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Objective@#To evaluate the clinical value and outcomes of technical improvement of hybrid operatical clipping for large paraclinoid internal carotid artery aneurysms.@*Methods@#A review was conducted on 18 cases of large paraclinoid internal carotid artery aneurysm which were clipped by balloon non-fluoroscopic occlusion of the parent artery via a micro-bone window frontolateral approach in hybrid operating room at Neurosurgery Department of Tianjin Medical University General Hospital from June 2014 to December 2017. There were 8 males and 10 females with age of (63±4) years. There were 6 cases of unruptured aneurysm and 12 cases of ruptured aneurysm of subarachnoid hemorrhage (6 cases of grade Ⅱ, 4 cases of grade Ⅲ and 2 cases of grade Ⅳ in Hunt-Hess classification). Frontolateral approach incision (average length of about 5 cm) and bone window about 3 cm×3 cm were performed. No incision of the neck was needed to expose the internal carotid artery for temporary occlusion. In the operation, the balloon was slowly pushed to the preset position of the internal carotid artery under non-fluoroscopy. The balloon was expanded to block the blood flow of internal carotid artery. Then aneurysm was clipped. The balloon was loosened and retraced to the guiding catheter after clipping. The clipping condition was examined by cerebral angiography. If there was residual aneurysm neck or stenosis of the parent artery, the balloon was pushed under non-fluoroscopy again to temporary occlusion and the clip was adjusted until the aneurysm neck was clamped satisfactorily.@*Results@#Eighteen aneurysms were successfully clipped in hybrid operating room. Fourteen aneurysms showed complete occlusion of the aneurysm neck and no stenosis of the parent artery. Four cases showed residual aneurysm neck after clipping by intraoperative angiography, then aneurysms were clipped satisfy by adjusting the aneurysm clip. The patients were followed up for 3 months to 1 year. Ten patients recovered well (modifed Rankin score (mRS): 0), and 3 patients had no obvious disability (mRS: 1). Two patients with Hunt-Hess grade Ⅲ were slightly disabled (mRS: 2). 1 patients with Hunt-Hess grade Ⅲ were moderately disabled (mRS: 3). 1 patients with Hunt-Hess grade Ⅳ were severely disabled (mRS: 4). One elderly patients with Hunt-Hess grade Ⅳ were seriously disabled (mRS: 5).@*Conclusions@#Application of balloon non-fluoroscopic occlusion clipping for large paraclinoid internal carotid artery aneurysm via a micro-bone window frontolateral approach is safe, effective and minimally invasive.
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Objective To investigate the application of diffusion tensor imaging (DTI) in evaluating prognosis of patients with moderate and severe diffuse axonal injury (DAI).Methods A prospective cohort study was made on 35 patients with moderate and severe DAI,who were enrolled from June 2013 to December 2015 as study group.There were 21 males and 14 females,with age of (55.1 ± 11.6) years.The Glasgow coma scale (GCS) was (8.2 ± 2.9)points on admission.Moderate DAI was seen in 20 patients and severe DAI in 15 patients.Other 15 healthy volunteers were selected as control group.Fractional anisotropy (FA) was measured by DTI in three areas of interests as follows:the corpus callosum,thalamus,and brainstem areas.Glasgow outcome scale (GOS) was adopted to assess the prognosis of DAI patients at 6 months after the injury.The FA values of the three areas between study group and control group as well as FA values of patients when they were admitted to hospital and 6 months after injury were measured.In this way,the relationship between the FA values of different areas of interests in DAI patients on admission and the prognosis 6 months after injury was analyzed.Results The FA values of the corpus callosum,thalamus and brainstem area in study group were all lower than those in control group (P ＜ 0.05).Further,FA values of the corpus callosum,thalamus,brainstem area in severe DAI patients were lower than those of moderate DAI patients (P ＜ 0.05).FA values of the corpus callosum,thalamus,brain stem areas in DAI patients at 6 months after injury were lower than those of corresponding areas when DAI patients were admitted to hospital (P ＜ 0.05).FA values of the corpus callosum,thalamus and brain stem on admission were significantly positively correlated with GOS at 6 months after injury (P ＜ O.05).Conclusions Lower FA values of the corpus callosum,thalamus and brainstem area in patients with moderate and severe DAI are associated with more severe injury and worse prognosis.DTI scans can be used as a valuable tool to evaluate the prognosis of DAI patients.
