Non-redundant functions of group 2 innate lymphoid cells.

Protective immunity relies on the interplay of innate and adaptive immune cells with complementary and redundant functions. Innate lymphoid cells (ILCs) have recently emerged as tissue-resident, innate mirror images of the T cell system, with which they share lineage-specifying transcription factors and effector machinery1. Located at barrier surfaces, ILCs are among the first responders against invading pathogens and thus could potentially determine the outcome of the immune response2. However, so far it has not been possible to dissect the unique contributions of ILCs to protective immunity owing to limitations in specific targeting of ILC subsets. Thus, all of the available data have been generated either in mice lacking the adaptive immune system or with tools that also affect other immune cell subsets. In addition, it has been proposed that ILCs might be dispensable for a proper immune response because other immune cells could compensate for their absence3-7. Here we report the generation of a mouse model based on the neuromedin U receptor 1 (Nmur1) promoter as a driver for simultaneous expression of Cre recombinase and green fluorescent protein, which enables gene targeting in group 2 ILCs (ILC2s) without affecting other innate and adaptive immune cells. Using Cre-mediated gene deletion of Id2 and Gata3 in Nmur1-expressing cells, we generated mice with a selective and specific deficiency in ILC2s. ILC2-deficient mice have decreased eosinophil counts at steady state and are unable to recruit eosinophils to the airways in models of allergic asthma. Further, ILC2-deficient mice do not mount an appropriate immune and epithelial type 2 response, resulting in a profound defect in worm expulsion and a non-protective type 3 immune response. In total, our data establish non-redundant functions for ILC2s in the presence of adaptive immune cells at steady state and during disease and argue for a multilayered organization of the immune system on the basis of a spatiotemporal division of labour.

Antiplatelet Agents Inhibit Platelet Adhesion and Aggregation on Glass Surface Under Physiological Flow Conditions: Toward a Microfluidic Platelet Functional Assay Without Additional Adhesion Protein Modification.

ABSTRACT: As the pathogenesis of arterial thrombosis often includes platelet adhesion and aggregation, antiplatelet agents are commonly used to prevent thromboembolic events. Here, a new microfluidic method without additional adhesion protein modification was developed to quantify the inhibitory effect of antiplatelet drugs on the adhesion and aggregation behavior of platelets on glass surfaces under physiological flow conditions. Polydimethylsiloxane-glass microfluidic chips were fabricated by soft photolithography. Blood samples from healthy volunteers or patients before and after taking antiplatelet drugs flowed through the microchannels at wall shear rates of 300 and 1500 second -1 , respectively. The time to reach 2.5% platelet aggregation surface coverage (Ti), surface coverage (A 150s ), and mean fluorescence intensity (F 150s ) were used as quantitative indicators. Aspirin (80 µM) prolonged Ti and reduced F 150s . Alprostadil, ticagrelor, eptifibatide, and tirofiban prolonged Ti and reduced A 150s and F 150s in a concentration-dependent manner, whereas high concentrations of alprostadil did not completely inhibit platelet aggregation. Aspirin combined with ticagrelor synergistically inhibited platelet adhesion and aggregation; GPIb-IX-von Willebrand factor inhibitors partially inhibited platelet aggregation, and the inhibition was more pronounced at 1500 than at 300 second -1 . Patient administration of aspirin or (and) clopidogrel inhibited platelet adhesion and aggregation on the glass surface under flow conditions. This technology is capable of distinguishing the pharmacological effects of various antiplatelet drugs on inhibition of platelet adhesion aggregation on glass surface under physiological flow conditions, which providing a new way to develop microfluidic platelet function detection method without additional adhesive protein modification for determining the inhibitory effects of antiplatelet drugs in the clinical setting.

Impact of rise and fall phases of shear on platelet activation and aggregation using microfluidics.

