Interleukin-16 polymorphism is associated with an increased risk of ischemic stroke.

Clinical and experimental data have demonstrated that inflammation plays fundamental roles in the pathogenesis of ischemic stroke. Interleukin-16 (IL-16) is identified as a proinflammatory cytokine that is a key element in the ischemic cascade after cerebral ischemia. We aimed to examine the relationship between the IL-16 polymorphisms and the risk of ischemic stroke in a Chinese population. A total of 198 patients with ischemic stroke and 236 controls were genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing method. We found that the rs11556218TG genotype and G allele of IL-16 were associated with significantly increased risks of ischemic stroke (TG versus TT, adjusted OR = 1.88; 95% CI, 1.15-3.07; G versus T, adjusted OR = 1.54; 95% CI, 1.05-2.27, resp.). However, there were no significant differences in the genotype and allele frequencies of IL-16 rs4778889 T/C and rs4072111 C/T polymorphisms between the two groups, even after stratification analyses by age, gender, and the presence or absence of hypertension, diabetes mellitus, hypercholesterolemia, and hypertriglyceridemia. These findings indicate that the IL-16 polymorphism may be related to the etiology of ischemic stroke in the Chinese population.

[Changes of nitric oxide and endothelin serum level after carotid balloon denudation or stent assisted angioplasty: an experimental and clinical observation].

OBJECTIVE: To observe the changes of nitric oxide (NO) and endothelin (ET) serum level in the Guangxi BA-MA minipigs whose carotid arteries were injured by balloon denudation and in the patients with carotid stent assisted angioplasty. METHODS: Twelve Guangxi BA-MA minipigs were chosen. High fat/cholesterol feeding and endovascular balloon denudation were used to create a carotid artery atherosclerotic stenosis animal model. Blood samples were collected from peripheral veins before starting the procedure, and again, at 2 and 3 weeks after the procedure, respectively. Serum NO and ET concentrations of blood samples were tested. Nineteen patients with carotid artery stenosis who underwent stent assisted angioplasty were randomly selected, and their serum NO and ET were tested using the same methods as above. RESULTS: In the animal group, there was a significant decrease of mean NO concentration at 2 weeks after carotid injury (t-test, P < 0.05), however, no significant change of ET was observed. A very significant increase of ET was observed at 3 weeks after the procedure (t-test, P < 0.01). In the patient group, there were no significant differences among serum NO or ET concentration of peripheral vein blood before, immediately after, and 6 h after the endovascular treatment. CONCLUSIONS: In this study, a decrease of NO concentration and an increase of ET concentration of peripheral vein blood are found in BA-MA minipigs after carotid arteries are injured by balloon denudation, which might be a cue for the formation of atherosclerosis. However, no significant changes are observed in this group of patients who underwent carotid angioplasty treatment. Therefore, further studies are needed.

Regulatory effects of the long non­coding RNA RP11­543N12.1 and microRNA­324­3p axis on the neuronal apoptosis induced by the inflammatory reactions of microglia.

