The global nitrogen regulator GlnR is a direct transcriptional repressor of the key gluconeogenic gene pckA in actinomycetes.

In most actinomycetes, GlnR governs both nitrogen and non-nitrogen metabolisms (e.g., carbon, phosphate, and secondary metabolisms). Although GlnR has been recognized as a global regulator, its regulatory role in central carbon metabolism [e.g., glycolysis, gluconeogenesis, and the tricarboxylic acid (TCA) cycle] is largely unknown. In this study, we characterized GlnR as a direct transcriptional repressor of the pckA gene that encodes phosphoenolpyruvate carboxykinase, catalyzing the conversion of the TCA cycle intermediate oxaloacetate to phosphoenolpyruvate, a key step in gluconeogenesis. Through the transcriptomic and quantitative real-time PCR analyses, we first showed that the pckA transcription was upregulated in the glnR null mutant of Amycolatopsis mediterranei. Next, we proved that the pckA gene was essential for A. mediterranei gluconeogenesis when the TCA cycle intermediate was used as a sole carbon source. Furthermore, with the employment of the electrophoretic mobility shift assay and DNase I footprinting assay, we revealed that GlnR was able to specifically bind to the pckA promoter region from both A. mediterranei and two other representative actinomycetes (Streptomyces coelicolor and Mycobacterium smegmatis). Therefore, our data suggest that GlnR may repress pckA transcription in actinomycetes, which highlights the global regulatory role of GlnR in both nitrogen and central carbon metabolisms in response to environmental nutrient stresses. IMPORTANCE: The GlnR regulator of actinomycetes controls nitrogen metabolism genes and many other genes involved in carbon, phosphate, and secondary metabolisms. Currently, the known GlnR-regulated genes in carbon metabolism are involved in the transport of carbon sources, the assimilation of short-chain fatty acid, and the 2-methylcitrate cycle, although little is known about the relationship between GlnR and the TCA cycle and gluconeogenesis. Here, based on the biochemical and genetic results, we identified GlnR as a direct transcriptional repressor of pckA, the gene that encodes phosphoenolpyruvate carboxykinase, a key enzyme for gluconeogenesis, thus highlighting that GlnR plays a central and complex role for dynamic orchestration of cellular carbon, nitrogen, and phosphate fluxes and bioactive secondary metabolites in actinomycetes to adapt to changing surroundings.

Real Active Site Identification of Co/Co3O4 Anchoring Ni-MOF Nanosheets with Fast OER Kinetics for Overall Water Splitting.

Doping metals and constructing heterostructures are pivotal strategies to enhance the electrocatalytic activity of metal-organic frameworks (MOFs). Nevertheless, effectively designing MOF-based catalysts that incorporate both doping and multiphase interfaces poses a significant challenge. In this study, a one-step Co-doped and Co3O4-modified Ni-MOF catalyst (named Ni NDC-Co/CP) with a thickness of approximately 5.0 nm was synthesized by a solvothermal-assisted etching growth strategy. Studies indicate that the formation of the Co-O-Ni-O-Co bond in Ni NDC-Co/CP was found to facilitate charge density redistribution more effectively than the Co-O-Ni bimetallic synergistic effect in NiCo NDC/CP. The designating Ni NDC-Co/CP achieved superior oxygen evolution reaction (OER) activity (245 mV @ 10 mA cm-2) and robust long stability (100 h @ 100 mA cm-2) in 1.0 M KOH. Furthermore, the Ni NDC-Co/CP(+)||Pt/C/CP(-) displays pregnant overall water splitting performance, achieving a current density of 10 mA cm-2 at an ultralow voltage of 1.52 V, which is significantly lower than that of commercial electrolyzer using Pt/C and IrO2 electrode materials. In situ Raman spectroscopy elucidated the transformation of Ni NDC-Co to Ni(Co)OOH under an electric field. This study introduces a novel approach for the rational design of MOF-based OER electrocatalysts.

Minimal Coarse-Grained Models of Polar Solvent for Electrolytes: Stockmayer Versus Dumbbell.

