All patients underwent chest CT examination before operation and completed relevant 3D reconstruction based on images generated by IMR, iDose4 and FBP reconstruction algorithms, and the image quality of the three reconstruction algorithms was analyzed. The subjective evaluation score and objective evaluation indexes of image quality in each group were recorded and compared including costal cartilage CT value, noise, signal-to-noise ratio and contrast noise ratio. Results: The CT reconstruction images obtained by the three algorithms could all accurately assess the degree of costal cartilage calcification. Subjective evaluation of image quality showed that the IMR group obtained higher score and held obvious advantages, mainly reflected in the IMR reconstruction algorithm compared with iDose4 and FBP two algorithms in the costal cartilage calcification spot edge display and inhibition of liver, gastrointestinal content and weakened heart beating artifacts. Objective evaluation of costal cartilage CT value, SNR, contrast noise ratio and statistical analysis, the final statistical results showed that IMR, iDose4 and FBP reconstruction algorithm group showed significant statistical differences, IMR group showed obviously the best image quality. Conclusion: All three reconstruction algorithms of IMR, idose4 and FBP can accurately evaluate the presence and degree of calcification of autologous costal cartilage. Under the same scanning conditions, compared with the reconstruction algorithms of FBP and iDose4, IMR reconstruction algorithms can reducecan significantly reduce image noise, improve CNR and SNR, and improve the subjective image quality of costal cartilage. It is more acceptable to clinical surgeons and can be used as the preferred algorithm reconstruction technique for costal cartilage CT examination, which is most worthy of promotion.

As a class of novel biomaterials manufactured by synthetic biology technologies, recombinant collagens are candidates for a variety of medical applications. In this article, a regulatory scientific perspective on recombinant collagens and their medical devices is presented with a focus on the definition, translation, classification and technical review. Recombinant collagens are categorized as recombinant human collagen, recombinant humanized collagen and recombinant collagen-like protein, as differentiated by specific compositions and structures. Based on their intended uses and associated risks, recombinant collagen-based medical devices are generally classified as Class Ⅱ or Ⅲ in China. The regulatory review of recombinant collagen-based medical devices aims to assess their safety and efficacy demonstrated by scientific evidences generated from preclinical and clinical evaluations. Taken together, opportunities as well as challenges for their future clinical translation of recombinant collagen-based medical devices abound, which highlights the essential role of regulatory science to provide new tools, standards, guidelines and methods to evaluate the safety and efficacy of medical products.

Animal studies play a vital role in validating the concept, feasibility, safety, performance and efficacy of biomaterials products during their bench-to-clinic translation. This article aims to share regulatory considerations for animal studies of biomaterial products. After briefly emphasizing the importance of animal studies, issues of animal studies during biomaterial products’ translation are discussed. Animal studies with unclear purposes, flawed design and poor reporting quality could significantly reduce the translation efficiency and create regulatory challenges. Regulatory perspectives on the purpose, principle, quality and regulatory science of animal studies are also presented. Animal studies should have clear purposes, follow principles of 3R+DQ (replacement, reduction, refinement, design and quality) and execute under an efficiently operating quality management system. With the advancement of regulatory science, National Medical Products Administration of China has been developing a series of standards and guidance documents on animal studies of medical devices. Case studies of making decisions on whether to conduct animal studies are provided in the end with drug-eluting stents as examples. In summary, animal studies of biomaterial products should pay close attention to the rationale, design and quality in order to achieve their purposes.

Accurate snow cover data are critical for understanding the Earth’s climate system, and exploring hydrological processes and regional water resource management over High Mountain Asia (HMA). However, satellite-based remote sensing observations of snow cover have inevitable data gaps originating from cloud cover, sensor, orbital limitations and other factors. Here an effective cloud-gap-filled (CGF) method was developed to fully fill the data gaps in Moderate Resolution Imaging Spectroradiometer (MODIS) normalized difference snow index (NDSI) product. The CGF method combines the respective strengths of the cubic spline interpolation method and the spatio-temporal weighted method for generating the CGF Terra-Aqua MODIS NDSI product over HMA from 2000 to 2021. Based on the validation results of in situ snow-depth observations, the CGF NDSI product achieves a high range overall accuracy (OA) of 93.54–98.08%, a low range underestimation error (MU) of 0.15–3.49% and an acceptable range overestimation error (MO) of 0.84–5.77%. Based on the validation results of high-resolution Landsat images, this product achieves the OA of 88.52–92.40%, the omission error (OE) of 1.42–10.28% and the commission error (CE) of 5.97–17.58%. The CGF MODIS NDSI product can provide scientific support for eco-environment sustainable management in the high mountain region.

