Microdialysis, a technique for sampling the biochemical substances of extracellular fluid in vivo, has been widely utilized for physiological, pharmacological and pathological research. The principles and methodology of microdialysis and its applications in neurointensive care was reviewed.

Objective: To investigate the expression of pituitary tumor transforming gene (PTTG) and basic fibroblast growth factor (bFGF) in oral squamous cell carcinoma and their relation with each other, as well as the relationship of their expression with clinical-pathological indexes. Methods: The expression of PTTG and bFGF were examined among 55 oral squamous cell carcinoma tissues and 10 normal oral mucosal tissues by the streptavidin-biotinperoxidase (S-P) method. Results:The positive rate of PTTG and bFGF was 78.2% and 67.3% respectively in oral squamous cell carcinoma. The positive rate and grade of PTTG and bFGF were significantly higher than that in the normal mucosa tissues (P

Objective To investigate the expression and cellular localization of cyclin-dependent kinase 5 (Cdk5) in cerebral cortex after subarachnoid hemorrhage (SAH) in rats.Methods Fifty-two male Sprague Dawley (SD) rats were randomly divided into either a sham operation group (n =12) or a SAH group (n =40).The latter was randomly redivided into 6,12,24 h,and day 2 and 3 subgroups (n =8 in each group).A rat SAH model was induced by injecting fresh blood into the prechiasmatic cistern.Western blot and immunohistochemistry were used to detect the expression of Cdk5 in rat brain cortex.Double labeling immunofluorescence staining was used to detect the cellular localization of Cdk5 protein in cerebral cortex.Neuronal nuclear antigen labeled neurons,and glial fibrillary acidic protein labeled astrocytes.Results Western blot showed that the expression of Cdk5 protein was up-regulated at 12 hours after SAH (t =3.709,P =0.001),and it reached the peak on day 1 (t =3.475,P=0.002).Immunohistochemistry showed that the proportion of Cdk5 positive cell was also increased gradually after SAH,and the changes of time course were consistent with the results of Western blot,and it reached the peak on day 1 (t =4.320,P =0.000).Double labeling immunofluorescence showed that Cdk5 was mainly expressed in the neuronal cytoplasm in the sham operation group,and Cdk5 shifted to the neuronal nuclei in the SAH group.Cdk5 was mainly colocalized between astrocytes and neurons.Conclusions SAH up-regulates the expression of Cdk5 protein in cerebral cortex.Cdk5 may be involved in early brain injury after SAH.

Objective To explore the formation mechanism of peritumoral brain edema(PTBE)by vascular endothelial growth factor(VEGF).Methods 40 biopsies were obtained from 37 patients.Inmunohistochemical staining and Western blot were performed to detect the expression of VEGF protein.Reverse-transcriptase polymerase chain reaction(RT-PCR)was used to analyze the presence and quantity of VEGF mRNA.The extent of PTBE was estimated as an edema index(EI)based on preoperative magnetic resonance imaging.Results In VEGF-positive cases,a decreasing gradient of VEGF protein expression was observed with increasing distance from tumors(0.38±0.08,0.20±0.03,0.04±0.02).In meningiomas,the protein level and the mRNA level were congruent and the expression of both protein and mRNA had a significant correlation with EI(protein: r =0.892,RNA: r =0.875,P ＜ 0.05).However,in peritumoral areas,protein level were not consistent with the mRNA level.Protein results showed high correlation with EI(r =0.912,P ＜ 0.05),but mRNA almost was almost undetectable(0.06±0.02).Conclusion VEGF is impartant on PTBE.It is concluded that VEGF macromolecules are secreted by tumor tissue and enter peritumoral normal brain tissue to induce edemagenesis in meningiomas.

Objective Nrf2 is an important neuroprotective factor and the ubiquitin proteasome system ( UPS) , as a highly specific intracellular protein degradation pathway, plays an important role in maintaining gene and protein functions.This paper pres-ents a preliminary study on the relationship between Nrf2 and the ubiquitin proteasome system in the mouse model of traumatic brain in-jury ( TBI) . Methods Forty-two healthy male ICR mice were randomly divided into three groups: control, TBI +sulforaphane ( SFN) and TBI+vehicle, and 12 Nrf2-knockout mice were included in the TBI+Nrf2 -/-group.The animals of the TBI+SFN group were treated with SFN while those of the TBI+vehicle group with the same volume of 10%corn oil at 5 minutes after TBI.At 24 hours after TBI, brain samples were collected from the mice for determining the Nrf2 expression and ubiquitinated protein content by Western blot and the changes in the Nrf2 and ubiquitinated proteins were observed by immunohistochemistry and electron microscopy. Results Compared with the controls, the mice in the TBI+vehicle group showed significantly increased expressions of Nrf2 ( 0.09 ± 0.02 vs 0.66 ±0.09, P<0.05) and ubiquitinated proteins (3.27 ± 0.21 vs 10.58 ±0.75, P<0.05).In comparison with animals in the TBI+vehicle group, those in the TBI+SFN group exhibited a signifi-cant increase in the Nrf2 protein level (0.66 ±0.09 vs 1.22 ±0.14, P<0.05) but a decrease in the ubiquitinated protein level (10.58 ±0.75 vs 6.97 ±0.86, P<0.05), and those in the TBI+Nrf2 -/-group showed a markedly decreased expression of the Nrf2 protein (0.66 ±0.09 vs 0.17 ±0.02, P<0.05) but increased expression of the ubiquitinated protein (10.58 ±0.75 vs 14.35 ± 0.65, P<0.05).Similar results were observed by immunohistochemistry and electron microscopy. Conclusion Nrf2 played a neu-roprotective role in the mouse model of traumatic brain injury by regulating the ubiquitin proteasome system.

