Cathepsin K deficiency prevented stress-related thrombosis in a mouse FeCl3 model.

BACKGROUND: Exposure to chronic psychological stress (CPS) is a risk factor for thrombotic cardiocerebrovascular diseases (CCVDs). The expression and activity of the cysteine cathepsin K (CTSK) are upregulated in stressed cardiovascular tissues, and we investigated whether CTSK is involved in chronic stress-related thrombosis, focusing on stress serum-induced endothelial apoptosis. METHODS AND RESULTS: Eight-week-old wild-type male mice (CTSK+/+) randomly divided to non-stress and 3-week restraint stress groups received a left carotid artery iron chloride3 (FeCl3)-induced thrombosis injury for biological and morphological evaluations at specific timepoints. On day 21 post-stress/injury, the stress had enhanced the arterial thrombi weights and lengths, in addition to harmful alterations of plasma ADAMTS13, von Willebrand factor, and plasminogen activation inhibitor-1, plus injured-artery endothelial loss and CTSK protein/mRNA expression. The stressed CTSK+/+ mice had increased levels of injured arterial cleaved Notch1, Hes1, cleaved caspase8, matrix metalloproteinase-9/-2, angiotensin type 1 receptor, galactin3, p16IN4A, p22phox, gp91phox, intracellular adhesion molecule-1, TNF-α, MCP-1, and TLR-4 proteins and/or genes. Pharmacological and genetic inhibitions of CTSK ameliorated the stress-induced thrombus formation and the observed molecular and morphological changes. In cultured HUVECs, CTSK overexpression and silencing respectively increased and mitigated stressed-serum- and H2O2-induced apoptosis associated with apoptosis-related protein changes. Recombinant human CTSK degraded γ-secretase substrate in a dose-dependent manor and activated Notch1 and Hes1 expression upregulation. CONCLUSIONS: CTSK appeared to contribute to stress-related thrombosis in mice subjected to FeCl3 stress, possibly via the modulation of vascular inflammation, oxidative production and apoptosis, suggesting that CTSK could be an effective therapeutic target for CPS-related thrombotic events in patients with CCVDs.

Knocking Down HN1 Blocks Helicobacter pylori-Induced Malignant Phenotypes in Gastric Mucosal Cells and Inhibits Gastric Cancer Cell Proliferation, Cytoskeleton Remodeling, and Migration.

Helicobacter pylori (H. pylori) is implicated in the aberrant proliferation and malignant transformation of gastric mucosal cells, heightening the risk of gastric cancer (GC). HN1 is involved in the development of various tumors. However, precise mechanistic underpinnings of HN1 promoting GC progression in H. pylori remain elusive. The study collected 79 tissue samples of GC patients, including 47 with H. pylori-positive GC and 32 H. pylori-negative controls. Using human gastric epithelial cells (GES-1) and human gastric adenocarcinoma cells (HGC-27), the effect of overexpression / knocking down of HN1 and H. pylori infection was evaluated on cell function (proliferation, migration, apoptosis), cytoskeleton, and expression of cell malignant phenotype factors that promote the malignant biological behavior of cancer cells. The expression of HN1 in GC tissues is higher than that in paracancerous tissue and is closely related to infiltration, lymphatic metastasis, distant metastasis, survival, and H. pylori infection. Downregulation of HN1 effectively hinders the ability of H. pylori strains 26695 and SS1 to promote migration of GES-1 and HGC-27 cells, while lowering the expression of key indicators associated with malignant phenotype. Downregulated GSK3B, ß-catenin, and Vimentin after knockdown Integrinß1, but HN1 expression remained largely unchanged, when HN1 and Integrinß1 were knocked down, GSK3B, ß-catenin, and Vimentin expression were considerably reduced. Our research demonstrated the crucial role of HN1 in H. pylori-induced acquisition of a malignant phenotype in GES-1 cells. Knockdown of HN1 blocked the pathogenic mechanism of H. pylori-induced GC and downregulated the expression of GSK3Β, ß-catenin and Vimentin via Integrin ß1.

Extracellular vesicles from Fusobacterium nucleatum: roles in the malignant phenotypes of gastric cancer.

