MicroRNAs (miRNAs) are endogenous non-coding RNAs composed of 18-25 nucleotides,and they may play a role in gene regulation through completely or partially binding to target gene mRNA complementary sequences and make it degrade or prevent its translation.miRNAs play important roles in the onset and pathophysiological processes of ischemic stroke.This article reviews the roles of miRNAs in the etiology of ischemic stroke and pathophysiological mechanisms after stroke,and the potential application of miRNAs is prospected.

Objective To investigate the relationship between obstructive sleep apneahypopnea syndrome (OSAHS) and carotid stenosis in patients with ischemic cerebrovascular disease (ICVD) and to provide reference for developing the intervention strategy of carotid stenosis. Methods Eighty-seven patients with ICVD were screened from Nanjing Stroke Registry Program. The patients were divided into without (n=21), mild(n=24), moderate (n=27) and severe (n = 11) OSAHS groups according to the apnea-hypopnea index (AHI); in addition, the patients were divided into with (n =34) and without carotid stenosis (n=49) groups according to the results of digital subtraction angiography (DSA). The effects of the risk factors for cerebrovascular diseases and OSAHS on carotid stenosis in patients with ICDV were analyzed.Results There were significant differences in the proportions of alcohol consumption (χ2=8.56, P =0. 036), hypertension (χ2 = 13.20, P =0. 004) and carotid stenosis (χ2 =22.97, P =0. 006) between the no OSAHS and the mild, moderate and severe OSAHS groups. The univariate analysis showed that age (OR = 1. 066, 95% CI 1. 023- 1.112; P = 0. 003),hypertension (OR =3.587, 95% CI 1. 294- 9. 949; P =0. 014), alcohol consumption (OR =5.275,95% CI 1.855-15.001; P= 0.002) and OSAHS (OR= 1.073, 95% CI 1.033-1.115; P = 0. 000) were the risk factors for carotid stenosis. The multivariate logistic regression analysis showed that age (OR = 1. 113, 95% CI 1. 047-1. 182; P =0. 001), OSAHS (OR = 1. 096, 95% CI 1. 034-1. 160; P = 0. 000), and alcohol consumption (OR = 4. 292,95% CI 1. 217-15. 139; P = 0. 024) were the independent risk factors for carotid stenosis.Spearman rank correlation analysis suggested that the AHI levels were positively correlated with the degree of carotid stenosis (r = 0. 435, P = 0. 000). There were significant differences among the without stenosis (n =34), unilateral stenosis (n =22), and bilateral stenosis (n=27)groups (12.97 ± 10.04 vs. 21.40 ± 16.38 vs. 29.33 ± 13.81, F= 11.64, P<0.01).Conclusions OSAHS is an independent risk factor for carotid stenosis and it was positively correlated with the severity of carotid stenosis. AHI may reflect the degree of carotid stenosis and the range of neck vascular involvement to some extent.

BACKGROUND:Studies have shown that neural stem cel transplantation combined with exercise training can promote the recovery of hindlimb motor function from spinal cord injury in rats, but its mechanism of action has not been fuly elucidated. OBJECTIVE:To investigate the effects of early exercise training combined with neural stem cel transplantation on the recovery of hindlimb motor function in rats with spinal cord injury. METHODS:Sixty Sprague-Dawley rats with spinal cord injury were randomly divided into three groups: control group (n=20, given conventional treatment after injury), cel transplantation group (n=20, given neural stem cel transplantation after injury), experimental group, (n=20, given neural stem cel transplantation combined with early exercise training after injury). Recovery of the hindlimb motor function was assessed by Basso, Beattie and Bresnahan scale and inclined plane test before and at 1, 7, 14, 21 days after injury. Western blot assay was used to detect caspase-3 and myeloperoxidase expression. Hematoxylin-eosin staining was done at 21 days after injury to observe the structure changes of the injured spinal cord. RESULTS AND CONCLUSION:(1) Scores of Basso, Beattie and Bresnahan scale and inclined plane test were significantly better in the experimental group than the cel transplantation group folowed by the control group (P < 0.05). (2) In the control group, the expression of caspase-3 and myeloperoxidase was significantly increased at 14 days after injury. In the cel transplantation, the expression of caspase-3 and myeloperoxidase was significantly higher than the experimental group (P < 0.05). (3) Pathological inflammation was reduced most in the experimental group folowed by the cel transplantation group. In the experimental group, the structure of injured spinal cord was improved and became relatively clear and intact. These findings indicate that neural stem cell transplantation combined with early exercise training can effectively promote the recovery of hindlimb motor function from spinal cord injury in rats, by reducing the expression of caspase-3 and myeloperoxidase and alleviating secondary lesion of the spinal cord.

Objective To assess the effectiveness of initial Glasgow Coma Scale (GCS) and National Institutes of Health Stroke Scale (NIHSS) as predictors for clinical outcomes in patients with top of the basilar syndrome (TOBS).Methods A total of 64 patients with TOBS were selected from Nanjing Stroke Registration Program (NSRP). Initial GCS and NIHSS were retrospectively evaluated by reviewing patients' records for details of clinical presentation and outcomes at 30 days measured by modified Rankin Scale (mRS) score. Patients were categorized as favorable outcome group (mRS 0-3) and unfavorable outcome group (mRS 4-6).Results The mean GCS was lower in the cases with mRS of 4-6 compared with those with mRS of 0-3 (P<0.01) and the mean NIHSS score was higher in favorable outcome group compared with unfavorable outcome group (P=0.011). In multivariate logistic regression analysis, after adjusting for age, gender and treatment approaches, the GCS OR was 0.301(95% CI 0.167～0.542), NIHSS OR was 1.436(95% CI 1.147～1.796), and both of them turned out to be the independent predictors of outcome at 30 days. ROC curve analysis suggested that GCS score of 10 represented a good cut-off point for predicting the outcome with the prognostic sensitivity of 87.9% and specificity of 83.9%. NIHSS score of 14 could also serve as a good cut-off point with the prognostic sensitivity of 63.6% and specificity of 77.4%.Conclusions Conclusions Both GCS and NIHSS can predict outcomes in patients with acute TOBS with GCS score ≤10 and NIHSS score ≥14 as the cutoff points of poor outcome. GCS cutoff point is more strongly predictive of outcome than that of NIHSS.

