Knockdown of LCN2 Attenuates Brain Injury After Intracerebral Hemorrhage via Suppressing Pyroptosis.

Objective: The aims of this study are to screen novel differentially expressed genes (DEGs) for intracerebral hemorrhage (ICH) and reveal the role of Lipocalin-2 (LCN2) in ICH. Methods: We constructed the ICH model by injection of autologous whole blood into the right basal ganglia in rats. RNA-sequencing and bioinformatics analyses were performed to identify the DEGs between ICH and sham rats, and some important ones were confirmed using quantitative real-time PCR (qRT-PCR). LCN shRNA was used to knockdown of LCN2 in ICH rats. Pathological examination was carried out using 2,3,5-triphenyltetrazolium chloride (TTC) staining and Hematoxylin-eosin (HE) staining. Immunohistochemistry detected Caspase-3, and co-staining of Terminal dUTP nick end labeling (TUNEL) and NEUN staining were performed for neuron apoptosis assessment. Western blot analysis was performed to quantify pyroptosis-related proteins. Enzyme-linked immunosorbent assay (ELISA) was used to measure inflammatory cytokine levels. Results: ICH rats exhibited significant hematomas, higher brain water content, obvious interstitial edema, and inflammatory infiltration, as well as more apoptotic cells in brain tissues. RNA-seq analysis identified 103 upregulated and 81 downregulated DEGs. The expression of LCN2, HSPB1, CXCL10, and MEF2B were upregulated in ICH rats. ICH triggered the release of interleukin (IL)-1ß, tumor necrosis factor-α (TNF-α), and IL-18, and promoted the expression of pyroptosis-related proteins Caspase-1, GSDMD, NLRP3, and ASC. LCN2 knockdown attenuated the pathological characteristics of ICH, and also reduced pyroptosis in brain tissues. Conclusion: Inhibition of LCN2 attenuates brain injury after ICH via suppressing pyroptosis, which provide guidance for ICH management.

Bioinformatics and experimental analyses of glutamate receptor and its targets genes in myocardial and cerebral ischemia.

BACKGROUND: There is a mutual hemodynamic and pathophysiological basis between the heart and brain. Glutamate (GLU) signaling plays an important role in the process of myocardial ischemia (MI) and ischemic stroke (IS). To further explore the common protective mechanism after cardiac and cerebral ischemic injuries, the relationship between GLU receptor-related genes and MI and IS were analyzed. RESULTS: A total of 25 crosstalk genes were identified, which were mainly enriched in the Toll-like receptor signaling pathway, Th17 cell differentiation, and other signaling pathways. Protein-protein interaction analysis suggested that the top six genes with the most interactions with shared genes were IL6, TLR4, IL1B, SRC, TLR2, and CCL2. Immune infiltration analysis suggested that immune cells such as myeloid-derived suppressor cells and monocytes were highly expressed in the MI and IS data. Memory B cells and Th17 cells were expressed at low levels in the MI and IS data; molecular interaction network construction suggested that genes such as JUN, FOS, and PPARA were shared genes and transcription factors; FCGR2A was a shared gene of MI and IS as well as an immune gene. Least absolute shrinkage and selection operator logistic regression analysis identified nine hub genes: IL1B, FOS, JUN, FCGR2A, IL6, AKT1, DRD4, GLUD2, and SRC. Receiver operating characteristic analysis revealed that the area under the curve of these hub genes was > 65% in MI and IS for all seven genes except IL6 and DRD4. Furthermore, clinical blood samples and cellular models showed that the expression of relevant hub genes was consistent with the bioinformatics analysis. CONCLUSIONS: In this study, we found that the GLU receptor-related genes IL1B, FOS, JUN, FCGR2A, and SRC were expressed in MI and IS with the same trend, which can be used to predict the occurrence of cardiac and cerebral ischemic diseases and provide reliable biomarkers to further explore the co-protective mechanism after cardiac and cerebral ischemic injury.

Deferoxamine in intracerebral hemorrhage: Systematic review and meta-analysis.

