YTHDF2-regulated matrilin-3 mitigates post-reperfusion hemorrhagic transformation in ischemic stroke via the PI3K/AKT pathway.

Hemorrhagic transformation can complicate ischemic strokes after recanalization treatment within a time window that requires early intervention. To determine potential therapeutic effects of matrilin-3, rat cerebral ischemia-reperfusion was produced using transient middle cerebral artery occlusion (tMCAO); intracranial hemorrhage and infarct volumes were assayed through hemoglobin determination and 2,3,5-triphenyltetrazoliumchloride (TTC) staining, respectively. Oxygen-glucose deprivation (OGD) modeling of ischemia was performed on C8-D1A cells. Interactions between matrilin-3 and YTH N6-methyladenosine RNA binding protein F2 (YTHDF2) were determined using RNA immunoprecipitation assay and actinomycin D treatment. Reperfusion after tMCAO modeling increased hemorrhage, hemoglobin content, and infarct volumes; these were alleviated by matrilin treatment. Matrilin-3 was expressed at low levels and YTHDF2 was expressed at high levels in ischemic brains. In OGD-induced cells, matrilin-3 was negatively regulated by YTHDF2. Matrilin-3 overexpression downregulated p-PI3K/PI3K, p-AKT/AKT, ZO-1, VE-cadherin and occludin, and upregulated p-JNK/JNK in ischemic rat brains; these effects were reversed by LY294002 (a PI3K inhibitor). YTHDF2 knockdown inactivated the PI3K/AKT pathway, inhibited inflammation and decreased blood-brain barrier-related protein levels in cells; these effects were reversed by matrilin-3 deficiency. These results indicate that YTHDF2-regulated matrilin-3 protected ischemic rats against post-reperfusion hemorrhagic transformation via the PI3K/AKT pathway and that matrilin may have therapeutic potential in ischemic stroke.

Neurocircuitry underlying the antidepressant effect of retrograde facial botulinum toxin in mice.

BACKGROUNDS: Botulinum toxin type A (BoNT/A) is extensively applied in spasticity and dystonia as it cleaves synaptosome-associated protein 25 (SNAP25) in the presynaptic terminals, thereby inhibiting neurotransmission. An increasing number of randomized clinical trials have suggested that glabellar BoNT/A injection improves depressive symptoms in patients with major depressive disorder (MDD). However, the underlying neuronal circuitry of BoNT/A-regulated depression remains largely uncharacterized. RESULTS: Here, we modeled MDD using mice subjected to chronic restraint stress (CRS). By pre-injecting BoNT/A into the unilateral whisker intrinsic musculature (WIM), and performing behavioral testing, we showed that pre-injection of BoNT/A attenuated despair- and anhedonia-like phenotypes in CRS mice. By applying immunostaining of BoNT/A-cleaved SNAP25 (cl.SNAP25197), subcellular spatial localization of SNAP25 with markers of cholinergic neurons (ChAT) and post-synaptic membrane (PSD95), and injection of monosynaptic retrograde tracer CTB-488-mixed BoNT/A to label the primary nucleus of the WIM, we demonstrated that BoNT/A axonal retrograde transported to the soma of whisker-innervating facial motoneurons (wFMNs) and subsequent transcytosis to synaptic terminals of second-order neurons induced central effects. Furthermore, using transsynaptic retrograde and monosynaptic antegrade viral neural circuit tracing with c-Fos brain mapping and co-staining of neural markers, we observed that the CRS-induced expression of c-Fos and CaMKII double-positive neurons in the ventrolateral periaqueductal grey (vlPAG), which sent afferents to wFMNs, was down-regulated 3 weeks after BoNT/A facial pre-administration. Strikingly, the repeated and targeted silencing of the wFMNs-projecting CaMKII-positive neurons in vlPAG with a chemogenetic approach via stereotactic injection of recombinant adeno-associated virus into specific brain regions of CRS mice mimicked the antidepressant-like action of BoNT/A pre-treatment. Conversely, repeated chemogenetic activation of this potential subpopulation counteracted the BoNT/A-improved significant antidepressant behavior. CONCLUSION: We reported for the first time that BoNT/A inhibited the wFMNs-projecting vlPAG excitatory neurons through axonal retrograde transport and cell-to-cell transcytosis from the injected location of the WIM to regulate depressive-like phenotypes of CRS mice. For the limited and the reversibility of side effects, BoNT/A has substantial advantages and potential application in MDD.

