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Objective To explore the effect of Dexmedetomidine (Dex) on acute brain edema in mice in condition with targeted temperature management (TTM) following traumatic brain injury (TBI).Methods A total of 180 male C57BL/6J mice were divided into control group,sham operation group,TBI group,TBI + Dex group,TBI + TTM group,and TBI + Dex + TTM group according to the random number table (n =30 per group).The sham operation group only opened the bone window but did not hit it,and the control group did not open the bone window.The TBI + Dex,TBI + TIM,and TBI + Dex + TTM groups were intraperitoneally injected with Dex (60 μg/kg once every 2 h for 3 times) and/or hypothermia after TBI.The brain tissue injury volume,EB extravasation and brain water content of each group were determined by toluidine blue,Evans blue staining and dry-wet weight method at 24 hours after injury.Real-time quantitative PCR and Western blot were used to detect the expression of Claudin-5 in the injured brain tissue.At 24,48,and 72 hours after injury,the neurological deficiency degree was assessed using the modified neurological severity scores (mNSS).Results Compared with the sham operation group,TBI mice showed significant increase in brain tissue injury volume [(0.49 ± 0.04)mm3 vs.(1 1.57 ± 1.01)mm3],blood-brain barrier permeability [(16.4 ± 0.8) μg/g vs.(54.3 ± 1.7) μg/g],brain tissue water content [(76.7 ± 0.9) ％ vs.(83.1 ± 0.8) ％],and mNSS score [(1.6 ± 0.7) points vs.(13.4 ± 0.7) points] at 24 hour after TBI (all P ＜ 0.01).However,Dex or TTM treatment reduced brain tissue injury volume [(7.20±0.18)mm3 and (5.94 ±0.18)mm3],blood-brain barrier permeability [(32.7 ± 1.2) μg/g and (27.6 ± 1.0) μg,/g],brain tissue water content [(78.5 ± 0.4) ％ and (78.2 ± 0.6) ％],and neurological function [mNSS:(7.3 ± 1.1) points and (5.8 ± 1.3) points] (all P＜0.01).Moreover,Dex + TTM group showed better neuroprotection [reduced brain tissue injury volume:(3.92 ± 0.05) mm3,reduced BBB permeability:(21.6 ± 0.7) μg/g,reduced brain water content:(77.7 ±0.3)％,and reduced mNSS:(4.3 ± 1.2) points] compared with Dex or TTM alone (all P ＜ 0.01).Additionally,the mRNA expression of Claudin-5 (0.23 ± 0.01) decreased significantly at 24 hours after TBI compared with sham group (0.93 ± 0.04,P ＜ 0.01),but Dex or TTM could increase the expression of Claudin-5 (0.47 ± 0.01,and 0.54 ± 0.09) compared with TBI group (P ＜0.01),especially that of TBI + Dex + TTM group (0.64 ± 0.02,P ＜ 0.01).Furthermore,the protein expression of Claudin-5 was in accordance with the result of its mRNA expression.Conclusion Dex in condition with targeted temperature management can up-regulate Claudin-5 expression in early TBI,protect the integrity of blood-brain barrier,attenuate acute brain edema and neurological damage,and improve neurological function recovery.
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BACKGROUND: Hyaluronan-methylcellulose hydrogel cannot only be conjugated with short peptide sequences and growth factors to achieve sustained release, but also has a role in blocking dural defects and reducing inflammation. It is an ideal biomaterial for the treatment of spinal cord injury. OBJECTIVE: To investigate the effect of neurotrophin-3 modified hyaluronan-methylcellulose (HAMC-NT-3) hydrogel on the recovery of neurological function in rats with spinal cord injury. METHODS: Fifty-four female Sprague-Dawley rats (provided by the Experimental Animal Center of the Academy of Military Medical Sciences in China) were randomly divided into three groups (n=18 per group) . The sham group only underwent T10 laminectomy. In the model group and the experimental group, an aneurysm clip was used to establish spinal cord injury models after T10 laminectomy. The experimental group was locally injected with HAMC-NT-3 hydrogel. The Basso Beattie Bresnahan function scoring was performed at 1 day, 1, 2, 3, 4, 5, 6, 7, and 8 weeks after surgery. The inclined plane test was performed at 4, 6 and 8 weeks after surgery to evaluate the recovery of hindlimb motor function. ELISA was used to detect the concentrations of inflammatory factors in the spinal cord at 1 week after surgery. Immunohistochemical staining was used to observe the area of syringomyelia, glial fibrillary acidic protein expression and nerve regeneration at 8 weeks after surgery. RESULTS AND CONCLUSION: (1) The Basso Beattie Bresnahan scores of the model group and the experimental group were lower than those of the sham group at various time points after surgery (P < 0.05) . The Basso Beattie Bresnahan scores of the experimental group were higher than those of the model group at 4-8 weeks after surgery (P < 0.05) . (2) In the inclined plane test, the maximum inclined angles of the model group and the experimental group at each time point after surgery were lower than that of the sham group (P < 0.05) . The maximum inclined angles of the experimental group at 6 and 8 weeks after surgery were higher than those of the sham group (P < 0.05) . (3) The concentrations of tumor necrosis factor-α, interleukin-1β, interleukin-6 and interleukin-10 in the experimental group and the model group were higher than those in the sham group (P < 0.05) . The concentrations of tumor necrosis factor-α, interleukin-1β and interleukin-6 in the experimental group were lower than those in the model group (P < 0.05) . The concentration of interleukin-10 in the experimental group was higher than that in the model group (P < 0.05) . (4) Immunohistochemical staining showed that the expression levels of glial fibrillary acidic protein in the experimental group and the model group were higher than those in the sham group, while the expression of glial fibrillary acidic protein in the experimental group was lower than that in the model group. The area of syringomyelia in the experimental group was smaller than that in the model group (P < 0.05) . These results indicate that local injection of HAMC-NT-3 hydrogel can effectively inhibit inflammation as well as astrocyte activation and proliferation, reduce fibrous scar formation, and promote the protection of nerve tissue and the recovery of hindlimb motor function after spinal cord injury.