This study explores the potential of the dumbbell solvent as a minimal model for understanding electrolyte solutions in polar solvents. Our investigation involves a comparative analysis of the dumbbell model and the Stockmayer model, focusing on ion solvation and ion-ion correlations. We examine electrolytes containing symmetric monovalent salts dissolved in polar solvents while varying the ion density and solvent polarity. Both models predict an augmented solvent coordination number around ions as the solvent polarity increases, with the dumbbell solvent displaying a more pronounced effect. Notably, radial distribution functions (RDFs) between solvent and ions yield differing trends; Stockmayer models exhibit a nonmonotonic relationship due to strong dipole-dipole interactions at higher polarity, while RDFs for ions and dumbbell solvents consistently rise. In response to increased solvent polarity, Stockmayer solvents within the ion's solvation shell undergo continuous dipole orientation shifts, whereas the dumbbell solvent predominantly adopts pointing-away dipole orientations, diminishing pointing-to orientations. This underscores the significance of the interplay between the solvent molecular orientation and dipole rotation. Both models qualitatively predict ion pairing and clustering behaviors across varying solvent dipole strengths and salt concentrations. The Stockmayer solvent generally provides stronger electrostatic screening than the dumbbell solvent due to its neglect of the coupling between molecular orientation and dipole rotation. What's more, at a high dipole moment regime, ion-ion correlations in Stockmayer solvent can become stronger with increasing dipole moment due to stronger solvent-solvent correlations. This study underscores the effectiveness of the dumbbell solvent model in systematically elucidating the fundamental principles governing electrolytes and offers potential applications in the rational design of electrolyte systems.

Self-etching assembly of designed NiFeMOF nanosheet arrays as high-efficient oxygen evolution electrocatalyst for water splitting.

2D metal-organic frameworks (MOFs) have emerged as potential candidates for electrocatalytic oxygen evolution reactions (OER) due to their inherent properties like abundant coordination unsaturated active sites and efficient charge transfer. Herein, a versatile and massively synthesizable self-etching assembly strategy wherein nickel-iron foam (NFF) acts as a substrate and a metal ion source. Specifically, by etching the nickel-iron foam (NFF) surface using ligands and solvents, Ni/Fe metal ions are activated and subsequently reacted under hydrothermal conditions, resulting in the formation of self-supporting nanosheet arrays, eliminating the need for external metal salts. The obtained 33 % NiFeMOF/NFF exhibits remarkable OER performance with ultra-low overpotentials of 188/231 mV at 10/100 mA cm-2 , respectively, outperforming most recently reported catalysts. Besides, the built 33 % NiFeMOF/NFF(+) ||Pt/C(-) electrolyzer presents low cell voltages of 1.55/1.83 V at 10/100 mA cm-2 , superior to the benchmark RuO2 (+) ||Pt/C(-) , implying good industrialization prospects. The excellent catalytic activity stems from the modulation of the electronic spin state of the Ni active site by the introduction of Fe, which facilitates the adsorption process of oxygen-containing intermediates and thus enhances the OER activity. This innovative approach offers a promising pathway for commercial-scale sustainable energy solutions.

Apelin ameliorates sepsis-induced myocardial dysfunction via inhibition of NLRP3-mediated pyroptosis of cardiomyocytes.

Sepsis-induced myocardial dysfunction (SMD) is the major cause of death in sepsis. Nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3)-mediated pyroptosis contributes to the occurrence and development of SMD. Although Apelin confers direct protection against SMD, the potential mechanisms remain unclear. This study aimed to determine whether Apelin protects against SMD via regulation of NLRP3-mediated pyroptosis of cardiomyocytes. Experimental SMD was induced in wild-type (WT) control mice and Apelin knockout (Apelin-/-) mice by cecal ligation and puncture (CLP). Neonatal mouse cardiomyocytes (NMCs) were treated with lipopolysaccharide (LPS) to simulate the physiological environment of SMD in vitro. The expression of Apelin was greatly decreased in the plasma from septic patients and septic mouse heart. Knockout of Apelin aggravated SMD, evidenced by decreased cardiac function, and increased cardiac fibrosis and NLRP3 inflammasome and pyroptosis levels in CLP-treated Apelin-/- mice compared with WT mice. Overexpression of Apelin activated the AMPK pathway and thereby inhibited NLRP3 inflammasome-mediated pyroptosis of NMCs induced by LPS in vitro These protective effects were partially abrogated by AMPK inhibitor. In conclusion, Apelin attenuated SMD by inhibiting NLRP3-mediated pyroptosis via activation of the AMPK pathway. Apelin may serve as a promising therapeutic target for SMD.

