Subarachnoid hemorrhage (SAH) is a serious cerebrovascular disease with high mortality, and the mean age at morbidity is younger than in other types of stroke. Early brain injury (EBI) plays a key role in the poor prognoses of SAH. In EBI, multiple forms of cell death have been identified and well studied; however, the role of ferroptosis has not been elucidated. Hence, in this study, we developed an in vivo (SAH rat model) and in vitro model (SH-SY5Y oxyhemoglobin injury model) to understand the role of ferroptosis in EBI, then explored the protective mechanism of ferrostatin-1 (Fer-1). Firstly, we found that neurological scores, blood–brain barrier permeability, brain edema deteriorated after SAH in the in vivo model, cell viability was decreased after SAH in both cortex and SH-SY5Y cells. Further, iron content in cortex was increased after SAH, while transferrin receptor 1 and ferroportin (Fpn) were increased in oxyhemoglobin-treated in vitro model. Additionally, glutathione content and glutathione peroxidase 4 activity were reduced in SAH rats, and lipid peroxides were increased in the oxyhemoglobin-treated cells. Finally, administration of Fer-1 upregulated Fpn and decreased the iron content, then improved the lipid peroxidation and EBI. However, Fer-1 had no effect on the apoptosis. Our study indicated that the ferroptosis was involved in EBI of SAH, and the inhibitor Fer-1 provided neuroprotection against EBI by alleviating ferroptosis, the potential protective mechanism might be via suppressing lipid peroxidation.

Ischemic stroke (IS) constitutes the majority of stroke cases. Ferroptosis, a non-apoptotic form of programmed cell death, is an essential mechanism of IS. This study aimed to investigate a ferroptosis-related molecular mechanism in IS progression. IS model in vitro was induced by oxygen-glucose deprivation/reoxygenation (OGD/R) in SH-SY5Y cells. RT-qPCR and Western blotting were performed for detection of NOP2/Sun RNA methyltransferase family member 2 (NSUN2), nuclear receptor coactivator 4 (NCOA4), and Krueppel-like factor 6 (KLF6). Cell functions were assessed by MTT assay, flow cytometry, and enzyme-linked immunosorbent assay. Mitochondrial membrane potential and other ferroptosis indicators were examined using kits. Molecular binding was detected using methylated RNA immunoprecipitation (MeRIP), RIP, dual-luciferase reporter assay and chromatin immunoprecipitation (ChIP) assay. IS animal model was established by middle cerebral artery occlusion (MCAO). OGD/R-induced apoptosis, inflammation, and ferroptosis of SH-SY5Y cells were suppressed by NCOA4 knockdown. NSUN2 could promote NCOA4 mRNA stability by mediating 5-methylcytosine (m5C) methylation modification of NCOA4, and YBX1 served as a reader protein. NSUN2 contributed to OGD/R-induced SH-SY5Y cell injury via upregulating NCOA4. KLF6 acted as a transcription factor to activate NSUN2 transcription, and then facilitated nerve injury of OGD/R-induced SH-SY5Y cells. Animal assay showed that silencing NSUN2 inhibited infarct volume, tissue injury, neurological function, and neuroinflammation in MCAO rats. These current findings affirmed that KLF6-activated NSUN2 could contribute to ferroptosis of OGD/R-induced SH-SY5Y cells via inducing NCOA4 m5C modification, providing a novel insight into the mechanism of ferroptosis in IS.

Acute myeloid leukemia (AML) is a heterogeneous hematologic malignancy associated with poor clinical outcomes. RHOBTB2, an atypical member of the Rho-GTPase family, contains a conserved GTPase domain at its N-terminus. While previous studies have implicated RHOBTB2 in AML progression, its underlying mechanisms remain inadequately defined. In this study, bioinformatics analyses revealed that RHOBTB2 is markedly upregulated in AML and correlates with unfavorable prognosis. To elucidate its functional role, we overexpressed or silenced RHOBTB2 in human AML cell lines KG-1 and MOLM-13. Functional assays, including CCK-8, Transwell, and Annexin V/PI staining, demonstrated that RHOBTB2 overexpression enhanced proliferation and migration, suppressed apoptosis, and shortened G0/G1 phase. Conversely, RHOBTB2 silencing exerted opposing effects. In addition, a direct interaction between RHOBTB2 and KLHL13 was identified through STRING database predictions and validated by co-immunoprecipitation. Western blot analysis confirmed that RHOBTB2 upregulates KLHL13 protein expression levels. Notably, KLHL13 downregulation induced by RHOBTB2 knockdown was reversed upon treatment with the proteasome inhibitor MG132, indicating that RHOBTB2 stabilizes KLHL13 by inhibiting its proteasomal degradation. Collectively, our findings highlight the RHOBTB2/KLHL13/Hippo pathway as a critical regulatory mechanism in AML malignancy and suggest RHOBTB2 as a potential therapeutic target.