With the improvement in people’s living standards, the development and application of smart textiles are receiving increasing attention. In this study, a carbon nanosurface was successfully coated with a SiO2 layer to form C@SiO2 nanomaterials, which improved the dispersion of carbon nanomaterials in an aqueous solution and enhanced the absorption of light by the carbon nanoparticles. C@SiO2 nanoparticles were coupled on the surface of silk fabric with the silane coupling agent KH570 to form C@SiO2 nanosilk fabric. The silk fabric that was subjected to such surface modification was endowed with a special photothermal function. The results obtained with scanning electron microscopy (SEM), energy dispersive spectrometer (EDS), and infrared spectroscopy (FTIR) showed that C@SiO2 nanoparticles were successfully modified on the surface of the silk fabric. In addition, under the irradiation of near-infrared light with a power of 20 W and a wavelength of 808 nm, the C@SiO2 nanosilk fabric experienced rapid warming from 23 °C to 60 °C within 30 s. After subjecting the functional fabric to hundreds of photothermal experiments and multiple washes, the photothermal efficiency remained largely unchanged and proved to be durable and stable. In addition, the thermogravimetric (TG) analysis results showed that the C@SiO2 nanoparticles contributed to the thermal stability of the silk fabric. The UV transmittance results indicated that C@SiO2 nanofabric is UV-resistant. The silk modification method developed in this study is low-cost, efficient, and environmentally friendly. It has some prospects for future applications in the textile industry.

Abstract Background Soluble triggering receptor expressed on myeloid cells 2 (sTREM2) in cerebrospinal fluid (CSF) has been described as a biomarker for microglial activation, which were observed increased in a variety of neurological disorders. Objective Our objective was to explore whether genetically determined CSF sTREM2 levels are causally associated with different neurological diseases by conducting a two-sample Mendelian randomization (MR) study. Methods Single nucleotide polymorphisms significantly associated with CSF sTREM2 levels were selected as instrumental variables to estimate the causal effects on clinically common neurological diseases, including stroke, Alzheimer’s diseases, Parkinson’s diseases, amyotrophic lateral sclerosis, multiple sclerosis, and epilepsy and their subtypes. Summary-level statistics of both exposure and outcomes were applied in an MR framework. Results Genetically predicted per 1 pg/dL increase of CSF sTREM2 levels was associated with higher risk of multiple sclerosis (OR = 1.038, 95%CI = 1.014–1.064, p = 0.002). Null association was found in risk of other included neurological disorders. Conclusions These findings provide support for a potential causal relationship between elevated CSF sTREM2 levels and higher risk of multiple sclerosis.

Improved treatment of glioblastoma (GBM) can be achieved by inducing differentiation of glioblastoma stem cells (GSCs). Here, the authors show that zinc finger protein 117 (ZNF117) is a regulator of GSC differentiation via Notch signaling through interaction with JAG2, and can be targeted for therapy.

Abstract Glioblastoma (GBM) is the most common and aggressive primary malignant brain tumor. The unregulated expression of Claudin-4 (CLDN4) plays an important role in tumor progression. However, the biological role of CLDN4 in GBM is still unknown. This study aimed to determine whether CLDN4 mediates glioma malignant progression, if so, it would further explore the molecular mechanisms of carcinogenesis. Our results revealed that CLDN4 was significantly upregulated in glioma specimens and cells. The inhibition of CLND4 expression could inhibit mesenchymal transformation, cell invasion, cell migration and tumor growth in vitro and in vivo. Moreover, combined with in vitro analysis, we found that CLDN4 can modulate tumor necrosis factor-α (TNF-α) signal pathway. Meanwhile, we also validated that the transforming growth factor-β (TGF-β) signal pathway can upregulate the expression of CLDN4, and promote the invasion ability of GBM cells. Conversely, TGF-β signal pathway inhibitor ITD-1 can downregulate the expression of CLDN4, and inhibit the invasion ability of GBM cells. Furthermore, we found that TGF-β can promote the nuclear translocation of CLDN4. In summary, our findings indicated that the TGF-β/CLDN4/TNF-α/NF-κB signal axis plays a key role in the biological progression of glioma. Disrupting the function of this signal axis may represent a new treatment strategy for patients with GBM.