Objective To investigate the relationship between obstructive sleep apnea-hypopnea syndrome(OSAHS)and plasma homocysteine(Hcy)levels in patients with ischemie cerebrovascular disease(ICVD).Methods Seventy-six patients with ICVD were monitored with polysomnography(PSG) for 7-8 hours during sleeping.The levels of fibrinogen(FBG),high density lipoprotein cholesterol(HDLC),low-density lipoprotein cholesterol(LDL-C),triglyceride(TG)and plasma Hey were assessed.Results Of the 76 ICVD patients,57 showed apnea symptom during sleeping,of which 53(69.7%)were diagnosed with OSAHS and 4 with central sleep apnea,According to the mean apnea-hypopnea index (AHI),the severity of apnea was classified as no apnea symptom(19 cases with mean AHI of 2.38±0.96 and Hey level of(8.78 ±2.01)μmol/L),mild apnea(21 cages with mean AHI of 14.14 ±4.37 and Hcy level of(12.91 ±3.00)μmol/L),moderate apnea(24 cases with mean AHI of 29.62±5.81 and Hcy level of(14.85 ±4.15)μmol/L)and severe apnea(8 cases with mean AHI of 46.75±2.82 and Hcy level of(19.30±4.82)μmol/L).The level of Hcy was statistical significant among these 4 groups(F=40.32,P<0.01)and correlated with the mean AHI(r=0.598,P<0.01).Conclusion Patients with ICVD have a high morbidity of OSAHS,mainly suffering from mild to moderate apnea;The plasma Hcy level elevates in the ICVD patients with OSAHS and is correlated with the severity of apnea.

Objective:To establish and validate a fluorescence focus assay (FFA) for rapid titration of Japanese encephalitis virus (JEV) and to evaluate its application in the production of Japanese encephalitis vaccine.Methods:Recombinant JEV non-structural protein 1 (NS1) was expressed in a prokaryotic expression system. After purification, JEV-NS1 was used to immunize rabbits to induce polyclonal antibody. FFA was established with the polyclonal antibody to titer JEV. The accuracy of FFA was validated by comparing with plaque assay, and the specificity, precision, linearity, range and robustness of FFA were also validated. Twenty-eight batches of live-attenuated Japanese encephalitis vaccine were titrated with FFA and plaque assay to analyze the relationship between the two assays.Results:FFA established with polyclonal antibody against JEV-NS1 could be used to titrate JEV, and there was no cross reaction with other viruses (tick-borne encephalitis virus, yellow fever virus, coxsackievirus A2, coxsackievirus A4). Results of the validation tests showed that FFA met the requirement of quality control for live-attenuated Japanese encephalitis vaccine. FFA was more consistency than plaque assay.Conclusions:The established FFA could be used for virus titration in the production of live-attenuated Japanese encephalitis vaccine.

Objective To test the immune therapeutic effects of PICKCa adjuvant rabies vaccine for the post-exposure mice and beagle dogs and immune memory effect for human.Methods For the mice and beagle dogs first to infect with rabies wild virus and then immunize with PICKCa rabies vaccine.For volunteers, first to immunize with PICKCa rabies vaccine after 2.5 years to take peripheral blood mononuclear cell and then to check cells secreted interferon-gwith flow cytometry. Results In 3 independent assays of post-explosure immunization of mice and beagle dogs PICKCa rabies vaccine were much better then commercial adjuvant-free rabies vaccines, the protective rate from 20%-30%to 70%-100%Also, in the assay of cells secreted IFN-γby peripheral blood mononuclear cells from volunteers immunized by PICKCa rabies vaccine the IFN-γcell levels were much higher than control.Conclusions Compared with commercial adjuvant-free rabies vaccines the PICKCa rabies vaccine had significant immune therapeutic effects in the mice and dogs.Also PICKCa rabies vaccine had good immune memory effect in human.

The present study employed UPLC-MS/MS to analyze and identify compounds in the raw powder of Tongluo Shenggu capsules. An HPLC method for the determination of vitexin content was established. The analysis of this drug was performed on a 30 ℃ thermostatic Acquity UPLC® BEH C18 (2.1 mm×100 mm,1.7 μm) column, with the mobile phase comprising 0.2% formic acid-methanol flowing at 0.3 mL /min in a gradient elution manner. Mass spectrometry was detected by ESI sources in both positive and negative ion modes for qualitative identification of chemical constituents. 12 flavonoid and 3 stilbenes compounds in the raw powder of Tongluo Shenggu capsules were successfully identified. Additionally, an HPLC method for the determination of vitexin content was established using a XBridge C18 column (4.6 mm × 250 mm, 5 µm) with a mobile phase of 0.05% glacial acetic acid in methanol for gradient elution, at a column temperature of 30 °C, a flow rate of 1.0 mL/min, and an injection volume of 20 μL. The method demonstrated good linearity in the concentration range of 10 µg/mL to 40 µg/mL (R=1.000) with an average recovery rate of 96.7%. The establishment of these methods provides a scientific basis for the quality control and development of the raw powder of Tongluo Shenggu capsules.