The increase of the Fusobacterium nucleatum level has been previously identified in various cancers including gastric cancer (GC), but how the F. nucleatum exerts its carcinogenic role in GC remains unclear. Several studies revealed that F. nucleatum contributes to cancer progression via its secretion of extracellular vehicles (EVs). Hence, it's designed to reveal the influence of F. nucleatum-derived EVs (Fn-EVs) in GC progression. The tumor and adjacent tissues were collected from 30 GC patients, and the abundance of F. nucleatum was found to be highly expressed in tumor samples. The ultracentrifugation was employed to isolate EVs from F. nucleatum and Escherischia coli (E. coli), which were labeled Fn-EVs and E. coli-EVs, respectively. After treating GC cells with Fn-EVs and E. coli-EVs, cell counting kit 8, colony formation, wound healing as well as transwell assay were performed, which revealed that Fn-EVs effectively enhanced oxaliplatin resistance, and facilitated cell proliferation, migration, invasion, and stemness in GC cells while E. coli-EVs exert no significant effect on GC cells. Besides, the stemness and DNA repair of GC cells were also enhanced by Fn-EVs, as revealed by the sphere-forming assay and the detection of stemness- and DNA repair-associated proteins by western blotting. In vivo analyses demonstrated that Fn-EVs administration not only promoted GC tumor growth and liver metastasis but also conferred GC tumor resistance to oxaliplatin resistance. This study first revealed the contributive role of F. nucleatum in GC development via Fn-EVs, which provided a better perspective for manipulating F. nucleatum in treating GC patients with malignant phenotypes.

The many roles of cathepsins in restenosis.

Drug-eluting stents (DES) and dual antiplatelet regimens have significantly improved the clinical management of ischemic heart disease; however, the drugs loaded with DES in clinical practice are mostly paclitaxel or rapamycin derivatives, which target symptoms of post implantation proliferation and inflammation, leading to delayed re-endothelialization and neo-atherosclerosis. Along with the treatments already in place, there is a need for novel strategies to lessen the negative clinical outcomes of DES delays as well as a need for greater understanding of their pathobiological mechanisms. This review concentrates on the function of cathepsins (Cats) in the inflammatory response and granulation tissue formation that follow Cat-induced damage to the vasculature scaffold, as well as the functions of Cats in intimal hyperplasia, which is characterized by the migration and proliferation of smooth muscle cells, and endothelial denudation, re-endothelialization, and/or neo-endothelialization. Additionally, Cats can alter essential neointima formation and immune response inside scaffolds, and if Cats are properly controlled in vivo, they may improve scaffold biocompatibility. This unique profile of functions could lead to an original concept for a cathepsin-based coronary intervention treatment as an adjunct to stent placement.

Cathepsin S activity controls chronic stress-induced muscle atrophy and dysfunction in mice.

Exposure to chronic psychological stress (CPS) is an intractable risk factor for inflammatory and metabolic diseases. Lysosomal cysteinyl cathepsins play an important role in human pathobiology. Given that cathepsin S (CTSS) is upregulated in the stressed vascular and adipose tissues, we investigated whether CTSS participates in chronic stress-induced skeletal muscle mass loss and dysfunction, with a special focus on muscle protein metabolic imbalance and apoptosis. Eight-week-old male wildtype (CTSS+/+) and CTSS-knockout (CTSS-/-) mice were randomly assigned to non-stress and variable-stress groups. CTSS+/+ stressed mice showed significant losses of muscle mass, dysfunction, and fiber area, plus significant mitochondrial damage. In this setting, stressed muscle in CTSS+/+ mice presented harmful alterations in the levels of insulin receptor substrate 2 protein content (IRS-2), phospho-phosphatidylinositol 3-kinase, phospho-protein kinase B, and phospho-mammalian target of rapamycin, forkhead box-1, muscle RING-finger protein-1 protein, mitochondrial biogenesis-related peroxisome proliferator-activated receptor-γ coactivator-α, and apoptosis-related B-cell lymphoma 2 and cleaved caspase-3; these alterations were prevented by CTSS deletion. Pharmacological CTSS inhibition mimics its genetic deficiency-mediated muscle benefits. In C2C12 cells, CTSS silencing prevented stressed serum- and oxidative stress-induced IRS-2 protein reduction, loss of the myotube myosin heavy chain content, and apoptosis accompanied by a rectification of investigated molecular harmful changes; these changes were accelerated by CTSS overexpression. These findings demonstrated that CTSS plays a role in IRS-2-related protein anabolism and catabolism and cell apoptosis in stress-induced muscle wasting, suggesting a novel therapeutic strategy for the control of chronic stress-related muscle disease in mice under our experimental conditions by regulating CTSS activity.