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Objective To investigate the neuroprotective effect of Ghrelin on traumatic brain injury (TBI) in mice.Methods TBI model of C57BL/6 mice was established by electronic cortical impact instrument (eCCI).According to the random figure table method,twenty-four mice were randomly divided into sham group(Sham group),TBI group and Ghrelin intervention group(Ghrelin group) with 8 mice in each group.The model of TBI was established in TBI group and Ghrelin group.The mice in Ghrelin group was injected intraperitoneally 0.5 g/kg before and 1 h after injury respectively.And the mice Sham group and TBI group were injected with the same amount of normal saline.The changes of cerebral blood perfusion (CBP)were monitored in real time by laser speckle contrast analysis(LSCI),the changes of neuroelectrophysiology were observed by monitoring motor evoked potential (MEP),and the status of neurological deficit was evaluated by modified neurological deficit score (mNSS).Results Compared with Sham group,the mice in TBI group had significantly lower cerebral blood perfusion(CBP) (t=-12.36,P＜0.01),longer latency and lower amplitude of motor evoked potential (MEP) (t=5.03,-11.55,all P＜0.01),and significantly higher mNSS scores (t=9.34,P＜0.01).However,compared with the TBI group,the cerebral blood perfusion(CBP) of Ghrelin group increased significantly at 12 h after TBI((196.87± 17.36) PU/mm2 vs (123.62±8.04)PU/mm2,t=10.45,P＜0.01),while the latency of MEP decreased((5.30±0.33) ms vs (6.80±0.97) ms,t =-5.01,P＜0.01),the amplitude of MEP increased ((2.21 ± 0.16) mV vs (1.27± 0.27) mV,t =9.65,P＜0.01).And compared with the TBI group,the neurological deficit score of Ghrelin group decreased significantly at 24 h after TBI((4.9±1.2) vs (8.4±2.6),t=-3.87,P＜0.01).Conclusion Ghrelin exhibits a significant neuroprotective role by increasing cerebral blood flow perfusion,reducing the degree of neurological deficit and promoting motor function recovery in TBI mice.
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Objective@#To investigate the neuroprotective effect of Ghrelin on traumatic brain injury (TBI) in mice.@*Methods@#TBI model of C57BL / 6 mice was established by electronic cortical impact instrument (eCCI). According to the random figure table method, twenty-four mice were randomly divided into sham group(Sham group), TBI group and Ghrelin intervention group(Ghrelin group) with 8 mice in each group. The model of TBI was established in TBI group and Ghrelin group.The mice in Ghrelin group was injected intraperitoneally 0.5 g/kg before and 1 h after injury respectively. And the mice Sham group and TBI group were injected with the same amount of normal saline. The changes of cerebral blood perfusion (CBP) were monitored in real time by laser speckle contrast analysis(LSCI), the changes of neuroelectrophysiology were observed by monitoring motor evoked potential (MEP), and the status of neurological deficit was evaluated by modified neurological deficit score (mNSS).@*Results@#Compared with Sham group, the mice in TBI group had significantly lower cerebral blood perfusion(CBP) (t=-12.36, P<0.01), longer latency and lower amplitude of motor evoked potential (MEP) (t=5.03, -11.55, all P<0.01), and significantly higher mNSS scores (t=9.34, P<0.01). However, compared with the TBI group, the cerebral blood perfusion(CBP) of Ghrelin group increased significantly at 12 h after TBI((196.87±17.36) PU/mm2 vs (123.62±8.04)PU/mm2, t=10.45, P<0.01), while the latency of MEP decreased((5.30±0.33)ms vs (6.80±0.97)ms, t=-5.01, P<0.01), the amplitude of MEP increased((2.21±0.16)mV vs (1.27±0.27)mV, t=9.65, P<0.01). And compared with the TBI group, the neurological deficit score of Ghrelin group decreased significantly at 24 h after TBI((4.9±1.2) vs (8.4±2.6), t=-3.87, P<0.01).@*Conclusion@#Ghrelin exhibits a significant neuroprotective role by increasing cerebral blood flow perfusion, reducing the degree of neurological deficit and promoting motor function recovery in TBI mice.