The present study aimed to examine how the long non­coding RNA (lncRNA) RP11­543N12.1 interacted with microRNA (miR)­324­3p to modify microglials (MIs)­induced neuroblastoma cell apoptosis, which may pose benefits to the treatment of Alzhemier's disease (AD). The cell model of AD was established by treating SH­SY5Y cells with amyloid ß (Aß)25­35, and MI were acquired using primary cell culture technology. The lncRNAs that were differentially expressed between SH­SY5Y and control cells were screened through a microarray assay and confirmed via polymerase chain reaction. In addition, overexpression of RP11­543N12.1 and miR­324­3p was established by transfection of SH­SY5Y cells with pcDNA3.1(+)­RP11­543N12.1 and miR­324­3p mimics, respectively, while downregulation of RP11­543N12.1 and miR­324­3p was achieved by transfection with RP11­543N12.1­small interfering RNA (siRNA) and miR­324­3p inhibitor, respectively. The interaction between RP11­543N12.1 and miR­324­3p was confirmed with a dual­luciferase reporter gene assay. The results revealed that the expression levels of total and phosphorylated tau in SH­SY5Y cells were significantly elevated following Aß25­35 treatment (P<0.05), and RP11­543N12.1 was found to be differentially expressed between the control and Aß25­35­treated cells (P<0.05). Furthermore, the targeted association of RP11­543N12.1 and miR­324­3p was predicted based on miRDB4.0 and PITA databases, and then validated via the dual­luciferase reporter gene assay. SH­SY5Y cells transfected with siRNA or inhibitor, and treated with Aß25­35 displayed cellular survival and apoptosis that were similar to the normal levels (P<0.05). Finally, co­culture of MI and SH­SY5Y cells transfected with RP11­543N12.1­siRNA/miR­324­3p inhibitor significantly enhanced cell apoptosis (P<0.05). In conclusion, RP11­543N12.1 targeted miR­324­3p to suppress proliferation and promote apoptosis in the AD cell model, suggesting that RP11­543N12.1 and miR­324­3p may be potential biomarkers and therapeutic targets for AD.

Granulocyte colony-stimulating factor regulates JNK pathway to alleviate damage after cerebral ischemia reperfusion.

BACKGROUND: Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a potent hematopoietic growth factor that both enhances the survival and drives the differentiation and proliferation of myeloid lineage cells. Recent studies have suggested that GM-CSF has a neuroprotective effect against cerebral ischemia injury, but the molecular mechanisms have been unclear. This study aimed to investigate the influences of a short-acting (half-life 3.5 hours) G-CSF and a long-acting (half-life 40 hours) pegylated G-CSF on the JNK signaling pathway after cerebral ischemia reperfusion. METHODS: A total of 52 Sprague-Dawley rats were randomly divided into four groups: a sham group (n = 4), a vehicle with saline (n = 16), a short-acting G-CSF treatment group (n = 16) and a long-acting G-CSF treatment group (n = 16). The cerebral ischemia reperfusion model was established for the sham group and G-CSF treatment groups by middle cerebral artery occlusion (MCAO). Five days post reperfusion, rats were sacrificed and the brains were removed. Changes in neurological function after cerebral ischemia reperfusion was evaluated according to Neurological Severity Score (NSS) and the lesion volume and infarct size were measured by 2,3,5-triphenyltetrazolium chloride staining. The numbers of apoptotic neurons in these ischemic areas: left cerebral cortex, striatum and hippocampus were calculated by TUNEL assay, and expression of JNK/P-JNK, c-jun/P-c-jun in these areas was detected by Western blotting. RESULTS: Compared with the saline vehicle group ((249.68±23.36) mm(3), (19.27±3.37)%), G-CSF-treated rats revealed a significant reduction in lesion volume (long-acting: (10.89±1.90)%, P < 0.01; short-acting G-CSF: (11.69±1.41)%, P < 0.01) and infarct size (long-acting: (170.53±18.47) mm3, P < 0.01; short-acting G-CSF: (180.74±16.93) mm3, P < 0.01) as well as less neuron functional damage (P < 0.01) and a smaller number of apoptotic neurons in ischemic areas (P < 0.01). The activity of P-JNK and P-c-jun in the cerebral ischemia reperfusion-damaged cortex and hippocampus was significantly decreased in all G-CSF-treated rats (P < 0.05). However, between the long-acting and short-acting G-CSF sets, there were no significant differences found in the activity of P-JNK and P-c-jun in the cortex, hippocampus and striate body (P > 0.05). CONCLUSIONS: Hypodermic injection of 50 µg/kg G-CSF attenuated the damage caused by cerebral ischemia reperfusion in rats, which might be associated with down-regulated activation of the P-JNK and P-c-jun pathway after cerebral ischemia reperfusion. Long-acting G-CSF may be a novel choice for both clinical and basic research in treating cerebral ischemia.