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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.

An attractive and convenient strategy for the direct acylation/annulation of indoles has been developed using Pd(0) as an efficient catalyst. The main feature of this protocol is the use of acyl chlorides as the acylating agents with the primary amine as the directing group. A variety of indolo[1,2-a]quinoxalines were readily obtained in reasonable efficiency and satisfactory yields with good functional group tolerance. Based on control experiments, a tentative catalytic mechanism was proposed.

Mycoviruses are viruses that infect fungi, and hypovirulence-associated mycoviruses have the potential to control fungal diseases. However, it is unclear how mycovirus-mediated hypovirulent strains live and survive in the field, and no mycovirus has been applied for field crop protection. In this study, we found that a previously identified small DNA mycovirus (SsHADV-1) can convert its host, Sclerotinia sclerotiorum, from a typical necrotrophic pathogen to a beneficial endophytic fungus. SsHADV-1 downregulates the expression of key pathogenicity factor genes in S. sclerotiorum during infection. When growing in rapeseed, the SsHADV-1-infected strain DT-8 significantly regulates the expression of rapeseed genes involved in defense, hormone signaling, and circadian rhythm pathways. As a result, plant growth is promoted and disease resistance is enhanced. Field experiments showed that spraying DT-8 at the early flowering stage can reduce the disease severity of rapeseed stem rot by 67.6% and improve yield by 14.9%. Moreover, we discovered that SsHADV-1 could also infect other S. sclerotiorum strains on DT-8-inoculated plants and that DT-8 could be recovered from dead plants. These findings suggest that the mycoviruses may have the ability to shape the origin of endophytism. Our discoveries suggest that mycoviruses may influence the origin of endophytism and may also offer a novel strategy for disease control in which mycovirus-infected strains are used to improve crop health and release mycoviruses into the field.

Introduction The aim of this study was to explore whether the antibrain edema of hypertonic saline (HS) is associated with alleviating ischemic blood‐brain barrier (BBB) permeability by downregulating astrocyte‐derived vascular endothelial growth factor (VEGF), which is mediated by microglia‐derived NOD‐like receptor protein 3 (NLRP3) inflammasome. Methods The infarct volume and BBB permeability were detected. The protein expression level of VEGF in astrocytes in a transient focal brain ischemia model of rats was evaluated after 10% HS treatment. Changes in the NLRP3 inflammasome, IL‐1β protein expression, and the interleukin‐1 receptor (IL1R1)/pNF‐кBp65/VEGF signaling pathway were determined in astrocytes. Results HS alleviated the BBB permeability, reduced the infarct volume, and downregulated the expression of VEGF in astrocytes. HS downregulates IL‐1β expression by inhibiting the activation of the NLRP3 inflammasome in microglia and then downregulates VEGF expression by inhibiting the phosphorylation of NF‐кBp65 mediated by IL‐1β in astrocytes. Conclusions HS alleviated the BBB permeability, reduced the infarct volume, and downregulated the expression of VEGF in astrocytes. HS downregulated IL‐1β expression via inhibiting the activation of the NLRP3 inflammasome in microglia and then downregulated VEGF expression through inhibiting the phosphorylation of NF‐кBp65 mediated by IL‐1β in astrocytes., Hypertonic saline inhibits activation of NLRP3 inflammasome in microglia to alleviate ischemic blood‐brain barrier permeability by downregulating astrocyte‐derived VEGF.

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.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, RLRL 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).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.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.

This study investigates the microscopic morphology of a hydraulic servo spool valve caused by particle erosion. The microscopic morphology of spool working edges and solid particle characteristics in oil of a typical hydraulic servo system were experimentally studied. The formation mechanism was analyzed using dynamic impact calculation. The results reveal that the erosion micro-morphology of the spool working edge was mainly characterized by surface peeling, notches, grooves, and chamfers. The contour of the working edge was rounded. The average and variance of the radius of the old spool’s working edges were obviously higher than those of the new spool. The main distribution range of solid particle size in typical servo-hydraulic system oil was 5–50 μm, and the number of particles decreased sharply with an increase in particle size, up to 90.89%. The actual shape of particles was close to an ellipsoid with strong erosion destructive power. When a single solid particle collided with the working edge, a large amount of stress was generated at the collision center point, and the stress can reach 1500 MPa when the pressure differential at the valve orifice was 3.5 MPa. The working edges were subjected to cyclic stress of high-frequency particle pulses, resulting in fatigue erosion.