Restoration of energy homeostasis under oxidative stress: Duo synergistic AMPK pathways regulating arginine kinases.

Rapid depletion of cellular ATP can occur by oxidative stress induced by reactive oxygen species (ROS). Maintaining energy homeostasis requires the key molecular components AMP-activated protein kinase (AMPK) and arginine kinase (AK), an invertebrate orthologue of the mammalian creatine kinase (CK). Here, we deciphered two independent and synergistic pathways of AMPK acting on AK by using the beetle Tribolium castaneum as a model system. First, AMPK acts on transcriptional factor forkhead box O (FOXO) leading to phosphorylation and nuclear translocation of the FOXO. The phospho-FOXO directly promotes the expression of AK upon oxidative stress. Concomitantly, AMPK directly phosphorylates the AK to switch the direction of enzymatic catalysis for rapid production of ATP from the phosphoarginine-arginine pool. Further in vitro assays revealed that Sf9 cells expressing phospho-deficient AK mutants displayed the lower ATP/ADP ratio and cell viability under paraquat-induced oxidative stress conditions when compared with Sf9 cells expressing wild-type AKs. Additionally, the AMPK-FOXO-CK pathway is also involved in the restoration of ATP homeostasis under oxidative stress in mammalian HEK293 cells. Overall, we provide evidence that two distinct AMPK-AK pathways, transcriptional and post-translational regulations, are coherent responders to acute oxidative stresses and distinguished from classical AMPK-mediated long-term metabolic adaptations to energy challenge.

Characterization of Glutamate-Gated Chloride Channel in Tribolium castaneum.

The glutamate-gated chloride channels (GluCls) play essential roles in signal transduction by regulating fast inhibitory synaptic transmission in the nervous system of invertebrates. While there is only one GluCl subunit in the insect, the diversity of insect GluCls is broadened by alternative splicing. In the present study, three TcGluCl variant genes were cloned from the red flour beetle Tribolium castaneum. Analysis of the characteristics of TcGluCls including sequence features, genomic structures, and alternative splicing revealed that TcGluCls had the typical structural features of GluCls and showed high homologies with the GluCls from other insect orders. The TcGluCl-encoding gene consists of nine exons and three variants (TcGluCl-3a, TcGluCl-3b, and TcGluCl-3c) were generated by the alternative splicing of exon 3, which was a highly conserved alternative splicing site in insect GluCls. Homology modeling of TcGluCl-3a showed that the exon 3 coding protein located at the N-terminal extracellular domain, and there were no steric clashes encountered between the exon 3 coding region and ivermectin/glutamate binding pocket, which indicated that the alternative splicing of exon 3 might have no impact on the binding of GluCls to glutamate and insecticide. In addition to the head tissue, TcGluCl-3a and TcGluCl-3c also had high expressions in the ovary and testis of T. castaneum, whereas TcGluCl-3b showed high expression in the midgut, suggesting the diverse physiological functions of TcGluCl variants in T. castaneum. The total TcGluCl and three variants showed the highest expression levels in the early stage larvae. The expressions of TcGluCl, TcGluCl-3b, and TcGluCl-3c were significantly increased from the late-stage larvae to the early stage pupae and indicated that the TcGluCl might be involved in the growth and development of T. castaneum. These results are helpful to further understand the molecular characteristics of insect GluCls and provide foundations for studying the specific function of the GluCl variant.

Cathepsin S deficiency improves muscle mass loss and dysfunction via the modulation of protein metabolism in mice under pathological stress conditions.