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Objective@#To investigate the effects of Casein kinase 2-interacting protein 1 (CKIP-1) gene silencing on the proliferation of glioma cells U87-MG.@*Methods@#The recombinant lentiviral vectors targeting CKIP-1 gene or negative control were constructed and then used to infect glioma U87-MG cell line.The effects of knock-down on the mRNA or protein expression of CKIP-1 were evaluated by real-time qPCR and western blotting.Cell cycle was detected by the flow cytometry assay, and cell proliferation changes were evaluated by cell counting, MTT, and BrdU assay, respectively.Lastly, the colony formation was used to investigate the effect of CKIP-1 knock-down on the clone formation.@*Results@#Compared with the group of Ctrl, CKIP-1 siRNA was observed to significantly inhibit CKIP-1 expression at the mRNA levels (Ctrl (1.01±0.13) vs CKIP-1 siRNA (0.23±0.02), P<0.01) and protein levels in the U87-MG cells.Also, CKIP-1 suppression mediated by RNAi decreased the ratio of G0/G1 phase (Ctrl (69.64±0.79) vs CKIP-1 siRNA(62.64±0.66), P<0.01), increased that of G2/M phase (Ctrl (8.36±0.52) vs CKIP-1 siRNA(13.87±2.90), P<0.05), and significantly inhibited the cell proliferation and clone formation (Colony number: Ctrl (25±2) vs CKIP-1 siRNA(2±1), P<0.05) of transfected U87-MG cells.@*Conclusion@#Knocking down the expression of CKIP-1 significantly inhibit cell proliferation in human U87-MG glioma cells, indicating that CKIP-1 is involved in the development of gliomas and could promote the cell proliferation.
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Objective To investigate the effect of Ghrelin on gastrointestinal motility after traumatic brain injury (TBI).Methods A total of 72 adult male SD rats were randomly divided into sham operation group (n =8),TBI group (n =32) and Ghrelin group (n =32),according to the random number table.In the sham operation group,the scalp was sutured after craniotomy and sterilization,without any strike.In the TBI group,after intraperitoneal anesthesia,the skull was opened and the electric cortical contusion impactor was used to strike the center of bone window at the depth of 3 mm and the rate of 5 m/s.The duration of hitting the lowest point was 200 ms.In the Ghrelin group,20 μg/kg of Ghrelin was injected into the rat via the tail vein 30 minutes after injury.The modified neurologicalseverity score (mNSS),percentage of water content in feces and percentage of gastric contents in body weight at 6,24,48 and 72 hours after operation in each group were measured.The stomach,the small intestine 15 cm from ileocecal junction,ileocecal junction (about 3 cm in the proximal ileal loops,about 3 cm in the distal ileal loops,and 3 cm colon loops) were taken out to prepare the electron microscopy section and observe the microscopic changes of the gastrointestinal mucosa.Results The mNSS in the TBI and Ghrelin groups was higher than that in the sham operation group after 24,48 and 72 hours (P ＜0.01).The mNSS in the TBI group was higher than that in the Ghrelin group after 24,48 and 72 hours (P ＜0.01).In the sham operation group,the intestinal wall was pink.In the TBI group,gastric dilatation and thinner wall with pale or dark red color were seen,and small intestine cavity expansion with dark color and even congestion were observed.There was much mucus in the intestinal wall.The Ghrelin group improved obviously than the TBI group after 6 hours.Compared with the Ghrelin group,the percentage of fecal water content in the TBI group decreased significantly after 24 hours (P ＜ 0.05),and the decrease rate dropped with time.Obvious delayed gastric emptying occurred (P ＜ 0.05),and the percentage of gastric contents in body weight demonstrated downtrend.The changes of gastric mucosa were as follows:the chief cells in the gastric glands were observed 72 hours after TBI in the TBI group,and scattered short microvilli were seen in the cell surface.The cytoplasm protruded into the glandular cavity,and a large number of rough endoplasmic reticulum could be seen in the cytoplasm,with irregular arrangement.Medullary bodies could be seen inside the mitochondria which swelled locally.Abundant endocrine granules were seen in the cytoplasm.Mitochondria were scattered and swollen,and mitochondria cristae became shorter and fewer,which contained medullary bodies.The Ghrelin group improved obviously than TBI group after 72 hours in terms of gastric mucosa changes.With respect to cecum mucosa,in the TBI group 72 hours after TBI,severe edema of the cecum absorption epithelium,obvious dilation of the rough endoplasmic reticulum,expansion of the free water gap inside the cell,and local decrease of the microvilli at the top of the cell were observed.Abundant microvilli were seen in the cecum absorption epithelium and cell top.The connection complex composed of tight connections,intermediate connections,and bridging particle connections could be seen between cells.The Ghrelin group improved obviously than TBI group after 72 hours in terms of cecum mucosa changes.Conclusions Ghrelin can improve gastrointestinal motility and protect gastrointestinal mucosa in rats after TBI.