Surface topography plays an important role in determining the performance of hydraulic components, which often leads to component failure. The dual nozzle electro-hydraulic servo valve is composed of torque motor, pilot nozzle flapper valve and servo spool valve, whereas the failure is at the level of servo spool valve. In more details, the degradation of the static characteristic parameters of the hydraulic servo valve, which is experimentally studied by artificially controlling the fillet radius of the spool working edges, is one of these failures. In addition, the microstructure characteristics of the MOOG J761 valve working edges and the circumferential distribution law of the fillet radius are obtained. The research shows that valve orifice erosion caused fillet on the working edges and increases the dispersion of the circumferential distribution. The erosion micromorphology of the working edge is mainly characterized by surface peeling, notches, grooves, and chamfers. The average and variance of the fillet of the spool working edge at the high-pressure oil exceed those of the working edge of the return oil. When the average fillet radius of the working edge increases by 2.71 times and the average fillet variance increases by 2.55 times, the maximum leakage increases by 5.08 times, the flow gain decreases, and the positive and negative flow gains are asymmetric, decreasing by 7.78% and 1.75%, respectively. The pressure gain decreases significantly by 40.30%. The relation between the microstructure of the orifice working edge and the performance degradation provides a new idea for the performance evaluation and manufacture of hydraulic spool servo valve.

Dental follicle stem cells (DFSCs), which are the stem cells present in the dental follicle tissue of the tooth germ and are derived from the neural crest, are direct precursor cells of periodontal tissues and can form the periodontal ligament, cementum, and alveolar bone proper in the late stage of tooth development. DFSCs have the ability to achieve osteogenic, adipogenic, chondrogenic, neural, and cardiomyocytic differentiation in a specific induced environment. Recently, isolated dental follicle epithelial stem cells from DFSCs were also found to have the ability to form salivary gland cells and ductal cells. In recent years, DFSCs have also been reported to play an active role in the treatment of inflammatory diseases and autoimmune diseases in animal models. Therefore, DFSCs could not only be used as a good seed source for tissue regeneration but also provide new treatment strategies for autoimmune diseases. The study of DFSCs in tissue regeneration and immunoregulation has developed from in vitro experiments to animal experiments. In this review, we will discuss recent advances in our understanding of DFSCs in tissue regeneration and immunomodulation. We expect DFSCs to be considered for stem cell-based therapies in the future.

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.

Background: In patients receiving anti-cancer chemotherapy, polyethylene glycolated recombinant human granulocyte-colony stimulating factor (PEG-rhG-CSF) was used for prophylaxis of chemotherapy-induced neutropenia. However, the side effect of PEG-rhG-CSF use on fasting blood glucose (FBG) level remains unclear. Materials and Methods: Cancer patients receiving chemotherapy and PEG-rhG-CSF were enrolled in our study. Baseline glucose (Glucose 1) was measured before PEG-rhG-CSF use, a second FBG test (Glucose 2) was performed after PEG-rhG-CSF use. Mean glucose levels were compared using t test. Results: The time interval between PEG-rhG-CSF use and the second glucose test was 2.4±1.5 days. The mean Glucose 1 was 5.18±0.53 mmol/L, and Glucose 2 was 3.80±1.13 mmol/L. Statistical analysis showed a significant difference between Glucose 1 and 2 existed (P