Objective:To explore the relevant factors affecting the prognosis of subarachnoid hemorrhage. Methods:284 patients with subarachnoid hemorrhage who were hospitalized in our hospital from January 1, 2022 to June 30, 2024 were selected and divided into a good prognosis group and a poor prognosis group according to the modified Rankin Scale (mRS) score. The general clinical data of the patients were also collected, and the independent risk factors affecting the poor prognosis of the patients were screened by univariate logistic regression analysis. Results:Patients with a favorable prognosis had a lower incidence rate of rebleeding (4.72% vs 17.65%; P =0.001), electrolyte disturbances (21.46% vs 41.18%; P <0.001), lower respiratory tract infection (5.58% vs 35.29%; P <0.001), urinary tract infection (1.72% vs 15.69%; P <0.001) and gastrointestinal infection (2.15% vs 11.76%; P <0.001) than patients with an unfavorable prognosis. Therefore, coinfection is an independent risk factor for prognosis. After adjusting for covariates, logistic regression analysis identified the prognosis of subarachnoid hemorrhage was related to coinfections (adjusted odds ratio =2.057; 95% CI: 1.516~2.791; P<0.001). Conclusion:Coinfection is a very important independent risk factor affecting prognosis, and clinical care should focus on how to reduce coinfection during hospitalization in patients with subarachnoid hemorrhage and treat it aggressively to reduce mortality and disability and improve patient prognosis.

The mammalian target of rapamycin (mTOR) was reported to regulate cell autophagy and outcomes of several neurological diseases. Mitochondria, which serve as critical organelles in neurons. are also involved in the pathology of neurological diseases. However, the role of mTOR in mitochondrial morphology has not been clarified especially in subarachnoid hemorrhage (SAH). In this study, we established SAH models both in vivo and in vitro. Rapamycin and 3-methyl adenine (3-MA) were then administered to alter mTOR activity. Post-SAH assessment included SAH grading, neurological evaluation, blood–brain barrier (BBB) permeability, brain water content, mitochondrial membrane potential (MMP), mitochondrial morphology, ATP content, cell viability, cytotoxicity, and expression of proteins related to apoptosis and mitochondrial fission. The results showed that (1) neurological deficits, BBB permeability, and brain edema were increased after SAH and that cell viability was exacerbated in brain tissue. (2) Excessive mitochondrial fission was evident based on changes in mitochondrial morphology, while MMP and ATP content were decreased in neurons after SAH. (3) Administration of rapamycin improved the excessive mitochondrial fission and restored mitochondrial function, which subsequently reduced apoptosis. (4) 3-MA showed an adverse effect on mitochondria and aggravated excessive mitochondrial fission and dysfunction in SAH. Neurological deficits and neuronal viability were also exacerbated following the administration of 3-MA. Therefore, our study suggests that mTOR inhibition has neuroprotective effects against neuronal injury after SAH via alleviating excessive mitochondrial fission.

This study found that the level of neuroepithelial cell-transforming gene 1 protein (NET1) was significantly increased in a mouse cardiac fibrosis model. Moreover, the expression level of NET1 was increased in cardiac fibrosis induced by TGF-β1, suggesting that NET1 was involved in the pathological process of cardiac fibrosis. Overexpression of NET1 promoted β-catenin expression in the nucleus and significantly increased the proliferation and migration of cardiac fibroblasts. NET1 may form a complex with β-catenin through GSK3β. Knockdown of β-catenin alleviated the effects of NET1 overexpression on collagen production and cell migration. In the heart of NET1 knockout mice, NET1 knockout can reduce the expression of β-catenin, α-SMA, and collagen content induced by MI. In conclusion, NET1 may regulate the activation of Wnt/β-catenin and TGF/Smads signaling pathway, promote collagen synthesis in fibroblasts, and participate in cardiac fibrosis. Thus, NET1 may be a potential therapeutic target in cardiac fibrosis.