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The refractory epilepsy refers to the epilepsy whose seizures couldn't be cured after using two kinds of correctly selected antiepileptic drugs which can be tolerated and enough dosage and duration of monotherapy or combination therapy.The pathogenesis of intractable epilepsy is complex,and there is no established theory at home and abroad.Recently,more and more attention has been paid to the correlation between mitochondrial dysfunction caused by oxidative stress and intractable epilepsy.Based on the summary of common treatment of refractory epilepsy and by searching the related literature at home and abroad,further investigation would be made to explore the diagnosis and treatment of intractable epilepsy theory in order to provide new strategies for diagnosis and treatment of intractable epilepsy.
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Objective To screen altered proteins of hippocampus in the stress-induced depression (STRID) rat model, and explore the potential molecular mechanism. Methods Twenty Sprague-Dawley rats were randomly divided into the control group and STRID group, 10 rats in each group. Chronic unpredictable mild stress (CUMS) methods including fasting for solids and liquids, electric foot-shock, reversing day and night, cold water swimming, cage tilt, scare stimulation and tail pinch were conducted on STRID rats with no repeats for 28 days to make up the depression animal model. The control group was normally fed during this period. After the stress stimulation, the hippocampus protein samples were used for two dimensional electrophoresis to screen the differentially expressed protein, and then mass spectrum identification and function analyze were conducted. Results Compared with the control group, 34 proteins were altered in STRID group. Among which, 18 were up-regulated, and 16 were down-regulated. The differentially expressed proteins mainly located in cytoplasm, mitochondrion, extracellular exosome and myelin sheath. The involved signaling pathways included metabolic pathway, oxidative phosphorylation pathway, and Alzheimer's disease, Parkinson's disease and Huntington's disease pathways. Conclusion The altered proteins and dysfunction of nerve signaling, and the excess of oxidative phosphorylation in hippocampus of STRID rats may be one of the pathogenesises.
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Objective To explore the effects of different doses of dexmedetomidine on acute brain edema in mice with traumatic brain injury (TBI).Methods A total of 132 male C57BL/6J mice were randomly divided into six groups:control group (group C),sham-operation group (group Sham),traumatic brain injury group (group TBI),Dex 20 μg/kg (group D20),40 μg/kg (group D40),and 60 μg/kg (group D60),n=22 in each group.The TBI animal model was established by electric controlled cortical impactor (eCCI),then intraperitoneal injected by the administration of different doses of dexmedetomidine at 0,2 and 4 h after TBI.Twenty-four hours post-TBI,brain water content was measured by the dry-wet method,histological observation was performed using HE staining,and aquaporin 4 (AQP4) and NF-κB expression were detected using Western blot assay,respectively.Then,the modified neurological scale scores (mNSS) on 1,2,3,and 7 d and Morris water maze (MWM) test on 4,5,6 and 7 d post-TBI were used to evaluate the neurologic deficit of TBI mice.Results After traumatic brain injury,the mNSS scores,the escape latency,the brain water content and the expression of AQP4 and NF-κB increased significantly in group TBI (P＜0.01).Different doses of dexmedetomidine significantly reduced the mNSS scores,the escape latency,the brain water content and the expression of AQP4 and NF-κB (P ＜ 0.05 or P ＜ 0.01).And meanwhile dexrnedetomidine can lessen neuronal degeneration,and inflammation response.Additionally,the effect was remarkably in group D60 compared with group D20 (P ＜ 0.05 or P ＜ 0.01).Conclusion Dexmedetomidine can lessen brain edema and cognition impairment induced with traumatic brain injury,which is a dose-effect relationship within 20-60 μg/kg,and this effect may be related to the downregulation of AQP4 and NF-κB expression.