Objective: To investigate the optimal dosage of contrast agent in three-dimensional dynamic enhanced magnetic resonance angiography (3D CE-MRA) in the imaging of arterial lesions of both lower limbs. Methods: 45 patients who underwent 3D CE-MRA angiography of lower extremity arteries in our hospital were randomly divided into three groups A, B and C. The gadolinium dosages of the three groups were respectively 0.1 mmol/kg, 0.2 mmol/kg and 0.3 mmol/kg. They were injected intravenously with saline at the ratio of 1:1. The images of different dose scanning schemes were scored subjectively and objectively. Subjective scoring: the lower limb arteries were divided into femoral artery, popliteal artery and calf artery (posterior tibial artery, anterior tibial artery and common peroneal artery). The quality of MIP reconstruction images was evaluated, and the overall venous overlap scoring of calf and foot vessels was also carried out. Finally, the differences of the three scanning schemes were compared by Wilcoxon test.The objective scoring is to measure and calculate the signal intensity Si, signal-to-noise ratio SNR and contrast-to-noise ratio of the original enhanced images of the three groups, compare the horizontal signal intensity differences of femoral artery, popliteal artery and calf artery, and compare the differences of the three scanning schemes by t-test. Results: When the contrast medium dosage was 0.1 mmol/kg, the image quality scores of femoral artery, popliteal artery and calf artery were (3.35±0.25), (2.97±0.25), (2.35±1.15) respectively; When the dosage of contrast agent was 0.2 mmol/kg, the scores of the above parts were (3.75±0.35), (3.55±0.32), (2.97±0.70) respectively; When the dosage of contrast agent was 0.3 mmol/kg, the scores of the above parts were (3.90±0.41), (3.83±0.52), (3.10±0.75) respectively. 33.33% of the images in group A did not meet the diagnostic needs, while all the images in group B and C met the diagnostic needs. There was significant difference between group A and group B, and there was no significant difference between group B and group C. B The Si, SNR and CNR of femoral artery and popliteal artery in group C were significantly higher than those in group A, and there was no significant difference between group B and C. The images of group B and C were better than those of group A. Conclusion: Appropriate increase of the contrast medium dosage is helpful to improve the 3D CE-MRA imaging quality of both lower limb arteries. The contrast medium dosage of 0.2 mmol/kg is reliable and accurate for the evaluation of blood vessels, and can provide accurate and reliable imaging basis for the formulation of treatment plan for patients with peripheral artery diseases.

Abstract Background Ferroptosis plays a key role in human cancer, but its function and mechanism in glioma is not clear. P62/SQSTM1 was reported to inhibit ferroptosis via the activation of NRF2 signaling pathway. In this study we reveal a dual role of p62 in ferroptosis of glioblastoma (GBM) according to p53 status. Method Lipid peroxidation analysis, transmission electron microscopy (TEM), GSH assay were performed to determine the level of ferroptosis. Western blot and qPCR were obtained to detect the expression of ferroptosis markers. Construction of mutant plasmids, immunoprecipitation, luciferase assay and rescue-experiments were performed to explore the regulatory mechanism. Results P62 overexpression facilitates ferroptosis and inhibits SLC7A11 expression in p53 mutant GBM, while attenuates ferroptosis and promotes SLC7A11 expression in p53 wild-type GBM. P62 associates with p53 and inhibits its ubiquitination. The p53-NRF2 association and p53-mediated suppression of NRF2 antioxidant activity are diversely regulated by p62 according to p53 status. P53 mutation status is required for the dual regulation of p62 on ferroptosis. In wild-type p53 GBM, the classical p62-mediated NRF2 activation pathway plays a major regulatory role of ferroptosis, leading to increased SLC7A11 expression, resulting in a anti-ferroptosis role. In mutant p53 GBM, stronger interaction of mutant-p53/NRF2 by p62 enhance the inhibitory effect of mutant p53 on NRF2 signaling, which reversing the classical p62-mediated NRF2 activation pathway, together with increased p53’s transcriptional suppression on SLC7A11 by p62, leading to a decrease of SLC7A11, resulting in a pro-ferroptosis role. Conclusion Together, this study shows novel molecular mechanisms of ferroptosis regulated by p62; the mutation status of p53 is an important factor that determines the therapeutic response to p62-mediated ferroptosis-targeted therapies in GBM.