Blood flow disorders are often the result of the non-physiological narrowing of blood arteries caused by atherosclerosis and thrombus. The blood then proceeds through rising-peak-decreasing phases as it passes through the narrow area. Although abnormally high shear is known to activate platelets, the shear process that platelets undergo in small arteries is complex. Thus, understanding how each shear phase affects platelet activation can be used to improve antiplatelet therapy and decrease the risk of side effects like bleeding. Blood samples were sheared (68.8 ms,5200 s-1) in vitro by the microfluidic technique, and platelet activation levels (P-selectin and integrin αIIbß3) and von Willebrand factor (vWF) binding to platelets were analyzed by flow cytometry. Post-stenosis platelet aggregation was dynamically detected using microfluidic technology. We studied TXA2, P2Y12-ADP, and integrin αIIbß3-fibrinogen receptor pathways by adding antiplatelet drugs, such as acetylsalicylic acid (ASA, an active ingredient of aspirin that inhibits platelet metabolism), ticagrelor (hinders platelet activation), and tirofiban (blocks integrin αIIbß3 receptor) in vitro, respectively, to determine platelet activation function mediated by transient non-physiological high shear rates. We demonstrated that platelets can be activated under transient pathological high shear rates. The shear rise and fall phases influenced shear-induced platelet activation by regulating the binding of vWF to platelets. The degree of platelet activation and aggregation increased with multiple shear rise and fall phases. ASA did not inhibit shear-mediated platelet activation, but ticagrelor and tirofiban effectively inhibited shear-mediated platelet activation. Our data demonstrated that the shear rise and fall phases play an important role in shear-mediated platelet activation and promote platelet activation and aggregation in a vWF-dependent manner. Blocking integrin αIIbß3 receptor and hindering P2Y12-ADP were beneficial to reducing shear-mediated platelet activation.

The rapid change of shear rate gradient is beneficial to platelet activation.

Fluid shear plays a key role in hemostasis and thrombosis, and the purpose of this study was to investigate the effect of shear gradient change rate (SGCR) on platelet reactivity and von Willebrand factor (vWF) activity and its mechanism. In this study, we developed a set of microfluidic chips capable of generating different shear gradients and simulated the shear rate distribution in the flow field by COMSOL Multiphysics software. Molecular markers of platelet activation (P-selectin, activated GPIIb/IIIa, phosphatidylserine exposure, and monocyte-platelet aggregate formation) were analyzed by flow cytometry. Platelet aggregation induced by shear gradient was studied by a microfluidic experimental platform, and plasma vWF ristocetin cofactor (vWF: RCO) activity was investigated by flow cytometry. The expression of p-Akt was studied by Western blotting. The results showed that the faster the SGCR, the higher the expression of platelet p-Akt, and the stronger the platelet reactivity and vWF activity. This indicates that fluid shear stress can activate platelets and vWF in a shear gradient-dependent manner through the PI3K/AKT signal pathway, and the faster the SGCR, the more significant the activation effect.

What is the context? Recent studies have shown that fluid shear stress plays a key role in platelet activation and thrombosis. However, its mechanism and effect have not been fully elucidated.The development of microfluidic chip technology enables people to study platelet function in a precisely controlled flow field environment.Previous studies have shown that the PI3K-AKT signal pathway may be a mechanically sensitive signal transduction pathway.What is new?In this study, we designed a microfluidic model with different narrow geometry, and controlled the injection pump to perfuse fluid at the same flow rate, so that the platelets flowing through the model experienced the flow field environment of different shear gradients.We studied the activities of platelets and von Willebrand factor in different flow fields and explored their signal transduction pathways.What is the impact? Our results suggest that vascular stenosis does increase platelet activity and the risk of thrombosis. However, its ability to activate platelets is not only related to the peak shear rate and shear time, but also closely related to the decreasing rate of shear gradient. Even if the peak shear rate at the stenosis is the same, the faster the shear rate decreases, the higher the reactivity of platelets and von Willebrand factor, which may be mediated by the PI3K-AKT signal pathway. This study not only helps clinicians to judge the risk of thrombosis in patients with atherosclerosis or percutaneous coronary intervention, but also helps us to better understand the mechanism of shear-induced platelet activation.

Evaluating the impact of transient shear stress on platelet activation, adhesion, and aggregation with microfluidic chip technique.

