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1.
Article in Chinese | WPRIM | ID: wpr-776519

ABSTRACT

OBJECTIVE@#To investigate the effects of optical genetic techniques on new neurons through the Wnt/β-Catenin pathway.@*METHODS@#Neural stem cells (ESCs)were extracted from the cerebral cortex of fetal rat and transfected by lentivirus carrying DCX-ChR2-EGFP gene and the expression of DCX of newborn neurons differentiated from neural stem cells were observed. All cells were divided into 3 groups(n=9): control group, NSCs+EGFP and NSCs+ChR2 groups. The control group was normal cultured NSCs (NSCs group); the neural stem cells in NSCs+EGFP group were transfected with lentivirus carrying EGFP gene. The neural stem cells in NSCs+ChR2 group were infected with lentivirus carrying DCX-ChR2-EGFP gene. After 48 hours of lentivirus infection, 470 nm blue laser irradiation was performed for 3 consecutive days. NeuN positive cell density(the maturation of neural stem cells)and the ratio of NeuN/Hoechst in each group were observed. Western blot was used to detect the expression levels of MAP2, NeuN, Neurog2, NeuroD1 and GluR2. Western blot was used to detect the expressions of β-catenin and TCF4 associated with Wnt/β-catenin signaling channel. Verapamil (100 μmol/L, L-type calcium channel blockers) and Dkk1 (50 μg/ml, β-catenin inhibitor) were used to treat stem cells of the NSCs+ChR2 group and then the expressions of MAP2, NeuN, Neurog2, NeuroD1 and GluR were detected by Western blot.@*RESULTS@#After 3 days of 470 nm blue laser irradiation, NeuN positive cell density(the maturation of neural stem cells)and the ratio of NeuN/Hoechst, the expression levels of the protein MAP2, NeuN, Neurog2, NeuroD1, GluR and the protein β-catenin and TCF4 associated with Wnt/β-catenin signaling channel detected by Western blot were significantly increased in the group of NSCs+ChR2, compared with NSCs and NSCs+EGFP groups. The expressions of MAP2, NeuN, Neurog2, NeuroD1 and GluR were remarkably decreased after treated by verapamil and Dkk1 in the group of NSCs+ChR2. It was proved that the opening of ChR2 channel producing cationic influx promoted the maturation of neural stem cells and induced by the Wnt/β-catenin signaling pathway.@*CONCLUSION@#Optical genetic promoted the maturation of newborn neurons through the Wnt/β-catenin signaling pathway.


Subject(s)
Animals , Cells, Cultured , Neural Stem Cells , Cell Biology , Neurons , Cell Biology , Optogenetics , Rats , Transfection , Wnt Signaling Pathway
2.
Article in Chinese | WPRIM | ID: wpr-776518

ABSTRACT

OBJECTIVE@#To study the effect of exendin-4(Ex-4) on the differentiation of neural stem cells(NSCs) in adult mouse subventricular zone(SVZ)and its mechanism .@*METHODS@#NSCs in the SVZ were derived from 5-week C57BL/6J mice and the expression of nestin was detected by immunofluorescence. The cell morphology was observed after the cells treatmed with 100 nmol/L Ex-4 for 14 days.The expressions of nestin and glucagon-like peptide-1 receptor (GLP-1R) were detected by immunofluorescence. GLP-1R was knocked down by using shRNA and the study was divided into four groups: control group, Ex-4 group, GLP-1R knockdown group, GLP-1R knockdown + Ex-4 group. After treatment with 100 nmol/L Ex-4 for 14 d, β-tublin III and glial fibrillary acidic protein (GFAP) were labeled by immunofluorescence and then the proportion of β-tublin III positive cells were counted. Western blot was used to detect the activation of cAMP-response element binding protein (CREB) in NSCs. In order to further study the effects of Ex-4 on mitogen-activated protein kinase(MAPK) and phosphatidylinositol 3-hydroxy kinase (PI3K) pathways, the cells were pretreated with MAPK inhibitor U0126 at a concentration of 0.07 μmol/L for 30 min or PI3K inhibitor LY294002 at 50 μmol for 2 h, respectively. The study was divided into six groups: control group, Ex-4 group, U0126 group, U0126 + Ex-4 group, LY294002 group, LY294002 + Ex-4 group. The activation of CREB in each group was detected by Western blot. The experiment was repeated three times independently.@*RESULTS@#NSCs were successfully extracted from SVZ of C57BL/6J mice. Immunofluorescence showed that nestin and GLP-1R were positive in NSCs. Compared with the control group, the proportion of neurons differentiated from Ex-4 group was higher. The percentage of neurons in GLP-1R knockdown + Ex-4 group was basically the same as that in control group (P<0.01). The positive cells of beta-tublin III showed positive activation of GLP-1R and CREB. Western blot showed that CREB was significantly activated in the Ex-4 group, and knockdown of GLP-1R abolished its activation (P<0.01). U0126 did not affect Ex-4-mediated CERB activation, and LY294002 significantly reduced Ex-4-mediated CREB activation (P<0.01).@*CONCLUSION@#Ex-4 promotes the differentiation of NSCs into neurons in SVZ of adult mice through GLP-1R receptor, which may be achieved through PI3K/CREB pathway.