Cathepsin S (CTSS) is a widely expressed cysteinyl protease that has garnered attention because of its enzymatic and non-enzymatic functions under inflammatory and metabolic pathological conditions. Here, we examined whether CTSS participates in stress-related skeletal muscle mass loss and dysfunction, focusing on protein metabolic imbalance. Eight-week-old male wildtype (CTSS+/+ ) and CTSS-knockout (CTSS-/- ) mice were randomly assigned to non-stress and variable-stress groups for 2 weeks, and then processed for morphological and biochemical studies. Compared with non-stressed mice, stressed CTSS+/+ mice showed significant losses of muscle mass, muscle function, and muscle fiber area. In this setting, the stress-induced harmful changes in the levels of oxidative stress-related (gp91phox and p22phox ,), inflammation-related (SDF-1, CXCR4, IL-1ß, TNF-α, MCP-1, ICAM-1, and VCAM-1), mitochondrial biogenesis-related (PPAR-γ and PGC-1α) genes and/or proteins and protein metabolism-related (p-PI3K, p-Akt, p-FoxO3α, MuRF-1, and MAFbx1) proteins; and these alterations were rectified by CTSS deletion. Metabolomic analysis revealed that stressed CTSS-/- mice exhibited a significant improvement in the levels of glutamine metabolism pathway products. Thus, these findings indicated that CTSS can control chronic stress-related skeletal muscle atrophy and dysfunction by modulating protein metabolic imbalance, and thus CTSS was suggested to be a promising new therapeutic target for chronic stress-related muscular diseases.

Dauricine exhibits anti-inflammatory property against acute ulcerative colitis via the regulation of NF-κB pathway.

We aim to investigate the therapeutic effect of dauricine on ulcerative colitis (UC). Our results indicated that dauricine attenuated the reduction of colonic length, weight loss, disease activity index, colonic tissue damage, and inflammatory cytokine levels in dextran sulfate sodium mice. In addition, dauricine reduced lipopolysaccharide-induced inflammation in HT-29 cells. Mechanically, dauricine docked with p65, a member of nuclear transcription factor kappaB (NF-κB) family, through which reduced the inflammatory cytokine release from HT-29 cells. Together, the above results inferred that dauricine had therapeutic effect for UC by suppressing NF-κB pathway, which provided a promising mean for UC treatment.

Targeting the mitochondrial calcium uniporter inhibits cancer progression and alleviates cisplatin resistance in esophageal squamous cell carcinoma.

Cisplatin is the standard chemotherapeutic drug used for the treatment of esophageal squamous cell carcinoma (ESCC). Acquired cisplatin resistance is the primary obstacle to prolonging patient survival time. Here, the therapeutic effects of mitochondrial calcium uniporter (MCU) inhibition on tumor growth and cisplatin resistance in ESCC were assessed. MCU was stably overexpressed or knocked down in three ESCC cell lines and three cisplatin­resistant ESCC cell lines. Then, proliferation, migration, and mitochondrial membrane potential (MMP) were measured by colony formation, wound healing, Transwell, and JC­1 staining assays. MCU, MICU2, MICU1, and PD­L1 levels were detected through western blotting and immunofluorescence. ESCC and cisplatin­resistant ESCC xenograft mouse models were established. After MCU knockdown, tumor volume was measured. The expression levels of proliferation markers (CyclinD1 and Ki­67), MICU1/2, PD­L1, epithelial-mesenchymal transition (EMT) markers (vimentin, ß­catenin, and E­cadherin), and the angiogenesis marker CD34 were detected through western blotting, immunohistochemistry, or immunofluorescence. The results showed that MCU overexpression significantly promoted proliferation, migration, and MMP in ESCC cells and cisplatin­resistant ESCC cells. However, proliferation, migration, and MMP were suppressed following MCU knockdown. In ESCC cells, MCU overexpression markedly increased MICU2, MICU1, and PD­L1 levels, and the opposite results were observed when MCU was stably knocked down. Similarly, MCU inhibition decreased MICU2, MICU1, and PD­L1 expression in cisplatin­resistant ESCC cells. Moreover, MCU knockdown substantially decreased tumor growth, EMT, and angiogenesis in ESCC and cisplatin­resistant ESCC xenograft mice. Collectively, targeting MCU may inhibit cancer progression and alleviate cisplatin resistance in ESCC.

CTSS Modulates Stress-Related Carotid Artery Thrombosis in a Mouse FeCl3 Model.