Hepato-pancreato-biliary (HPB) cancer is a serious category of cancer including tumors originating in the liver, pancreas, gallbladder and biliary ducts. It is limited by two-dimensional (2D) cell culture models for studying its complicated tumor microenvironment including diverse contents and dynamic nature. Recently developed three-dimensional (3D) bioprinting is a state-of-the-art technology for fabrication of biological constructs through layer-by-layer deposition of bioinks in a spatially defined manner, which is computer-aided and designed to generate viable 3D constructs. 3D bioprinting has the potential to more closely recapitulate the tumor microenvironment, dynamic and complex cell-cell and cell-matrix interactions compared to the current methods, which benefits from its precise definition of positioning of various cell types and perfusing network in a high-throughput manner. In this review, we introduce and compare multiple types of 3D bioprinting methodologies for HPB cancer and other digestive tumors. We discuss the progress and application of 3D bioprinting in HPB and gastrointestinal cancers, focusing on tumor model manufacturing. We also highlight the current challenges regarding clinical translation of 3D bioprinting and bioinks in the field of digestive tumor research. Finally, we suggest valuable perspectives for this advanced technology, including combination of 3D bioprinting with microfluidics and application of 3D bioprinting in the field of tumor immunology.

Prediction of dam behavior based on monitoring data is important for dam safety and emergency management. It is crucial to analyze and predict the seepage field. Different from the mechanism-based physical models, machine learning models predict directly from data with high accuracy. However, current prediction models are generally based on environmental variables and single measurement point time series. Sometimes point-by-point modeling is used to obtain multi-point prediction values. In order to improve the prediction accuracy and efficiency of the seepage field, a novel multi-target prediction model (MPM) is proposed in which two deep learning methods are integrated into one frame. The MPM model can capture causal temporal features between environmental variables and target values, as well as latent correlation features between different measurement points at each moment. The features of these two parts are put into fully connected layers to establish the mapping relationship between the comprehensive feature vector and the multi-target outputs. Finally, the model is trained for prediction in the framework of a feed-forward neural network using standard back propagation. The MPM model can not only describe the variation pattern of measurement values with the change of load and time, but also reflect the spatial distribution relationship of measurement values. The effectiveness and accuracy of the MPM model are verified by two cases. The proposed MPM model is commonly applicable in prediction of other types of physical fields in dam safety besides the seepage field.

BackgroundRegulatory T cells (Treg) have been identified as a key modulator of neuroinflammation in stroke. However, little is known about the association of Treg subpopulations with clinical outcome in patients with acute ischemic stroke (AIS).MethodsPatients within 1 week from stroke onset were prospectively enrolled in this study. Healthy controls were sex-and age-matched 1:1 to AIS patients. The frequencies of Treg and Treg subsets were analyzed by flow cytometry and compared with nonstroke control. Univariate and multivariate logistic regression analysis was performed to investigate the prognostic value of Treg subsets in stroke outcomes.ResultsA total of 328 patients and 328 controls were included in the study. Compared with controls, patients with AIS had higher levels of Treg frequency and memory Treg (mTreg) frequency, but lower levels of naïve Treg (nTreg) frequency and nTreg/mTreg ratio. One hundred twenty-six (38.4%) patients experienced unfavorable outcome (modified Rankin score 2–6). Multivariate regression analysis showed that nTreg/mTreg ratio was negatively associated with unfavorable 90-day outcome (the highest tertile versus the lowest tertile: odds ratio 0.13, 95% confidential interval [CI] 0.05–0.35). The risk estimation of unfavorable 90 day outcome can be significantly improved by adding nTreg/mTreg ratio to the conventional clinical parameters (continuous net reclassification improvement 91.26, 95% CI 69.04–113.5%, p