Accumulating evidence suggests that neuronal apoptosis plays a critical role in early brain injury (EBI) after subarachnoid hemorrhage (SAH), and the inhibition of apoptosis can induce neuroprotective effects in SAH animal models. c-Abl has been reported to promote neuronal apoptosis in Alzheimer's disease and cerebral ischemia, but its role in SAH had not been illuminated until now. In the present study, the effect of c-Abl on neuronal apoptosis induced by SAH was investigated. c-Abl protein levels and neuronal apoptosis were markedly increased 24 h after SAH, and the inhibition of endogenous c-Abl reduced neuronal apoptosis and mortality and ameliorated neurological deficits. Furthermore, c-Abl inhibition decreased the expression of cleaved caspase-3 (CC-3) after SAH. These results demonstrate the proapoptotic effect of c-Abl in EBI after SAH. Additionally, c-Abl inhibition further enhanced the SAH-induced phosphorylation of Akt and glycogen synthase kinase (GSK)3β. LY294002 abrogated the beneficial effects of targeting c-Abl and exacerbated neuronal apoptosis after SAH. SAH decreased LRP-1 levels and downregulated LRP-1 by RAP, and LRP-1 small interfering RNA (siRNA) induced a dramatic decrease in Akt/GSK3β activation in the presence of c-Abl siRNA. This is the first report showing that the c-Abl tyrosine kinase may play a key role in SAH-induced neuronal apoptosis by regulating the LRP-1-dependent Akt/GSK3β survival pathway. Thus, c-Abl has the potential to be a novel target for EBI therapy after SAH.

Fibroblast growth factor 21 (FGF21) and β-Klotho (KLB) play an important role in preventing and treating overweight and obesity. However, it is unclear what conditions promote FGF21 and KLB expression in different tissues. Therefore, we studied expression of FGF21 and KLB with respect to two exercise regimes: moderate-intensity continuous training (MICT) and high-intensity interval training (HIIT) (two popular strategies in weight loss). Mice were randomly divided into three groups (n = 8 per group): MICT, HIIT, and sedentary lifestyle (SED). All mice were fed a high-fat diet (HFD) for 12 weeks to induce obesity. The exercise was performed on a motorized treadmill for another eight weeks and the diet continued in each group. We found that both MICT and HIIT had positive effects on the loss of HFD-induced body weight increase and serum FGF21 levels. HIIT promoted decrease of the body weight and serum triglyceride (TG) levels, while MICT was more effective at enhancing FGF21 and KLB expression in the liver, brown adipose tissue (BAT), and muscle at the mRNA and protein levels.

Apelin, an endogenous ligand for the orphan G-protein-coupled receptor APJ, possesses anti-apoptotic and neuroprotective properties. It has been shown to be a protective factor for different types of central nervous system insults, such as ischemia and traumatic brain injury. Here, we investigated the effects of apelin-13 on early brain injury (EBI) following subarachnoid hemorrhage (SAH), and the underlying molecular mechanisms involved. Apelin-13 was delivered to rats via intracerebroventricular administration. Neurological scores, brain water content and neuronal apoptosis were measured 24 h after SAH. The PI3K/Akt inhibitor LY294002 or GLP-1R siRNA were injected into the lateral cerebral ventricle before induction of SAH. Changes in the expression of p-Akt, GLP-1R and apoptosis-associated proteins (Bax, Bcl-2, cleaved caspase-3) were then investigated. Results showed that the levels of GLP-1R in neurons increased significantly, reaching a peak at 24 h after the induction of SAH. Treatment with apelin-13 improved neurological deficits, as well as alleviated brain edema and apoptotic cell death. Apelin-13 was also able to increase the levels of p-Akt, GLP-1R and Bcl-2, while inhibiting the expression levels of Bax and cleaved caspase-3. The anti-apoptotic and neuroprotective effects of apelin-13 were partially reversed by addition of LY294002 or GLP-1R siRNA. These results provide evidence that apelin-13 attenuates EBI following SAH via suppressing neuronal apoptosis, and that this effect may act partially via the activation of the GLP-1R/PI3K/Akt signaling pathway.