BACKGROUND: Exposure to chronic psychological stress is a risk factor for metabolic cardiovascular disease. Given the important role of lysosomal CTSS (cathepsin S) in human pathobiology, we examined the role of CTSS in stress-related thrombosis, focusing on inflammation, oxidative stress, and apoptosis. METHODS: Six-week-old wild-type mice (CTSS+/+) and CTSS-deficient mice (CTSS-/-) randomly assigned to nonstress and 2-week immobilization stress groups underwent iron chloride3 (FeCl3)-induced carotid thrombosis surgery for morphological and biochemical studies. RESULTS: On day 14 poststress/surgery, stress had increased the lengths and weights of thrombi in the CTSS+/+ mice, plus harmful changes in the levels of PAI-1 (plasminogen activation inhibitor-1), ADAMTS13 (a disintegrin and metalloproteinase with thrombospondin type 13 motifs), and vWF (von Willebrand factor) and arterial tissue CTSS expression. Compared to the nonstressed CTSS+/+ mice, the stressed CTSS-/- mice had decreased levels of PAI-1, vWF, TNF (tumor necrosis factor)-α, interleukin-1ß, toll-like receptor-4, cleaved-caspase 3, cytochrome c, p16INK4A, gp91phox, p22phox, ICAM-1 (intercellular adhesion molecule-1), MCP-1 (monocyte chemoattractant protein-1), MyD88 (myeloid differentiation primary response 88), and MMP (matrix metalloproteinase)-2/-9 and increased levels of ADAMTS13, SOD (superoxide dismutase)-1/-2, eNOS (endothelial NO synthase), p-Akt (phospho-protein kinase B), Bcl-2 (B-cell lymphoma-2), p-GSK3α/ß (phospho-glycogen synthase kinases alpha and beta), and p-Erk1/2 (phospho-extracellular signal-regulated kinase 1 and 2) mRNAs and/or proteins. CTSS deletion also reduced the arterial thrombus area and endothelial loss. A pharmacological inhibition of CTSS exerted a vasculoprotective action. In vitro, CTSS silencing and overexpression, respectively, reduced and increased the stressed serum and oxidative stress-induced apoptosis of human umbilical vein endothelial cells, and they altered apoptosis-related proteins. CONCLUSIONS: CTSS inhibition appeared to improve the stress-related thrombosis in mice that underwent FeCl3-induction surgery, possibly by reducing vascular inflammation, oxidative stress, and apoptosis. CTSS could thus become a candidate therapeutic target for chronic psychological stress-related thrombotic events in metabolic cardiovascular disease.

Mechanisms underlying the effects of low concentrations of chlorantraniliprole on development and reproduction of the fall armyworm, Spodoptera frugiperda.

It is well known that sublethal dose of insecticides induces life history trait changes of both target and non-target insect species, however, the underlying mechanisms remain not well understood. In this study, the effects of low concentrations of the anthranilic diamide insecticide chlorantraniliprole on the development and reproduction of the fall armyworm (FAW), Spodoptera frugiperda, were evaluated, and the underlying mechanisms were explored. The results showed that exposure of FAW to LC10 and LC30 chlorantraniliprole prolonged the larvae duration, decreased the mean weight of the larvae and pupae, and lowered the pupation rate as well as emergence rate. The fecundity of female adults was also negatively affected by treatment with low concentrations of chlorantraniliprole. Consistently, we found that exposure of FAW to LC30 chlorantraniliprole downregulated the mRNA expression of juvenile hormone (JH) esterase (SfJHE), leading to the increase of JH titer in larvae. We also found that treatment with low concentrations of chlorantraniliprole suppressed the expression of ribosomal protein S6 kinase1 (SfS6K1) in female adults, resulting in the downregulation of the gene encoding vitellogenin (SfVg). These results provided insights into the mechanisms underlying the effects of low concentrations of insecticides on insect pests, and had applied implications for the control of FAW.

Comparative phosphoproteomics analysis provides insights into the responses of Chilo suppressalis to sublethal chlorantraniliprole exposure.