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Objective To explore the effect and mechanism of necroptosis related proteins in middle cerebral artery occlusion (MCAO) induced brain ischemia/reperfusion injury in mice.Methods C57BL/6 mice were used to establish the brain ischemia/reperfusion injury model induced by MCAO.MCAO mice were treated with z-VAD.fmk (zVAD,1.1 g/kg),GSK'872 (0.7 g/kg) and combined intervention of zVAD and GSK'872,and neurological defect was evaluated by mNSS while brain infarct volume was measured by TTC staining.Western blot and immunofluorescence assay were used to detect protein expression and location of RIP1,RIP3 and MLKL,respectively.Results Neurological defect and brain infarction were caused by MCAO.Compared with MCAO group,zVAD,GSK'872 and the combined intervention alleviated neurological defect and reduced brain infarct volume significantly (P＜0.05 or P＜0.01).The protein levels of RIP3 and RIP1 MLKL were increased in mice of MCAO group,while GSK'872 and the combined intervention obviously downregulated the aforementioned protein expression [RIP1 (GSK'872:0.64± 0.02 vs MCAO:1.28±0.02,P＜0.01);RIP3 (GSK'872:1.08±0.02 vs MCAO:1.45±0.02,P＜0.01);MLKL (GSK'872:0.54±0.01 vs MCAO:1.00±0.01,P＜0.01)].However,zVAD only slightly reduced protein expression of MLKL (P＜0.05) but didn't change the protein expression of RIP1 and RIP3 (P＞0.05).Conclusion RIP1,RIP3 and MLKL are involved in the execution of necroptosis and contribute to the pathological progress of brain ischemia/reperfusion injury.
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Objective To investigate the signaling pathway and the key signal molecules of protein kinase (RIPK)3 in SH-SY5Y cells. Methods SH-SY5Y cells were transfected with RIPK3 expression plasmid vector to upregulate intracellular RIPK3, while the SH-SY5Y cells were transfected with empty vector plasmid, which was considered as control group. Western blot assay was used to check the expression of exogenous RIPK3 in cells. The proliferation rate of SH-SY5Y cells was determined by MTT assay at designated time to detect exogenous RIPK3 activity. Whole transcriptome sequencing (RNAseq) was used to detect the transcription of genes. Whole-transcriptomic gene transcription was measured by following Ingenuity Pathway Analysis (IPA) to obtain downstream signaling pathways and the key molecule, which were partly confirmed by following droplet digital PCR (ddPCR). Results Exogenous RIPK3 showed biological activity in SH-SY5Y, which inhibited the proliferation of cells. IPA showed that znic finger protein 36 (ZFP36) was significantly up-regulated as compared with that of the control group. The tran?scription levels of ZFP36 downstream genes such as tumor necrosis factor (TNF), brain derived neurotrophic factor (BDNF), vascular endothelial growth factor (VEGF) and mRNA-decapping enzyme 2 (DCP2) were affected at the same time. Conclusion Within the limitations of this study, it seems that RIPK3 is notable for the development, inflammation and tumorigenesis of the nervous system as an independent regulator of ZFP36 gene and downstream effectors.
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Objective To investigate the role of three-dimensional scaffolds seeded with NeuroD1-modified neural stem cells (NSCs) for repair of spinal cord injury in rats.Methods A new three-dimensional bio-printer was used to make bionic spinal cord scaffolds.NSCs are transduced with retrovirus vectors encoding NeuroD1 to express transgenes in high levels.Forty healthy female SD rats were divided into control group,scaffold group,scaffold + NSCs-green fluorescent protein (GFP) group and scaffold + NSCs-NeuroD1 group with 10 rats per group,according to the random number table.Spinal cord injury in rats was induced using the electric controlled cortical impactor.A week later,the control group was excised 3 mm spinal cord at the injury site under microscope.RT-PCR was used to confirm the construction of NeuroD1 overexpressing NSCs.Survival and differentiation of transplanted NSCs were detected with fluorescent staining.Rat neurological motor function was evaluated with BBB score at postoperative 1,2,4,6 and 8 weeks.Rat electrophysiological changes were observed by monitoring motion evoked potential and sensory evoked potential at 8 weeks.Results RT-PCR results confirmed the successful reconstruction of NeuroD1-overexpressing NSCs.BBB score in scaffold + NSCs-NeuroD1 group was the highest and had significant differences compared to other three groups (P ＜ 0.05).Electrophysiological results showed the motor and sensory in scaffold + NSCs-NeuroD1 group had the shortest latencies and highest amplitudes,which revealed significant differences compared to other three groups (P ＜0.05).Immunofluorescence staining showed GFP cells in scaffold + NSCs-NeuroD1 group at 8 weeks,which differentiated into neurons and astrocytes.GAP-43 was positively stained,and myelin formation was detected.Conclusion Three-dimensional scaffolds seeded with NeuroD1-modified NSCs can promote nerve loop reconstruction in spinal cord injury rats,and accelerate recovery of motor and sensory function.