(+)-Borneol inhibits the generation of reactive oxygen species and neutrophil extracellular traps induced by phorbol-12-myristate-13-acetate.

Background and purpose: Neutrophil extracellular traps (NETs) are special web-like structures that can be generated in both infectious and noninfectious diseases. Previous studies showed that reactive oxygen species (ROS) were crucial in the formation of NETs (NETosis). The purpose of this study is to evaluate the effect of (+)-borneol, an antioxidant, on NETosis. Methods: Human neutrophils were stimulated with phorbol-12-myristate-13-acetate (PMA) to induce NETosis in vitro. Neutrophils treated with (+)-borneol at three different time points (-30 min, 0, and 30 min) associated with PMA stimulation were used to examine the effect of (+)-borneol on the formation of NETs. The ROS generation of neutrophils was also measured to explore the potential mechanism of the inhibitory effect of (+)-borneol on NETosis. Results: (+)-Borneol pretreatment inhibited NETosis induced by PMA. Immunofluorescence staining visualized and confirmed the inhibitory effect. (+)-Borneol inhibited the burst of ROS in neutrophils caused by PMA. Suppressing NADPH oxidase or protein kinase C (PKC) eliminated the effect of (+)-borneol on NETosis. Moreover, inhibiting Toll-like receptor 2 (TLR2) led to increased NETosis which can be inhibited by (+)-borneol. Conclusion: (+)-Borneol decreases the ROS level in activated neutrophils and inhibits NETosis triggered by PMA stimulation in vitro. (+)-Borneol therapy may be effective in some NET-dependent conditions.

Enamel Matrix Derivative Enhances the Odontoblastic Differentiation of Dental Pulp Stem Cells via Activating MAPK Signaling Pathways.

The odontoblastic differentiation of dental pulp stem cells (DPSCs) contributes to pulp-dentin regeneration. Enamel matrix derivative (EMD) is considered to be a critical epithelial signal to induce cell differentiation during odontogenesis and has been widely applied to clinical periodontal tissue regeneration. The purpose of this study was to explore the effect of EMD on DPSCs proliferation and odontoblastic differentiation, as well as the underlying mechanisms. We conducted in vitro and in vivo researches to get a comprehensive understanding of EMD. In vitro phase: cell proliferation was assessed by a cell counting kit-8 (CCK-8) assay; then, alkaline phosphatase (ALP) activity and staining, alizarin red staining, real-time RT-PCR, and western blot analysis were conducted to determine the odontoblastic potential and involvement of MAPK signaling pathways. In vivo phase: after ensuring the biocompatibility of VitroGel 3D-RGD via scanning electron microscopy (SEM), the hydrogel mixture was subcutaneously injected into nude mice followed by histological and immunohistochemical analyses. The results revealed that EMD did not interfere with DPSCs proliferation but promoted the odontoblastic differentiation of DPSCs in vitro and in vivo. Furthermore, blocking the MAPK pathways suppressed the EMD-enhanced differentiation of DPSCs. Finally, VitroGel 3D-RGD could well support the proliferation, differentiation, and regeneration of DPSCs. Overall, this study demonstrates that EMD enhances the odontoblastic differentiation of DPSCs through triggering MAPK signaling pathways. The findings provide a new insight into the mechanism by which EMD affects DPSCs differentiation and proposes EMD as a promising candidate for future stem cell therapy in endodontics.

Nitrous oxide-related neurological disorders: Clinical, laboratory, neuroimaging, and electrophysiological findings.