BACKGROUND: When nonphysiological stenosis occurs, the transient high shear stress formed in vessels increases the risk of thrombosis and is a potential factor for cardiovascular diseases. But the platelet adhesion and aggregation behavior at nonphysiological post-stenosis and its affecting factors are not fully understood yet. METHODS: In this experiment, platelet aggregation on collagen and fibrinogen at different shear stresses and different hematocrits were observed by microfluidic technology. Platelet activation (P-selectin, glycoprotein IIb/IIIa) and monocyte-platelet aggregate (MPA) levels under different shear stresses were analyzed by flow cytometry. RESULTS: On fibrinogen, platelets aggregate more at higher shear stress conditions. While on collagen, it becomes more difficult for platelets to form stable aggregation at higher shear stress conditions. If platelets adhere initially at low shear stress, stable platelet aggregation can be formed at subsequent high shear stress. Moreover, when the shear stress increases, platelet activity markers (P-selectin, glycoprotein IIb/IIIa and MPAs) increase significantly. Hematocrit affects the degree of platelet aggregation, and the influence of hematocrit is obvious at high shear stress. CONCLUSION: Transient high shear stress (46 ms) can effectively activate platelets. Platelet aggregation behavior was different for coated fibrinogen and collagen protein. Stable platelet adhesion at post-stenosis is more dependent on fibrinogen and platelet aggregation is stable on both fibrinogen and collagen. Hematocrit can significantly affect the formation of platelet aggregation.

Clarifying the relationship between insecure attachment and problematic social media use across platforms: a network analysis.

BACKGROUND: Despite evidence suggesting that insecure attachment is a significant risk factor for Problematic Social Media Use (PSMU), there remains a lack of comprehensive studies exploring this relationship, and a unified understanding of its role has yet to be established. METHODS: We employed network analysis to construct an integrated model for examining the complex interrelations between negative emotions, trait and state attachment, motives, and PSMU across three platforms (i.e., WeChat, Sina Weibo, and TikTok), as well as for identifying potential mediating variables between attachment and PSMU. Data were collected from 685 young adults via online self-reported questionnaires. RESULTS: We found that negative emotions are positively correlated with insecure trait and state attachment but have a negligible direct relationship with PSMU. The conformity motive and state attachment security emerged as important central nodes when measured by strength, closeness, and betweenness. Moreover, attachment states and motives were found to be clustered. Such strong interrelationships were also evident between insecure attachment and PSMU, while trait attachment anxiety and avoidance were observed to be related to PSMU across various platforms. CONCLUSIONS: Our findings promote a deeper understanding of the relationship between insecure attachment and PSMU from a cross-platform perspective and offer novel insights into the mechanisms underlying their co-occurrence, which may guide the development of effective interventions for healthier social media engagement.

Association of night-time sleep and daytime napping with painful temporomandibular disorder.

BACKGROUND: Painful temporomandibular disorder (TMD) is the common cause of chronic oro-facial pain, which may interfere with sleep. Previous studies have documented an association between sleep and TMD. OBJECTIVES: This study aimed to further explore the association of night-time sleep and daytime napping with painful TMD. METHODS: A total of 419 patients (aged 31.88 ± 11.54 years with women forming 85.4%) from a TMD/Orofacial Pain center were enrolled. Patients' sleep conditions were evaluated with the Pittsburgh Sleep Quality Index (PSQI) questionnaire, and information on night-time sleep duration, napping duration and napping frequency was interviewed. TMD was diagnosed according to the Diagnostic Criteria for TMD protocol and stratified into myalgia (muscle pain), arthralgia (joint pain) and combined (muscle and joint pain) subgroups. The severity of TMD was measured with the Fonseca Anamnestic Index (FAI) questionnaire. Restricted cubic spline (RCS) regression models were established to explore relationships between sleep and painful TMD subgroups. RESULTS: Patients with poor sleep quality (PSQI≥6) had higher FAI scores (median 60, p < .001) and higher proportions of painful TMDs. The myalgia subgroup had higher PSQI scores (median 8, p < .001) than the arthralgia subgroup. The RCS models indicated a non-linear relationship between night-time sleep duration and myalgia (p < .001), which was not observed in arthralgia. However, there were no significant findings concerning napping and painful TMD subgroups. CONCLUSION: This study found that the association between sleep and TMD is mainly related to painful TMD conditions, which are associated with night-time sleep duration.

Orthodontic extrusion with fixed appliances for treatment of intrusive luxation injuries: A prospective study of 28 permanent maxillary incisors.