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ObjectiveTo analyze and compare the difference and prognosis between vascular embolization and craniotomy occlusion in patients suffering from aneurysmal subarachnoid hemorrhage (aSAH) with Hunt-Hess levelⅢ-Ⅳ, and acute postoperative hydrocephalus.Methods A retrospective study was conducted on 767 patients who had undergone vascular embolization (vascular embolization group,n = 403) or craniotomy occlusion operation (craniotomy occlusion operation group,n = 364), and the patients with postoperative acute hydrocephalus were screened. The clinical data of patients of both groups was analyzed. By judging short-term prognosis in patients with hydrocephalus with Glasgow outcome scale (GOS) score estimated at discharge, the advantages and disadvantages of two surgical procedures were compared.Results The number of cases with postoperative hydrocephalus in vascular embolization group was 56 (13.90%), while that in craniotomy occlusion group was 33 (9.07%). The difference between the two groups of incidence of hydrocephalus was statistically significant (χ2= 4.350,P = 0.037 ). In 767 patients with aSAH, the incidence of hydrocephalus among the patients after the hematoma removal operation was significantly lower than that of patients without hematoma removal [3.07% (11/358) vs. 19.07% (78/409),χ2 = 47.635,P = 0.000]. The incidence of hydrocephalus among the patients after ventricular drainage was significantly lower than that of patients without the drainage [2.77% (19/685) vs. 85.37% (70/82),χ2 = 487.032,P = 0.000]. In 403 cases of vascular embolization group, the incidence of hydrocephalus in the patients after the hematoma removal operation was lower than that of patients without it [8.06% (5/62) vs. 14.96% (51/341),χ2 = 2.082,P = 0.168]. The incidence of hydrocephalus in the patients after the ventricular drainage was lower than that of patients without drainage [2.59% (9/347) vs. 83.93% (47/56),χ2 = 266.599,P = 0.000]. In 364 cases of craniotomy occlusion operation group, the incidence of hydrocephalus in the patients after hematoma removal operation was significantly lower than that of patients did not receive [2.03% (6/296) vs. 39.71% (27/68),χ2 = 95.226,P = 0.000]. The incidence of hydrocephalus among the patients after the ventricular drainage was significantly lower than that of patients without drainage [2.96% (10/338) vs. 88.46% (23/26),χ2 = 203.852,P = 0.000]. The difference in incidence of hydrocephalus between the patients who had hematoma removal surgery between vascular embolization group and craniotomy occlusion operation group was statistically significant [8.06% (5/62) vs. 2.03% (6/296),χ2 = 4.411,P = 0.027], while no statistically difference was present in ventricular drainage patients [2.59% (9/347) vs. 2.96% (10/338),χ2 = 0.085,P = 0.819]. There were 23 patients (41.07%) with good outcome (GOS score 4-5), while 33 (58.93%) with poor outcome (GOS score 1-3) in 56 patients undergone vascular embolization operation. Good result (GOS score 4-5) was shown in 21 (63.64%) and 12 (36.36%) with poor outcome (GOS score 1-3) among 33 patients with hydrocephalus after craniotomy occlusion operation, and the difference was statistically significant (χ2 = 4.230,P = 0.039).Conclusions Hematoma is one of the main factor contributing to the differences in the incidence of postoperative hydrocephalus of Hunt-Hess gradeⅢ-Ⅳ patients either receiving vascular embolization or craniotomy occlusion operation. Lateral ventricle drainage may not be the factor that contributes to the difference in incidence of hydrocephalus formation between the vascular embolization and craniotomy occlusion operation groups in Hunt-Hess levelⅢ-Ⅳ patients. The short term prognosis in the craniotomy occlusion operation group is superior to that of endovascular intervention embolization group.
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Objective To investigate the effects of synuclein-γ (SNCG) gene silencing on the proliferation and apoptosis of glioma U87-MG cells.Methods Five small hairpin RNA templates targeting SNCG and a negative control were synthesized and cloned into the lentiviral vector system and all the constructs were sequenced.Then the recombinant lentiviral vectors were used to infect U87-MG cells.The lentiviruses which can effectively inhibit protein expression levels of SNCG were selected by RT-PCR for further study.Colony formation and flow cytometry assay were used to investigate the effects of SNCG downregulation by RNA interference on the clony formation,proliferation,and apoptosis of U87-MG cells,respectively.Results The lentiviral vectors carrying 5 shRNAs targeting the SNCG gene were successfully constructed,and SNCG siRNA3 and siRNA5 showed higher interfering efficiency than other vectors.In comparison with the group of negative control,SNCG siRNA3 and siRNA5 were observed to significantly inhibit SNCG expression at the mRNA levels (the relative mRNA levels:siRNA3 (0.17± 0.01)％,siRNA5 (0.13±0.01)％ vs (1.00±0.10)％,P＜0.05).Also,SNCG suppression mediated by RNAi significantly inhibited the clone formation (colony number:siRNA3 (66± 12),siRNA5 (1 ± 1) vs (80± 5),P＜0.05),and the proliferation (ratio of cells in S phase:siRNA3 (41.2±0.7) ％,siRNA5 (39.9±0.5) ％ vs (47.6±2.2) ％,P ＜0.05),but promoted the apoptosis (cell apoptosis:siRNA3 (22.9± 0.4) ％,siRNA5 (28.6± 0.9) ％ vs (1.1 ± 0.1) ％,P＜0.01) of transfected U87-MG cells.Conclusion SNCG suppression at the mRNA level mediated by RNAi can inhibit the proliferation and the clony formation,but induce the apoptosis of glioma U87-MG cells in vitro,suggesting that SNCG suppression mediated by an RNAi strategy may become a novel approach for treating human gliomas.
