Current Neurovascular Research - Volume 16, Issue 3, 2019

Background: Retinal degeneration and related eye disorders have limited treatment interventions. Since stem cell therapy has shown promising results, ciliary epithelium (CE) derived stem cells could be a better choice given the fact that cells from eye niche can better integrate with the degenerating retina, rewiring the synaptic damage. Objective: To test the effect of human fetal pigmented ciliary epithelium-derived neurospheres in the mouse model of laser-induced retinal degeneration. Methods: C57 male mice were subjected to retinal injury by Laser photocoagulation. Human fetal pigmented ciliary epithelium was obtained from post-aborted human eyeballs and cultured with epidermal growth factor (rhEGF) and fibroblast growth factor (rhFGF). The six day neurospheres were isolated, dissociated and transplanted into the subretinal space of the laser injured mice at the closest proximity to Laser shots. Mice were analyzed for functional vision through electroretinogram (ERG) and sacrificed at 1 week and 12 week time points. Retinal, Neurotropic, Apoptotic and proliferation markers were analysed using real-time polymerase chain reaction (PCR). Results: The CE neurospheres showed an increase in the expression of candidate genes analyzed in the study at 1 week time point, which sustained for longer time point of 12 weeks. Conclusion: We showed the efficacy of human CE cells in the regeneration of retinal degeneration in murine model for the first time. CE cells need to be explored comprehensively both in disease and degeneration.

Background: The present study aimed at determining pericytes, a missing component in the previously proposed living neurovascular unit (NVU) of the sexually dimorphic nucleus of the preoptic area (SDN-POA) in rats. Materials and Methods: Calbindin D28K-immunoreactivities (CB28-irs) were used to delineate the SDN-POA in which CD13-immunoreactivities (CD13-irs) or alpha-smooth muscle actinimmunoreactivities (αSMA-irs), two pericyte biomarkers serving the indexes of pericytes, were tagged using two adjacent brain sections (90-micron intervals). In addition, the nestinimmunoreactive (nestin-ir) cells in the SDN-POA were counted as pericytes referring to additional standards: location and nucleic and cellular morphology. Male SDN-POA volume (5.0±0.3x10-3 mm3) was significantly larger than the female (1.7±0.3x10-3 mm3). Within the SDN-POA, the CD13-irs were characterized as dots, densely packed and net-like in distribution, while the αSMAirs, excluding pipe-like or circular structures, appeared as short rod-like structures that were sparsely distributed. Results: The immunoreactive counts of alpha-smooth muscle actin were 353±57/mm2 in males and 124±46/mm2 in females (p<0.05). On the other hand, densities of the dot-like CD13-irs were similar between males (4009±301/mm2) and females (4018±414/ mm2). There was no difference between the male and the female in the nestin-ir pericyte count in the SDN-POA. Conclusion: In conclusion, the present study adds new information concerning pericytes to the living NVU of the SDN-POA. There is a difference of sex in the count of the αSMA-irs in the living NVU of the SDN-POA. However, why such a difference exists warrants further investigations.

Background: Moyamoya disease (MMD) is a rare cerebrovascular disease. The difference of hemorrhagic patterns in adult patients with bilateral and unilateral MMD is still unclear. Objective: For a better understanding of their characteristics, we compared the patterns of acute intracranial hemorrhage in adult patients with bilateral and unilateral MMD. Methods: Adult MMD patients with acute intracranial hemorrhage were retrospectively included. Clinical and radiological characteristics of adult patients with bilateral and unilateral MMD were collected and analyzed. Chi-square test, t-test, or rank sum test were used for statistical analyses. Results: A total of 107 patients were included. Among 74 patients with bilateral MMD, 9 (12.2%) were at Suzuki Stage 2, 48 (64.9%) were at Stage 3, 16 (21.6%) were at Stage 4, and another (1.4%) was at Stage 5. However, in patients with unilateral MMD, 8 (24.2%) were at Stage 2, 23 (69.7%) were at Stage 3, and 2 (6.1%) were at Stage 4. Intraparenchymal hemorrhage was found in 40 (54.1%) patients with bilateral MMD and 16 (48.5%) patients with unilateral MMD (P=0.594). Intraventricular hemorrhage was shown in 65 (87.8%) patients with bilateral MMD and 19 (57.6%) patients with unilateral MMD (P<0.001). Subarachnoid hemorrhage was observed in 17 (23.0%) patients with bilateral MMD and 18 (54.5%) patients with unilateral MMD (P=0.001). Conclusion: Unilateral MMD patients with acute intracranial hemorrhage are at the earlier Suzuki stage than the bilateral MMD patients. Intraventricular hemorrhage occurs more frequently in bilateral MMD, while subarachnoid hemorrhage is more frequent in unilateral MMD.