Mitochondrial dysfunction is considered a crucial therapeutic target for early brain injury following subarachnoid hemorrhage (SAH). Emerging evidence indicates that docosahexaenoic acid (DHA), an essential omega-3 fatty acid, protects mitochondria in various chronic diseases. This study aimed to investigate the neuroprotective effects of DHA on mitochondrial dynamic dysfunction after EBI using in vivo and in vitro approaches. For in vivo experiments, the rat endovascular perforation SAH model was performed, whereby DHA was administered intravenously 1 h after induction of SAH. Primary cultured neurons treated with oxyhemoglobin (OxyHb) for 24 h were used to mimic SAH in vitro. Our results demonstrated that DHA improved neurological deficits and reduced brain edema in rats with SAH, and attenuated OxyHb-induced neuronal death in primary cultured cells. DHA reduced the amount of reactive oxygen species-positive cells and improved cell viability when compared to the SAH + vehicle group in vitro. DHA attenuated malondialdehyde levels and superoxide dismutase stress, increased Bcl2 and Bcl-xl, and decreased Bax and cleaved caspase-3 in vivo. Additionally, DHA ameliorated mitochondrial dysfunction, upregulated the mitochondrial fusion-related protein Optic Atrophy 1, and downregulated the mitochondrial fission-related protein Dynamin-Related-Protein 1 (Drp1) and Serine 616 phosphorylated Drp1 after SAH both in vitro and in vivo. Taken together, our current study demonstrates that DHA might prevent oxidative stress-based apoptosis after SAH. The characterization of the underlying molecular mechanisms may further improve mitochondrial dynamics-related signaling pathways.

Here, we report a case of a 64-year-old female with acute-onset vertigo, nausea, and vomiting. In an emergency imaging examination, the results of computed tomography (CT) and diffusion weighted imaging (DWI) were negative. However, on 1 day post-hospital admission, a small acute infarct in the posterolateral aspect of the left medulla was detected by DWI. Extra attention should be payed to the false-negative imaging results to avoid diagnosis and treatment delay.

Oxoglutarate dehydrogenase like (OGDHL) is involved in tricarboxylic acid cycle and has been reported as a candidate tumor suppressor in some other tumors. We first explored the role of OGDHL in human pancreatic ductal adenocarcinoma (PDAC) progression. OGDHL was frequently down-regulated in human PDAC and predicted poor prognosis. OGDHL suppressed PDAC growth though G1 cell cycle arrest and also inhibited migration and invasion ability of PDAC both in vivo and in vitro. Compared with non-tumor tissues, PDAC tissues showed down-regulation of OGDHL and up-regulation of miR-214 and TWIST1. The results showed that OGDHL was a target gene of miR-214 and always negatively regulated by miR-214 and the decreased expression level of OGDHL was on account of the increased expression level of miR-214 in PDAC. In addition, TWIST1 was frequently up-regulated in PDAC and induces miR-214 expression. However OGDHL could inhibit TWIST1 expression via both promoting ubiquitin-mediated proteasomal degradation of HIF1a and regulating AKT pathways. The effect of OGDHL/HIF1a/TWIST1/miR-214 signaling pathway in pancreatic carcinogenesis and metastasis was also determined both in vivo and in vitro. A combination of down-regulation OGDHL and over-expression miR-214 and TWIST1 predicts a poorer overall survival in PDAC patients. Finally, we demonstrated that the relationship of expression among OGDHL, miR-214 and TWIST1 may be a significant predictor of prognosis in PDAC patients. It is a novel pathway in OGDHL-regulated inhibition of PDAC tumorgenesis and metastasis. It may be a brand new targeted therapy in PDAC through OGDHL, TWIST1, miR-214, and HIF1a for prevention, treatment and prognosis.

The purpose of this study was to evaluate the neuroprotective effects of astaxanthin on early brain injury (EBI) caused by subarachnoid hemorrhage (SAH) in rats and to explore possible molecular mechanisms. Experimental SAH model was introduced in adult male SD rats by injecting autologous arterial blood into the prechiasmatic cistern. Astaxanthin (75 mg/kg bodyweight) or olive oil was administered by oral gavage at 3 h after SAH. Our results showed that astaxanthin attenuated SAH-induced cerebral vasospasm and reduced neuronal apoptosis. Astaxanthin inhibited mitochondria-associated neuron apoptosis in the prefrontal cortex after SAH: increased mitochondrial membrane potential, decreased Bax/Bcl-2 ratio, inhibited cytochrome C release in cytoplasm, and suppressed caspase-3 enzyme activity. Furthermore, the cerebral expression levels of synaptic proteins (Synapsin-1, postsynaptic density-95 and growth-associated protein-43) and nerve growth and neuronal differentiation factors (brain-derived neurotropic factor and purine-rich binding protein-alpha) were reduced following SAH. Astaxanthin partly restored their expression. In conclusion, our current work demonstrates that astaxanthin attenuates SAH-induced EBI, possibly by improving neuronal survival and mitochondrial function.