BackgroundThe NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) inflammasome has been identified as an important mediator of blood–brain-barrier disruption in sepsis-associated encephalopathy (SAE). However, no information is available concerning the critical upstream regulators of SAE.MethodsLipopolysaccharide (LPS) was used to establish an in vitro model of blood–brain barrier (BBB) disruption and an in vivo model of SAE. Disruption of BBB integrity was assessed by measuring the expression levels of tight-junction proteins. NLRP3 inflammasome activation, pro-inflammatory cytokines levels, and neuroapoptosis were measured using biochemical assays. Finally, the FITC-dextran Transwell assay and Evan’s blue dye assay were used to assess the effect of Maf1 on LPS-induced endothelial permeability in vitro and in vivo.ResultsWe found that Maf1 significantly suppressed the brain inflammatory response and neuroapoptosis induced by LPS in vivo and in vitro. Notably, Maf1 downregulated activation of the NF-κB/p65-induced NLRP3 inflammasome and the expression of pro-inflammatory cytokines. In addition, we found that Maf1 and p65 directly bound to the NLRP3 gene promoter region and competitively regulated the function of NLRP3 in inflammations. Moreover, overexpression of NLRP3 reversed the effects of p65 on BBB integrity, apoptosis, and inflammation in response to LPS. Our study revealed novel role for Maf1 in regulating NF-κB-mediated inflammasome formation, which plays a prominent role in SAE.ConclusionsRegulation of Maf1 might be a therapeutic strategy for SAE and other neurodegenerative diseases associated with inflammation.

To explore the mechanism of resveratrol on ameliorating the cognitive dysfunction induced by sepsis associated encephalopathy (SAE) in rats.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×kgCompared 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 P0.01], 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 P0.01], the expression of MMP-9 protein was increased (MMP-9/β-actin: 0.73±0.01 vs. 0.24±0.01, P0.01), the protein expressions of Occludin and Claudin-5 were decreased (Occludin/β-actin: 0.45±0.02 vs. 0.86±0.04, Claudin-5/β-actin: 0.62±0.03 vs. 0.96±0.05, both P0.01). 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 P0.01]. 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 P0.01], 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 P0.01], the expression of MMP-9 protein was decreased (MMP-9/β-actin: 0.25±0.01 vs. 0.73±0.01, P0.01), the protein expressions of Occludin and Claudin-5 were increased (Occludin/β-actin: 0.82±0.03 vs. 0.45±0.02, Claudin-5/β-actin: 0.92±0.04 vs. 0.62±0.03, both P0.01). 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 P0.01].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.

To investigate the effect of hypercapnia at admission on the clinical prognosis and the severity of infection in patients with severe community-acquired pneumonia (SCAP).The clinical data of 219 SCAP patients admitted to the department of emergency and critical care medicine of Guangdong Provincial People's Hospital from December 2017 to November 2019 were retrospectively analyzed. Based on the partial pressure of arterial carbon dioxide (PaCOCompared with the HO group (n = 68) and NC group (n = 72), the HC group (n = 79) had higher proportion of preexisting comorbid chronic obstructive pulmonary disease (COPD) and PSI score, lower PCT, CRP, IL-6, and pH values. Compared with the HO group and NC group, there were smaller improvement trends on the levels of WBC, PCT, CRP, IL-6, PaOHypercapnia at admission was associated with higher proportion of IMV and 28-day mortality in patients with SCAP, which may be related to its early suppression of inflammation and then increment of infection.

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 IL‑1β and IL‑18 secretion by activating the NOD‑like receptor protein 3 (NLRP3) inflammasome in microglia of ischemic adult rats. Sprague‑Dawley 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 Caspase‑1, gasdermin D‑N domains (GSDMD‑N), IL‑1β and IL‑18 in microglial cells. ICP levels were significantly increased, which was accompanied by ROS overproduction, in the brain tissue following ischemia‑reperfusion (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 oxygen‑glucose deprivation (OGD) treatment resulted in increased ROS production in BV‑2 microglial cells compared with those subjected to OGD treatment alone in vitro. Elevated pressure upregulated the expression of Caspase‑1, GSDMD‑N, IL‑18 and IL‑1β in IR‑treated or OGD‑treated microglia both in vivo and in vitro. More importantly, Caspase‑1, GSDMD‑N, IL‑18 and IL‑1β expression in microglia was significantly downregulated when elevated pressure was reduced or removed. These results suggested that elevated ICP‑induced IL‑1β and IL‑18 overproduction via activation of the NLRP3 inflammasome by ischemia‑activated microglia may augment neuroinflammation.