Subject(s)
Animals , Cell Differentiation , Cells, Cultured , Cyclic AMP Response Element-Binding Protein , Metabolism , Exenatide , Pharmacology , Gene Knockdown Techniques , Glucagon-Like Peptide-1 Receptor , Genetics , Metabolism , Lateral Ventricles , Cell Biology , Mice , Mice, Inbred C57BL , Neural Stem Cells , Cell Biology , Phosphatidylinositol 3-Kinases
3.
Tianjin Medical Journal ; (12): 523-527, 2018.
Article in Chinese | WPRIM | ID: wpr-698057

ABSTRACT

Objective To study the clinical characteristics of brain contusion in children and discuss the corresponding treatment strategies.Methods From February 2013 to December 2017,the clinical data of 32 patients(aged 1-14 years)with cerebral contusion and treated in the department of brain of the affiliated hospital of armed police logistics college were collected,including 22 males and 10 females.The age of patients ranged from 1 to 13.5 years,with the average of(6.03 ± 3.96)years.Patients were divided into low age group(1-4 years old,n=14)and high age group(5-14 years old,n=18)according to their age.Data of the brain damage location and distribution,Glasgow Coma Scale(GCS)/Children's Coma Scale (CCS), intracranial pressure (ICP), cerebral edema, airway condition, traumatic epilepsy and electroencephalogram (EEG) were summarized and analyzed. Results The duration of hospitalization was 17-57 d, with the average of (34.2 ± 11.5)d.All the children were treated with hyperbaric oxygen for 14-51 d,with an average of(36.1±5.1)d.There were no significant differences in the depth of brain damage and the location of brain injury between the two age groups(P>0.05). The coma scores at the admission and the discharge were significantly lower in low age group than those of the high age group (P<0.05). The duration of ICP monitoring was 6-15 d, with the average of (7.5±2.0) d. The ICP level and the resonance index(RI)value of cerebral edema monitoring at the admission was higher in the low age group than those in the high age group(P<0.05).The abnormal time of RI was less in the low age group than that in the high age group(P<0.05).There were no significant differences in seizures and EEG classification between two groups (P>0.05). There were 10 cases in each group received the transnasal intubation,and the average days with intubation were longer in the low age group than those of the high age group(P<0.05).Conclusion The clinical manifestations,imaging features,ICP,the degree of brain edema, seizures and airway management in children of different ages are different. The treatment should be carried out as soon as possible,actively and comprehensively,especially in the clinical management for the children of low age.

4.
Tianjin Medical Journal ; (12): 60-64, 2018.
Article in Chinese | WPRIM | ID: wpr-697973

ABSTRACT

Objective To study the findings and clinical application of the diffusion tensor imaging (DTI) of the trigeminal nerve root in patients with trigeminal neuralgia (TN). Methods Forty-nine patients with unilateral TN caused by vascular compression and surgically confirmed were collected in this study. The subjects were divided into group A (contact, n=22), group B (displacement, n=11) and group C (atrophy, n=16), according to the relationship between blood vessels and nerves. Conventional MRI and horizontal DTI were performed to measure cross-sectional area (CSA), mean diffusivity (MD) value, fractional anisotropy (FA) value, axial diffusivity (AD) value and radial diffusivity (RD) value in nerve roots of affected side and unaffected side. The trigeminal indexes of both affected and unaffected trigeminal nerves and data of the affected trigeminal indexes of TN patients were compared between the groups. Results The CSA and FA values were significantly lower, MD and RD values of trigeminal nerves were significantly higher in affected side than those of unaffected side ( P<0.05). But there was no significant difference in AD value between affected side and unaffected side of TN patients ( P>0.05). Values of CSA and FA were significantly higher in group A than those of group C. The value of RD was significantly lower in group A than that of group C. FA value was significantly higher in group B than that of group C (P<0.05). Conclusion The magnetic resonance DTI sequence can reflect changes in trigeminal nerve microstructure, suggesting that trigeminal demyelination is caused by vascular compression, and altered diffusion indexes of trigeminal nerve are reliable basis for the pathogenesis and the severity evaluation of TN.

