Subarachnoid Hemorrhage Group Research Articles

Blocking VEGF by Bevacizumab Attenuates VEGF-Induced Vasospasm After Experimental Subarachnoid Hemorrhage in Rabbits

Vasospasm after subarachnoid hemorrhage (SAH) plays a vital role in the development of delayed cerebral ischemia. Anti- vascular endothelial growth factor (VEGF) antibodies, like bevacizumab (BEV), may attenuate VEGF-stimulated angiogenesis, reduced vascular cell proliferation, and improve vasospasm after SAH. Thirty-two adult male New Zealand white rabbits were randomly divided into 4 groups of 8 rabbits in each group: group 1 (control); group 2 (SAH); group 3 (SAH+ vehicle); and group 4 (SAH+ BEV). BEV (5 mg/kg, intraperitoneally) was administered 5 minutes after the intracisternal blood injection and continued for 72 hours once per day in the same dose for group 4. Animals were sacrificed 72 hours after SAH. Basilar artery cross-sectional areas, arterial wall thicknesses, and hippocampal degeneration scores were evaluated in all groups. VEGF is associated with the narrowing of the basilar artery. Treatment with BEV statistically significantly increased the cross-sectional area of the basilar artery when compared with the SAH and the vehicle groups. Basilar artery wall thicknesses in the BEV group was statistically significant smaller than in the SAH and vehicle groups. The hippocampal degeneration scores for the BEV and control groups were similar and significantly lower than those for the SAH and vehicle groups. Cellular proliferation and subsequent vessel wall thickening is a reason to delay cerebral ischemia and deterioration of the neurocognitive function. Intraperitoneal administration of BEV was found to attenuate cerebral vasospasm and prevent delayed cerebral ischemia and improve neurocognitive function after SAH in rabbits.

Nix Plays a Neuroprotective Role in Early Brain Injury After Experimental Subarachnoid Hemorrhage in Rats.

Nix is located in the outer membrane of mitochondria, mediates mitochondrial fission and implicated in many neurological diseases. However, the association between Nix and subarachnoid hemorrhage (SAH) has not previously been reported. Therefore, the present study was designed to evaluate the expression of Nix and its role in early brain injury (EBI) after SAH. Adult male Sprague-Dawley (SD) rats were randomly assigned to various time points for investigation after SAH. A rat model of SAH was induced by injecting 0.3 ml of autologous non-heparinized arterial blood into the prechiasmatic cistern. The expression of Nix was investigated by Western blot and immunohistochemistry. Next, Nix-specific overexpression plasmids and small interfering RNAs (siRNAs) were separately administered. Western blot, neurological scoring, Morris water maze, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining and fluoro-jade B (FJB) staining were performed to evaluate the role of Nix in EBI following SAH. We found that Nix was expressed in neurons and its expression level in the SAH groups was higher than that in the Sham group, which peaked at 24 h after SAH. Overexpression of Nix following SAH significantly decreased the expression of translocase of outer mitochondrial membrane 20 (TOMM20, a marker of mitochondria), ameliorated neurological/cognitive deficits induced by SAH, and reduced the total number of apoptotic/neurodegenerative cells, whereas siRNA knockdown of Nix yielded opposite effects. Taken together, our findings demonstrated that the expression of Nix is increased in neurons after experimental SAH in rats, and may play a neuroprotective role in EBI following SAH.

The Impact of Propolis Factor Caffeic Acid Phenethyl-Ester on the Cerebral Vasospasm and Early Brain Damage in the Experimentally Induced Subarachnoid Hemorrhage on Rats

Caffeic acid phenethyl ester (CAPE), a phenolic compound, besides being 1 of the biologically active components of propolis, is a compound with antioxidant, antiinflammatory, antiviral, reperfusion damage prevention, immune stimulant, and carcinostatic, anticancer properties. The aim of this study was to investigate the possible effects of CAPE on cerebral vasospasm and early brain injury, which were experimentally administered intraperitoneally in rats with subarachnoid hemorrhage. Thirty-two Wistar Albino rats weighing 200 to 300 g were used in our study. The rats divided into 3 groups: the control group (n= 10), subarachnoid hemorrhage group (n= 11), and subarachnoid hemorrhage+ CAPE group (n= 11). These groups were evaluated according to the Ischemia index in hippocampal CA3 regions and the morphometric analysis of basilar artery diameter after being sacrificed at the end of 72nd hour. A significant difference was found between group 1 and group 2 for the CA-3 region, it was concluded that early brain damage occurred after subarachnoid hemorrhage. When the neuronal damage in CA-3 region was evaluated between group 2 and group 3, a statistically significant difference was found between the groups. There was a statistically significant difference between group 1 and group 3 in terms of ischemia detection. It was shown that CAPE has a preventive effect on early brain injury after subarachnoid hemorrhage and has a positive effect on reducing cerebral vasospasm. Our study is the first study in the literature showing that CAPE inhibits ischemic brain injury following subarachnoid hemorrhage.