Precise Anchoring of Fe Sites by Regulating Crystallinity of Novel Binuclear Ni-MOF for Revealing Mechanism of Electrocatalytic Oxygen Evolution.

Bimetallic metal-organic framework (BMOF) exhibits better electrocatalytic performance than mono-MOF, but deciphering the precise anchoring of foreign atoms and revealing the underlying mechanisms at the atomic level remains a major challenge. Herein, a novel binuclear NiFe-MOF with precise anchoring of Fe sites is synthesized. The low-crystallinity (LC)-NiFe0.33 -MOF exhibited abundant unsaturated active sites and demonstrated excellent electrocatalytic oxygen evolution reaction (OER) performance. It achieved an ultralow overpotential of 230 mV at 10 mA cm-2 and a Tafel slope of 41 mV dec-1 . Using a combination of modulating crystallinity, X-ray absorption spectroscopy, and theoretical calculations, the accurate metal sequence of BMOF and the synergistic effect of the active sites are identified, revealing that the adjacent active site plays a significant role in regulating the catalytic performance of the endmost active site. The proposed model of BMOF electrocatalysts facilitates the investigation of efficient OER electrocatalysts and the related catalytic mechanisms.

The expression of CD86 in CD3+CD56+ NKT cells is associated with the occurrence and prognosis of sepsis-associated encephalopathy in sepsis patients: a prospective observational cohort study.

The role of CD3+CD56+ natural killer T (NKT) cells and its co-signaling molecules in patients with sepsis-associated encephalopathy (SAE) is unknown. In this prospective observational cohort study, we initially recruited 260 septic patients and eventually analyzed 90 patients, of whom 57 were in the SAE group and 37 were in the non-SAE group. Compared to the non-SAE group, 28-day mortality was significantly increased in the SAE group (33.3% vs. 12.1%, p = 0.026), while the mean fluorescence intensity (MFI) of CD86 in CD3+CD56+ NKT cells was significantly lower (2065.8 (1625.5 ~ 3198.8) vs. 3117.8 (2278.1 ~ 5349), p = 0.007). Multivariate analysis showed that MFI of CD86 in NKT cells, APACHE II score, and serum albumin were independent risk factors for SAE. Furthermore, the Kaplan-Meier survival analysis indicated that the mortality rate was significantly higher in the high-risk group than in the low-risk group (χ2 = 14.779, p < 0.001). This study showed that the decreased expression of CD86 in CD3+CD56+ NKT cells is an independent risk factor of SAE; thus, a prediction model including MFI of CD86 in NKT cells, APACHE II score, and serum albumin can be constructed for diagnosing SAE and predicting prognosis.

Design and discovery of new selective and potent VEGF receptor 2 tyrosine kinase inhibitors.

A series of novel substituted 4-anilinoquinazolines and their related compounds were designed and prepared by 3D modeling as potential inhibitors of VEGFR-2. Evaluation of VEGFR inhibitory activities suggested that compound I10 was a more potent (IC50 = 0.11 nM) VEGFR-2 inhibitor than most of the listed drugs. Kinase panel assays demonstrated that compound I10 was the selective VEGFR-2 inhibitor. The prediction of 3D modeling unveiled a unique binding mode of this lead compound to VEGFR-2. Compound I10 exhibited remarkable anti-angiogenesis and anti-proliferation in HUVEC at low nanomolar concentrations. PK studies indicated that the lead compound possessed adequate oral bioavailability in various species. In vivo subcutaneous tumor model demonstrated that oral administration of I10 demonstrated potent efficacy in inhibiting tumor growth and angiogenesis. All these results suggested compound I10 is a potential drug candidate for cancer treatment.

Efficacy of invisible advancement correction for mandibular retraction in adolescents based on Pancherz analysis.