Rifamycin derivatives, such as rifampicin, have potent antibiotic activity and have long been used in the clinic as mainstay components for the treatment of tuberculosis, leprosy, and AIDS-associated mycobacterial infections. However, the extensive usage of these antibiotics has resulted in the rapid development of bacterial resistance. The resistance mechanisms mainly include mutations of the rifamycin target RNA polymerase of bacteria and enzymatic modifications of rifamycin antibiotics. One modification is the recently characterized rifamycin degradation catalyzed by Rox enzymes, which belong to the widely occurring flavin monooxygenases. Intriguingly, our recent sequence analysis revealed the rifamycin producers also encode Rox homologs that are not yet characterized. In this work, we expanded the study of the Rox-catalyzed rifamycin degradation. We first showed that the Rox proteins from rifamycin producers have the enzymatic rifamycin SV-degrading activity. Then we used the structurally diverse rifamycin compounds rifampicin and 16-demethylrifamycin W to probe the substrate scope and found that they each have a slightly different substrate scope. Finally, we demonstrated that Rox proteins can also catalyze the transformation of 16-demethylsalinisporamycin to 16-demethylsaliniketal A. Since 16-demethylsalinisporamycin and 16-demethylsaliniketal A are the counterpart analogs of salinisporamycin and saliniketal A, our biochemical findings not only uncover a previously uncharacterized self-resistance mechanism in the rifamycin producers, but also bridge the gap between the biosynthesis of the potential antitumor compound saliniketal A.

Background Sepsis is a serious systemic inflammatory response syndrome caused by infection, with an extremely high mortality rate. Peripheral blood mononuclear cells (PBMCs) played a key role in the immune response against infection, whose components and functions were altered radically in Sepsis. Here, we wondered to characterize the alteration of PBMCs in sepsis at the single-cell transcriptional level. Methods We isolated PBMCs from seven septic patients and four donors. Based on BD Rhapsody, PBMCs were generated by single-cell RNA sequencing, and cell types were clustered and named by unsupervised clustering and annotation analysis. Results PBMCs were profiled for 6 kinds of cell types, the biological properties of T cell and monocytes were shown in a detailed manner. We noticed that monocytes could be clustered into 6 subsets, with great heterogeneity in the alteration of composition, gene profile, and signaling pathways driven by sepsis. Moreover, the expression of representative genes was high associated with septic clinical indicators in clusters of monocytes, such as NEAT1. Conclusions Although the study was preliminary, we revealed sepsis-specific alteration of PBMCs and associated pathways. These results give a panoramic picture of PBMCs in composition, genes profiles, and pathway signatures that are driven by sepsis, which offers a unique perspective to understand disease progression or treatment in clinical practice.

Background The total survival rate in patients with acute aortic dissection (AAD) has been greatly improved because of surgical technique advances. However, the pre-operative mortality rate remained high. In this study, we sought to evaluate the effects of dexmedetomidine (DEX) on heart rate control and preoperative outcome in AAD. Methods Retrospectively enrolled 461 patients who were diagnosed with AAD during the first 7-day after admission and divided into two groups according to the use of intravenous DEX: DEX group (91 patients) and Control group (370 patients). The heart rate and systolic blood pressure (SBP) level in both groups were recorded, and the incidence of aortic dissection rupture and pre-operative survival rates within 7 days were considered as the primary clinical outcomes. Results Compared to the Control group, heart rate of DEX group in the early 3 hours was significantly higher (P=0.009), and the 24-hour heart rate fluctuation was smaller (P=0.012). There was no difference in the systolic blood pressure (SBP) between the two groups, but the 24-hour fluctuation of SBP in DEX group was less (P=0.003). We performed a propensity-matched analysis to minimize selection bias and found that there were 7 (7.9%) patients in the DEX group occurred acute pulmonary edema, 17 (19.1%) patients in the Control group (P=0.047). And the pre-operative survival rates within 7 days were significantly improved in DEX group (P=0.004). And the pre-operative survival rates within 7 days were significantly improved in DEX group (P=0.004). Conclusions DEX can be beneficial to facilitate heart rate control, keep SBP more steady, and reduce the incidence of pre-operative aortic rupture in patients with AAD.