BACKGROUND: Limited evidence exists on the treatment options of tooth repositioning after intrusive luxation. AIM: The study aimed to investigate the outcomes and complications of orthodontic extrusion in treating intruded maxillary permanent incisors. DESIGN: A prospective study was conducted involving 28 intruded maxillary permanent incisors treated with orthodontic extrusion, compared with a retrospective control group of 29 teeth that underwent spontaneous re-eruption. The success rate of tooth repositioning, as well as pulp condition, periodontal healing, and root development were assessed and compared. RESULTS: The success rate of orthodontic extrusion was 96.4%, excluding one tooth that was ankylosed before treatment. There were no significant differences in pulp condition between the orthodontic extrusion and control groups for teeth with immature root development. Teeth with mature root development in the orthodontic group, however, showed a significantly higher rate of pulp necrosis (100%, p < .05). Periodontal healing outcomes were similar across both groups, regardless of the maturity of root development. The root length continued increasing during orthodontic extrusion treatment. CONCLUSIONS: Orthodontic extrusion treatment could effectively reposition moderately to severely intrusive permanent incisors, without increasing the risk of complications compared with spontaneous re-eruption.

Clinical features of non-syndromic late developing supernumerary teeth: a sign of the third dentition?

OBJECTIVES: This study aimed to summarize the clinical features of non-syndromic late developing supernumerary teeth (LDST) and comparisons with common supernumerary teeth (ST) and explore the association between LDST and the third dentition. MATERIALS AND METHODS: This study retrospected cone-beam computed tomography (CBCT) and medical history of 41,903 consecutive patients from January to December 2021. Comparisons between ST and LDST were evaluated by Chi-square test or Fisher exact test. Correlation between chronological age and dental stage age was evaluated by Spearman's rank correlation coefficient. Binary logistic regression analysis was used to explore the features of LDST originating from the third dentition. RESULTS: Sixty patients with 126 non-syndromic LDST and 1602 patients with 1988 non-syndromic ST were identified. The prevalence of ST and LDST was 3.82% and 0.14%, respectively, with a male-female ratio of 1.78:1 and 1.31:1. LDST patients mainly had LDST in multiple (58.33%) and bilaterally (41.67%), with an average of 2.1/patient. Most LDST were normal-shaped (84.13%), vertically oriented (71.43%), located in the mandible (80.16%), and distributed in the premolar region (82.54%). The study also indicated that the development of LDST was correlated with permanent teeth, with LDST developing 6.48 to 10.45 years later. In this study, 72.22% of LDST met the clinical criteria for the third dentition. CONCLUSIONS: LDST manifested different clinical features from common ST. LDST might be closely related to the third dentition. CLINICAL RELEVANCE: This work would help to comprehend LDST from a clinical perspective, and may be complementary to the criteria of the third dentition.

DENTAL AND SKELETAL CHANGES OF LONG-TERM USE OF MANDIBULAR ADVANCEMENT DEVICES FOR THE TREATMENT OF ADULT OBSTRUCTIVE SLEEP APNEA: A SYSTEMATIC REVIEW AND META-ANALYSIS.

BACKGROUND: Mandibular advancement devices (MADs) are indicated for use in patients with mild to moderate obstructive sleep apnea (OSA). Long-term use of MADs has been found to be associated with dental and skeletal changes. This study aims to conduct a systematic review and meta-analysis to improve knowledge about the dental and skeletal changes of long-term (>1 year) use of MADs for the treatment of OSA. MATERIAL AND METHODS: Electronic databases were systematically searched. Two reviewers conducted screening, quality assessment, and data extraction independently. Thirty-four studies were included in the systematic review and 23 in the meta-analysis. RESULTS: The mean change of overjet and overbite was -0.77mm (95%CI -1.01 to -0.53, P < .00001) and -0.64mm (95%CI -0.85 to -0.43, P < .00001), with progressive change over the treatment duration. The inclination of the upper incisor (U1/SN) and the lower incisor (L1/MP) showed a mean change of retroclined -2.10° (95%CI -3.93 to -0.28, P = .02) and proclined 1.78° (95%CI 0.63 to 2.92, P = .002), respectively. The mean change of the anteroposterior position of the mandible (SNB) was -0.33° posteriorly (95%CI -0.65 to -0.02, P = .04). CONCLUSIONS: The meta-analysis showed a gradual decrease in overjet and overbite with treatment duration with long-term use of MADs for the treatment of OSA. Upper and lower incisors retroclined and proclined, respectively. The skeletal changes might include the mandibular position. Patients treated with MADs need to be continuously monitored over time.