BACKGROUND: Intracerebral hemorrhage (ICH) is a stroke with a high morbidity and mortality rate. Deferoxamine (DFX) is thought to be effective in treating Intracerebral Hemorrhage. In our study, we performed a meta-analysis to evaluate the treatment effects of DFX. METHODS: We systematically searched PubMed, Embase, Web of Science, the Cochrane Central Register of Controlled Trials, and Chinese Biomedical Literature Database in Jan 2022 for studies on DFX for ICH patients. Outcome measures included relative hematoma volume, relative edema volume, good neurological functional outcome and adverse events. Odds risk (OR) and weighted mean difference (WMD) were used to evaluate clinical outcomes. RESULTS: After searching 636 articles, 4 RCTs, 2 NRCTs, and 1cohort study were included. We found that DFX was effective in hematoma absorption on day 7 after onset, but the difference was not significant on day 14. DFX could suppress edema expansion on days 3, 7, and 14 after onset. DFX did not contribute to better outcomes after 3 and 6 months when used the modified Rankin Scale and the Glasgow Outcome Scale to evaluate neurological prognosis. The pooled results showed no statistically significant difference in Serious adverse events between the experimental and control groups. CONCLUSIONS: DFX could limit edema expansion on days 3, 7, and 14 after commencement and facilitate hematoma absorption at week 1 without significantly increasing the risk of adverse events, but it did not improve neurological prognosis.

Peroxiredoxin-3 plays a neuroprotective role in early brain injury after experimental subarachnoid hemorrhage in rats.

Subarachnoid hemorrhage (SAH), a type of hemorrhagic stroke, is a neurological emergency associated with a high morbidity and mortality rate. After SAH, early brain injury (EBI) is the leading cause of poor prognosis in SAH patients. Peroxiredoxins (PRDXs) are a family of sulphhydryl-dependent peroxidases. Peroxiredoxin-3 (PRDX3) is mainly located in the mitochondria of neurons, which can remove hydrogen peroxide (H2O2); however, the effect of PRDX3 on EBI after SAH remains unclear. In this study, an endovascular perforation model was used to mimic SAH in Sprague Dawley rats in vivo. The results revealed that after SAH, PRDX3 levels decreased in the neurons. PRDX3 overexpression by neuron-specific adeno-associated viruses upregulated PRDX3 levels. Furthermore, PRDX3 overexpression improved long- and short-term behavioral outcomes and alleviated neuronal impairment in rats. Nissl staining revealed that the upregulation of PRDX3 promoted cortical neuron survival. PRDX3 overexpression decreased the H2O2 content and downregulated caspase-9 expression. In conclusion, PRDX3 participates in neuronal protection by inhibiting the neuronal mitochondria-mediated death pathway; PRDX3 may be an important target for EBI intervention after SAH.

Effects of PAK1/LIMK1/Cofilin-mediated Actin Homeostasis on Axonal Injury after Experimental Intracerebral Hemorrhage.

Intracerebral hemorrhage (ICH) is a hemorrhagic stroke with a high mortality and disability rate. Neurological impairment after ICH is closely associated with neuronal axon damage. Serine/threonine-protein kinase p21 activated kinase 1 (PAK1) participates in cytoskeletal remodeling and regulates the F-actin and G-actin ratio in neuronal axons, but the function of PAK1 after ICH remains unclear. We established an in vivo rat ICH model by autologous whole blood injection into the right basal ganglia and an in vitro neuron oxyhemoglobin intervention. The results showed that the phosphorylated PAK1 level increased while the PAK1 expression level unchanged after ICH, After PAK1 knockdown, PAK1 and phosphorylated PAK1 levels were both reduced. Meanwhile, downstream proteins LIMK1 and cofilin expression levels were unchanged while phosphorylated LIMK1 and phosphorylated cofilin were decreased. F-actin/G-actin ratio decreased after PAK1 knockdown. Moreover, PAK1 knockdown improved short- and long-term neurobehavioral impairments in rats. In vitro, phosphorylated PAK1 expression increased after oxyhemoglobin intervention. After PAK1 knockdown, the axon length of neurons increased while F-actin/G-actin ratio decreased. Spersman correlation analysis showed a negative correlation between phospho-PAK1 fluorescence intensity and neuronal axon length. Knockdown of PAK1 increased the live/dead cell ratio and promoted neurons survival. Our study showed that PAK1 is involved in ICH early secondary brain injury by affecting F-actin/G-actin ratio through the PAK1/LIMK1/cofilin pathway. PAK1 may be an essential target for early secondary brain injury intervention after ICH.

Efficacy and Safety of Glucocorticoids Versus Placebo as an Adjuvant Treatment to Surgery in Chronic Subdural Hematoma: A Systematic Review and Meta-Analysis of Randomized Controlled Clinical Trials.