BACKGROUND: Sublethal exposure to insecticides causes changes in insect behaviors and physiologies including feeding, mobility, communication, hormone homeostasis, development and fecundity, however, the underlying molecular mechanisms were largely unclear. Our previous studies revealed that sublethal chlorantraniliprole exposure disturbed the hormone homeostasis, reduced the weight and longevity and prolonged the developmental duration of Chilo suppressalis. In the present study, the potential phosphorylation modification regulation mechanisms in C. suppressalis in response to sublethal chlorantraniliprole exposure were explored using comparative and quantitative phosphoproteomics. RESULTS: A total of 2640 phosphopeptides belonging to 1144 phosphoproteins were identified, among which 446 phosphopeptides derived from 303 unique phosphoproteins were differentially phosphorylated between the chlorantraniliprole-treated and control larvae. The phosphorylation levels of differentially phosphorylated phosphopeptides were further validated using parallel reaction monitoring (PRM). Functional classification and protein-protein interaction of the differentially phosphorylated proteins (DPPs) were analyzed. Generalized analysis of the DPPs and the differentially expressed genes (DEGs) identified in our previous study showed that sublethal chlorantraniliprole exposure significantly changed the transcription and phosphorylation levels of genes/proteins associated with carbohydrate and lipid metabolism, cytoskeleton, signal transduction, transcription, translation and post-translational modification, leading to the dysfunctions of energy metabolism, transcription regulation, protein synthesis and modification, and signal transduction in C. suppressalis. Further analysis of the phosphorylation motifs in DPPs revealed that the MAPKs, CDKs, CaMK II, PKA, PKC and CK II protein kinases might be directly responsible for the phosphoproteomics response of C. suppressalis to chlorantraniliprole treatment. CONCLUSION: Our results provide abundant phosphorylation information for characterizing the protein modification in insects, and also provide valuable insights into the molecular mechanisms of insect post-translational modifications in response to sublethal insecticide exposure. © 2023 Society of Chemical Industry.

The pleiotropic AMPK-CncC signaling pathway regulates the trade-off between detoxification and reproduction.

The association of decreased fecundity with insecticide resistance and the negative sublethal effects of insecticides on insect reproduction indicates the typical trade-off between two highly energy-demanding processes, detoxification and reproduction. However, the underlying mechanisms are poorly understood. The energy sensor adenosine monophosphate-activated protein kinase (AMPK) and the transcription factor Cap "n" collar isoform C (CncC) are important regulators of energy metabolism and xenobiotic response, respectively. In this study, using the beetle Tribolium castaneum as a model organism, we found that deltamethrin-induced oxidative stress activated AMPK, which promoted the nuclear translocation of CncC through its phosphorylation. The CncC not only acts as a transcription activator of cytochrome P450 genes but also regulates the expression of genes coding for ecdysteroid biosynthesis and juvenile hormone (JH) degradation enzymes, resulting in increased ecdysteroid levels as well as decreased JH titer and vitellogenin (Vg) gene expression. These data show that in response to xenobiotic stress, the pleiotropic AMPK-CncC signaling pathway mediates the trade-off between detoxification and reproduction by up-regulating detoxification genes and disturbing hormonal homeostasis.

Molecular Characterization of Spodoptera frugiperda Heme Oxygenase and Its Involvement in Susceptibility to Chlorantraniliprole.

The mammalian heme oxygenase (HO) plays an important role in cytoprotection against oxidative-stress-induced cell damage; however, functional characterization of insect HO is still limited. In this study, cDNA encoding a HO, named SfHO, was cloned from Spodoptera frugiperda. Analysis of the transcription level and enzymatic activity showed that exposure of the LC30 concentration of chlorantraniliprole to the third instar larvae significantly upregulated both the mRNA level and enzymatic activity of SfHO at 24 h after treatment. Further injection of the HO activator, hemin, into the third instar larvae led to the upregulation of SfHO as well as decreased susceptibility of S. frugiperda to chlorantraniliprole. Consistently, overexpression of SfHO increased the Sf9 cell viability under chlorantraniliprole treatment. Strikingly, both RNAi and the dual-luciferase reporter assay in Sf9 cells revealed that, unlike mammalian HO that is regulated by the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2), SfHO was not subject to the regulation by cap 'n' collar isoform C (CncC), the Nrf2 homologue in insects. These data provide insights into the function and regulatory mechanism of insect HOs and had applied implications for the control of S. frugiperda.

NLRP3 inflammasome activation contributes to the cognitive decline after cardiac surgery.

