ABSTRACT
Objective:To explore the action mechanism of suppressing expression of mitogen- activated protein kinase 14（MAPK14）to alleviate glutamate excitatory toxicity and its neuronal protection effect.Methods:Lentivirus-mediated MAPK14 interference vector was synthetized by Shanghai Jikai Gene Chemical Technology Co.，Ltd. Astrocytes were obtained from SD rats 48 hours after birth，which were cultured in vitro and transfected by lentivirus-mediated transfection. According to the random number table，the cells were divided into three groups：（1）un-transfected group（normal group）with normal astrocytes and the cells were cultured in regular medium composed of Dulbecco's?modified Eagle's?medium（DMEM）；（2）negative control group with astrocytes transfected by MAPK14 no-loaded interference vector；（3）lentivirus transfected group with astrocytes transfected by MAPK14 interference vector. Seventy-two hours after transfection，astrocytes were co-cultured with neurons for 48 hours，and then they were cultured in a medium containing glutamate for 2 hours. The detection indexes included the optimal multiplicity of infection（MOI）value for astrocytes transfected by lentivirus vector，mRNA levels of MAPK14 and glial glutamate transporter 1（GLT-1）detected by rPCR 72 hours after transfection，protein levels of MAPK14 and GLT-1 detected by Western blot 72 hours after transfection，level of lactate dehydrogenase（LDH）and mortality of neurons measured by spectrophotometry and flow cytometry 2 hours after culturing in the medium with glutamate. Results:（1）The optimal MOI value for lentivirus transfecting astrocytes was 30，and astrocytes grew well after transfection.（2）Seventy-two after transfection，the mRNA level of MAPK14 in lentivirus transfected group（0.005 7±0.000 6）was significantly decreased as compared with un-transfected group（0.013 1±0.001 1）and negative control group（0.013 9±0.001 0）（ P<0.01），the mRNA level of GLT-1 in lentivirus transfected group（0.009 1±0.001 2）was not significantly changed as compared with un-transfected group（0.008 7±0.000 3）and negative control group（0.008 9±0.001 1）（ P>0.05）.（3）Seventy-two hours after transfection，the protein level of MAPK14 in lentivirus transfected group（0.29±0.04）was significantly decreased as compared with non-transfected group（0.61±0.05）and negative control group（0.63±0.01）（ P<0.01），the protein level of GLT-1 in lentivirus transfected group（0.73±0.06）was significantly increased as compared with un-transfected group（0.20±0.03）and negative control group（0.23±0.09）（ P<0.01）.（4）After astrocytes were co-cultured with neurons and subsequently cultured in the medium containing glutamate for 2 hours，the level of LDH in lentivirus transfected group［（109.67±2.40）U/L］was significantly lower than that in un-transfected group［（141.52±3.88）U/L］and negative control group［（141.29±3.61）U/L］（ P<0.01）. The mortality of neurons in lentivirus transfected group［（38.72±0.26）%］was significantly lower than that in un-transfected group［（52.94±1.36）%］and negative control group［（54.30±1.23）%］（ P<0.01）. Conclusions:The transfection with lentivirus-mediated MAPK14 interference vector can increase expression of GLT-1 in astrocytes to increase glutamate re-uptake and relieve the glutamate excitatory toxicity in neurons，which may provide a new experimental basis for future use of astrocyte gene regulation to alleviate neuronal injury caused by glutamate excitatory toxicity after traumatic brain injury.