BACKGROUND: Chronic subdural hematoma (CSDH) is a common neurosurgical disease with a high recurrence rate, especially among the elderly. Glucocorticoids have been tested for the treatment of CSDH in observational studies and randomized clinical trials. METHODS: We systematically searched the PubMed, Embase, and Cochrane Central Register of Controlled Trials database for randomized trials from the earliest date available to May 23, 2021 that had compared glucocorticoids and placebo as a postoperative treatment of CSDH. Trials were included if the study participants were aged ≥18 years and had had CSDH after surgery. The relative risk (RR) was used to evaluate the clinical outcomes. RESULTS: We included 5 eligible randomized controlled trials with a total of 1251 patients. The findings showed that the use of adjuvant glucocorticoid therapy can effectively reduce the recurrence risk of CSDH compared with placebo (RR, 0.40; 95% confidence interval [CI], 0.28-0.58; P < 0.001). No significant differences were found between the glucocorticoid and placebo groups regarding favorable neurological outcomes (RR, 1; 95% CI, 0.93-1.08; P = 0.92). We found that the use of adjuvant glucocorticoids resulted in a significant increase in psychiatric symptoms (RR, 3.22; 95% CI, 1.83-5.64; P < 0.001). No significant differences were found for infection between the 2 groups (RR, 1.86; 95% CI, 0.56-6.14; P = 0.31). CONCLUSIONS: Glucocorticoid therapy can effectively reduce the recurrence risk of CSDH after surgery without an increase in the postoperative infection rate. However, significantly increased psychiatric symptoms were reported in the glucocorticoid group.

Endovascular coiling versus surgical clipping for aneurysmal subarachnoid hemorrhage: A meta-analysis of randomized controlled trials.

BACKGROUND: Aneurysmal subarachnoid hemorrhage is a relatively rare cause of stroke, carrying a bad prognosis of mortality and disability. The current standard procedure, neurosurgical clipping, has failed to achieve satisfactory outcomes. Therefore, endovascular detachable coils have been tested as an alternative. This meta-analysis was aimed to compare the outcomes of surgical clipping and endovascular coiling in aneurysmal subarachnoid hemorrhage. MATERIALS AND METHODS: Relevant randomized trials up to June 2018 were identified from Medline, Central, and Web of Science. Data for poor outcomes (Modified Rankin Scale [mRS] scores 3 to 6) at 2-3 months, 1 year, and 3-5 years were extracted and analyzed as odds ratios (ORs) with 95% confidence intervals (CIs), using RevMan software. RESULTS: Five studies (2780: 1393 and 1387 patients in the coiling and clipping arms, respectively) were included in the current analysis. The overall effect estimate favored endovascular coiling over surgical clipping in terms of reducing poor outcomes (death or dependency, mRS > 2) at 1 year (OR = 0.67, 95% CI: 0.57-0.79) and 3-5 years (OR = 0.8, 95% CI: 0.67-0.96). Moreover, coiling was associated with a significantly lower rate of cerebral ischemia (OR = 0.37, 95% CI: 0.16-0.86). Postprocedural mortality (OR = 0.79, 95% CI: 0.6-1.05) and rebleeding (OR = 1.15, 95% CI: 0.75-1.78) rates were comparable between the two groups. However, technical failure was significantly more common with coiling interventions than with clipping surgeries (OR = 2.84, 95% CI: 1.86-4.34). CONCLUSION: Our analysis suggests that coiling can be a better alternative to clipping in terms of surgical outcomes. Further improvements in the coiling technique and training may improve the outcomes of this procedure.

Association Between Meteorological Factors and the Rupture of Intracranial Aneurysms.

Background Both meteorological factors and morphological factors are important factors to predict intracranial aneurysm rupture. This study investigated the relationship between meteorological factors and aneurysmal subarachnoid hemorrhage (aSAH). Additionally, the morphological differences between ruptured and unruptured aneurysms under these high-risk meteorological conditions were assessed. Methods and Results The records of 1751 patients with aSAH with 2124 intracranial aneurysms were retrospectively analyzed. Spearman rank correlation analysis was used to assess the risks of incident aSAH on the basis of daily meteorological data. Morphological parameters were analyzed using 1-way ANOVA tests, and significant parameters (P<0.05) were further examined using a multivariable logistic regression analysis. Daily aSAH incidence had significant negative correlations with daily mean, maximum, and minimum temperature (P<0.001) and a significant positive correlation with daily mean atmospheric pressure (P<0.001). Additionally, 58 patients with multiple aneurysms were assessed to determine morphological differences. There were significant differences in the mean values for aneurysm size, neck width, length, height, width, parent artery diameter, shape of the aneurysm, aspect ratio, size ratio, and bottleneck factor (P<0.05). The multivariable logistic regression analysis showed that aspect ratio (ß=1.277, odds ratio=3.585, 95% CI, 1.588-8.090; P=0.002) was an independent risk factor for aneurysm rupture. Receiver operating characteristic curve analysis indicated that the ruptured aneurysm threshold of size was 3.45 mm and aspect ratio was 1.05. Conclusions Lower daily mean, maximum, and minimum temperatures and a higher daily mean atmospheric pressure were associated with an increased rate of aSAH. Additionally, under these meteorological conditions, the aneurysm size and aspect ratio thresholds for predicting rupture of an aneurysm may be lower.