Background: Perioperative neurocognitive disorders (PND) are a common complication of cardiac surgery in elderly patients. The etiopathogenesis of PND is not clear. Nod-like receptor family pyrin domain containing 3 (NLRP3) inflammasome, a macromolecular protein complex, regulates inflammation by inducing the release of proinflammatory cytokines interleukin (IL)-1ß and IL-18. Studies have demonstrated a close link between the NLRP3 inflammasome and central nervous system diseases. Nevertheless, the involvement of NLRP3 inflammasome in the causation of PND occurring after cardiac surgery is unclear. This study aimed to investigate the association of serum NLRP3 level with PND. Methods: We performed a retrospective study, enrolled 75 patients undergoing elective cardiac surgery and evaluated their cognitive functions one day before and 7 days after surgery. PND were determined according to the International Study of Postoperative Cognitive Dysfunction studies. Demographics and perioperative parameters were recorded. Perioperative serum NLRP3 protein, IL-1ß, and IL-18 levels were monitored. Results: The PND incidence in our cohort was 33.33%. NLRP3 protein levels were significantly increased in all patients at each postoperative time-point after general anesthesia and cardiac surgery under cardiopulmonary bypass. Patients showing cognitive dysfunction had higher serum NLRP3 protein, caspase-1, IL-1ß, and IL-18 levels immediately after the operation. Variables associated with the incidence of early PND were included in the regression models. After adjusting for confounding variables, high serum NLRP3 protein level at the end of the operation and old age were identified as independent predictors of PND. Conclusions: High serum NLRP3 protein level at the completion of cardiac surgery was associated with a higher risk of PND seven days after surgery. Trial registration: The study was registered at Clinicaltrials.gov (registration number: NCT04191642).

RNAi-mediated knockdown of arginine kinase genes leads to high mortality and negatively affect reproduction and blood-feeding behavior of Culex pipiens pallens.

BACKGROUND: Arginine kinase (AK) is one of the crucial enzymes involved in energy metabolism in invertebrates, and has been proposed as the target for RNA interference (RNAi)-based control of agricultural insect pests. While there is only one AK gene in most insects, two AK genes were identified in Culex pipiens pallens, the primary vector of lymphatic filariasis and epidemic encephalitis. METHODS: The full-length cDNA sequences of CpAK1 and CpAK2 genes were obtained by reverse transcription PCR(RT-PCR) and rapid amplification of cDNA ends (RACE). The expression levels of CpAK1 and CpAK2 in different developmental stages and tissues were detected by reverse transcription quantitative PCR (RT-qPCR). The role of CpAK1 and CpAK2 in the reproduction and blood feeding behavior was analyzed using RNA interference (RNAi). RESULTS: Full-length cDNAs of CpAK1 and CpAK2 were isolated from Cx. pipiens pallens. Analysis of the expression pattern revealed that the mRNA level of CpAK1 was significantly higher than CpAK2 in all development stages and tissues examined, and the expressions of both CpAK1 and CpAK2 were upregulated in response to blood feeding. The co-knockdown of CpAK1 and CpAK2 mediated by RNAi led to high mortality (74.3%) of adult female mosquitoes and decreased hatchability (59.9%). Remarkably, the blood feeding rate and the engorgement rate of the female mosquitoes were negatively affected by co-injection of dsRNAs targeting CpAK1 and CpAK2. CONCLUSION: CpAK1 and CpAK2 were detected in all developmental stages and tissues, but showed divergence in expression level. RNAi-mediated knockdown of AK genes leads to high mortality and negatively affect blood-feeding behavior of Cx. pipiens pallens, suggesting that AK could be used for the target of RNAi-based mosquito control in the future.

Young bone marrow transplantation prevents aging-related muscle atrophy in a senescence-accelerated mouse prone 10 model.