First definition of burned choroid plexus in acidic cerebrospinal fluid-filled brain ventricles during subarachnoid hemorrhage: Experimental study.

Blood and cerebrospinal fluid (CSF) acidosis is the most troubling complication in subarachnoid hemorrhage (SAH) if carotid body (CB) networks are disrupted. However, histopathological examination of the choroid plexus (CP) in acidic CSF has not been evaluated so far. In this study, we aimed to investigate the CP in acidic CSF following SAH. Twenty-eight rabbits were used. Five rabbits were used to analyze CB network (control group; n = 5); seven rabbits were injected 1 mL of saline (Sham group; n = 7); and the rest 16 rabbits were given 1 mL of autologous arterial blood inject into the cisterna magna to create SAH (SAH group; n = 16). Blood and CSF pH values were recorded before/during/after the experimental procedures. Nuclear darkening, cellular shrinkage and pyknosis suggested the presence of apoptosis of epithelial cells of CP. The densities of normal and degenerated epithelial cells of CPs were estimated using stereological methods. The relationship between the pH values and degenerated epithelial cell densities of CPs were statistically compared by Mann-Whitney U-test. The pH values of blood were estimated as 7.359 ± 0.039 in the control group, 7.318 ± 0.062 in the Sham group, 7.23 ± 0.013 in the SAH group. CSF pH values were 7.313 ± 0.028 in the control group, 7.296 ± 0.045 in the Sham group, and 7.224 ± 0.012 in the SAH group. Degenerated epithelial cell density of CP was 25 ± 7 in the control group, 226 ± 64 in the Sham group, and 2115 ± 635 in the SAH group. There was a considerable link between CSF pH values and degenerated epithelial cells of CP (P < 0.0001). This study shows that CB insult causes acidosis of CSF as well as cellular degeneration of CP during SAH. This is the first description of this in the literature.

Abstract WP308: Role of Microthrombi and Vasospasm in the Development of Delayed Neurological Deficits After Subarachnoid Hemorrhage in Mice

Rationale: Microthrombosis has been suggested as a major factor contributing to delayed neurological deterioration in patients after subarachnoid hemorrhage (SAH). However, experimental studies on the role of microthrombi in delayed deficits after SAH has not been investigated. Our hypothesis is that, following SAH, mice which develop delayed neurological deficits have a greater number of microthrombi than mice which do not develop delayed neurological deficits. Methods: SAH was induced in adult male and female C57BL/6 mice via endovascular perforation. Mice were randomly assigned into sham (n=6/sex) or SAH groups (n=22-24/sex). Neurobehavior was performed on days 1-3, 5, and 7 post-SAH using a composite neuroscore. Animals were sacrificed on the day of delayed deficits or 7 days post-SAH. Microthrombi count and vessel diameters (for vasospasm) were measured using H&amp;E stained brain slices. All outcomes were performed and all data were analyzed by a blinded investigator. Results: Seventeen percent (4/24) of male mice and thirty-six percent (8/22) of female mice developed delayed deficits on days 3-5 post-SAH (Figures 1A and 1B). Those mice which developed delayed deficits had significantly more microthrombi in their brains than mice which did not develop delayed deficits; vasospasm did not correlate with delayed deficits. Additionally, female SAH mice develop delayed deficits at a higher frequency than males (Figure 1C). Conclusions: This work shows for the first time delayed deficits in a SAH mouse model. Further, microthrombi correlated with delayed deficits, whereas no correlation was between delayed deficits and vasospasm. The data within this study suggests that preventing microthrombi may improve functional recovery and reduce the risk of delayed deficits.