BACKGROUND: Mandibular retraction is the main etiological mechanism of class II malocclusion in China and the subsequent distal molar relationship can cause functional discomfort in mastication, breathing and the temporomandibular joint. The use of mandibular advancement (MA) devices has recently emerged as an adolescent mandibular retraction treatment; however, current studies regarding the effect thereof are relatively few, and there is lack of sufficient clinical support. AIM: To investigate the clinical effect of invisalign MA on the treatment of mandibular retraction in adolescents. METHODS: This study included 30 adolescent patients who underwent treatment with the MA appliances from December 2017 to June 2021. The lateral cephalometric data before and after treatment were collected and imported into Dolphin Imaging software. The changes were measured by linear measurement superimposed with lateral cephalometric trajectory based on the Pancherz technology. RESULTS: There was no significant difference in the length and position of maxilla before and after the treatment. The position of the mandible moved 3.13 mm, the length increased 4.14 mm, the mandibular ramus length increased 4.09 mm, the body length increased 4.25 mm, and the position of the condyle moved 1.03 mm forward after treatment. Additionally, changes in the incisor sagittal position and labial inclination were observed. The position of the upper incisor point moved back 1.33 mm, without statistical difference, the inclination and tooth angle decreased by 3.44° and 4.06°, respectively; the position of the lower incisor point was moved 2.98 mm, and the inclination and tooth angle increased by 2.62° and 1.23°, respectively. Furthermore, changes in the incisor overjet and molar relationship were seen. Overjet decreased by 4.31 mm, of which 1.78 mm was due to dental factors, accounting for 41.3% of the effect as opposed to 58.7% due to skeletal factors. Molar relationship improved 3.87 mm, with 1.34 mm due to dental factors, and dental and skeletal factors were accounted for 34.6% and 65.4% of the effect, respectively. CONCLUSION: For adolescent patients with mandible retraction, invisalign MA can effectively promote the mandible growth, and it was proven to be mainly due to skeletal effects.

Assessment of mandibular retromolar space in adults with regard to third molar eruption status.

OBJECTIVE: The aim of this study was to compare the difference in length and width of the mandibular retromolar space (RMS) stratified by the different eruption and impaction statuses of the third molars in patients with skeletal Class I malocclusion. MATERIALS AND METHODS: The right mandibular RMS in 186 adult patients categorized according to the different statuses of the third molar was analyzed by using cone-beam computed tomography (CBCT). The shortest distances between the inner lingual cortex of the mandibular body and second molar root were measured parallel to the posterior occlusal line (POL) at depths of 2, 4, 6, 8, and 10 mm (mandibular retromolar space length in root level, RLin2,4,6,8,10) on the axial slices with the cementoenamel junction (CEJ) as the reference level. The width of the RMS and second molar root was measured vertical to the POL at the terminal point of the molar distalization at depths of 2, 4, 6, 8, and 10 mm (width of the mandibular retromolar space, BW2,4,6,8,10/ width of the second molar distal root, TW2,4,6,8,10) from the CEJ. RESULTS: RL in different measurement planes was 2.72 ± 2.22 ~ 3.74 ± 2.26 for Group A, 5.27 ± 1.68 ~ 9.10 ± 2.04 for Group B, 1.94 ± 2.34 ~ 5.71 ± 4.37 for Group C, 1.83 ± 2.95 ~ 5.05 ± 4.24 for Group D, and 5.93 ± 3.97 ~ 10.52 ± 2.16 for Group E. The BW measurement results for A ~ E group were 9.71 ± 1.41 ~ 10.51 ± 1.81, 9.83 ± 1.39 ~ 12.55 ± 2.11, 9.96 ± 1.21 ~ 12.17 ± 1.62, 9.82 ± 1.47 ~ 12.28 ± 2.77, and 10.02 ± 1.20 ~ 12.75 ± 0.82, respectively. There was no significant difference between men and women in any measurements (P > 0.05). Patients with normal third molars erupted and those vertically impacted possessed larger RMS lengths than those in which the third molars were missing, horizontally impacted or mesially impacted (P < 0.05). In each measurement plane, TW was significantly smaller than BW (P < 0.05). CONCLUSIONS: Sex had no effect on the length or width of the mandibular RMS. Different statuses of third molars can also differentially affect the mandibular RMS. The mandibular RMS width is not a limit for mandibular molar distalization. CLINICAL RELEVANCE: When considering the distalization of mandibular molars, more attention should be directed to the lingual cortex of the mandible, and CBCT scans are recommended for patients who require significant mandibular molar distalization. The mandible buccal shelf and retromolar area maybe a safe zone to insert the miniscrew for molar distalization.