BACKGROUND: Recreational N2 O abuse is an important etiology of neurological impairment in young patients, which may easily be ignored clinically. Few current studies have investigated the characteristics or the effects experienced by its users. We aimed to explore any correlation between the clinical severity and biomarkers and spinal magnetic resonance imaging (MRI) abnormalities, identify independent factors associated with spinal MRI abnormalities, and ascertain factors affecting depression/anxiety in patients with N2 O-related neurological disorders. METHODS: Patients with N2 O-related neurological disorders were enrolled retrospectively between February 2017 and July 2020. Their demographic, clinical, laboratory, neuroimaging, electrophysiological, and neuropsychological findings were analyzed. Correlation analyses were conducted using Spearman's or Pearson's correlation and linear regression analysis. Independent factors associated with spinal MRI abnormalities were identified using univariate and multivariate analyses. RESULTS: The principal clinical manifestations of N2 O-related neurological disorders (n = 63; 38 men, 25 women; mean age ± SD: 22.60 ± 4.46 years) were sensory disturbance, followed by gait disturbance and pyramidal tract damage. A significant negative correlation existed between serum vitamin B12 levels and clinical severity (r = -0.309, p = .014), which disappeared after linear regression. An interval of less than 6 months between initial N2 O abuse and hospitalization was independently associated with spinal MRI abnormalities (39.47% vs. 72.00%, respectively; χ2  = 6.40, p = .01). Thirty-eight (60.32%) and 40 (63.49%) patients experienced anxiety and depression, respectively. Moreover, the higher the clinical scores/serum homocysteine levels, the greater the severity of anxiety/depression (r = 0.442, p < .01; r = 0.346, p < .01; r = 0.477, p < .01; r = 0.324, p < .01). CONCLUSIONS: The significant inverse correlation between initial vitamin B12 levels and clinical severity could aid prognosis prediction in patients with N2 O-related neurological disorders. Spinal MRI abnormalities were not related to clinical severity but depended on the time interval between initial N2 O abuse and hospitalization. Anxiety and depression were common comorbidity in these patients, and their severity increased with the intensity of clinical impairment and/or serum homocysteine levels.

Serum uric acid and risk of stroke: a dose-response meta-analysis.

The relationship between serum uric acid and risk of stroke is still controversial. Therefore, we conducted a meta-analysis based on the cohort study to explore the relationship between serum uric acid and risk of stroke, and further illuminate whether there is a linear or non-linear relationship between them. We manually searched the database including Cochrane, PubMed, Embase, Web of Science, and selected cohort studies focusing on the relationship between serum uric acid and stroke risk. Random effect model was used for statistical analysis. Twenty-one cohort studies involving 818,098 participants were included. The pooled relative risk for the high-vs-low categories was 1.22 (95% CI: 1.15-1.30). In addition, there was a non-linear dose-response relationship between uric acid and stroke risk. Serum uric acid was in the range of 3-5 mg/dl, with the lowest risk of stroke. In conclusion, high serum uric acid level increases the risk of stroke, with a non-linear dose-response relationship.

Association between daytime napping and stroke: A dose-response meta-analysis.

Daytime napping is common in many regions around the world and has been an important part of people's daily life. Daytime napping has attracted increasing attention in recent years. Thus, we conducted a meta-analysis to evaluate the relationship between daytime napping and stroke, and help reduce the risk of stroke by improving living habits. The Embase, PubMed, Web of Science and PsycINFO databases were searched for cohort studies published before October 2020 and eight eligible studies with 524,408 participants and 5,875 stroke cases were included in the final analysis. The pooled relative risk (RR) of stroke was 1.47 (95% confidence interval [CI]: 1.24-1.74; p < .001) with significant heterogeneity (I2  = 58%, p for heterogeneity = 0.02). However, the heterogeneity decreased when the study in which adjusting for sleep duration and stratifying the results based on sleep duration was not performed was excluded (RR: 1.38; 95% CI: 1.19-1.60, I2  = 44%, p for heterogeneity = 0.10). In dose-response analysis, the linear trend indicated that for every 10-min increase in daytime napping, the risk of stroke increased by 3%. Further well-designed large studies are needed to explore the effects of daytime napping on stroke and the underlying biological mechanisms.