Abstract Background Glioblastoma (GBM) is the most common and fatal primary tumor in the central nervous system (CNS). Due to the existence of blood–brain barrier (BBB), most therapeutics cannot efficiently reach tumors in the brain, and as a result, they are unable to be used for effective GBM treatment. Accumulating evidence shows that delivery of therapeutics in form of nanoparticles (NPs) may allow crossing the BBB for effective GBM treatment. Methods Betulinic acid NPs (BA NPs) were synthesized by the standard emulsion approach and characterized by electron microscopy and dynamic light scattering analysis. The resulting NPs were characterized for their anti-tumor effects by cell viability assay, EdU-DNA synthesis assay, cell cycle assay, mitochondrial membrane potential, and PI-FITC apoptosis assay. Further mechanistic studies were carried out through Western Blot and immunostaining analyses. Finally, we evaluated BA NPs in vivo for their pharmacokinetics and antitumor effects in intracranial xenograft GBM mouse models. Results BA NPs were successfully prepared and formed into rod shape. BA NPs could significantly suppress glioma cell proliferation, induce apoptosis, and arrest the cell cycle in the G0/G1 phase in vitro. Furthermore, BA NPs downregulated the Akt/NFκB-p65 signaling pathway in a concentration dependent manner. We found that the observed anti-tumor effect of BA NPs was dependent on the function of CB1/CB2 receptors. Moreover, in the intracranial GBM xenograft mouse models, BA NPs could effectively cross the BBB and greatly prolong the survival time of the mice. Conclusions We successfully synthesized BA NPs, which could cross the BBB and demonstrated a strong anti-tumor effect. Therefore, BA NPs may potentially be used for effective treatment of GBM. Graphical Abstract

BackgroundThe prognostic value of the tumor burden score (TBS) in patients with combined hepatocellular-cholangiocarcinoma (cHCC-CCA) remains unknown. This study aimed to investigate the impact of TBS on long-term outcomes after surgery.MethodsPatients who underwent radical-intent resection between June 2013 and December 2019 were retrospectively reviewed. Kaplan–Meier curves were used to analyze patient survival, and disease-free survival (DFS) and overall survival (OS) were examined in relation to TBS.ResultsA total of 178 patients were included in this study, with 119 in the training cohort and 59 in the validation cohort. Kaplan–Meier curves showed that TBS was a strong prognostic indicator in patients with cHCC-CCA. Elevated TBS was associated with poorer DFS and OS (both P-value < 0.001) and was identified as an independent prognostic indicator. In addition, the prognostic value of TBS outperformed tumor size and number alone, microvascular invasion, and lymph node invasion. The prognostic significance of TBS was confirmed by the internal validation cohort.ConclusionsThe present study suggested the significance of tumor morphology in assessing the prognosis of patients with cHCC-CCA who undergoing curative resection. The TBS is a promising prognostic index in patients with cHCC-CCA. Elevated TBS was related to a lower long-term survival rate and was identified as an independent risk factor for poor DFS and OS. Further research is needed to verify our results.

ObjectiveCarotid artery stenting (CAS) performed perioperatively with high-dose atorvastatin may reduce the incidence of new ischemic brain lesions, but more high-level evidence is needed. Furthermore, the optimal dose and course of perioperative statin therapy remain uncertain.MethodsA single-center, prospective, parallel controlled, pilot randomized clinical trial was conducted at Beijing Hospital. The study includes a total of 130 patients with CAS. The patients were randomly assigned to receive a high-dose of 80 mg/day atorvastatin (n = 65) or a standard-dose of 20 mg/day atorvastatin (n = 65) 3 days before and 3 days after planned CAS. The primary endpoint event was the cumulative incidence of silent new ischemic cerebral lesions (sNICL) on post-CAS cerebral diffusion-weighted magnetic resonance imaging (DW-MRI), transient ischemic attack (TIA), or ischemic stroke within 30 days after CAS.ResultsAmong the 130 patients, 123 completed the study, of which 63 were in the high-dose group and 60 were in the standard-dose group. The incidence of major endpoint events was 69.8% (44 cases) and 78.3% (46 cases) in the high-dose and standard-dose groups, respectively. There was no significant difference between the two groups (HR, 0.705; 95% CI, 0.315–1.576; p = 0.393). According to the stratified analysis results, the sNICL incidence was significantly different between the two groups in the symptomatic patients (HR, 0.263; 95% CI, 0.70–0.984; p = 0.04).ConclusionAmong patients with CAS, a periprocedural high-dose of atorvastatin did not reduce the rate of periprocedural ischemic brain damage. However, high-dose statins can reduce the incidence of sNICL after CAS in patients with symptomatic carotid stenosis.Clinical Trial RegistrationClinicaltrials.gov, identifier NCT03079115.