Surgery-first treatment of a skeletal Class III malocclusion with a completely digital workflow.

Completely digital design/completely digital manufacturing (CDD/CDM) workflows have been widely used in orthodontic and orthognathic treatments. This case report introduces a CDD/CDM workflow consisting of clear aligners and virtual planning for a surgery-first approach (SFA) in a patient with a skeletal Class III malocclusion. Following a shortened treatment time of 5 months, the patient's facial appearance improved significantly, and well-balanced occlusion was obtained. SFAs with clear aligners can enable patients to achieve complete esthetic satisfaction during the therapeutic period. The CDD/CDM workflow provided accurate results, improved the clinical outcome, and reduced treatment time.

Stenting with high-intensity focused ultrasound ablation for distal biliary obstruction caused by pancreatic carcinoma: a meta-analysis.

Introduction: Between 42% and 77% of patients with distal malignant biliary obstruction (MBO) suffer from pancreatic carcinoma (PC). Aim: To analyse the clinical efficacy of stenting accompanied by high-intensity focused ultrasound (HIFU) ablation in patients with distal MBO from PC. Material and methods: Relevant articles published through March 2021 were identified in the Pubmed, Cochrane Library, Embase, Wanfang, VIP, and CNKI databases. RevMan v5.3 and Stata v12.0 were used for the meta-analysis. Results: Twenty-nine articles were initially identified, and 5 of these were eventually included. These articles described 142 patients who underwent biliary stenting alone and 132 patients who underwent biliary stenting with HIFU ablation. The pooled Δ total bilirubin (TBIL) values were comparable between the 2 treatment groups (p = 0.10). The pooled stent dysfunction rate was significantly greater in the group with stenting alone (p = 0.03), and the pooled HR for the stent patency duration indicated that the duration of stent patency was increased in the stenting with HIFU ablation group (p < 0.0001). Overall survival rates were significantly longer in the stenting with HIFU ablation group (p < 0.0001). HIFU ablation was associated with an 80% pooled clinical response rate. The pooled cholangitis (p = 0.47) and pancreatitis (p = 0.56) rates were comparable between the 2 groups. Funnel plots did not reveal any significant evidence of endpoint-associated publication bias. Conclusions: Stenting with HIFU ablation increased both stent patency and overall survival in patients with distal MBO caused by PC compared to stenting alone.

Combining a digital design-mediated surgery-first approach and clear aligners to treat a skeletal Class III defect for aesthetic purposes: a case report.

This case report introduces digital surgery-first approach orthognathic surgery assisted by three-dimensional virtual planning and combined with invisible orthodontic treatment for a 21-year-old female patient with a skeletal Class III high-angle gummy smile malocclusion. We explored the clinical significance of the widespread application of digital technology for rapid development of the orthodontic/orthognathic field. The regional acceleratory phenomenon and clear aligners were used to achieve fast and aesthetic tooth movement after surgery. The treatment lasted only 8 months, and the patient was satisfied with the aesthetic results. The results remained stable after 1 year of follow-up. This case report highlights the advantages of combining a digital design and a surgery-first approach to produce accurate, rapid, safe, stable, and fulfilling cosmetic results. The combination of the surgery-first approach and clear aligners can facilitate patient-oriented surgical orthodontic treatment.

NanoZnO-modified titanium implants for enhanced anti-bacterial activity, osteogenesis and corrosion resistance.

Titanium (Ti) implants are widely used in dentistry and orthopedics owing to their excellent corrosion resistance, biocompatibility, and mechanical properties, which have gained increasing attention from the viewpoints of fundamental research and practical applications. Also, numerous studies have been carried out to fine-tune the micro/nanostructures of Ti and/or incorporate chemical elements to improve overall implant performance. Zinc oxide nanoparticles (nano-ZnO) are well-known for their good antibacterial properties and low cytotoxicity along with their ability to synergize with a variety of substances, which have received increasingly widespread attention as biomodification materials for implants. In this review, we summarize recent research progress on nano-ZnO modified Ti-implants. Their preparation methods of nano-ZnO modified Ti-implants are introduced, followed by a further presentation of the antibacterial, osteogenic, and anti-corrosion properties of these implants. Finally, challenges and future opportunities for nano-ZnO modified Ti-implants are proposed.