5.
Neurology Asia ; : 49-58, 2017.
Article in English | WPRIM | ID: wpr-625433

ABSTRACT

Background & Objective: Radiotherapy and temozolomide are the standard therapy for newly diagnosed glioblastoma multiforme (GBM). However, it is unclear whether adding another agent to the commonly used radiotherapy-temozolomide (RT + TMZ) benefits newly diagnosed GBM patients. The present network meta-analysis aimed to assess the efficacy of combining other agents with RT + TMZ for GBM treatment. Methods: A comprehensive literature search was conducted on PubMed, EMBASE.com, Web of Science, and the Cochrane Central Register of Controlled Trials from inception to September 23, 2014, to include all randomized controlled trials of RT + TMZ-based therapy in GBM patients. Pairwise and network meta-analyses were performed to compare the therapeutic regimens. Results: Seventeen studies involving 4,148 patients were identified. The results of pairwise meta-analysis indicated no significant differences among most comparison groups, except for bevacizumab + RT + TMZ versus RT + TMZ for progression-free survival (hazard ratio [HR] = 0.71, 95% confidence interval [CI]: 0.59–0.86; P = 0.000) and RT + TMZ versus RT alone for overall survival (HR = 0.71, 95% CI: 0.58–0.88; P = 0.001). The results of network meta-analysis also showed no significant differences in most comparisons; however, adverse events were more common among patients receiving additional therapeutic agents other than RT + TMZ. The ranking probability analysis indicated that bevacizumab + RT + TMZ and nimustine + cisplatin + RT + TMZ were associated with the best progression-free and overall survival, but they also caused the most adverse events in GBM patients. RT + bevacizumab + irinotecan had the highest probability of being the best regimen for minimizing adverse events. Conclusions: The addition of other targeted agents, particularly bevacizumab and nimustine, to RT + TMZ could be slightly effective for the treatment of newly diagnosed GBM patients; however, adverse events remained common.


Subject(s)
Glioblastoma
6.
Chinese Journal of Traumatology ; (6): 179-181, 2016.
Article in English | WPRIM | ID: wpr-235753

ABSTRACT

The presence of large segmental defects of the diaphyseal bone is challenging for orthopedic surgeons. Free vascularized fibular grafting (FVFG) is considered to be a reliable reconstructive procedure. Stress fractures are a common complication following this surgery, and hypertrophy is the main physiological change of the grafted fibula. The exact mechanism of hypertrophy is not completely known. To the best of our knowledge, no studies have examined the possible relationship between stress fractures and hypertrophy. We herein report three cases of patients underwent FVFG. Two of them developed stress fractures and significant hypertrophy, while the remaining patient developed neither stress fractures nor significant hypertrophy. This phenomenon indicates that a relationship may exist between stress fractures and hypertrophy of the grafted fibula, specifically, that the presence of a stress fracture may initiate the process of hypertrophy.


Subject(s)
Adult , Female , Fibula , Pathology , Transplantation , Fractures, Stress , Pathology , Humans , Hypertrophy , Male , Middle Aged , Tibia , General Surgery , Tibial Fractures , General Surgery
7.
Article in Chinese | WPRIM | ID: wpr-243381

ABSTRACT

<p><b>OBJECTIVE</b>To simulate the chemical microenvironment of injured brain tissue, and to explore the effect of this chemical microenvironment on temperature sensitive umbilical cord mesenchymal stem cells (tsUC).</p><p><b>METHODS</b>Rat models of traumatic brain injury (TBI) were made by fluid percussion injury, and then the brain tissue extracts of the injured regions were acquired. Human umbilical cord mesenchymal stem cells (UC) were isolated and cultured, and the tsUC were obtained through the infection of temperature-sensitive Simian 40 Large T- antigen (ts-SV40LT) retrovirus. After that, both the two kinds of cells were cultured on the polyacrylamide gels which mimicking the elastic modulus of brain. Four groups were included: UC cultured under normal temperature (UC group), UC cultured added brain tissue extract under normal temperature (UC plus extract group), tsUC cultured under mild hypothermia (tsUC group), and tsUC added brain tissue extract under mild hypothermia for 3 days, then normal temperature for 4 days (tsUC plus extract group). After 24 hours, the apoptosis level was checked. Cell growth and morphological changes in each group were given dynamic observation. Seven days later, cell immunofluorescences were implemented for examining neural differentiation level.</p><p><b>RESULTS</b>Compared with UC plus extract group, the apoptosis and proliferation in UC plus extract group were significantly reduced (P < 0.01) and increased (P < 0.01) respectively. Cell immunofluorescence showed that the both GFAP and Neuron positive cells were significantly enhanced in UC plus extract group than those in tsUC plus extract group.</p><p><b>CONCLUSION</b>tsUC combining with mild hypothermia could significantly reverse injury induced cell apoptosis, improve cell proliferation and neural differentiation under chemical microenvironment after brain injury, which confirmed the adaptation and resistance of tsUC under mild hypothermia after TBI.</p>