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Objective To investigate the clinical therapeutic effect of mild hypothermia in elderly patients with severe traumatic brain injury.Methods 72 cases of elderly patients with severe traumatic brain injury(GCS ≤ 8) were divided into mild hypothermia therapy group(36 patients)and control group(36 patients) according to the random number table method.Mild hypothermia therapy group received mild hypothermia treatment while control group received normal treatment.The clinical prognosis was analyzed between the two groups.Results After 24h treatment,both mild hypothermia therapy group and control group intracranial pressure began to rise.But the intracranial pressure of the mild hypothermia therapy group(24 h:(13.0±4.5)mmHg,3 d:(16.6±4.0) mmHg,5 d:(19.9±3.9) mmHg,1 mmHg=0.133 kPa) were significantly lower than those of the control group (24 h:(16.6± 3.8) mmHg,3 d:(20.4±4.8) mmHg;5 d:(24.1 ± 6.2) mmHg),and the difference was statistically significant (t=2.225,2.260,2.192,P=0.035,0.033,0.039).The rate of good recovery to the control group and the mild hypothermia therapy group were 22.22％ and 47.22％ respectively while the mortality were 30.56％ and 13.89％ respectively,and the differences were statistically significant (x2 =4.936,5.675,P=0.047,0.035).Conclusion Mild hypothermia treatment can inhibit the increase of intracranial pressure and reduce disability rate and mortality in elderly patients with severe traumatic brain injury,which can increase the survival rate and improve the long-term prognosis.
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Objective:To explore the effect of bone marrow mesenchymal stem cell transplantation on silicosis fibrosis in different time windows in rats.Methods:BMSCs were isolated and cultured from male 3-week-old SD rats in vitro.Fifty healthy female SD rats were randomly divided into 5 groups:control group,silicosis model group,early treatment group,middle treatment group,late treatment group(n=10).The silicosis model was made by one-time infusion of silica dust suspension using the non-exposed tracheal in-tubation(50 μg/ml),and 1 d,14 d,28 d of BMSCs were given for intervention therapy (1 ×106 ml-1 ).Rats in each group were sacrificed 14 days after treatment.The BMSCs identified by flow cytometry;the morphological changes of the lung tissues were observed by HE staining;the expression of MMP-9,collagen type Ⅰ and collagen type Ⅲ were detected by immunocytochemistry and Western blot analysis.Results: BMSCs in early silicosis ( 1 d ) and the middle silicosis ( 14 d ) compared to silicosis model group can significantly alleviate the pathological process of silicosis fibrosis (P0.05).Conclusion:Exogenous BMSCs transplantation on rat silicosis early pathological processes play a role in delaying , late treatment effect is not obvious.
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Objective To observe expression of autophagy-related proteins LC-3 and Beclin-1 in alveolar macrophages in the silicosis model of rats , and to investigate the molecular mechanisms of silicosis formation from cells autophagy perspective .Methods Fifty adult male SD rats were randomly divided into control and model groups , 25 rats for each group .The silicosis model was made by one-time infusion of silica dust suspension through the trachea without exposed.The rats were sacrificed on the 1st, 3rd, 7th, 14th or 28th day after the modeling .Alveolar macrophages were obtained from bronchoalveolar lavage and used for subsequent research after culture and purification .Morphological characteristics of alveolar macrophages were observed by HE staining and transmission electron microscope ;The expression of LC-3 and Beclin-1 was detected by means of immunocytochemistry and Western blotting in each group .Results Compared with the control group , alveolar macrophages of the model group had larger volume and abundant cytoplasm , the phagocytic silica dust particles were observed in some cells , and autophagosomes were detected by transmission electron microscope .The expressions of LC-3 and Beclin-1 were increased at all time points in the model group ( P<0.05 ) .Both LC-3 and Beclin-1 began to increase at the 1st day.As the extension of time the expression gradually enhanced , peaked at the 14th day(P<0.05), and decreased at the 28th day, but higher than the basal expression .Conclusion Autophagy is activated in alveolar macrophages of the silicosis model , and alveolar macrophages autophagy is involved in the pathological process of silicosis in the rat .
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Objective To explore the influence of injury severity on transplantation of embryonic neural stem cells (NSCs) after traumatic brain injury (TBI).Methods The NSCs were isolated from the hippocampus of fetal rats aged at from 12-14 days.The cells were cultured and proliferated in the serum-free medium and identified in vitro.The animals received transplants in the bilateral hippocampal areas at day 3 following mild or moderate TBI separately.Conventional histology,TUNEL and immunohistology were examined to detect BrdU,NSE,GFAP,GalC,NGF and BDNF at day 14 post-implantation.Results BrdU-labeled positive cells in the bilateral hippocampus in the mild TBI group were more than those in the moderate TBI group at day 14 post-implantation.Significant differentiation of the astrocytes recognized as GFAP positive cells in the bilateral hippocampus was found at day 14 post-implantation.The expression of NGF and BDNF proteins was increased following TBI,the most evident in the mild TBI group.Conclusion The influence of injury severity on transplantation may be associated with the change of the microenvironment after TBI.