Lipid levels and the risk of hemorrhagic stroke: A dose-response meta-analysis.

BACKGROUND AND AIMS: Hemorrhagic stroke (HS) could damage human health and impose heavy social and economic burden around the world. An accumulating number of studies revealed the effect of lipid levels on HS, whereas the results were inconsistent. Therefore, we conducted a dose-response meta-analysis to evaluate the relationship between lipid levels and HS. METHODS AND RESULTS: We searched the databases for relative cohort studies, which were published before April 2020. We pooled adjusted effect size and performed the dose-response analysis by random-effect model. 31 eligible studies with 2,291,643 participants and 12,147 hemorrhagic stroke cases were included. An inverse association was observed between the risk of hemorrhagic stroke and total cholesterol (TC) (RR: 0.72; 95% CI: 0.64-0.82) or low-density lipoprotein cholesterol (LDL-C) (RR: 0.69; 95% CI: 0.53-0.89). Additionally, in dose-response analysis, the non-linear trend was also found between TC, high-density lipoprotein cholesterol (HDL-C), and risk of HS. When the level of TC and HDL-C was about 6 and 1.3 mmol/L separately, the risk of HS was decreased to the lowest. And we found a linear trend that for every 1 mmol/L triglyceride (TG) increase, the risk of HS decreased by 7%. CONCLUSION: TC and LDL-C were both inversely related to the risk of HS. In dose-response analysis of TG, we also found the inverse linear trend. Furthermore, the non-linear trend suggested the level of TC and HDL-C was about 6 and 1.3 mmol/L separately could lead to the lowest risk of HS.

Association between skipping breakfast and risk of cardiovascular disease and all cause mortality: A meta-analysis.

BACKGROUND & AIMS: Previous studies on the association between skipping breakfast and risk of cardiovascular disease and all cause mortality have drawn controversial conclusions. Therefore, we carried out a meta-analysis to illuminate this association. METHODS: Studies about the association between skipping breakfast and risk of cardiovascular disease and all cause mortality were identified by searching Pubmed, Embase, Cochrane, and Web of Science databases until June 2019. Then we screened articles for eligibility, extracted data, and pooled the results using a random-effects model. RESULTS: Seven cohort studies concerning a total of 221,732 participants were included in this meta-analysis. Skipping breakfast was associated with elevated risk of cardiovascular disease (relative risk 1.22 95% confidence interval 1.10-1.35) and all cause mortality (relative risk 1.25 95% confidence interval 1.11-1.40) compared with eating breakfast regularly. CONCLUSION: Skipping breakfast increases the risk of cardiovascular disease and all cause mortality. Eating breakfast regularly may promote cardiovascular health and decrease all cause mortality.

25-Hydroxyvitamin D level, vitamin D intake, and risk of stroke: A dose-response meta-analysis.

BACKGROUND & AIMS: A growing number of studies have shown that vitamin D are related to the risk of stroke, however, the dose-response association between vitamin D and the risk of stroke is still unclear. Accordingly, we conducted a dose-response meta-analysis to evaluate the relationships between 25-hydroxyvitamin D [25(OH)D] level, vitamin D intake, and the risk of stroke by summarizing cohort studies. METHODS: PubMed, Embase, Cochrane and the Web of Science database were searched for related studies. Cohort studies examining the influence of 25(OH)D level and vitamin D intake on stroke risk were summarized. Dose-response relationships were determined using a random-effect model. RESULTS: Twenty cohort studies involving 217,235 participants were included. The pooled relative risk for the high-versus-low categories was 0.74 (95% CI: 0.66-0.83) for 25(OH)D level, and 0.75 (95% CI: 0.57-0.98) for vitamin D intake. In addition, there were non-linear relationships between 25(OH)D level, vitamin D intake, and stroke risk. The incidence of stroke was reduced to its lowest point, with a reduction of about 20%, when 25(OH)D level was about 50 nmol/L or vitamin D intake was about 12 µg/day. CONCLUSION: 25(OH)D level and vitamin D intake were both inversely related to stroke risk, with a non-linear dose-response relationship.