Subject(s)
Animals , Apoptosis , Brain , Cell Biology , Pathology , Brain Injuries , Pathology , Cell Proliferation , Humans , Mesenchymal Stem Cells , Chemistry , Neurons , Cell Biology , Rats , Temperature , Umbilical Cord , Cell Biology
8.
Article in Chinese | WPRIM | ID: wpr-255040

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the influence of therapeutic bloodletting at Jing-well points and hypothermia on acute cerebral edema after traumatic brain injury (TBI) in rats.</p><p><b>METHODS</b>Seventy-five SD rats were randomly divided into sham-operation group (Sham), TBI group (TBI), bloodletting group (BL), mild-induced hypothermia group (MIH), and bloodletting plus MIH group (BL + MIH) (n = 15). The model of TBI was established by electric controlled cortical impactor (eCCI). The rats of BL group were bloodletting at Jing-well points immediately after injury, twice daily. While the MIH group was settled on a hypothermia blanket promptly after TBI for 6 hours, so that the temperature dropped to 32 degrees. Each of measurement was performed after 48 hours. Magnetic resonance imaging (MRI) was used to evaluate the dynamic impairment of cerebral edema after TBI (n = 3). In addition, mNSS score, measurements of wet and dry brain weight, and Evans Blue assay were performed to investigate the neurologic deficit, cerebral water content (n = 8), and blood-brain barrier permeability (BBB), (n = 4), respectively.</p><p><b>RESULTS</b>MRI analysis showed that the cerebral edema, hematoma and midline shifting of rats in TBI group was more serious than other treatment group. Meanwhile compared with TBI group, the mNSS scores of every treatment group were meaningfully lower (all P < 0.05). Furthermore, treatment with BL+ MIH group was superior to the separated BL and MIH group (all P < 0.01). In addition, brain water content of each intervention group reduced to varying degrees (all P < 0.05), especially that of MIH group and BL + MIH group (P <0.01). BBB permeability of each treatment group was also significantly improved (all P < 0.01), and the improvement in MIH group and BL + MIH group was much better than the BL alone group (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>Our major finding is that bloodletting at Jing-well points and MIH can reduce cerebral edema and BBB dysfunction and exert neuroprotective effects after TBI. The results suggest that the combination of BL and MIH is more effective than other treatment being used alone.</p>


Subject(s)
Animals , Blood-Brain Barrier , Bloodletting , Brain , Pathology , Brain Edema , Brain Injuries , Therapeutics , Hypothermia, Induced , Rats , Rats, Sprague-Dawley
9.
Article in Chinese | WPRIM | ID: wpr-254986

ABSTRACT

<p><b>OBJECTIVE</b>To explore the mechanisms of acupuncture treatment promoting the motor function recovery of neonate rats with cerebral palsy.</p><p><b>METHODS</b>The improved hypoxic-ischemic encephalopathy (HIE) means was performed to establish the model of neonate rats with cerebral palsy. All neonate rats were randomly divided into 3 groups: sham group, model group and acupuncture group (n = 20). We observed and scored motor function of rats, measured the levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in serum, and also measured the expression of synaptophysin (SYP) and growth associated protein-43 (GAP-43) in the diseased region of cerebral tissue.</p><p><b>RESULTS</b>The motor function scores (11.3 +/- 0.29) and the serum level of SOD (147.1 +/- 12.7) U/ml in acupuncture treatment group were higher than those of model group ( P < 0.05). The serum level of MDA was lower in acupuncture treatment group than that of model group (P < 0.05). The expression of SYP and GAP-43 in the diseased region of cerebral tissue of acupuncture treatment group were higher than those of model group ( P < 0.05) .</p><p><b>CONCLUSION</b>Acupuncture-therapy could improve the motor function of neonate rats with cerebral palsy by decreasing the content of MDA in serum, increasing the contents of SOD in serum, and prolonging the upregulation of SYP and GAP-43 expressions in hmin tissue.</p>