Mechanically stretched skeletal muscle undergoes dramatic shifts in structure, mass, and function. In vitro tensile strain models have demonstrated that myogenic progenitor cells, including satellite cells and myoblasts, are highly mechanosensitive cells, and respond to mechanical strain in a wide variety of aspects. However, the experimental results from different researchers and laboratories are not always in support of each other. Moreover, some specific molecules or signaling pathways were reported to play distinct roles in stretched myogenic cells, according to the statements of different studies. The purpose of this review is to integrate the researches conducting in vitro culture of satellite cells or myoblasts and exploring their mechanoresponses using in vitro stretching apparatus. These responses will be categorized into several groups, such as activation, proliferation, myogenic differentiation, cellular damage or apoptosis, properties of plasma membrane, transdifferentiation, reorientation, etc. In addition, detailed experimental designs like culturing conditions and straining regimens will be displayed and compared, to interpret some contradictory statements in different studies. Furthermore, the currently known interconnections among some mechanosensitive pathways will be pictured to give a better understanding about the complex regulations of myogenic cell responses to mechanical stretch. Hopefully, by summarizing the published studies about mechanoresponses of myogenic progenitor cells, future directions, and perspectives would be made clearer to researchers in this field.

Botrytis cinerea, a ubiquitous necrotrophic plant-pathogenic fungus, is responsible for grey mold and rot disease in a very wide range of plant species. Subtilisin-like proteases (or subtilases) are a very diverse family of serine proteases present in many organisms and are reported to have a broad spectrum of biological functions. Here, we identified two genes encoding subtilisin-like proteases (Bcser1 and Bcser2) in the genome of B. cinerea, both of which contain an inhibitor I9 domain and a peptidase S8 domain. The expression levels of Bcser1 and Bcser2 increased during the sclerotial forming stage, as well as during a later stage of hyphal infection on Arabidopsis thaliana leaves, but the up-regulation of Bcser1 was significantly higher than that of Bcser2. Interestingly, deletion of Bcser1 had no effect on the fungal development or virulence of B. cinerea. However, deletion of Bcser2 or double deletion of Bcser1 and Bcser2 severely impaired the hyphal growth, sclerotial formation and conidiation of B. cinerea. We also found that ∆Bcser2 and ∆Bcser1/2 could not form complete infection cushions and then lost the ability to infect intact plant leaves of Arabidopsis and tomato but could infect wounded plant tissues. Taken together, our results indicate that the subtilisin-like protease Bcser2 is crucial for the sclerotial formation, conidiation, and virulence of B. cinerea.

Botrytis cinerea, a ubiquitous necrotrophic plant-pathogenic fungus, is responsible for grey mold and rot disease in a very wide range of plant species. Subtilisin-like proteases (or subtilases) are a very diverse family of serine proteases present in many organisms and are reported to have a broad spectrum of biological functions. Here, we identified two genes encoding subtilisin-like proteases (Bcser1 and Bcser2) in the genome of B. cinerea, both of which contain an inhibitor I9 domain and a peptidase S8 domain. The expression levels of Bcser1 and Bcser2 increased during the sclerotial forming stage, as well as during a later stage of hyphal infection on Arabidopsis thaliana leaves, but the up-regulation of Bcser1 was significantly higher than that of Bcser2. Interestingly, deletion of Bcser1 had no effect on the fungal development or virulence of B. cinerea. However, deletion of Bcser2 or double deletion of Bcser1 and Bcser2 severely impaired the hyphal growth, sclerotial formation and conidiation of B. cinerea. We also found that ∆Bcser2 and ∆Bcser1/2 could not form complete infection cushions and then lost the ability to infect intact plant leaves of Arabidopsis and tomato but could infect wounded plant tissues. Taken together, our results indicate that the subtilisin-like protease Bcser2 is crucial for the sclerotial formation, conidiation, and virulence of B. cinerea.