Objective: This study aims to analyze the clinical characteristics, treatment and prognosis of children with cutaneous polyarteritis nodosa (CPAN), in order to improve the understanding of this disease. Methods: Data of 14 children with CPAN, who were hospitalized in the Beijing Children's Hospital of Capital Medical University from January 2006 to December 2016, were collected. The clinical characteristics of all patients were summarized, the antistreptolysin-O (ASO)-positive and ASO-negative groups were compared, and the follow-up results were analyzed. X2-test, Fisher’s exact probability test, t-test and Mann-Whitney test were used for statistical analysis. Results: Among these 14 CPAN patients, nodular rash was the most common manifestation (14/14). The ASO-positive group had more nodules in the lower limbs and the ASO-negative group appeared more in the upper limbs, which were statistically significant (p<0.05). ASOpositive children were more likely to have joint symptoms (P<0.05), and were more prone to elevated white blood cells (P<0.05). Follow-ups were performed on nine patients, and the prognoses were all good. The occurrence of systemic polyarteritis nodosa was not observed. Conclusion: The main clinical manifestation of children with CPAN is skin nodules, which rarely affects the internal organs. Streptococcal infection is often the main cause. Anti-infection treatment should be simultaneously considered.

Background: Studies have previously shown greater arterial and venous extracranial vascular changes in persons with multiple sclerosis (PwMS) when compared to healthy controls (HCs). Objectives: To determine the change in the number and size of secondary neck vessels in PwMS and HCs over a 5-year follow-up period. Methods: Both at baseline and follow-up, 83 PwMS and 25 HCs underwent magnetic resonance angiography (MRA) imaging and analysis. The number and cross-sectional area (CSA) of all secondary neck vessels (excluding the common/internal carotid, vertebral artery, and internal jugular vein) measured at levels from C2-T1 were determined by semi-automated edge detection/ contouring software. The longitudinal change in the number and CSA of the secondary neck vessels from the PwMS and HCs were analyzed by non-parametric Wilcoxon repeated measure. Benjamini-Hochberg procedure adjusted for false discovery rate (FDR). Results: For over 5 years, PwMS demonstrated a consistent longitudinal decrease in both the number of secondary neck vessels (Z-change between -3.3 and -5.4, q=0.001) and their CSA (Zchange between -2.9 and -5.2, q=0.004). On the contrary, the HCs did not demonstrate a significant longitudinal change in secondary neck vessels over the follow-up period. Due to the longitudinal decrease, the PwMS showed a lower number of secondary neck vessels when compared to HCs measured at follow-up (p<0.029, except for C4 with trending p=0.071). The PwMS changes were also corroborated within each MS phenotype. Conclusion: PwMS demonstrate a significant mid-term decrease in the number and the size of the secondary neck vessels. The clinical relevance of these findings and the effect on intracranial blood flow are currently unknown.

Background: The vascular morphology and the characteristics of atherosclerotic plaques in the middle cerebral artery (MCA) have not been fully studied with high-resolution magnetic resonance imaging (HR-MRI). Objective: HR-MRI was applied to investigate vascular morphology and atherosclerotic plaque in patients with symptomatic MCA stenosis. Materials and Methods: A total of 343 patients with symptomatic MCA stenosis were enrolled in this study. All the patients were examined by HR-MRI to analyze the morphology of MCA and the M1 segment (MCA-M1), the characteristics and the location of the plaques. Results: The proportion of L-shaped MCA-M1 decreased, while the proportion of S-shaped MCAM1 increased with age. The anterior plaques were the most common in all the patients. The superior plaques were relatively common in patients with L-shaped and U-shaped MCA-M1, while the inferior plaques were relatively common in patients with inverted U-shaped and S-shaped MCAM1. Among all the plaques, the majority were isointense or heterogeneous. The MCA-M1 morphology had no direct relationship with the common risk factors of atherosclerosis and the clinical outcomes of the patients after 12 months of follow up. Conclusion: The morphology of MCA-M1 is not directly related to the plaque burden or the degree of stenosis in patients with symptomatic MCA stenosis. The morphology of MCA-M1 is not associated with the risk factors of atherosclerosis, or the clinical outcomes of the patients.