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AIM: To investigate the effect of extracellular signal-regulated kinase 1/2 signaling pathway after severe diffuse brain injury (DBI) in rats, and to provide base for treatment. METHODS: Male Sprague-Dawley rats were randomly divided into four groups: control group, traumatic group, low dose of inhibitor U0126 treatment group and high dose of inhibitor U0126 treatment group. DBI rat model was established according to the description of Marmarou's diffused brain injury. At 30 min and 1 h, 6 h, 24 h, 48 h and 72 h after injury, morphological changes were observed under light and electronic microscopes. The ERK1/2 phosphorylation and c-Fos were measured by Western blotting. Apoptosis was measured with TUNEL method. Learning and memory function were performed with Morris water maze from 3 days to 7 days after injury. RESULTS: After trauma, some neurons displayed histopathologic changes of necrosis and apoptosis, axon myelin sheath internalization and disconnection. ERK1/2 phosphorylation protein was apparently increased at 30 min after injury, approached peak at 6 h and continued to 24 h. c-fos protein was markedly increased at 30 min after injury, approached peak at 6 h and returned to bottom at 24 h. The number of apoptotic nerve cells increased at 6 h after and approached peak at 72 h. Latencies of searching safety island prolonged. Rats treated with U0126 had reduction in ERK1/2 activity, c-Fos protein, neuronal apoptosis and searching safety island latencies. CONCLUSION: The activated ERK1/2 signaling pathway plays an important role in processing of nerve cell apoptosis after severe DBI.
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Objective To study the mechanism of interaction between neuronal nitric oxide syn-thase (nNOS) and inducible nitric oxide synthase (iNOS) following traumatic brain injury (TBI) in rats. Methods A total of 250 male Wistar rats were randomly divided into five groups, ie, sham oper-ation group, trauma group, 7-nitroindazole (7-NI) treatment group, aminoguanidine (AG) treatment group and combined AG and 7-NI treatment group. Severe closed TBI was made by using Marmarou meth-od. Protein expressions of nNOS and iNOS in hippocampus CAI were detected by means of immunohisto-chemical staining at 1,3, 6, 12 hours and at days 1,3, 7 and 14 after TBI. Results The expression of nNOS reached a peak at 6 hour after injury in all groups, with no statistical difference between groups (P > 0. 05), when there was no statistical difference between 7-NI treatment group and trauma group (P > 0. 05) but statistical difference in AG treatment group and combined AG and 7-NI treatment group compared with trauma group at 12 hours after TBI (P <0.05). The expression of iNOS reached maximal level at day 3 after TBI, with lower level in 7-NI group, AG treatment group and combined AG and 7-NI treatment group compared with trauma group (P < 0.05). Conclusions After TBI, nNOS interacts with iNOS by means of the feedback of nitric oxide. The enhanced expression of nNOS is initial factor for increase of iNOS expression, which can down regulate the expression of iNOS.
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Objective To optimize human acellular dermal matrix(ADM) and evaluate its biological characters. Methods Human skin was treated with hypertonic saline followed by NaOH maceration(group A), hypertonic saline followed by sodium dodecyl sulfate (SDS) detergent(group B) or Dispase Ⅱ followed by Triton X-100(group C), the resulting ADM were sectioned, and then were stained by special immunohistochemistry method. The cytotoxicity of them were evaluated by methyl thiazolyl tetrazolium (MTT) colorimetry and then cell compatibility was analyzed by cell culture;The optimized ADM resulted was choosen for use. Fibrablasts(FBs)were transfected with adenovirus vector encoding green fluorescent protein gene(Ad-GFP)and the growth of them on the optimized ADM was observed by fluorescent microscopy. Results Collagen and elastic fibers can still be observed in three kinds of ADM. The cells in dermis can be disintegrated both in group A and C, but not in group B. The cytotoxicity scores of the ADM prepared in group A and B were grade 0 or grade 1, while that of group C was more than grade 1.The ADM prepared by NaCl-NaOH maceration had good biocompatibility. There was statistical difference in adhering number of NIH3T3 cells in group A and B. NIH3T3 cells grew well in group A and the resulted ADM was optimized. FBs transfected with Ad-GFP grew well in the optimized ADM. Conclusion The ADM prepared by NaCl-NaOH maceration was a good tissue engineering biomaterial with a little cytotoxicity and rich in resouce.
