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Objective Constructed Pseudomonas aeruginosa PA14 PA0745 mutant and verified its pathogenicity.Methods Constructed Pseudomonas aeruginosa PA0745 gene deletion and point mutant by homologous recombination.Pseudomonas aeruginosa mutant strains E126A,E146A and △PA0745 were screened by plasmid resistance and auxotrophy.Analysed mutant PA0745,PA14 growth characteristics and pathogenic changes.Results PA14 PA0745 mutant strains E126A,E146A and △PA0745 were successfully constructed.The results of growth curve showed that PA14 PA0745 mutation did not affect PA14 normal growth.The results of cell infection showed that the infection ability of E126A,E146A and △PA0745 decreased by 29.51%,60.66% and 79.34% respectively compared with PA14.Conclusion PA14 PA0745 mutant was successfully constructed in this study,and the function of PA0745 was confirmed,which had some guiding significance for the development of antibacterial drugs.
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AIM: To investigate the effect of microRNA (miR)-30c on the viability and migratory ability of human umbilical vein endothelial cells (HUVECs) by targeting plasminogen activator inhibitor-1 (PAI-1).METHODS:The HUVECs were transfected with miR-30c mimic and inhibitor or negative control (NC), and then the expression levels of miR-30c, PAI-1 mRNA and protein were detected by RT-qPCR and Western blot.The viability and migratory ability of HUVECs were measured by CCK-8 assay and wound healing test .After bioinformatic analysis, the assessment of miR-30c binding to PAI-1 3’-UTR was carried out using a luciferase reporter gene assay .RESULTS:miR-30c directly down-regu-lated PAI-1 levels by binding to the 3’ UTR seed sequence of PAI-1 mRNA.Furthermore, transfection of a miR-30c mimic down-regulated the expression of PAI-1 at mRNA and protein levels, leading to enhanced migratory ability and viability of the HUVECs.However, transfection of a miR-30c inhibitor up-regulated the expression of PAI-1 at mRNA and protein le-vels, leading to decreased migratory ability and viability .CONCLUSION:Regulation of miR-30c level changes the migra-tory ability and viability of HUVECs by affecting the PAI-1 expression, indicating the involvement of miR-30c in modulating endothelial function .
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Objective To investigate the instant clinical efficacy of intra-arterial infusion of fasudil combined with routine anti-vasospasm for symptomatic cerebral vasospasm (SCVS). Methods The clinical data of 21 patients with subarachnoid hemorrhage (SAH) due to ruptured aneurysm, who were admitted to authors' hospital during the period from May 2010 and February 2014, were retrospectively analyzed. The lesions included Fisher gradeⅡ(n=2), gradeⅢ (n=16) and gradeⅣ (n=3). Endovascular embolization of the aneurysm was carried out within 48 hours after the confirmation of the diagnosis with total cerebral DSA;no bleeding occurred during the operation and routine anti-vasospasm therapy was given. Within 4-9 days after the onset of the disease, all 21 patients presented SCVS. Half dose systemic heparinization, superselective intra-arterial infusion of fasudil (30 mg fasudil+250 ml saline, lasting for 30 min) were adopted. Reexamination of angiography performed at 15 min after fasudil infusion was employed, and the results were evaluated with NIHSS score by comparing the preoperative findings. Results Imaging examination performed after the treatment showed that significant improvement was obtained in 15 patients and no obvious changes in 6 patients. Clinical symptoms were remarkably improved in 11 patients, partially improved in 4 patients and remained unchanged in 6 patients. The mean NIHSS score was improved from preoperative 28.6 to postoperative 21.2. Conclusion For the treatment of symptomatic cerebral vasospasm, superselective intra-arterial infusion of fasudil is effective and safe, and it has good clinical application value.
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Objective To observe the cilioretinal artery and its relationship with central visual loss in central retinal artery occlusion(CRAO) patients.Methods A total of 140 CRAO patients (140 eyes) were enrolled in this study.The patients included 83 males and 57 females.The age was ranged from 42 to 75 years old,with an average of (55.70 ± 22.20) years.All the patients were affected unilaterally,including 79 right eyes and 61 left eyes.The disease duration was from 1 to 10 days,with a mean of (4.7±3.9)hours.Central vision and fluorescence fundus angiography were measured for all patients.The central visual loss was divided into 3 types:mild (≥0.1),moderate (finger counting to 0.08) and severe (no light perception to hand movement).The number,length and location of cilioretinal artery were observed.The correlation between cilioretinal artery and central visual loss was analyzed.Results There were 41 eyes (29.3%) with cilioretinal artery,which including 13 eyes (31.7%) with ≥3 cilioretinal arteries,23 eyes (56.1%) with 2 cilioretinal arteries,5 eyes (12.2%) with 1 cilioretinal arteries.The cilioretinal artery was within 1 disk diameter (DD) in length and not reached the macular area in 37 eyes (90.2%),was more than 1DD in length and reached the macular foveal area in 4 eyes (9.8%).The cilioretinal artery located in the temporal side of optic disk in 29 eyes (70.7%),and in other quadrant in 12 eyes (29.3%).The distribution of central visual loss degree as follow:mild in 15 eyes (10.7%),moderate in 50 eyes (35.7%),severe in 75 eyes (53.6%).The difference of central visual loss in the eyes with or without cilioretinal arteries was not significant (x2=0.16,P>0.05).Conclusions Cilioretinal artery exists in 29.3% CRAO eyes.There was no close correlation between cilioretinal artery and central visual loss.
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The characteristics of non-arteritic anterior ischemic optic neuropathy (NAION) include sudden decline of visual acuity, sector-shaped visual field defect connected with the physiological blind spot or hemi-visual field defect, and optic disc edema. Early angiographic abnormalities of optic disc and peripapillary choroidal circulation are important cues for the diagnosis. Angiography combined with visual field test can make the diagnosis more accurate. The pathological mechanism and process of NAION and ischemic cardiovascular/cerebrovascular disease are essentially the same. There are several critical issues in this field we need focus on, including strengthening the cooperation with relevant professional disciplines; improving the ocular ischemia through drugs, intervention and other means; preventing more serious cardiovascular events and intervening in a number of high-risk populations to reduce the incidence of AION.
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Objective To evaluate the application of CT myelography (CTM) in detecting the site of spontaneous cerebrospinal fluid (CSF) leaks and analyze it's imaging features.Methods Six patients (3 women and 3 men) with spontaneous intracranial hypotension (SIH) were included, who met the criteria of the International Headache Classification (2nd edition, 2004). Five patients subsequently underwent whole spine MRI and all 6 patients underwent CTM. Autologous blood mixed with omnipaque (300 mg/ml) was injected followed by selective puncture at the leak site indicated by CTM. Results MRI was failed to find leak site in the 5 patients, whereas CTM successfully found leak sites in all 6 patients. There were 1 to 7 leak sites respectively with an average of 4.2 sites (totally 25 points). Leak sites at cervical (12 sites) and thoracic (12 sites) were more frequent than at lumbar (1 site). CTM was featured by linear leakage of the contrast medium along the spinal nerve roots, paraspinal collections of hyper-density contrast medium and beak-like enlargement of the nerve sleeves. All patients responded well to the treatment, with complete resolution of symptoms. Conclusion CTM has been shown to be a study of choice to accurately define the location and extent of a CSF leak.