Abstract TMP23: Inflammasome-derived Caspase-1 is Elevated in the Cerebrospinal Fluid of Subarachnoid Hemorrhage Patients and Correlated to Outcome

Introduction: The role of neuroinflammation following aneurysmal subarachnoid hemorrhage (SAH) and its relationship to outcome is the subject of many ongoing studies. The proteolytic enzyme, caspase-1, activated by the inflammasome complex, is known to contribute to numerous downstream pro-inflammatory effects. In this study, we investigated caspase-1 activity in the cerebrospinal fluid (CSF) of SAH patients and its association to outcome. Methods: SAH patients were recruited from a regional stroke referral center. CSF samples from 18 SAH subjects were collected via an external ventricular drain and obtained within 72 hours of the onset of symptoms. For control subjects, we collected the CSF from 9 patients undergoing lumbar puncture with normal CSF and normal brain MRI. Caspase-1 activity was measured using commercially available luminescence assays. SAH subjects were categorized at hospital discharge into those with good outcomes (Glasgow Outcome Scale, GOS, of 4-5) and poor outcomes (GOS of 1-3). The levels of caspase-1 activity in various groups were analyzed using Mann-Whitney and Pearson correlation tests. Caspase-1 activity was also adjusted by initial severity of bleed using analysis of covariance (ANCOVA). Results: Caspase-1 levels from SAH patients were significantly higher than that measured from the control group (mean 1.06x10-2 vs 1.90x10-3 counts per second (CPS)/μl*min), p = 0.0002). Within the SAH group, 10 patients (55.6%) had good outcomes and 8 patients (44.4%) had poor outcomes. Caspase-1 activity was significantly higher in the poor outcome group (mean 1.54x10-2 vs 1.60x10-3 CPS/μl*min), p = 0.0012). Additionally, caspase-1 activity had a statistically significant correlation with GOS score (r = -0.60; p = 0.0100). When adjusted for initial severity of bleed, the difference in caspase-1 activity in good vs. poor outcome remained significant (adjusted mean 7.10x10-3 vs. 2.54x10-2 CPS/μl*min, p=0.004). Conclusions: The inflammasome-dependent protein caspase-1 is elevated in CSF early after SAH and higher in those with poor functional outcome. Inflammasome activity therefore may serve as a novel biomarker to predict outcome shortly after aneurysm rupture.

Variation of membrane particle-bound CD133 in cerebrospinal fluid of patients with subarachnoid and intracerebral hemorrhage.

Previous studies have demonstrated that human CSF contains membrane particles carrying the stem cell antigenic marker CD133 (prominin-1). Here, the authors analyzed the variation of the amount of these CD133-positive particles in the CSF of patients with subarachnoid hemorrhage (SAH) and intracerebral hemorrhage (ICH). Consecutive CSF samples from 47 patients with SAH or ICH were compared to 14 healthy control patients. After differential ultracentrifugation of CSF, the membrane particle fraction was separated on gel electrophoresis and its CD133 content was probed by immunoblotting using the mouse monoclonal antibody 80B258 directed against human CD133. The antigen-antibody complexes were detected by chemiluminescence reagents and quantified using human Caco-2 cell extract as positive control with a standardized curve. As compared to healthy controls (6.3 ± 0.5 ng of bound CD133 antibody; n = 14), the amount of membrane particle-associated CD133 immunoreactivities was significantly elevated in patients with SAH and ICH (38.2 ± 6.6 ng and 61.3 ± 11.0 ng [p < 0.001] for SAH [n = 18] and ICH [n = 29], respectively). In both groups the CD133 level dropped during the first 7 days (i.e., day 5-7: SAH group, 24.6 ± 10.1 ng [p = 0.06]; ICH group, 25.0 ± 4.8 ng [p = 0.002]). Whereas changes in the amount of CD133-positive membrane particles between admission and day 5-7 were not associated with clinical outcomes in patients with ICH (modified Rankin Scale [mRS] scores 0-3, -30.9 ± 12.8 ng vs mRS scores 4-6, -21.8 ± 10.7 ng; p = 0.239), persistent elevation of CD133 in patients with SAH was related to impaired functional outcome 3 months after ictus (mRS scores 0-2, -29.9 ± 8.1 ng vs mRS scores 3-6, 7.6 ± 20.3 ng; p = 0.027). These data are expressed as the mean ± standard error of the mean (SEM). Levels of membrane particle-associated CD133 in the CSF of patients with SAH and ICH are significantly increased in comparison to healthy patients, and they decline during the hospital stay. Specifically, the persistent elevation of CD133-positive membrane particles within the first week may represent a possible surrogate measure for impaired functional outcome in patients with SAH.

