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Objective:To investigate the expression and role of chemokine CCL21 and its receptor CCR7 in ischemic brain white matter lesion (WML).Methods:Permanent bilateral common carotid artery occlusion (BCCAo) was used to prepare an ischemic WML model in C57Bl/6 mice. Evans blue perfusion staining was used to assess the permeability of the blood-brain barrier. Immunofluorescence staining was used to detect the expression of CCL21 and its receptor CCR7 in ischemic WML tissues. Cerebral microvascular endothelial cells were subjected to oxygen glucose deprivation (OGD) to prepare ischemic WML model in vitro. The expression level of CCL21 protein after OGD 6 hours and reoxygenation 24 hours was detected by Western blot analysis. Primary cultured oligodendrocyte precursor cells (OPCs) were treated with recombinant CCL21, and then inflammatory reaction was induced by lipopolysaccharide. The mRNA expression level of myelin basic protein (MBP), a marker of mature oligodendrocytes, was detected by quantitative polymerase chain reaction. Results:Compared with the sham operation group, the exudation of Evans blue stain in the brain tissue was significantly increased after BCCAo ( P<0.05), the expression level of CCL21 in cerebral vascular endothelial cells and CCR7 in OPCs was significantly down-regulated ( P<0.05). After cerebral microvascular endothelial cells were subjected to OGD, the expression level of CCL21 protein were significantly down-regulated compared with the control group ( P<0.05). After OPCs were treated with recombinant CCL21, the expression level of MBP mRNA was significantly up-regulated ( P<0.05). Conclusions:The expression of CCL21/CCR7 was significantly down-regulated after WML. The application of recombinant CCL21 may promote the differentiation and maturation of OPCs cells, suggesting that it may play an important role in ischemic WML.
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Objective:To investigate the correlation between white matter hypertensities (WMHs) and the overall burden of cerebral small vessel disease (CSVD) and clinical outcome of patients with acute ischemic stroke.Methods:From November 2018 to June 2019, patients with acute ischemic stroke hospitalized in the Department of Neurology, the Affiliated Jiangning Hospital of Nanjing Medical University were enrolled prospectively. Their demographic and clinical data were collected. The National Institutes of Health Stroke Scale (NIHSS) was used to evaluate the baseline severity of stroke. The total burden of CSVD was evaluated according to the head MRI findings. The severity of WMHs was assessed based on Fazekas scale. The modified Rankin Scale (mRS) was used to evaluate the outcomes at 90 d after onset. The mRS score 0-1 was defined as good outcome, and ≥2 was defined as poor outcome. Multivariate logistic regression analysis was used to determine independent risk factors for poor outcomes. Results:A total of 153 consecutive patients with acute ischemic stroke were enrolled, of which 126 (82.35%) had a good outcome and 27 (17.65%) had a poor outcome. There were significant differences in age, baseline NIHSS score, lipoprotein-associated phospholipase A 2, total Fazekas score, periventricular WMHs score, and deep WMHs score between the two groups, while there was no significant difference in the total burden of CSVD. Multivariate logistic regression analysis revealed that the baseline NIHSS score (odds ratio 1.245, 95% confidence interval 1.023-1.515; P=0.028) and the total Fazekas score (odds ratio 1.635, 95% confidence interval 1.049-2.549; P=0.030) were the independent risk factors for poor outcomes at 90 d after the onset in patients acute ischemic stroke. Conclusions:The overall burden of CSVD is not associated with the short-term outcomes in patients with acute ischemic stroke. WMHs and baseline NIHSS score are the independent risk factors for poor short-term outcomes in patients with acute ischemic stroke.