Benign pathologies results from lung nodule percutaneous biopsies: How to differentiate true and false benign?

OBJECTIVES: The objective was to identify predictors of true negatives in lung nodules (LNs) with computed tomography-guided percutaneous biopsy (CTPB)-based benign pathological results. MATERIALS AND METHODS: We included 90 total patients between January 2013 and December 2017 that had CTPB-based nonspecific benign pathologies and used these patients as a training group to accurately identify true-negative predictors. A validation group of 50 patients from January 2018 to June 2019 to confirm predictor reliability. RESULTS: CTPB was conducted on 90 LNs from the training group. True-negative and false-negative CTPB-based pathologies were obtained for 79 and 11 LNs, respectively. CTPB-based benign results had a negative predictive value of 87.8% (79/90). Univariate and multivariate analyses revealed younger age (P = 0.019) and CTPB-based chronic inflammation with fibroplasia (P = 0.010) to be true-negative predictors. A predictive model was made by combining these two prognostic values as follows: score = -7.975 + 0.112 × age -2.883 × CTPB-based chronic inflammation with fibroplasia (0: no present; 1: present). The area under receiver operator characteristic (ROC) curve was 0.854 (P < 0.001). To maximize sensitivity and specificity, we selected a cutoff risk score of -0.1759. The application of this model to the validation group yielded an area under the ROC curve of 0.912 (P < 0.001). CONCLUSIONS: Our predictive model showed good predictive ability for identifying true negatives among CTPB-based benign pathological results.

Hierarchically hybrid biocoatings on Ti implants for enhanced antibacterial activity and osteogenesis.

Titanium (Ti) is widely applied as bone-anchoring implants in dental and orthopedic applications owing to its superior mechanical characteristics, high corrosion resistance, and excellent biocompatibility. Nevertheless, Ti-based implants with the deficiencies of insufficient osteoinduction and associated infections can result in implant failure, which significantly limits its applications in some cases. In this work, hierarchically hybrid biocoatings on Ti implants are developed by gradual incorporation of polydopamine (PDA), ZnO nanoparticles (nZnO), and chitosan (CS)/nanocrystal hydroxyapatite (nHA) via oxidative self-polymerization, nanoparticle deposition, solvent casting and evaporation methods for enhancing their antibacterial activity and osteogenesis. The modification of PDA on porous reticular Ti substrates greatly reduces the surface roughness, wettability, protein adsorption, and provides high adhesion to the deposited nZnO. Further, incorporating nZnO on PDA-coated Ti surfaces affects the surface structure and wettability, significantly inhibits the growth of both Staphylococcus aureus and Escherichia coli. Moreover, the CS/nHA-doped coating on the nZnO-modified Ti surfaces remarkably improves cytocompatibility and enhances the osteogenic differentiation of MC3T3-E1 cells by upregulating the protein expression of alkaline phosphatase. This work offers a promising alternative for developing Ti implants with long-lifetime bioactivity to achieve strong antibacterial ability and enhanced bone formation for potential dental/orthopedic applications.

Elevated intracranial pressure induces IL­1ß and IL­18 overproduction via activation of the NLRP3 inflammasome in microglia of ischemic adult rats.

Elevated intracranial pressure (ICP) is one of the most common complications following an ischemic stroke, and has implications for the clinical and neurological outcomes. The aim of the present study was to examine whether elevated ICP may increase IL­1ß and IL­18 secretion by activating the NOD­like receptor protein 3 (NLRP3) inflammasome in microglia of ischemic adult rats. Sprague­Dawley rats that underwent middle cerebral artery occlusion were used for assessment of ICP. Reactive oxygen species (ROS) production was detected, and western blotting and immunofluorescence staining were used to determine the expression levels of Caspase­1, gasdermin D­N domains (GSDMD­N), IL­1ß and IL­18 in microglial cells. ICP levels were significantly increased, which was accompanied by ROS overproduction, in the brain tissue following ischemia­reperfusion (IR) injury in rats. Treatment with 10% hypertonic saline by intravenous injection significantly reduced the ICP and ROS levels of the rats. Furthermore, high pressure (20 mmHg) combined with oxygen­glucose deprivation (OGD) treatment resulted in increased ROS production in BV­2 microglial cells compared with those subjected to OGD treatment alone in vitro. Elevated pressure upregulated the expression of Caspase­1, GSDMD­N, IL­18 and IL­1ß in IR­treated or OGD­treated microglia both in vivo and in vitro. More importantly, Caspase­1, GSDMD­N, IL­18 and IL­1ß expression in microglia was significantly downregulated when elevated pressure was reduced or removed. These results suggested that elevated ICP­induced IL­1ß and IL­18 overproduction via activation of the NLRP3 inflammasome by ischemia­activated microglia may augment neuroinflammation.