Effects of apple polyphenols on oxidative stress and cerebral vasospasm after subarachnoid hemorrhage in a rabbit double hemorrhage model

Subarachnoid hemorrhage (SAH) is almost always caused by ruptured cerebral aneurysms, and the most serious complication after SAH is cerebral vasospasm. Oxidative stress due to free radicals released from subarachnoid hemorrhage clots causes cerebral vasospasm, as proven by animal experiments. Apple polyphenols have already been demonstrated to strongly suppress oxidized low-density lipoprotein (ox-LDL) and lectin-like oxidized LDL receptor-1 (LOX-1) derived from reactive oxygen species (ROS), and to improve cerebral vasospasm. However, it is unclear how apple polyphenol acts downstream of ROS. The purpose of this study is to reveal a more detailed mechanism of functioning for apple polyphenols in suppressing oxidative stress and cerebral vasospasm. Using the double-hemorrhage rabbit subarachnoid hemorrhage (SAH) model, we investigated the effect of apple polyphenols by measuring the basilar artery diameter, endothelial NO synthase (eNOS) expression, diacron reactive oxygen metabolites (d-ROM), biological antioxidant potential (BAP), and malondialdehyde (MDA). The mean diameter of the basilar artery in the apple polyphenol treatment group was significantly larger than in the SAH group (p < 0.01). The expression of endothelial nitric oxide synthase (eNOS), evaluated by immunohisto-chemistry in the apple polyphenol treatment group, was higher than in the SAH group (p < 0.05). The BAP test in the apple polyphenol treatment group showed a significant difference, while there was no significant difference in the values of d-ROM and MDA. The results of the present study revealed that apple polyphenol significantly improved cerebral vasospasm and eNOS. In spite of a significant increase in antioxidant power expressed by increased BAP, oxidative stress measured by the d-ROM value was not significantly suppressed. Either the number of experimental animals was not sufficient to reach statistical significance, or the eNOS increase due to apple polyphenols and suppression of vasospasm are attributable to pathways other than lipid oxidation detected by MDA.

Upregulation of Connexin 40 Mediated by Nitric Oxide Attenuates Cerebral Vasospasm After Subarachnoid Hemorrhage via the Nitric Oxide-Cyclic Guanosine Monophosphate-Protein Kinase G Pathway

The present study was performed to elucidate the role of nitric oxide (NO) and connexin 40 (Cx40) in the induction of cerebral vasospasm after subarachnoid hemorrhage (SAH) invivo. A SAH rat model was established using the double-bleed method. A total of 108 Sprague-Dawley rats weighing 250-300 g were randomly divided into 6 groups: SAH; SAH plus diethylenetriamine (DETA)/NO (exogenous NO donor); SAH plus 8-bromoadenosine (8-Br)-cyclic guanosine monophosphate (cGMP; protein kinase G [PKG] activator); SAH plus DETA/NO plus KT5823 (PKG inhibitor); SAH plus DETA/NO plus 40Gap27 (Cx40 inhibitor); and sham. The changes in the diameter of the branch microvessels in the middle cerebral artery were recorded. The neurological score was evaluated using the Garcia scoring system. Basilar artery (BA) tension was measured using the Danish Myo Technology myograph system. Cx40 protein expression was analyzed using immunofluorescence and Western blotting. Endothelial NO synthase, soluble guanylate cyclase, and PKG protein expression were measured by Western blotting. A considerable narrowing of the cerebral vessels was detected in the SAH group compared with that in the sham group. Moreover, compared with the sham group, the SAH group showed a marked decrease in Cx40, endothelial NO synthase, soluble guanylate cyclase, and PKG expression. The expression of Cx40 and PKG were obviously higher in the SAH plus DETA/NO and SAH plus 8-Br-cGMP groups than in the SAH group. However, Cx40 was lower in the SAH plus DETA/NO plus KT5823 and SAH plus DETA/NO plus 40Gap27 groups than in the SAH plus ETA/NO group. The BAs showed significant vasodilation in the SAH plus DETA/NO and SAH plus 8-Br-cGMP groups. However, the vasodilation response of BAs was inhibited in the SAH plus DETA/NO plus KT5823 and SAH plus DETA-NO plus 40Gap27 groups. The NO-cGMP-PKG pathway alleviated cerebral vasospasm via Cx40 upregulation.

Correlation between Arteriole Membrane Potential and Cerebral Vasospasm after Subarachnoid Hemorrhage in Rats.