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Objective To study the mechanism of dermal fibroblasts as a feeder layer to support the growth of human keratinocytes. Methods Human dermis fibroblasts were isolated and cultured and then treated with mitomycin-C. The expression of type Ⅰand type Ⅲ precollagen mRNA and relevant protein in feeder layer were examined by RT-PCR and Immunohistochemistry. KCs were cultured both on FB and NIH3T3 feed layer as control, the adhering numbers and the time of fusion were recorded. Results RT-PCR showed an increase of type Ⅰprecollagen mRNA in FB feeder layer as compared with that of normal fibroblasts (P
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<p><b>OBJECTIVE</b>To explore the correlation between cognition disorder and morphologic change of hippocampal neurons after traumatic brain injury (TBI).</p><p><b>METHODS</b>Wistar rat models with severe TBI were made by Marmarou's method. The histopathological change of the neurons in the hippocampus area were studied with hematoxylin-eosin (HE) staining and terminal deoxynucleotidyl transferase-mediated X-dUPT nick end labeling (TUNEL), respectively. The cognitive function was evaluated with the Morris water maze test.</p><p><b>RESULTS</b>The comprehensive neuronal degeneration and necrosis could be observed in CA2-3 regions of hippocampus at 3 days after injury. Apoptotic positive neurons in CA2-4 regions of hippocampus and dentate gyrus increased in the injured group at 24 hours following TBI. They peaked at 7 days and then declined. Significant impairment of spatial learning and memory was observed after injury in the rats.</p><p><b>CONCLUSIONS</b>The rats have obvious disorders in spatial learning and memory after severe TBI. Meanwhile, delayed neuronal necrosis and apoptosis can be observed in the neurons in the hippocampus area. It suggests that delay ed hippocampal cell death may contribute to the functional deficit.</p>
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Animals , Male , Rats , Apoptosis , Brain Injuries , Pathology , Cognition Disorders , Pathology , Hippocampus , Pathology , In Situ Nick-End Labeling , Memory Disorders , Pathology , Necrosis , Rats, WistarABSTRACT
Objective To explore the mechanism of neurotoxic effects of neuron nitric oxide synthase(nNOS)and inducible nitric oxide synthase(iNOS),and the therapeutic effects of 7-nitroindazole(7-NI)and aminoguanidine(AG),in traumatic brain injury(TBI)rats.Methods Two hundred and fifty adult male Wistar rats were randomly divided into 5 groups:sham operation group,traumatic group,AG group,7-NI group and AG+7-NI group.Animals in each group were divided into 8 subgroups according to the time after trauma(1,3,6,12 hours and 1,3,7,14 days).Severe diffused brain injury model was made with Marmarou method.Expressions of nNOS and iNOS in hippocampus CA1 region were determined by immunohistochemistry,nerve cells apoptosis in hippocampus CA1 region was observed by TUNEL methods,and the relationship between apoptosis and NOS was observed by double staining.Results In trauma group,the expression of nNOS in hippocampus CA1 region peaked at 6h post-trauma,the expression of iNOS and apoptosis of nerve cells in hippocampus CA1 region both peaked at 3d post-trauma,while the apoptosis was alleviated in AG group,7-NI group and AG+7-NI group.The number of both TUNEL staining and nNOS immunostaining positive cells increased at 6h post-trauma in trauma group,significantly higher than that in 7-NI group.The number of both TUNEL staining and iNOS immunostaining positive cells increased at 3d post-trauma in trauma group,significantly higher than that in AG group.Conclusions The over-expression of nNOS and iNOS has toxic effects to neural tissues of brain,serves as one of the factors inducing nerve cell apoptosis in hippocampus CA1 region.7-NI and AG can inhibit the expression of nNOS and iNOS,reduce the nerve cell apoptosis,and play an important role in neuroprotective effect.
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OBJECTIVE: To investigate the relation betwe en necrosis and apoptosis in the hippocampus of exogenous bFGF on this process. METHODS: With Marmarou's method we produced a severe diffuse brain injury and studied the changes in the hippocampus by adapting a modified T dT-mediated dUTP-biotin nick end labeling (TUNEL) method. At the same time we observed the effect of exogenous bFGF on neuronal necrosis and apoptosis. RESULTS: We found that together with cell necrosis there was an increase in the number of apoptotic neurons in the hippocampus CA2-3 sectors a s early as 4 h after injury, with numbers reaching a maximum at 7 d. Exogenous b FGF resulted in a definite reduction in the amount of necrosis and apoptosis. CONCLUSIONS: Neuronal necrosis and apoptosis occur in combinati on after brain injury and that one of the causes may be the insufficience expres sion of the bFGF gene in the hippocampus after severe injury. Exogenous bFGF and similar substance may prove clinically useful after brain injury by reducing ce ll necrosis and apoptosis.