Early brain injury following subarachnoid hemorrhage (SAH) strongly determines the prognosis of patients suffering from an aneurysm rupture, and apoptosis is associated with early brain injury after SAH. This study was designed to explore the role of X-linked inhibitor of apoptosis (XIAP) in early brain injury following SAH. The expression of XIAP was detected using western blotting and real-time RT-PCR in an autologous blood injection model of SAH. We also studied the role of XIAP in early brain injury and detected apoptosis-related proteins. The results showed that XIAP was significantly up-regulated in the cortex and hippocampus and that XIAP was mainly expressed in neuronal cells following SAH. The inhibition of endogenous XIAP aggravated blood-brain barrier disruption, neurological deficits and brain edema. Recombinant XIAP preserved the blood-brain barrier, improved the neurological scores and ameliorated brain edema. Recombinant XIAP treatment also decreased the expression of cleaved caspase-3, caspase-8 and caspase-9, whereas there was no effect on the expression of p53, apoptosis-inducing factor or cytochrome c. These results show that XIAP acts as an endogenous neuroprotective and anti-apoptotic agent following SAH. The effects of XIAP on early brain injury was associated with the inhibition of the caspase-dependent apoptosis pathway.

Anaplastic thyroid carcinoma (ATC) is one of the most lethal carcinoma with a poor prognosis; however, molecular mechanisms underlying the aggressiveness of ATC remain unclear. Our goal was to examine the expression of X-linked inhibitor of apoptosis protein (XIAP) in ATC, as well as its role in ATC tumorigenesis. This is a retrospective study of ATC patients from the Second Affiliated Hospital of Harbin Medical University during June 2003 to October 2013. The expression of XIAP in tumor specimens of ATC patients was examined by immunohistochemical staining. The roles of XIAP in proliferation, migration, invasion, and chemoresistance were investigated by shRNA mediated-knockdown of XIAP in human ATC cell lines. The effect of XIAP on tumorigenesis was evaluated using a xenograft tumor model with nude mice. XIAP expression was significantly higher in the invasive area of ATC samples, whereas XIAP expression was negative in either normal thyroid follicular epithelial cells or the differentiated papillary thyroid carcinoma. XIAP-depleted ATC cells showed a remarkable decrease in the proliferation, migration, and invasion compared with the scramble group. Knockdown of XIAP expression significantly enhanced the chemosensitivity of WRO and SW1736 cells to docetaxel or taxane. Moreover, knockdown of XIAP significantly suppressed ATC tumorigenesis in vivo. XIAP is highly expressed in ATC cells and tumors. XIAP play important roles in tumor behaviors and chemosensitivity of ATC cells. XIAP may function in ATC aggressiveness and may serve as a potential therapeutic target for ATC treatment.

The present research was designed to investigate whether endothelin-1 (ET-1) secretion can be induced by oxyhemoglobin and whether nuclear factor kappa B (NF-kappa B) is involved in the regulation of ET-1 transcription in cerebrovascular muscle cells. Cerebrovascular muscle cells isolated from a rabbit basilar artery were stimulated by oxyhemoglobin (OxyHb) and ET-1 production was increased significantly in the supernatant. Inhibition of NF-kappa B with pyrrolidine dithiocarbamate and small interfering RNA decreased the expression of ET-1. Nuclear translocation of NF-kappa B and the degradation of IkB-alpha was observed with the stimulation of OxyHb. The supernatant obtained from cerebrovascular muscle cells stimulated by OxyHb produced contractions in arterial rings and was blocked by the ET-1 receptor antagonist (BQ-123). The time course of the OxyHb-induced contractions of the basilar artery rings correlated with the time course of the OxyHb-induced ET-1 secretion. The contraction of the basilar artery rings induced by OxyHb was attenuated when the artery rings were preincubated with pyrrolidine dithiocarbamate and SN50 (20 and 10 mu M, respectively). These results indicate that cerebrovascular muscle cells may be an important source of ET-1 production after subarachnoid hemorrhage. NF-kappa B was involved in the expression of ET-1 and the inhibition of the NF-kappa B pathway may be beneficial for the treatment of cerebral vasospasm. Copyright (C) 2016 Wolters Kluwer Health, Inc. All rights reserved.