Microvessel constriction plays an important role in delayed cerebral ischemia after aneurismal subarachnoid hemorrhage (SAH). This constriction has been demonstrated in both animal model and clinical operation. The present study examined the time-related membrane potential (Em) alteration of arterioles isolated from SAH model rats and the correlation between the potential alteration of arterioles and the diameter of basilar artery. Sprague-Dawley rats (n = 90), weighing 300 g to 350 g, were divided into t control, sham, and SAH groups. In the SAH group, blood was injected into the prechiasmatic cistern of the rats. The Em of arterioles and basilar artery diameter was measured using whole-cell clamp recordings and pressure myograph, respectively, 1, 3, 5, 7, and 14 days after SAH. The correlation was evaluated using Pearson correlation coefficients. The Em of arterioles in the SAH group depolarized on days 3, 5, and 7, and peaked on day 7. The diameters significantly decreased on days 1, 3, 5, 7, and 14, and the smallest diameter was observed on day 7. A significant correlation between potential alteration of arterioles and diameter of basilar artery was found. Similar to the artery, arteriole constriction is also involved in the pathophysiological events of delayed cerebral ischemia.

Subarachnoid haemorrhage with negative tomographic angiography without vasospasm: is it necessary to perform angiographic controls?

In about 15% of cases, the lesion causing subarachnoid haemorrhage (SAH) is not detectable by imaging studies, and some controversy exists as regards how the diagnosis should be performed in these patients. To analyse the diagnostic performance of digital subtraction angiography (DSA) carried out on a significant cohort of patients with SAH and negative tomographic angiography, without cerebral vasospasm, based on the distribution of bleeding in a simple CT scan. A retrospective analysis is performed of the centre's radiological database over an eight-year period. The inclusion criteria were patients with SAH of a perimesencephalic or diffuse distribution with a negative tomographic angiography result, without cerebral vasospasm, with at least one DSA control of the brain vessels. A total of 125 patients met the inclusion criteria: 61 had a perimesencephalic SAH and 64 had a diffuse SAH. The first DSA revealed vascular injury in seven patients (5.6%): one was in the perimesencephalic SAH group (1.6% yield) and six in the diffuse SAH group (9.4% yield). A second DSA was performed on 86 patients, a third on 28 and a fourth on five of them. The absence of vasospasm in the first DSA performed in patients with SAH and negative tomographic angiography could be a useful criterion for establishing an optimal follow-up protocol in these patients, without exposing them to unnecessary risks and radiation.

Predestinating Role of Cardiac Ganglia on Heart Life Expectancy in Rabbits After Brain Death Following Subarachnoid Hemorrhage: An Experimental Study

BackgroundCardiac ganglia are rechargeable batteries of the heart. The essential role of cardiac ganglia on cardiac life expectancy has not been examined following brain death. The aim of this study was to determine cardiac ganglia numbers and neuron density following subarachnoid hemorrhage (SAH). MethodsTwenty-five hybrid rabbits were grouped as control (n = 5), sham (n = 5), and SAH (n = 15). The SAH groups’ animals were subjected to injections of lethal dose of 2.00 cc autologous blood into their cisterna magna until linear EEG was obtained. The hearts of all animals were extracted following intracardiac formalin injection and examined. Cardiac ganglia and normal/degenerated neuron densities of cardiac neurons were recorded. ResultsThe mean volume of normal neuron density of ganglia was 6.980 ± 830/mm3, and the degenerated neuron density of ganglia was 3 ± 1/mm3 in the control group, 6134 ± 712/mm3; 23 ± 9/mm3 in the sham group, 3456 ± 589; 1161 ± 72/mm3 in the surviving group; and 1734 ± 341/mm3, 4259 ± 865/mm3 in the dead animals in the SAH group. The algebraic results of heart work capacity (Wh) were estimated as 1375 ± 210 Wh in the control group, 1036 ± 225 in the sham group, 800 ± 110 Wh in the surviving group, and < 100 ± 20 in the dead animals in the SAH group. Degenerated cardiac neuron density/Wh correlation is statistically meaningful between the dead in the SAH group versus the SAH-surviving, sham, and control groups (P < .0005). ConclusionsNormal cardiac ganglia numbers and/or cardiac ganglia neuron density may be related to cardiac survival following brain death after subarachnoid hemorrhage.

MEL Ameliorates Post-SAH Cerebral Vasospasm by Affecting the Expression of eNOS and HIF1α via H19/miR-138/eNOS/NO and H19/miR-675/HIF1α