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OBJECTIVE: To study the effect of aquaporin 4(AQP4) in regulating the permeability of blood-brain barrier(BBB) induced by subacute 1,2-dichloroethane(1,2-DCE) inhalation. METHODS: Specific pathogen free healthy CD-1 male Aqp4 genetically engineered mice(Aqp4~(+/+)and Aqp4~(-/-)) were randomly divided into control and low-, medium-and high-dose groups. The mice were exposed to 1,2-DCE at the dosages of 0.00, 100.00, 350.00 and 700.00 mg/m~3 for 6 hours per day for consecutive 28 days by systemic dynamic inhalation. After the end of 1,2-DCE exposure, the BBB permeability was evaluated by Evans blue staining. Real-time fluorescence quantitative polymerase chain reaction method was used to detect the mRNA expression of genes related to BBB tight junction protein(Tjp)1, Tjp2, Tjp3, claudin(Cldn)3, Cldn5, Cldn11, occludin(Ocln), matrix metalloproteinase(Mmp)2, Mmp9 and Na-K-Cl cotransporter-1(Nkcc1). RESULTS: The BBB permeability in mice showed significant change with 1,2-DCE dose and Aqp4 genotype(P<0.01). The BBB permeability of Aqp4~(+/+) genotype mice was higher in low-, medium-and high-dose groups than that of control group(all P values were <0.05). The permeability of BBB was lower in Aqp4~(+/+) genotype mice in the control group than that of Aqp4~(-/-) genotype mice in the same group(P<0.05), but BBB permeability was higher in Aqp4~(+/+) genotype mice in the three dose groups than that of Aqp4~(-/-) genotype mice in the same group(all P values were <0.05). The Cldn3 and Olcn mRNA relative expression in the brain cortex had statistical difference in mice with different genotype(all P values were <0.01). The mRNA relative expressions of Cldn3 and Olcn in the brain cortex were higher in Aqp4~(-/-) genotype mice than that of Aqp4~(+/+) genotype mice(all P values were <0.01). The relative mRNA expression levels of Tjp1, Tjp2, Tjp3, Cldn5, Cldn11, Mmp2, Mmp9 and Nkcc1 in the cerebral cortex of mice were not statistically significant in aspect of 1,2-DCE exposure dose and genotype(all P values were >0.05). CONCLUSION: Exposure to 1,2-DCE can increase BBB permeability in mice, and the mechanism may be associated with 1,2-DCE-induced down-regulation of Aqp4 and up-regulation of mRNA expression of the cerebral cortex TJP-related molecules Cldn3 and Ocln.
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OBJECTIVE: To explore the molecular mechanism underlying 1,2-dichloroethane(1,2-DCE) induced apoptosis by screening differentially expressed proteins in human astrocytes( HAs). METHODS: HAs were cultured in complete medium with 1,2-DCE at various concentrations of 0-80 or 0-40 mmol/L. After 24 hours,apoptosis of HAs was evaluated using flow cytometry and staining with annexin Ⅴ-fluoresce in isothiocyanate and propidium iodide. An AAH-APO-1-2 protein chip was used to screen differentially expressed proteins and quantitative real-time polymease chain reaction(qRT-PCR) was used to verify related differentially expressed genes(DEGs). RESULTS: At 1,2-DCE concentrations of0-80 mmol/L,the total apoptosis rate of HAs increased with 1,2-DCE concentrations in a dose-dependent manner( P <0. 01). Seven different kinds of proteins were screened out by apoptotic protein chip. Among them,the expression of insulin-like growth factor-binding protein( IGFBP)-1,IGFBP-4 and cytochrome C( Cyto C) were up-regulated,while the expression of P27,cysteine aspartic acid specific protease-3( Caspase-3),B-cell lymphoma-2 interacting mediator of cell death( BIM) and BH3 interacting domain death agonist( BID) were down-regulated compared with the control group. The result of DEGs verified by qRT-PCR showed that the expression of mRNA of IGFBP-1,IGFBP-4 and Cyto C at 1,2-DCE concentrations of 40 mmol/L was up-regulated. This result was in consistent with the trend of target expression in the protein chip. The mRNA expression of Caspase-3,BIM and BID was also up-regulated. CONCLUSION: 1,2-DCE induces apoptosis of HAs through mitochondrial pathway.
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Objective To study the effect of Shaoyaozhitong Mixture on stageⅢ-Ⅳendometriosis, and explore its mechanism. Methods Ninty-five cases of stageⅢ-Ⅳ endometriosis were randomly divided into three groups. The traditional Chinese medicine (TCM) group (31 cases) was treated with Shaoyaozhitong Mixture, the GnRHa group (31 cases) was treated with Triptorelin Acetate for Injection, while the expectant group (33 cases) received no medications. The changes of CA125, TNF-α and VEGF levels, visual pain score and SF-36 score, total effective rate, pregnancy and recurrence were compared. Results There were significant increases in CA125, TNF-α and VEGF levels, visual analogue pain score and SF-36 score in all the three groups after treatment (P0.05). Conclusion Shaoyaozhitong Mixture is effective in treating endometriosis by inhibiting the growth of ectopic endometrium.