Background: The inflammatory response to acute cerebral ischemia is a major factor in stroke pathobiology and patient outcome. In the clinical setting, no effective pharmacologic treatments are currently available. Phenothiazine drugs, such as chlorpromazine and promethazine, (C+P) have been widely studied because of their ability to induce neuroprotection through artificial hibernation after stroke. The present study determined their effect on the inflammatory response. Methods: Sprague-Dawley rats were divided into 4 groups: (1) sham, (2) stroke, (3) stroke treated by C+P without temperature control and (4) stroke treated by C+P with temperature control (n=8 per group). To assess the neuroprotective effect of C+P, brain damage was measured using infarct volume and neurological deficits. The expression of inflammatory response molecules tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and nuclear factor kappa light chain enhancer of activated B cells (NF-ΚB) was determined by real-time PCR and Western blotting. Results: TNF-α, IL-1β, ICAM-1, VCAM-1, and NF-ΚB mRNA and protein expressions were upregulated, and brain damage and neurological deficits were increased after stroke. These markers of cerebral injury were significantly reduced following C+P administration under drug-induced hypothermia, while C+P administration under normal body temperature reduced them by a lesser degree. Conclusion: This study showed an inhibitory effect of C+P on brain inflammation, which may be partially dependent on drug-induced hibernation, as well as other mechanisms of action by these drugs. These findings further suggest the great potential of C+P in the clinical treatment of ischemic stroke.

Objective: Recent studies suggest that not only is constipation a clinical marker of premotor phase in Parkinson’s Disease (PD), but is also correlated with the duration and severity. Some reports indicated that inflammatory from gut dysbiosis might be involved in the pathogenesis of PD, but the correlation between them remains poorly understood. This study aims to investigate how the presence of constipation affects the dopamine level of nigrostriatal system and whether gastrointestinal (GI) inflammation is involved in the brain-gut axis. Methods: Clinical materials, serum inflammatory factors, and datum of dopamine level including 84 cases and 83 controls, were collected consecutively and randomly from November 1, 2017 to October 31, 2018. Dopamine levels of nigrostriatal system were detected by [18F]-DTBZ radiotracer (18F-AV-133). Data analysis was conducted by variance, covariance analysis, bicorrelation, partial correlation, chi-square analysis and logistic regression. Results: The mean age of cases was older than that of controls, and male predominance was also observed (P<0.05). The mean scores of Hoehn-Yahr and unified Parkinson’s disease rating scale III (UPDRS-III) were of significantly different duration between two groups (P<0.05). The total dose of levodopa was not different between two groups (P>0.05). The dopamine levels of putamen and caudate nucleus, especially in the dorsal part of putamen, were significantly decreased in cases than that in controls (P<0.05). There were significant differences of complement 3 (C3) and complement 4 (C4) between cases and controls (P<0.05). Dopamine levels in putamen and caudate nucleus were negatively correlated with serum concentrations of immunoglobulin A (IgA), immunoglobulin G (IgG) and C3 in cases (P<0.05). But we did not observe similar negative correlations in controls (P>0.05). Conclusion: The presence of constipation may increase the severity of motor symptoms and decrease dopamine levels of nigrostriatal system in PD. Inflammatory factors may be involved in the brain-gut axis of PD.

Background and Purpose: Recurrent ischemic strokes increase the risk of disability and mortality. The role of conventional risk factors in recurrent strokes may change due to increased awareness of prevention strategies. The aim of this study was to explore the potential risk factors besides conventional ones which may help to affect the advances in future preventive concepts associated with one-year stroke recurrence (OSR). Methods: We analyzed 6,632 adult patients with ischemic stroke. Differences in clinical characteristics between patients with and without OSR were analyzed using multivariate logistic regression and classification and regression tree (CART) analyses. Results: Among the study population, 525 patients (7.9%) had OSR. Multivariate logistic regression analysis revealed that male sex (OR 1.243, 95% CI 1.025 – 1.506), age (OR 1.015, 95% CI 1.007 - 1.023), and a prior history of ischemic stroke (OR 1.331, 95% CI 1.096 – 1.615) were major factors associated with OSR. CART analysis further identified age and a prior history of ischemic stroke were important factors for OSR when classified the patients into three subgroups (with risks of OSR of 8.8%, 3.8%, and 12.5% for patients aged > 57.5 years, ≤ 57.5 years/with no prior history of ischemic stroke, and ≤ 57.5 years/with a prior history of ischemic stroke, respectively). Conclusion: Male sex, age, and a prior history of ischemic stroke could increase the risk of OSR by multivariate logistic regression analysis, and CART analysis further demonstrated that patients with a younger age (≤ 57.5 years) and a prior history of ischemic stroke had the highest risk of OSR.