Anger, hostility and risk of stroke: a meta-analysis of cohort studies.

OBJECTIVE: A growing number of studies have been conducted on the relationship between anger and hostility and the risk of stroke, and their conclusions are not consistent. Accordingly, we performed a meta-analysis to evaluate the relationship between anger and hostility and the risk of stroke. METHODS: We searched the PubMed and Embase databases for cohort studies, focusing on the relationship between anger and hostility and risk of stroke. Then studies were selected according to the inclusion and exclusion criteria. Study results were pooled using a random effects model. RESULTS: Ten studies from seven articles involving 52,277 participants were included in this meta-analysis. No significant association was found between anger and hostility level and risk of stroke (hazard ratio 1.08; 95% confidence interval 0.79-1.47). However, a positive association was seen when people with high socioeconomic status were excluded (hazard ratio 1.30; 95% confidence interval 1.06-1.59). CONCLUSION: A higher level of anger and hostility is not associated with elevated risk of stroke. However, the association is positive among people with lower socioeconomic status.

25-Hydroxyvitamin D Levels and the Risk of Dementia and Alzheimer's Disease: A Dose-Response Meta-Analysis.

Background and Purpose: Conclusions of previous cohort studies on the relationship between 25-hydroxyvitamin D level and the risk of dementia and Alzheimer's disease were not consistent. Thus, we performed a dose-response meta-analysis to evaluate this relationship by summarizing cohort studies. Methods: Pubmed, Embase, Cochrane, and Web of Science databases were searched for relevant studies. Cohort studies concerning the association between 25-hydroxyvitamin D level and dementia or Alzheimer's disease were included. Results of studies were pooled and the dose-response relationship was determined using a random-effect model. Results: Ten cohort studies, with 28,640 participants were included. A significant inverse relationship was found between 25-hydroxyvitamin D level and the risk of dementia and Alzheimer's disease. In addition, we found a linear dose-response relationship in that a 10 nmol/L increase in 25-hydroxyvitamin D level may lead to a 5% decrease in the risk of dementia (relative risk, 0.95; 95% confidence interval, 0.93-0.98) and 7% in the risk of Alzheimer's disease (relative risk, 0.93; 95% confidence interval, 0.89-0.97). Conclusion: Plasma or serum 25-hydroxyvitamin D concentration was inversely related to the risk of dementia and Alzheimer's disease, consistent with a linear dose-response relationship.

Loop replacements with gut-binding peptides in Cry1Ab domain II enhanced toxicity against the brown planthopper, Nilaparvata lugens (Stål).

Bacillus thuringiensis (Bt) Cry toxins have been used widely in pest managements. However, Cry toxins are not effective against sap-sucking insects (Hemiptera), which limits the application of Bt for pest management. In order to extend the insecticidal spectrum of Bt toxins to the rice brown planthopper (BPH), Nilaparvata lugens, we modified Cry1Ab putative receptor binding domains with selected BPH gut-binding peptides (GBPs). Three surface exposed loops in the domain II of Cry1Ab were replaced with two GBPs (P2S and P1Z) respectively. Bioassay results showed that toxicity of modified toxin L2-P2S increased significantly (~9 folds) against BPH nymphs. In addition, damage of midgut cells was observed from the nymphs fed with L2-P2S. Our results indicate that modifying Cry toxins based on the toxin-gut interactions can broaden the insecticidal spectrum of Bt toxin. This method provides another approach for the development of transgenic crops with novel insecticidal activity against hemipteran insects and insect populations resistant to current Bt transgenic crops.