Background: Secreted protein acidic and rich in cysteine-like 1 (SPARCL1) was a kind of extracellular matrix glycoprotein. SPARCL1 was strongly inhibited in most cancers. However, the potential functions of SPARCL1 in the pan-cancer cohort have not been widely studied.Methods: We evaluated the transcriptional level and the prognostic value of SPARCL1 in 33 types of cancer and revealed the relationship between genetic alterations of SPARCL1 and the tumor mutation burden. Meanwhile, we assessed the correlations between SPARCL1 and tumor-infiltrating lymphocytes across cancers.Results: The transcriptional level of SPARCL1 was inhibited in most cancers. Although SPARCL1 was down-regulated in most cancers, SPARCL1 might play a protective or detrimental role in different cancers. We demonstrated that mutation count was elevated in the altered SPARCL1 group in several cancers. Additionally, we found a significant positive correlation between SPARCL1 and macrophage infiltration levels in most cancers. Especially, marker sets of M2 macrophages were strongly related to SPARCL1 in cholangiocarcinoma, colon adenocarcinoma, rectum adenocarcinoma, and pancreatic adenocarcinoma.Conclusion: Our study found that SPARCL1 might work as a biomarker for prognosis and immune infiltration in pan-cancer analysis.

Due to its low cost and convenience, direct seeding is an efficient technique for the production of rice in different rice growing areas. However, anaerobic conditions are a major obstacle to the direct seeding of rice and result in poor seedling establishment, which leads to yield losses. We constructed a collection of recombinant inbred lines (RIL) comprising 275 lines derived from the H335 and CHA-1 cross by the method of single seed descent. Via a genotyping-by-sequencing (GBS) strategy, a high-density genetic map containing 2498 recombination bin markers was constructed, the average physical distance between the markers was only 149.38 Kb. After anaerobic treatment, 12 phenotypes related to both the coleoptile at germination and seedling quality at budding were evaluated. There were no significant correlations between seedling and bud traits. Genetic mapping of quantitative traits was performed for these traits across two cropping seasons. A total of 20 loci were detected, named locus 1~20. Three of them were repeatedly detected across both seasons. Six loci overlapped with those in previous reports, and nine loci were associated with multiple traits at both stages. Notably, locus 3, which is located on chromosome 2 (26,713,837 to 27,333,897 bp), was detected for both the germination and bud traits. By focusing on the locus 3 interval and by combining gene annotation and expression analyses, we identified a promising candidate gene, trehalose-6-phosphate phosphatase (OsTPP1, LOC_Os02g44230). Furthermore, RILs (G289, G379, G403, G430 and G454) that have superior phenotypes and that pyramid elite alleles were recognized. The findings of present study provide new genetic resources for direct-seeding rice (DSR) varieties for molecular breeding strategies and expand our knowledge of genetic regulation of seedling establishment under anaerobic conditions.