ABSTRACT
Objective To compare the clinical and imaging characteristics of traumatic acute subdural hematoma acute subdural hematoma with rapid resolution and those without rapid resolution.Methods A retrospective case-control analysis was conducted on the clinical data of 60 traumatic acute subdural hematoma patients with hematoma thickness ≥ 5 mm admitted to Second Affiliated Hospital of Shantou University Medical College from January 2011 to May 2018.There were 37 males and 23 females,aged 18-80 years [(47.0±16.9)years].There were 27 patients in the rapid resolution group and 33 patients in the non-rapid resolution group.Coagulation function [prothrombin time(PT)and international normalized ratio(INR)] on admission,hospital stay,Glasgow outcome scale(GOS),and brain CT results were compared between the two groups.Results The PT and INR values in the rapid resolution group were(11.9±2.1)s and 1.1±0.2 respectively,while those in the non-rapid resolution group were(10.8±1.0)s and 1.0±0.1 respectively,with significant differences(P＜0.05).There was no significant difference in hospital stay and GOS between the two groups(P＞0.05).The thickness of subdural hematoma of the two groups in the first CT scanning was(8.2±2.3)mm and(7.3±1.8)mm,respectively,with no statistically significant difference(P＞0.05).In the second CT scanning,the hematoma thickness in the rapid resolution group was significantly lower than that in the non-rapid resolution group [(2.7±1.9)mm vs.(6.6±2.1)mm](P＜0.01).The incidence of low density zone between the hematoma and intracranial plate was statistically higher in rapid resolution group than that in non-rapid resolution group(93％vs.36％)(P＜0.01).The incidence of subarachnoid hemorrhage(SAH)increase wassignificantly higher in rapid resolution group than that in non-rapid resolution group in the second CT scan(74％vs.15％)(P＜0.01).The conversion rate of acute subdural hematoma to subacute or chronic subdural hematoma was 4％in the rapid resolution group,which was significantly lower than 18％in the non rapid resolution group(P＜0.05).Conclusions The abnormal coagulation function and low density zone indicated by CT are two important indicators of rapid resolution in patients with traumatic acute subdural hematoma.The risk of conversion from acute into subacute or chronic subdural hematoma is lower in rapid resolution of traumatic acute subdural hematoma,suggesting that rapid resolution may be one of the prognostic indicators of ASDH patients.
ABSTRACT
Objective@#To compare the clinical and imaging characteristics of traumatic acute subdural hematoma acute subdural hematoma with rapid resolution and those without rapid resolution.@*Methods@#A retrospective case-control analysis was conducted on the clinical data of 60 traumatic acute subdural hematoma patients with hematoma thickness≥5 mm admitted to Second Affiliated Hospital of Shantou University Medical College from January 2011 to May 2018. There were 37 males and 23 females, aged 18-80 years [(47.0±16.9)years]. There were 27 patients in the rapid resolution group and 33 patients in the non-rapid resolution group. Coagulation function [prothrombin time (PT) and international normalized ratio (INR)] on admission, hospital stay, Glasgow outcome scale (GOS), and brain CT results were compared between the two groups.@*Results@#The PT and INR values in the rapid resolution group were (11.9±2.1)s and 1.1±0.2 respectively, while those in the non-rapid resolution group were (10.8±1.0)s and 1.0±0.1 respectively, with significant differences (P<0.05). There was no significant difference in hospital stay and GOS between the two groups (P>0.05). The thickness of subdural hematoma of the two groups in the first CT scanning was (8.2±2.3)mm and (7.3±1.8)mm, respectively, with no statistically significant difference (P>0.05). In the second CT scanning, the hematoma thickness in the rapid resolution group was significantly lower than that in the non-rapid resolution group [(2.7±1.9)mm vs. (6.6±2.1)mm] (P<0.01). The incidence of low density zone between the hematoma and intracranial plate was statistically higher in rapid resolution group than that in non-rapid resolution group (93% vs. 36%) (P<0.01). The incidence of subarachnoid hemorrhage(SAH) increase was significantly higher in rapid resolution group than that in non-rapid resolution group in the second CT scan (74% vs. 15%) (P<0.01). The conversion rate of acute subdural hematoma to subacute or chronic subdural hematoma was 4% in the rapid resolution group, which was significantly lower than 18% in the non rapid resolution group (P<0.05).@*Conclusions@#The abnormal coagulation function and low density zone indicated by CT are two important indicators of rapid resolution in patients with traumatic acute subdural hematoma. The risk of conversion from acute into subacute or chronic subdural hematoma is lower in rapid resolution of traumatic acute subdural hematoma, suggesting that rapid resolution may be one of the prognostic indicators of ASDH patients.