BACKGROUND: Acute type A aortic dissection (TAAD) is cardiovascular emergency and requires surgical interventions. In-hospital mortality rate of surgical-treated TAAD patients remains high. We aim to examine the prognostic implications of peri-operative parameters to identify high-risk patient for in-hospital mortality. METHODS: A total of 264 surgically treated TAAD patients were included in this study. The association between in-hospital mortality and peri-operative parameters were examined. RESULTS: Thirty patients (11.36%) died during hospitalization. Patients with higher Apache II score had a significantly higher rate of in-hospital mortality when compared with patients scored ≤20 in unadjusted model [Score 21–25: HR =12.9 (1.7–100.8), P=0.0148; Score >25: HR =94.5 (12.6–707.6), P120 mmHg, Cr >200 mmol/L (both at admission and after surgery), BUN >8.2 mmol/L (both at admission and after surgery), AST >80 µ/L, aortic cross-clamping time >120 min and cardiopulmonary bypass time (CPBT) >230 min were also significantly related to higher rate of in-hospital mortality in univariate analysis. In multivariable analysis, APACHE II score [Score 21–25: HR =9.5 (1.2–74.4), P=0.032; Score >25: HR =51.0 (6.7–387.7), P=0.0001], AST >80 µmol/L [HR =2.3 (1.1–4.8), P=0.0251], aortic cross-clamping time >120 min (HR =2.9 (1.1–7.7), P=0.0315) remained significant in predicting TAAD in-hospital mortality. CONCLUSIONS: APACHE II score could be a useful tool to predict TAAD in-hospital mortality. AST >80 µ/L and aortic cross-clamping time >120 min were also independent predictors.

Aims To explore the involvement of NOD-like receptor protein 3 (NLRP3) inflammasome and M1 macrophage in root resorption (RR). Methods A rat RR model was established by excessive orthodontic force. After different force-loading time, the expression levels of NLRP3, caspase-1, and interleukin-1β (IL-1β) and distribution of M1 macrophages were analysed by immunohistochemistry and immunofluorescence staining in vivo. Then, the mechanism of NLRP3 activation was further verified by macrophage and human periodontal ligament cell (hPDLC) co-culture system in vitro. The production levels of NLRP3, caspase-1, pro-caspase-1, and IL-1β in M1 macrophages in the co-culture system were detected by Western blot, and the polarization of CD68+IL-1β+ M1 macrophages was detected by immunofluorescence staining. Results In the rat RR model, NLRP3, caspase-1, IL-1β, and M1 macrophages were expressed in periodontal ligament, mainly concentrated around RR areas. Force-pre-treated hPDLCs promoted M1 macrophage polarization and the production of NLRP3, caspase-1, and IL-1β in M1 macrophages in co-culture system. When MCC950, an inhibitor of NLRP3 inflammasome, was added, NLRP3 activation and M1 macrophage polarization were inhibited. Conclusions In periodontal tissues, hPDLCs stimulated by force promoted M1 macrophage polarization and increased IL-1β production by activating NLRP3 inflammasome in M1 macrophages, thus initiating the occurrence of RR.