Melatonin (MEL) has been demonstrated to exert a protective effect against subarachnoid hemorrhage (SAH), and nitric oxide (NO) has been shown to play an important role in the pathogenesis of vasospasm. This study aims to explore the underlying molecular mechanisms of MEL in the control of vasospasm following SAH. MEL administration attenuates SAH-induced vasospasm and neurobehavioral deficits. Expressions of H19, eNOS, and miR-675 are low in the SAH group, while expressions of miR-138 and HIF1α are high in the SAH group. Also, MEL treatment upon SAH rats completely restores the dysregulation of H19, eNOS, miR-675, miR-138, and HIF1α to their normal levels. Moreover, MEL dose dependently increases the luciferase activity of H19 promoter and hence the expression of H19. Additionally, H19 directly targets miR-675 and miR-138 to increase miR-675 expression and inhibit miR-138 expression. As virtual target genes of miR-675 and miR-138, respectively, HIF1α and eNOS are also regulated by the treatment with MEL. In particular, MEL treatment increases the expression of miR-675 and eNOS level while decreasing the expression of miR-138 and HIF1α in a dose dependent manner. Our study found that MEL ameliorates post-SAH vasospasm by regulating the expression of eNOS and HIF1α via the H19/miR-138/eNOS/NO and H19/miR-675/HIF1α signaling pathways.

Edaravone combined with cinepazide maleate on neurocyte autophagy and neurological function in rats with subarachnoid hemorrhage.

Effects of edaravone combined with cinepazide maleate on neurocyte autophagy and neurological function in rats with subarachnoid hemorrhage were investigated. Eighty SD rats were selected to establish subarachnoid hemorrhage (SAH) rat models, which were divided into sham operation group, SAH group, MCI group and combined group. Hippocampal tissue of each group was taken to observe the number of neurocytes. The expression levels of Beclin-1 and (light chain LC3)-II of rats in each group were detected by ELISA. Pearson's correlation factors were used to analyze the correlation between Beclin-1 and LC3-11, and neurological function tests were carried out on rats of each group 14 and 28 days after administration. The morphological and structural damage of nerve cells in the combined group was further alleviated, and the survival rate of neurons significantly increased at all time points (P<0.05). The expression levels of Beclin-1 and LC3-11 in combined group was significantly higher than those in SAH group and CMI group (P<0.05), and Beclin-1 was positively correlated with LC3-11 (r=0.9454). Longa score of the combined group was significantly lower than that of the other two groups, and muscle strength score was significantly higher than that of the other two groups (P<0.05). Edaravone combined with cinepazide maleate can effectively increase the survival rate of brain cells and promote the volatilization of neurological function in the treatment of hemorrhage in the subretinal space of the omentum, which is worthy of popularization and application.

Soluble receptor for advanced glycation end products as a biomarker of symptomatic vasospasm in subarachnoid hemorrhage.

The receptor for advanced glycation end products (RAGE) is a membrane protein associated with the induction of oxidative stress and inflammation in several pathological conditions. Previous studies have demonstrated that soluble RAGE (sRAGE) acts as a decoy for RAGE and protects cells against RAGE-mediated injury. The authors and other groups have reported that the expression of RAGE increases after brain ischemia and subarachnoid hemorrhage (SAH), and deletion of RAGE or overexpression of sRAGE improves neuronal survival. It has also been demonstrated that the plasma sRAGE level could be a predictor of the outcome after ischemic stroke. This study aimed to evaluate plasma sRAGE as a biomarker for symptomatic vasospasm (SVS) in SAH patients, as well as a rat model. The authors measured and compared plasma sRAGE levels in 27 SAH patients (7 with SVS and 20 without SVS) from day 5 to day 14 post-SAH. They also examined plasma sRAGE levels and expression of RAGE and heme oxygenase-1 (HO-1) in a rat SAH model. The relative plasma sRAGE levels were significantly lower in the SVS group than in the non-SVS group of patients. A cut-off value of 0.84 for predicting SVS was considered to be appropriate for the relative plasma sRAGE levels on day 7 versus day 5. In the rat SAH model, plasma sRAGE levels were significantly lower than those in sham-treated rats, and the expressions of RAGE and HO-1 were enhanced in the SAH group compared with the non-SAH group. Plasma sRAGE levels can be used as a potential biomarker for predicting SVS after SAH.

Incidence of hydrocephalus in traumatic brain injury: A nationwide population-based cohort study.