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Objective To investigate the therapeutic effect of Xuebijing injection on stroke associated pneu monia the changes of plasma C-reactive protein level .Methods 80 cases of post-stroke patients with pneumonia were randomly divided into Xuebijing injection treatment group 40 cases and control group of 40 cases, two groups were given conventional antibiotics and anti-inflammatory treatment ,treated with Xuebijing injection group was dealed with Xuebijing injection 50ml plus 0.9% sodium chloride solution 100ml intravenous drip on the basis of conventional therapy,2/d,for 7d,changes before and after treatment in the two groups were evaluated the body temperature ,periph-eral white blood cell count ,neutrophil percentage ,C-reactive protein index .Results The treatment group after treat-ment for 7d body temperature,blood routine,neutrophils and C-reactive protein were compared before treatment were significantly improved(t=9.99,24.09,12.44,43.98;all P<0.05),all of the indexes in the control group compared before treatment were significantly improved,the differences were significant(t=15.95,20.12,4.14,16.53;all P<0.05),after treatment the observation index except temperatrue decreased significantly ,with statistically significant differences compared with control group (t=4.83,6.15,7.93,all P<0.05).Conclusion Xuebijing injection syner-gistic effect of stroke-associated pneumonia antibiotic treatment significantly , more effective than antibiotic therapy alone,has the very good application and promotion of clinical value .
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Objective To estimate whether the ligation of bilateral internal carotid artery in combination with one vertebral artery can lead to chronic cerebral hypoperfusion. Methods The sham operation, 2?VO and 3?VO rat models were subjected to the matching operation. Four weeks after operation,the cortical blood flow was determined. The learning and memory abilities were measured with Morris water maze test eight weeks later,then the rats were sacrificed to observe the morphological change of hippocampal CA1 region. Results Compared with the sham operation group((47±8.797)ml·min-1·100 g-1),the cerebral blood flow of 2?VO((24.30±8.999)ml ·min-1·100 g-1) and 3?VO((9.870±2.208)ml·min-1·100 g-1) were significantly decreased (P<0.01). Compared with the sham operation group((8.33±4.88)s),escape latencies of Morris water maze of 2?VO group ((14.78±7.84)s) and 3?VO group((14.86±7.96)s) in the fifth days also presented significantly increased (P<0.01),but rare difference between the two groups. Compared with the sham operation group[ (37.20±9.21) s, (10.01.±2.91)times],the target quadrant swimming time and crossing times of 2?VO group((20.13±5.80)s, (6.60±3.19)times) and 3?VO group((20.05±5.76)s,(6.55±2.59)times) in the fifth days also presented signifi?cantly decreased (P<0.01). There were distinct pathomorphology changes in hippocampal CA1 region of the two groups. Conclusion The ligation of bilateral internal carotid artery in combination with one vertebral artery can lead to chronic cerebral hypoperfusion,and can make the similar ethology representation with the 2?VO models.
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Objective To establish the electric controlled cortical impact (eCCI)-induced traumatic brain injury (TBI) model in rats with different severity in degree,which may serve as a suitable platform to provide experimental evidence for the pathophysiological following TBI.Methods A total of 40 male Wistar rats were randomly divided into 3 experimental groups and sham group.TBI rats (n=10/group) were positioned beneath the controlled cortical impactor device (eCCI) and subjected to impact injury at 2 mm depth of penetration,for a sustained depression of 200 ms,at 4 m/s,5 m/s,6 m/s velocity for mild,moderate,and severe TBI,respectively.Sham-operated rats (n=10) underwent identical surgical procedures,including craniotomy,without receiving the cortical impact.Neurological function and regional cerebral flow (24 h after CCI),contusion volume,histopathological,and ultrastructural changes (48 h after CCI) were measured,respectively.Results The severity of the pathological changes in rats was increased as the injury aggravated.The eCCI device impacted the brain at 4 m/s,5 m/s,6 m/s velocity for mild,moderate,and severe TBI,respectively.TBI groups showed impaired neurological function,and decreased rCBF lower than that of sham-operated group (all P＜0.01).Furthermore,neuronal pathological abnormalities in TBI groups,including neuron shrinking,perineuronal vacuole,and structural abnormalities of mitochondria.Increased severity of injury was apparent following the increased level of the impacted velocity,and significant differences were observed between TBI groups (P＜0.05).Conclusion The TBI animal model with mild,moderate,and severe brain injury can be established successfully by 4 m/s,5 m/s,and 6 m/s of impact velocity respectively with the eCCI-6.3 device.The novel eCCI-induced TBI model in rats possibly serves as a novel useful approach in the development of TBI models.