Objective: This study aimed to examine whether DC101 (anti-VEGFR2 antibody)- modified micelles have applications as novel drug delivery devices, which allow small molecule antiangiogenic agents to deliver to angiogenic sites on a murine laser-induced choroidal neovascularization (CNV) model. Materials and Methods: CNV was induced by photocoagulation on the unilateral eye of each mouse under anesthesia. Immediately after laser coagulation, E7974-loaded DC101-modified micelles and motesanib-loaded DC101-modified micelles were intravitreally administrated. Two weeks after photocoagulation, CNV was visualized using fluorescein-conjugated dextran (MW=2,000 kDa), and the CNV area was measured in retinal pigment epithelium (RPE)-choroidal flat mounts. Results: Intravitreal administration of both DC101-modified micelles loaded with E7974 at 2 μM and motesanib at 2 μM significantly reduced CNV area in the murine laser-induced CNV model at a clearly lower concentration than the effective dose of each agent. Conclusion: These results suggest that DC101-modified micelle might be effective drug carrier system for treating CNV and other ocular angiogenic diseases.

Background and Purpose: Hemorrhagic transformation (HT) is a potentially serious complication in patients with acute ischemic stroke (AIS). Whether the ratio of low-density lipoprotein cholesterol to high-density lipoprotein cholesterol (LDL-C/HDL-C) is associated with HT remains unclear. Methods: Ischemic stroke patients within 7 days of stroke onset from January 2016 to November 2017 were included in this study. Lipid profiles were measured within 24h after admission. HT was determined by a second computed tomography or magnetic resonance imaging within 7 days after admission. Univariate and multivariate logistic regression analysis was used to assess the association between LDL-C/HDL-C and HT. Results: We enrolled 1239 patients with AIS (788 males; mean age, 64 ± 15 years), of whom 129 (10.4%) developed HT. LDL-C/HDL-C was significantly lower on admission in patients with HT than those without HT (2.00 ± 0.89 vs. 2.25 ± 1.02, P=0.009). The unadjusted odds ratio (OR) of low LDL-C/HDL-C for HT was 2.07 (95% confidence interval [CI] 1.42-3.01, P#156;0.001). After adjustment for possible confounders, lower LDL-C/HDL-C (≤1.52) was significantly associated with HT (OR 1.53, 95% CI: 1.02-2.31, P=0.046). Similar results were observed between lower LDL-C (≤ 4 mmol/L) and HT (OR 4.17, 95% CI: 1.25-13.90, P=0.02). However, no significant association was found between HT and high HDL-C, low triglycerides or low total cholesterol. Conclusion: Lower LDL-C/HDL-C and LDL-C were significantly associated with increased risk of HT after AIS. Further investigations are warranted to confirm these findings and then optimize lipid management in stroke patients with lower LDL/HDL-C or LDL-C.

Background: Ischemic stroke (IS) is a significant disease which threatens human health condition. Genome-wide association studies (GWAS) have demonstrated that two intergenic single- nucleotide polymorphisms (SNPs) rs11833579 and rs12425791 G>A on chromosome 12p13 are associated with IS susceptibility. However, later studies came to contradictory outcomes. Thus, we carried out a meta-analysis to identify the association between nerve injury-induced protein 2 (NINJ2) gene polymorphisms (rs11833579 and rs12425791) and the risk of IS. Methods: PubMed, Embase, Web of Science, CBM, Wanfang, VIP, and CNKI databases were searched until March 2019. Data was analyzed by RevMan 5.3 and STATA 12.0 software. The pooled odds ratio (OR) and 95% confidence interval (CI) were used to evaluate the strength of the association. Results: Eighteen qualified articles were selected in total. For rs12425791 and rs11833579, a total of 14055 cases with 13148 controls and 10635 cases with 10462 controls, respectively, were identified for the present study. Our meta-analysis found that rs12425791 was associated with IS for three genetic models (allele model: OR=1.04, 95% CI: 1.00-1.08, P=0.04; dominant model: OR=1.06, 95% CI: 1.01-1.12, P=0.01 and heterozygous model: OR=1.07, 95% CI: 1.01-1.12, P=0.02). Whereas rs11833579 polymorphism was not associated with IS among different genetic models. Conclusion: NINJ2 gene rs12425791 confers a susceptible factor for IS, while there is no association between NINJ2 gene rs11833579 and IS. Larger sample size studies should be performed to find the association between NINJ2 gene and IS.