Background:The aim of this study was to investigate the risk and peak time of post-traumatic hydrocephalus (PTH) in traumatic brain injury (TBI) patients with traumatic subarachnoid hemorrhage (SAH), compared to TBI patients without traumatic SAH.Methods:In this retrospective population-based cohort study, the data was extracted from Longitudinal Health Insurance Database from 2000 to 2010 in Taiwan. A total of 23,775 TBI patients who had a first event TBI during 2000 to 2010 were included and divided into TBI with SAH (TBI-S) group and TBI without SAH (TBI-NS) group. We focused on analyzing the PTH in both groups within 2 years after brain injury. Competing risk regression models were performed to assess the risk of developing PTH in the TBI-S group compared to the TBI-NS group.Results:Comparing to the TBI-NS group, there was a significantly higher cumulative incidence of PTH in the TBI-S group during the 2-year follow-up period. The adjusted hazard ratio (HR) of PTH in TBI-S group within 2 years was between 2.90–3.47, and the highest estimates were obtained within 6 months after injury (HR = 3.47, 95% CI: 2.43–4.94). The occurrence percentage of PTH was highest during 0–3rd month follow-up periods (1.95% in TBI-S group; 0.48% in TBI-NS group).Conclusions:The peak time of PTH occurrence was noted during 0–3rd month post brain injury. Traumatic SAH patients had an approximate 3-fold risk of developing PTH, comparing to TBI patients without traumatic SAH.

P3708Prognostic significance of electrocardiographic changes after subarachnoid hemorrhage

Abstract Background and purpose Many reports have examined electrocardiographic (ECG) changes after subarachnoid hemorrhage from long ago; however, there are few reports on the prognosis of patients who have an ECG abnormality. Therefore, in this study, the prognoses of patients with subarachnoid hemorrhage showing abnormal ECG findings were retrospectively analyzed. Methods Over 30 months from January 2014 to June 2016, 199 patients (mean age, 61.1±17.0 years; 46.7% male) admitted with a subarachnoid hemorrhage (SAH) were enrolled. To assess the mechanism underlying abnormal ECG in the SAH, predictor variables, such as demographics (age, sex, and body surface area), hemodynamics (heart rate and systolic blood pressure), blood biochemical results, neurological assessments (Glasgow Coma Scale), and computed tomography (CT) severity classification (World Federation of Neurosurgical Societies classification) were recorded. The subarachnoid hemorrhage was classified into either the cerebral aneurysm rupture group (N=132) or traumatic subarachnoid hemorrhage group (N=67) and analyzed. Results In the cerebral aneurysm rupture group, the QT prolongation was significantly increased compared with the traumatic subarachnoid hemorrhage group (424.8±87.7 ms vs. 400.5±95.8 ms, P&lt;0.05). There was a correlation between consciousness level (Glasgow Coma Scale) on admission and QT prolongation, and significant QT prolongation was observed in critical patients (Pearson's correlation coefficient test P=0.04). ST changes correlated with CT classification severity (WFNS classification), and the most severe group (WFNS Grade 5) showed significant ST changes. On admission, the cerebral aneurysm rupture group and 97.0% in the traumatic subarachnoid hemorrhage group. Conclusions Significantly prolonged QT and ST changes are noted in patients with severe aneurysmal subarachnoid hemorrhage. The cause of the ECG abnormality in subarachnoid hemorrhage patients is not yet understood. A small number of experimental animal models have shown that electrocardiogram abnormalities associated with subarachnoid hemorrhage develop in the brain stem, and an autonomic nerve abnormality may also be involved. A D-dimer is positively correlated with both aneurysm trauma and traumatic subarachnoid hemorrhage and may be used for auxiliary diagnosis of a subarachnoid hemorrhage.

Morphological Characteristics of Neuronal Death After Experimental Subarachnoid Hemorrhage in Mice Using Double Immunoenzymatic Technique.

Subarachnoid hemorrhage (SAH) is a devastating disease. Neuronal death is an important pathophysiology in the acute phase of SAH, but the histopathological features of dying neurons have been poorly studied. Using several staining methods including terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and microtubule-associated protein 2 (MAP-2) double immunolabeling, we investigated the morphological changes of nucleus and cytoskeleton in neurons and sought susceptible areas to neuronal death in filament perforation SAH mice under light microscope. TUNEL and MAP-2 double immunolabeling clearly showed morphological features of shrunken cytoplasm and sometimes curl-like fibers in dying neurons, besides nuclear abnormalities. More dying neurons were detected in the moderate SAH group than in the mild SAH group, and the temporal base cortex was the most susceptible area to neuronal death with deoxyribonucleic acid (DNA) damage among the cerebral cortices and hippocampus at 24 hr after SAH (p<0.01, ANOVA). Lesser hippocampal neuronal death was observed at 24 hr, but neuronal death was significantly increased in the CA1 region at 7 days after SAH (p<0.05, unpaired t-test). Using TUNEL and MAP-2 double immunolabeling, morphological features of not only the nucleus but also the cytoplasm in post-SAH neuronal death with DNA damage can be observed in detail under light microscope.