Refractory hypoxemia is the main symptom of acute respiratory distress syndrome (ARDS). Low tidal volume ventilation is routinely applied in clinical practice to correct hypoxemia, which aims to prevent ventilator‑induced lung injury. However, this ventilation strategy inevitably leads to hypercapnia. Our previous study demonstrated that hypercapnia aggravated cognitive impairment in hypoxemic rats; however, the underlying mechanism remains unclear. The aim of the present study was to investigate whether hypercapnia exacerbates the blood‑brain barrier (BBB) disruption through inducing interleukin (IL)‑1β overproduction in the blood of hypoxemic rats. The BBB permeability in a rat model of hypercapnia/hypoxemia was evaluated. The levels of IL‑1β in the blood of rats and human whole‑blood cultures were assessed. The expression of IL‑1 receptor 1 (IL‑1R1), phosphorylated IL‑1R1‑associated kinase (p‑IRAK‑1) and tight junctional proteins in cerebral vascular endothelial cells was examined in vitro and in vivo. In addition, IL‑1Ra, an IL‑1 receptor antagonist, was used to determine whether hypercapnia affects tight junctional protein expression in hypoxic cerebral vascular endothelial cells through inducing IL‑1β overproduction. It was observed that hypercapnia alone did not disrupt the BBB, but aggravated the damage to the BBB integrity in hypoxemic rats. Hypercapnia increased IL‑1β expression in the blood of hypoxemic rats as well as in hypoxic human whole‑blood cultures. IL‑1R1 and p‑IRAK‑1 expression was increased, while that of tight junctional proteins was reduced by hypercapnia in hypoxemic cerebral vascular endothelial cells in vitro and in vivo. Additionally, the expression of tight junctional proteins was markedly increased following treatment with IL‑1Ra. These results suggest that hypercapnia‑induced IL‑1β overproduction in the hypoxemic blood may decrease tight junctional protein expression in cerebrovascular endothelial cells via the IL‑1R1/p‑IRAK‑1 pathway, further disrupting BBB integrity, and eventually resulting in increased BBB permeability.

Hypercapnia in combination with hypoxemia is usually present in severe respiratory disease in the intensive care unit (ICU) and can lead to more severe cognitive dysfunction. Increasing evidence has indicated that the compromised blood–brain barrier (BBB) in the hippocampus in hypoxemia conditions can result in cognitive dysfunction. However, the role and underlying mechanism of hypercapnia in the BBB disruption remains poorly known. A rat model of hypercapnia was first established in this study by intubation and mechanical ventilation with a small-animal ventilator. After this, the cognitive function of the experimental rats was assessed by the Morris water maze test. The BBB permeability was evaluated by the Evans Blue (EB) test and brain water content (BWC). Western blot analysis was carried out to detect the protein expressions of total and nuclear hypoxia-inducible factor-1α (HIF-1α), matrixmetalloproteinase-9 (MMP-9) and Aquaporins-4 (AQP-4) in the hippocampus tissue. Double immunofluorescence further verified the protein expression of different biomarkers was localized in the astrocytes of the hippocampus. Hypercapnia alone did not disrupt the BBB, but it could further enhance the BBB permeability in hypoxemia. Concomitantly, up-regulation of nuclear HIF-1α, AQP-4, MMP-9 protein expression along with increased degradation of the occludin and claudin-5 proteins was found in the hypercapnia rat model, while the total HIF-1α remained unchanged. Interestingly, these changes were independent of the acidosis induced by hypercapnia. Of note, after premedication of 2-Methoxyestradiol (2ME2, an inhibitor of HIF-1α nuclear translocation), the disrupted BBB could be restored resulting in improvement of the cognitive impairment. Meanwhile, accumulation of nuclear HIF-1α, protein expression of AQP-4 and MMP-9 and protein degradation of the occludin and claudin-5 were decreased. Thus, our study demonstrated that hypercapnia can further disrupt the BBB through promoting HIF-1α nuclear translocation and up-regulation of AQP-4 and MMP-9 in hypoxemia. It is therefore suggested that the cascade of hypercapnia-induced nuclear HIF-1α protein translocation in hypoxia-activated astrocytes may be a potential target for ameliorating cognitive impairment. Electronic supplementary material The online version of this article (10.1007/s11064-020-03038-7) contains supplementary material, which is available to authorized users.

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/EBPβ 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.

Multi-unit parallel production system exists extensively in most enterprises, one repairman and spare unit are always matched to ensure the normal operation of the production system. However, it is a vital issue to optimize the preventive maintenance (PM) policy and operating unit number under the above condition. To this end, the paper oriented the multi-unit parallel production system and developed an optimization model that jointly considered the PM policy and the operating unit number. In the model, the spare unit is placed in an operational state during the period that one of the operating unit is preventively maintained. Then, a numerical example and some parameter sensitive analysises are also given to illustrate the proposed model. Results show the existence of the optimal results via minimizing the system long-running cost rate, indicate that a loss rate of the down time plays an important role for the optimal results and an optimal PM time exists which can minimize the system long-running cost rate.