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ObjectiveTo investigate the roles of 11 β-hydroxysteroid dehydrogenase type 1 ( 11 β-HSD1 )on hippocampus of rat with chronic unpredictable mild stress (CUMS).MethodsTwenty-four male SpragueDawley rats were randomly divided into control group and depressive model group. Chronic unpredictable mild stress (CUMS) was used to make up depressive animal model.Behavioral changes were recorded by body weight measuring,sucrose consumption test (SCT) and open field test (OFT),respectively.The mRNA transcription of 11β-HSD1 in hippocampus tissues of the rats were detected by real-time RT-PCR,and the protein expression of 11β-HSD1 were detected by western blot and immunofluorescence.ResultsBcforc starting CUMS protocol,the rats exhibited equivalent weight and sucrose consumption.Twenty-eight days after CUMS protocol,behavior parameters such as body weight,sucrose consumption,nunber of crossing,and number of rearing were significantly decreased in rats exposed to CUMS group compared with control group (P ＜ 0.05,P ＜ 0.01 ).Correspondingly,realtime RT-PCR assays showed the mRNA expression of 11 β-HSD1 in the hippocampus of CUMS group,which was (31 ±9) ％ lower than that of control group.Meanwhile,the protein expression of it in CUMS group was lower than that of control group (P ＜ 0.05 ).Inmunofluorescence revealed that the number of positive 11 3-HSD1 cells was high (223 ± 13) in the control group,while the number was decreased prominently (92 ± 11 ) in the CUMS group (P ＜ 0.01 ).ConclusionDepressive behavior of rats is induced and the expression of 11 β-HSD1 in the hippocampus is decreased prominently by CUMS,the mechanism of which is at least related to the low expression of 11β-HSD1 and disturbance of glucocorticoid metabolism caused by CUMS.
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Objective To study the effect of brain-derived neurotrophic factor (BDNF) on the environmental nutrition and neural differentiation of the transplanted stem cells under hypothermia.Methods The BDNF gene mediated by liposome was transfected into 293T cell line, and ELISA assay was applied to find the peak time of BDNF expression. When BDNF was highly expressed, the supernatant was collected for establishment of SD rat models of brain injury. The rats were divided into Group A (stem cell transplantation group) and Group B (stem cell transplantation and BDNF group). Rats in both groups were under hypothermia treatment for five days. Four and eight days later ( three days from rewarming), rat brain tissues were obtained to detect the expressions of proliferating cell nuclear antigen (PCNA), nestin, neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) by immunohistochemical method and to detect the apoptosis by in situ hybridization. Finally, the nerve function scores were obtained for evaluation of the nerve function. Results The ELISA showed that the high level of BDNF expression was at 48 to 60 hours after gene transfection. PCNA and nestin were highly expressed, while NES and GFAP showed nil or low level of expression in both groups at the fourth day after hypothermia, with little apoptotic cells especially in the Group B (P ＜0.05). The expressions of PCNA and nestin were decreased, but the expressions of NSE and GFAP were increased at the third day after rewarming. The positive rate of NSE expression in the Group B was much higher and the apoptotic cells were much less compared with the Group A ( P ＜ 0. 05 ). A better nerve score was obtained in the Group B. Conclusion BDNF can enhance the survival rate of the transplanted stem cells and induce their differentiation into neurons under hypothermia.
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Liver cancer is one of the most common neoplastic diseases with high mortality in China. Currently, antimicrotubule drugs such as paclitaxel (PTX) and vincristine (VCR), are used as the common agents in the clinical chemotherapy for liver cancer. However, the responses of patients to these drugs vary markedly. Successful identification of intracellular factors influencing liver cancer's sensitivity to antimicrotubule drugs would be of great clinical importance. In this study, by engineering human hepatoma cell HepG2 to overexpress synuclein-gamma (SNCG), we investigated if SNCG is a molecular factor associated with the sensitivity to antimicrotubule drug treatment. Real-time RT-PCR and Western blotting assays showed SNCG was successfully overexpressed in HepG2/ SNCG cells compared with HepG2/Neo cells. The overexpressed SNCG altered the proliferation activity in HepG2 cells, which was 66% higher than that of HepG2/Neo cells through MTT method. The overexpressed SNCG also reduced sensitivity of HepG2 cells to antimicrotubule drugs: after PTX or VCR treatment, the proportion of HepG2/SNCG cells in G2/M arrest was significantly lower than that in HepG2/Neo cells. Correspondingly, HepG2/SNCG cells showed significantly lower mitotic index than HepG2/Neo cells. Meanwhile, HepG2/SNCG cells showed higher resistance to PTX and VCR than HepG2/Neo cells, with resistance index 21 and 15 respectively. Our studies suggested that the overexpression of SNCG could confer resistance to antimicrotubule drugs in hepatoma cells; and it indicated that SNCG may be as a potential response marker for antimicrotubule drugs in liver cancer chemotherapy.