Study on the cerebral microcirculation of rats with subarachnoid hemorrhage by using laser speckle technique

Objective To study the value of laser speckle technique in measuring blood flow changes in the cerebral microcirculation of rats with subarachnoid hemorrhage (SAH). Methods Rat double blood injection method was used to establish the model of SAH. Rats were divided into control group, SAH 5-day group, SAH 7-day group, saline 5-day group and saline 7-day group. After sensory nerve function examination, cerebral cortical blood flow was measured by laser speckle technique, and apoptotic index and electron microscopy were detected. SPSS 17.0 statistical software was used to make statistical analysis by variance analysis or non-parametric test. Results SAH 5-day group showed significant sensory dysfunction. There were significant differences between SAH 5-day group (12.100±1.287) and control group (15), SAH 7-day group (14.700±0.675), saline 5-day group (14.900±0.316) and saline 7-day group (15) (Z values were -4.059, -3.655, -3.908, -4.059, respectively; respectively P<0.01); Electron microscopy showed that the morphology of neurons in the control group and saline injection groups was basically normal; neurons in the SAH 5-day group showed obvious edema and extensive neuronal degeneration or necrosis; neurons in the SAH 7-day group showed slight swelling and some neurons had degeneration or necrosis. There were obvious apoptotic cells in cerebral cortex of SAH groups. The apoptotic index of SAH 5-day group [(19.870±1.909)%] and control group [(1.840±0.474)%], SAH 7-day group [(13.770±1.924)%], saline 5-day group [(1.740±0.299)%], saline 7-day group [(1.790±0.407)%] had statistical differences (t values were 28.985, 7.117, 29.670, 29.291, respectively; respectively P<0.01). SAH 7-day group and control group, SAH 5-day group, saline 5-day group, saline 7-day group had statistical differences (t values were 19.037, -7.117, 19.537, 19.263, respectively; respectively P<0.01). Cerebral cortical blood flow in SAH groups decreased, and the most obvious decrease was found in SAH 5-day group (64.000±6.218), compared with control group (106.000±13.081), SAH 7-day group (75.700±8.274), saline 5-day group (105.100±13.169), saline 7-day group (104.100±10.609). There were significant differences (t values were -9.170, -3.575, -8.924, -10.312, respectively; successively P<0.01, 0.05, 0.01, 0.01). There were significant differences between SAH 7-day group and control group, SAH 5-day group, saline 5-day group and saline 7-day group (t values were -6.191, 3.575, -5.978 and -6.675, respectively; successively P<0.01, 0.05, 0.01, 0.01, 0.01). Conclusion Laser speckle technique is suitable for studying the blood flow changes of cerebral microcirculation in SAH rats. Key words: Subarachnoid hemorrhage; Cerebral microcirculation; Cerebral blood flow; Laser speckle

Describing a New Mechanism of Retinal Detachment Secondary to Ophthalmic Artery Vasospasm Following Subarachnoid Hemorrhage: An Experimental Study.

The pterygopalatine ganglion (PPG) and ophthalmic arteries (OpAs) have important roles in ocular autoregulation and retinal and visual functions. The relationship between PPG neuron density, OpA vasospasm, and retinal detachment in subarachnoid hemorrhage (SAH) has never been studied. This study was conducted on 25 rabbits. Five animals were in the control group (GI; n = 5), five in the sham group (GII; n = 5), and 15 in the study group (GIII; n = 15). After injection of 1 cc serum saline into the cisterna magna in the sham group, and autologous blood in the SAH group, the animals were followed for 3 weeks. All animals underwent a retinal examination five times a week for 3 weeks before and after the experiment. After the experiment, the neuron density of PPGs of the facial nerves, vasospasm index (VSI) of OpAs, and total basal surface values of the detached retinal parts (DRPs) were calculated. In the funduscopic examination, intravitreous hemorrhage ( Terson's syndrome) was detected in four animals in the SAH group. In the control groups, neuron density was 12,000 ± 1,240/mm3, VSI = 0.345 ± 0.076, and DRP = 0 to 1.5 mm2. Mean neuron density was 9,450 ± 940/mm3, VSI = 1.645 ± 0.940, and DRP = 6.23 ± 1.61 mm2 in the sham group (p < 0.05). Neuron density was 6,890 ± 932/mm3, VSI = 2.92 ± 0.97, and DRP = 9.43 ± 2.54 mm2 in SAH group. Mean neuron density, VSI of OpAs, and DRP values differed statistically significant between the SAH group and other groups (p < 0.005). There is an inverse relationship between PPG neurons and DRP. However, a direct relationship was observed between the mean VSI and DRP values.