Subject(s)
Acupuncture Therapy , Animals , Animals, Newborn , Cerebral Palsy , Therapeutics , Disease Models, Animal , GAP-43 Protein , Metabolism , Hypoxia-Ischemia, Brain , Therapeutics , Malondialdehyde , Metabolism , Rats , Superoxide Dismutase , Metabolism , Synaptophysin , Metabolism
10.
Article in English | WPRIM | ID: wpr-320331

ABSTRACT

<p><b>OBJECTIVE</b>The mechanism through which platelet activating factor (PAF) induces cardiac electrical activity and arrhythmia is not well understood and previous studies have suggested a potential involvement of ion channels in its action. The present study was aimed to clarify the role of PAF in fatal arrhythmias following acute myocardia infarction (AMI) and the underlying mechanism.</p><p><b>METHODS</b>(1) Blood PAF levels were measured among 72 AMI patients at the time of diagnosis with AMI and 48 h later, and their electrocardiogram (ECG) was recorded continuously. (2) Ischemia simulation and surface electrocardiogram were conducted in 20 pigs and their PAF levels were measured. (3) PAF perfusion and standard microelectrode recording were performed on guinea pig papillary muscles.</p><p><b>RESULTS</b>In both humans and pigs, elevated PAF levels were detected in AMI and simulated ischemia, respectively, and even higher PAF levels were found when fatal arrhythmias occurred. In guinea pig myocardium, PAF induced a shortening of action potential duration at 90% level of repolarization (APD90)under non-ischemic conditions and a more pronounced shortening under early simulated ischemic conditions.</p><p><b>CONCLUSION</b>AMI and ischemia are associated with increased PAF levels in humans and pigs, which are further raised when fatal arrhythmia follows. The effects of PAF on the myocardium may be mediated by multiple ion channels.</p>


Subject(s)
Animals , Arrhythmias, Cardiac , Blood , Electrocardiography , Female , Heart , Humans , Male , Middle Aged , Myocardial Ischemia , Blood , Platelet Activating Factor , Metabolism , Swine
11.
Chinese Journal of Cardiology ; (12): 598-601, 2013.
Article in Chinese | WPRIM | ID: wpr-261488

ABSTRACT

<p><b>OBJECTIVE</b>To analyze the clinical features, precaution and management of complete heart block (CHB) after transaortic extended septal myectomy operation (extended Morrow procedure) in patients with hypertrophic obstructive cardiomyopathy (HOCM).</p><p><b>METHODS</b>From October 1996 to December 2011, 10[6 men; mean age (45.4 ± 15.8) years, range 13-60 years] out of 160 consecutive HOCM patients underwent extended Morrow procedure developed CHB postoperatively. Their clinical data were retrospectively analyzed. Baseline transthoracic echocardiography showed that the left ventricular outflow tract (LVOT) gradients was from 68 to 149 (105.1 ± 25.9) mm Hg (1 mm Hg = 0.133 kPa), ECG showed right bundle branch block in 5 patients and atrial fibrillation, atrial premature beats or ST-T segment changes in other 5 patients. Besides extended Morrow procedure, concomitant surgical procedures included mitral valve replacement (MVR) in 2 (2/10) and MVR plus coronary artery bypass grafting in another 2 (2/10) patients. Follow-up data were obtained by subsequent clinic visits in outpatient department and telephone interviews.</p><p><b>RESULTS</b>The in-hospital mortality was 20% (these two patients died of low cardiac output syndrome and multiple organs failure). Four patients underwent MVR simultaneously survived the operation. Postoperative echocardiography demonstrated a reduced LVOT gradient[(13.6 ± 9.7) mm Hg, P < 0.001]. Permanent pacemakers were implanted in all 8 survived patients at 6 days to 7 months after operation. No other severe complications were observed. During follow-up [from 4 to 72 (19.4 ± 22.1) months], there was no death, 1 patient readmitted to our center at 71 months post operation to change the pacemaker because of low voltage of previously implanted pacemaker. Physical capacity and quality of life improved significantly post operation in these 8 patients. The NYHA functional class remained at I-II post operation and during follow up.</p><p><b>CONCLUSIONS</b>CHB is a severe complication after extended Morrow procedure for patients with HOCM and timely permanent pacemaker implantation is mandatory for patients with post procedure CHB.</p>


Subject(s)
Adolescent , Adult , Atrioventricular Block , Cardiomyopathy, Hypertrophic , General Surgery , Female , Humans , Male , Middle Aged , Postoperative Complications , Retrospective Studies , Treatment Outcome , Young Adult
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