Abstract Background The prognosis of lung cancer with synchronous brain metastasis (LCBM) is very poor, and patients often die within a short time. However, little is known about the early mortality and related factors in patients with LCBM. Methods Patients diagnosed with LCBM between 2010 and 2016 were enrolled from the Surveillance, Epidemiology, and End Result (SEER) database. Univariate and multivariate logistic regression analysis were used to identify significant independent prognostic factors, which were used to construct nomograms of overall and cancer-specific early death. Then, the prediction ability of the model was verified by receiver operating characteristic (ROC) curve. At last, the clinical application value of the model was tested through decision curve analysis (DCA). Results A total of 29,902 patients with LCBM were enrolled in this study. Among them, 13,275 (44.4%) patients had early death, and 11,425 (38.2%) cases died of lung cancer. The significant independent risk factors for overall and cancer-specific early death included age, race, gender, Gleason grade, histological type, T stage, N stage, bone metastasis, liver metastasis and marital status, which were used to construct the nomogram. The ROC curve demonstrated good predictive ability and clinical application value. The areas under the curve (AUC) of the training group was 0.793 (95% CI: 0.788–0.799) and 0.794 (95% CI: 0.788–0.799), in the model of overall and cancer-specific early death respectively. And the AUC of the validation group were 0.803 (95% CI: 0.788–0.818) and 0.806 (95% CI: 0.791–0.821), respectively. The calibration plots of the model showed that the predicted early death is consistent with the actual value. The DCA analysis indicated a good clinical application value of this model. Conclusions We established a comprehensive nomogram to predict early death in lung cancer patients with synchronous brain metastases. Nomograms may help oncologists develop better treatment strategies, such as clinical trials and hospice care.

Abstract Myocarditis is a disease characterized by localized or diffuse inflammation of the myocardium without efficient treatment. This study explored the regulatory mechanism of microRNA-133 (miR-133) secreted from bone marrow mesenchymal stem cell-derived exosome (BMSC-Exo) on myocardial fibrosis and epithelial–mesenchymal transition (EMT) in viral myocarditis (VMC) rats through regulating mastermind-like 1 (MAML1). BMSCs in rats were isolated and cultured to identify their immune phenotype and osteogenic and adipogenic ability, and BMSC-Exo were extracted and identified. Exosomes were obtained through ultracentrifugation, which were identified by transmission electron microscope and western blot analysis. The rats were injected with Coxsackie B3 virus for preparation of VMC model, and cardiomyocytes were isolated, cultured and grouped in the same way as animal experiments (NCExo, Ad-miR-133aExo, Adas-miR-133aExo). In vivo and in vitro experiments were conducted to figure out the roles of exosomal miR-133a and MAML1 in inflammation, apoptosis, EMT, fibrosis, and cell viability. The targeting relationship between miR-133a and MAML1 was verified by dual luciferase reporter gene assay. BMSC-Exo raised miR-133a expression in VMC rats and effectively improved the VMC rat cardiac function and myocardial fibrosis, increased cardiomyocyte viability and inhibited the EMT process. Elevated miR-133a in exosomes strengthened the improvements. Silenced miR-133a effectively reversed the effects of BMSC-Exo on VMC rats. miR-133a targeted MAML1. Inhibition of MAML1 improved cardiac function and myocardial fibrosis in VMC rats and could reverse the effect of miR-133a-silenced exosomes on VMC rats. Our study suggests that elevated exosomal miR-133a suppresses myocardial fibrosis and EMT in rats with VMC via down-regulating MAML1, thereby inhibiting the progression of myocarditis.

Studying the dynamics of snowline altitude at the end of the melting season (SLA-EMS) is beneficial in predicting future trends of glaciers and non-seasonal snow cover and in comprehending regional and global climate change. This study investigates the spatiotemporal variation characteristics of SLA-EMS in nine glacier areas of the Himalayas, utilizing Landsat images from 1991 to 2022. The potential correlations between SLA-EMS, alterations in temperature, and variations in precipitation across the Himalayas region glacier are also being analyzed. The results obtained are summarized below: (1) the Landsat-extracted SLA-EMS exhibits a strong agreement with the minimum snow coverage at the end of the melting season derived from Sentinel-2, achieving an overall accuracy (OA) of 92.6% and a kappa coefficient of 0.85. The SLA-EMS can be accurately obtained by using this model. (2) In the last 30 years, the SLA-EMS in the study areas showed an upward trend, with the rising rate ranging from 0.4 m·a−1 to 9.4 m·a−1. Among them, the SLA-EMS of Longbasaba rose fastest, and that of Namunani rose slowest. (3) The SLA-EMS in different regions of the Himalayas in a W-E direction have different sensitivity to precipitation and temperature. However, almost all of them show a positive correlation with temperature and a negative correlation with precipitation.