JAK/STAT signaling pathway-mediated microRNA-181b promoted blood-brain barrier impairment by targeting sphingosine-1-phosphate receptor 1 in septic rats.

BACKGROUND: Blood-brain barrier (BBB) impairment plays a significant role in the pathogenesis of sepsis-associated encephalopathy (SAE). However, the molecular mechanisms are poorly understood. In the present study, we aimed to investigate the regulatory relationship between the Janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathway, microRNA (miR)-181b and its target genes in sepsis in vivo and in vitro. METHODS: Four rat models (sham, sepsis, sepsis plus STAT3 inhibitor (Stattic), and sepsis plus miR-181b inhibitor [sepsis + anta-miR-181b]) were established. For the in vitro experiments, rat brain microvascular endothelial cells (rBMECs) and rat brain astrocytes (rAstrocytes) were cultured with 10% serum harvested from sham, sepsis, and sepsis + anta-miR-181b rats. Chromatin immunoprecipitation-quantitative polymerase chain reaction (ChIP-QPCR) analysis was carried out to detect the binding and enrichment of the JAK/STAT3 signal core transcription complex in the miR-181b promoter region. Dual-luciferase reporter gene assay was conducted to test miR-181b and its target genes. The cell adhesion rate of rBMECs was also measured. RESULTS: During our investigations, the expression levels of miR-181b, p-JAK2, p-STAT3, and C/EBPß were found to be significantly increased in the septic rats compared with the sham rats. STAT3 inhibitor halted BBB damage by downregulating the expression of miR-181b. In addition, miR-181b targeted sphingosine-1-phosphate receptor 1 (S1PR1) and neurocalcin delta (NCALD). The up-regulated miR-181b significantly decreased the cell adhesion rate of rBMECs. The administration of miR-181b inhibitor reduced damage to the BBB through increasing the expression of S1PR1 and NCALD, which again proved that miR-181b negatively regulates SIPR1 and NCALD to induce BBB damage. CONCLUSIONS: Our study demonstrated that JAK2/STAT3 signaling pathway induced expression of miR-181b, which promoted BBB impairment in rats with sepsis by downregulating S1PR1 and decreasing BBB cell adhesion. These findings strongly suggest JAK2/STAT3/miR-181b axis as therapeutic target in protecting against sepsis-induced BBB damage.

[Mechanism of resveratrol on ameliorating the cognitive dysfunction induced by sepsis associated encephalopathy in rats].