BACKGROUND: Young bone marrow transplantation (YBMT) has been shown to stimulate vascular regeneration in pathological conditions, including ageing. Here, we investigated the benefits and mechanisms of the preventive effects of YBMT on loss of muscle mass and function in a senescence-associated mouse prone 10 (SAMP10) model, with a special focus on the role of growth differentiation factor 11 (GDF-11). METHODS: Nine-week-old male SAMP10 mice were randomly assigned to a non-YBMT group (n = 6) and a YBMT group (n = 7) that received the bone marrow of 8-week-old C57BL/6 mice. RESULTS: Compared to the non-YBMT mice, the YBMT mice showed the following significant increases (all P < 0.05 in 6-7 mice): endurance capacity (>61.3%); grip strength (>37.9%), percentage of slow myosin heavy chain fibres (>14.9-15.9%). The YBMT also increased the amounts of proteins or mRNAs for insulin receptor substrate 1, p-Akt, p-extracellular signal-regulated protein kinase1/2, p-mammalian target of rapamycin, Bcl-2, peroxisom proliferator-activated receptor-γ coactivator (PGC-1α), plus cytochrome c oxidase IV and the numbers of proliferating cells (n = 5-7, P < 0.05) and CD34+/integrin-α7+ muscle stem cells (n = 5-6, P < 0.05). The YMBT significantly decreased the levels of gp91phox, caspase-9 proteins and apoptotic cells (n = 5-7, P < 0.05) in both muscles; these beneficial changes were diminished by the blocking of GDF-11 (n = 5-6, P < 0.05). An administration of mouse recombinant GDF-11 improved the YBMT-mediated muscle benefits (n = 5-6, P < 0.05). Cell therapy with young bone marrow from green fluorescent protein (GFP) transgenic mice exhibited GFP+ myofibres in aged muscle tissues. CONCLUSIONS: These findings suggest that YBMT can prevent muscle wasting and dysfunction by mitigating apoptosis and proliferation via a modulation of GDF-11 signalling and mitochondrial dysfunction in SAMP10 mice.

Research trends of artificial intelligence in pancreatic cancer: a bibliometric analysis.

Purpose: We evaluated the related research on artificial intelligence (AI) in pancreatic cancer (PC) through bibliometrics analysis and explored the research hotspots and current status from 1997 to 2021. Methods: Publications related to AI in PC were retrieved from the Web of Science Core Collection (WoSCC) during 1997-2021. Bibliometrix package of R software 4.0.3 and VOSviewer were used to bibliometrics analysis. Results: A total of 587 publications in this field were retrieved from WoSCC database. After 2018, the number of publications grew rapidly. The United States and Johns Hopkins University were the most influential country and institution, respectively. A total of 2805 keywords were investigated, 81 of which appeared more than 10 times. Co-occurrence analysis categorized these keywords into five types of clusters: (1) AI in biology of PC, (2) AI in pathology and radiology of PC, (3) AI in the therapy of PC, (4) AI in risk assessment of PC and (5) AI in endoscopic ultrasonography (EUS) of PC. Trend topics and thematic maps show that keywords " diagnosis ", "survival", "classification", and "management" are the research hotspots in this field. Conclusion: The research related to AI in pancreatic cancer is still in the initial stage. Currently, AI is widely studied in biology, diagnosis, treatment, risk assessment, and EUS of pancreatic cancer. This bibliometrics study provided an insight into AI in PC research and helped researchers identify new research orientations.

A Practical Risk-Based Model for Early Warning of Seafarer Errors Using Integrated Bayesian Network and SPAR-H.

Unsafe crew acts (UCAs) related to human errors are the main contributors to maritime accidents. The prediction of unsafe crew acts will provide an early warning for maritime accidents, which is significant to shipping companies. However, there exist gaps between the prediction models developed by researchers and those adopted by practitioners in human risk analysis (HRA) of the maritime industry. In addition, most research regarding human factors of maritime safety has concentrated on hazard identification or accident analysis, but not on early warning of UCAs. This paper proposes a Bayesian network (BN) version of the Standardized Plant Analysis Risk-Human Reliability Analysis (SPAR-H) method to predict the probability of seafarers' unsafe acts. After the identification of performance-shaping factors (PSFs) that influence seafarers' unsafe acts during navigation, the developed prediction model, which integrates the practicability of SPAR-H and the forward and backward inference functions of BN, is adopted to evaluate the probabilistic risk of unsafe acts and PSFs. The model can also be used when the available information is insufficient. Case studies demonstrate the practicability of the model in quantitatively predicting unsafe crew acts. The method allows evaluating whether a seafarer is capable of fulfilling their responsibility and providing an early warning for decision-makers, thereby avoiding human errors and sequentially preventing maritime accidents. The method can also be considered as a starting point for applying the efforts of HRA researchers to the real world for practitioners.