Anti-vascular endothelial growth factor in glioblastoma: a systematic review and meta-analysis.

Glioblastoma is one of the most common primary brain tumors in adults. The current treatment strategies failed to achieve satisfactory outcomes. Anti-vascular endothelial growth factor (anti-VEGF) agents have been proposed to enhance the survival and quality of life in these patients. To investigate this, different databases were searched in addition to hand searching. Relevant studies were screened and only ten randomized controlled trials (RCTs) met the eligibility criteria; six of them were considered for meta-analysis. Eligible RCTs were assessed regarding risk of bias using the Cochrane tool. Relevant data were extracted and meta-analysis was conducted using the random effects model analysis on RevMan software. One thousand seventy-eight patients in the anti-VEGF group and 946 patients in the control group were available for analysis. No statistically significant improvement in the overall survival (OS) was detected for anti-VEGF (OR 0.87, 95% CI 0.7-1.09, p = 0.23) or bevacizumab subgroup (OR 0.84, 95% CI 0.65-1.08, p = 0.17) compared to standard therapy alone. However, the progression-free survival (PFS) showed a significant improvement with both anti-VEGF (OR 0.76, 95% CI 0.65-0.89, p = 0.0007) and bevacizumab subgroup (OR 0.75, 95% CI 0.65-0.87, p = 0.0001). In conclusion, anti-VEGF agents can improve the PFS but not OS in glioblastoma patients. The current evidence is not satisfactory to declare a new therapeutic line. Further RCTs with sharply determined outcomes and assessment methods are required.

miR-484/MAP2/c-Myc-positive regulatory loop in glioma promotes tumor-initiating properties through ERK1/2 signaling.

Glioma is the most common intracranial malignant tumor. Cancer stem cells (CSCs) are resistant to chemotherapy and radiotherapy, and are closely related to cancer metastasis and recurrence. In this study, we aimed to explore the effect of miR-484 on glioma stemness and the underlying mechanism involved. miR-484 enhanced glioma tumor-initiating properties in vitro and in vivo. Moreover, miR-484 was shown to directly target MAP2, resulting in activation of ERK1/2 signaling and promotion of stemness in glioma. The ERK1/2 signaling facilitated the formation of a miR-484/MAP2/c-Myc positive feedback loop in glioma. High miR-484 expression predicted a poor prognosis for glioma patients, and high MAP2 expression predicted a good prognosis for glioma patients. Low miR-484 expression and high MAP2 expression was associated with the best prognosis in glioma. Our study suggests that miR-484 and MAP2 can be utilized as predictors for the clinical diagnosis and prognosis of glioma, and miR-484 and MAP2 are potential targets for the treatment of glioma.

Negative nuclear expression of CDKL2 correlates with disease progression and poor prognosis of glioma.

AIMS: This study aimed to investigate the nuclear expression status of cyclin dependent kinase like 2 (CDKL2) in glioma and its correlation with the characteristics of clinical pathology, including patient survival. METHODS AND RESULTS: In the present study, the expression of CDKL2 mRNA was detected by real-time QPCR in freshly collected glioma and para-carcinoma tissues. Moreover, immunohistochemistry was used to identified nuclear expression of CDKL2, and the characteristics of clinical pathology from glioma cases (n = 144) and non-cancerous brain tissues (n = 32) were counted. Low mRNA and nuclear protein expression of CDKL2 was observed in glioma tissues compared to non-cancerous tissues. Glioma patients with negative nuclear expression of CDKL2 were correlated with histologic type, clinical World Health Organization (WHO) grade, tumor location, and KI-67 expression status. Negative nuclear expression of CDKL2 in glioma patients predicted an observably shorter overall survival time than did positive expression. However, as demonstrated by multivariate analysis, nuclear expression of CDKL2 was not an independent prognostic biomarker for the survival of patients with glioma. CONCLUSIONS: Our study indicated that negative nuclear expression of CDKL2 may represent a potential unfavorable marker for progression and poor prognostic in glioma.