OBJECTIVE: To explore the mechanism of resveratrol on ameliorating the cognitive dysfunction induced by sepsis associated encephalopathy (SAE) in rats. METHODS: The 12 weeks old male Sprague-dawley (SD) male rats were randomly divided into sham group, sepsis group and resveratrol group, with 30 rats in each group. The rat model of sepsis was made by injecting LPS (10 mg/kg) into tail vein. The rats in sham group was given the same amount of normal saline (NS). After LPS injection, resveratrol (8 mg×kg-1×d-1) was intraperitoneally injected once daily for 2 days in the resveratrol group; the same amount of NS was given to the sepsis group and sham group. At 24 hours after model establishment, the cognitive function of the experimental rats was assessed by the Morris water maze test. The blood-brain barrier (BBB) permeability was evaluated by the brain water content (BWC) and Evans blue (EB) test. The protein expressions of matrix metalloproteinase 9 (MMP-9), Occludin and Claudin-5 in cortical tissue were detected by Western Blot. Double immunofluorescence was used to verify the co-localization of MMP-9 protein and the marker protein of astrocyte GFAP in the cortical tissue of rats. RESULTS: Compared with the sham group, the escape latency in the sepsis group was significantly longer [48-hour escape latency (s): 56.56±6.43 vs. 36.62±3.32, 72-hour escape latency (s): 57.72±7.23 vs. 26.46±4.24, both P < 0.01], the BWC and extravasation of EB were increased [BWC: (84.56±2.03)% vs. (76.82±2.22)%, EB (µg/g): 17.56±2.28 vs. 6.25±1.36, both P < 0.01], the expression of MMP-9 protein was increased (MMP-9/ß-actin: 0.73±0.01 vs. 0.24±0.01, P < 0.01), the protein expressions of Occludin and Claudin-5 were decreased (Occludin/ß-actin: 0.45±0.02 vs. 0.86±0.04, Claudin-5/ß-actin: 0.62±0.03 vs. 0.96±0.05, both P < 0.01). At the same time, the co-localization expression of MMP-9 protein and the astrocytes of the cortical were increased [MMP-9 fluorescence intensity (AU): 38.66±4.26 vs. 17.23±3.04, MMP-9 positive cells: (26.92±1.77)% vs. (12.82±1.46)%, both P < 0.01]. Compared with the sepsis group, the escape latency in resveratrol group was significantly shorter [48-hour escape latency (s): 41.42±6.27 vs. 56.56±6.43, 72-hour escape latency (s): 33.46±7.17 vs. 57.72±7.23, both P < 0.01], the BWC and extravasation of EB were decreased [BWC: (77.15±2.27)% vs. (84.56±2.03)%, EB (µg/g): 7.74±1.88 vs. 17.56±2.28, both P < 0.01], the expression of MMP-9 protein was decreased (MMP-9/ß-actin: 0.25±0.01 vs. 0.73±0.01, P < 0.01), the protein expressions of Occludin and Claudin-5 were increased (Occludin/ß-actin: 0.82±0.03 vs. 0.45±0.02, Claudin-5/ß-actin: 0.92±0.04 vs. 0.62±0.03, both P < 0.01). At the same time, the co-localization expression of MMP-9 protein and the astrocytes of the cortical were decreased [MMP-9 fluorescence intensity (AU): 19.44±4.37 vs. 38.66±4.26, MMP-9 positive cells: (13.11±1.29)% vs. (26.92±1.77)%, both P < 0.01]. CONCLUSIONS: Resveratrol can inhibit the expression of MMP-9 protein in the astrocytes of the cortical cortex of rats, and then reduce the degradation of tight junction proteins of Occludin and Claudin-5, thereby reducing BBB permeability and eventually ameliorate the cognitive dysfunction induced by SAE.

[Expression and significance of chemokines CCL21, E-selectins and Hsp90 in periodontal tissues of rats with experimental periodontitis].

PURPOSE: To investigate the expression and significance of chemokines CCL21, E-selectins and heat shock protein 90 (Hsp90) in periodontal tissues of rats with experimental periodontitis. METHODS: Forty 10-week-old male Wistar rats were significantly randomly divided into 4 groups with 10 rats in each group. Periodontitis models were established in groups A, B and C, and the rest were 10 blank control groups. Rats in group A, B and C were sacrificed at 4, 8 and 12 weeks after basic periodontal treatment, and the periodontal tissues of the first and second molars were taken for CCL21, E-selectins and Hsp90 protein expression detection. SPSS 25.0 software package was used to analyze the data. RESULTS: The levels of periodontal attachment in group A, B and C were higher than those in the control group(P<0.05). The levels of periodontal attachment, CCL21, E-selectins, Hsp90 mRNA and protein expression in periodontal tissues increased first and then decreased(P<0.05). The levels of periodontal attachment, CCL21, E-selectins, Hsp90 mRNA and protein expression in group B and C were significantly higher than those in group A(P<0.05). The levels of periodontal attachment, CCL21, E-selectins, Hsp90 and relative protein expression in periodontal tissues of group C were significantly lower than those of group B(P<0.05). CONCLUSIONS: The expression of CCL21, E-selectins and Hsp90 is up-regulated in periodontitis tissues. With local periodontal treatment, the expression level of CCL21, E-selectins and Hsp90 gradually decreases.