Objective Subarachnoid hemorrhage ( SAH) is a devastating disease with a high mortality.This study was to in-vestigate the effect of Nrf2 on secondary brain injury following SAH and its action mechanism in mice. Methods SAH models were established in wild-type ( WT) and Nrf2 knockout ( KO) ICR male mice by injecting fresh blood drawn from the femoral artery into the pre-chiasmatic cistern.The animals were divided into four groups, WT sham, WT SAH, KO sham, and KO SAH.At 24 hours after modeling, the expression levels of malondialdehyde ( MDA) , GSH/GSSG, TNF-αand IL-1β, the volume of brain water, and content of Evans blue were measured, the activity scores obtained, and cerebral vasospasm of the anterior and middle cerebral arteries ( ACA and MCA) detected. Results At 24 hours, the expressions of MDA, TNF-α, and IL-1βwere (3.299 ±0.335), (1.187 ± 0.436), and (59.330 ±21.787) mg/g in the WT sham group, (4.339 ±0.328), (2.432 ±0.434), and (121.584 ±21.675) mg/g in the WT SAH group, (3.488 ±0.634), (1.170 ±0.312), and (58.497 ±15.608) mg/g in the KO sham group, and (5.335 ±0.499), (3.132 ±0.548), and (171.117 ±50.479) mg/g in the KO SAH group, markedly increased in the SAH groups as compared with the sham controls (P<0.05), while the GSH/GSSG levels were significantly higher in the former two groups than in the latter (0.553 ±0.100 and 0.375 ±0.068 vs 0.714 ±0.091, 0.761 ±0.114, P<0.01).The contents of brain water and Evans blue were (0.784 ±0.005) and (7.055 ±1.046) μg/g in the WT sham group, (0.808 ±0.004) and (7.230 ±1.192) μg/g in the WT SAH group, (0.784 ±0.004) and (9.620 ±1.290) μg/g in the KO sham group, and (0.819 ±0.004) and (11.628 ±1.040)μg/g in the KO SAH group, remarkably increased in the SAH groups in comparison with the sham groups (P<0.05).The apoptosis rate 8.916 and 82.100 ±6.870 vs 70.833 ±8.750 and 51.767 ±13.006), ACA radius/wall thickness value (13.885 ±3.360 and 14.212 ±3.2545 vs 8.024 ±2.780 and 6.861 ±2.702), MCA radius/wall thickness value (18.648 ±2.893 and 19.435 ±2.775 vs 6.337 ±3.993 and 5.107 ±3.805), and activity score (2.733 ±0.450 and 2.767 ±0.430 vs 1.967 ±0.928 and 1.433 ±0.679) (all P<0.01). Conclusion Nrf2 knockout increases oxidative stress and inflammatory reaction following SAH and consequently aggravates secondary brain injury.Nrf2 has a protective effect against SAH-induced brain injury.

Objective Subarachnoid hemorrhage ( SAH) is a devastating disease with high fatality and morbidity and micro-glia/macrophage ( M/M) plays a vital role in SAH brain injury with complicated pathophysiological mechanism .This study was to ob-serve the effect of Nrf2 deficiency on M/M activation and M1 polarization after subarachnoid hemorrhage in mice . Meth ods We col-lected 70 wild-type ( WT) ICR mice and 35 Nrf2-knockout ( KO) mice to establish the SAH model by injecting fresh autologous blood into pre-chiasmatic cistern.WT mice were arranged into four groups: sham operation group, post operative day 1 (POD1) group, POD3 group and POD5 group.Then WT mice and Nrf2 Nrf2-knockout mice were divided into sham operation WT group , sham opera-tion KO group, SAH WT group and SAH KO group.Western blotting (WB) and immunohistochemistry (IHC) were applied to observe the activation and proliferation of M/M after SAH on WT mice .Difference in activation and M 1 polarization were observed by detecting Iba1 expression in WB and CD 16/32 +Iba1 +cells in immunofluorescence between WT and KO mice . Results Gray scale values of Iba1 expression by WB in WT mice are 0.491 ±0.039, 0.657 ± 0.069, 0.930 ±0.046 and 0.926 ±0.046;average optical intensity values of Iba1 expression by IHC in WT mice are 0.412 ±0.122, 0.625 ±0.135, 0.963 ±0.213 and 0.978 ±0.224.The data indica-ted that Iba1 expression increased in SAH KO group in comparison to SAH WT group on 1, 3, 5 day after SAH (P<0.05).Moreover, Nrf2 deficiency promoted the activation and polarization of M /M by increased Iba1 protein expression and CD16/32 +Iba1 +cells after SAH ( P<0.05). Conclusion SAH induces M/M activation and proliferation in mice, and Nrf2 deficiency promotes the activa-tion, proliferation and M1 polarization after SAH .

Objective The prognosis of traumatic brain injury is closely associated with the apoptosis of neural cells .This study investigated the anti-apoptosis effect of alpha lipoic acid (α-LA) and its possible mechanism in the mouse model of traumatic brain injury. Methods Seventy-two healthy male ICR mice were randomly divided into four groups of 18 each:sham operation +vehicle, sham operation +α-LA, trauma +vehicle, and trauma +α-LA.The model of traumatic brain injury was made by weight-dropping.The animals in the α-LA groups were treated with intragastric α-LA at 30 minutes after surgery, while those in the vehicle groups with oral dimethyl sulfoxide in corn oil .At 48 hours after treatment , brain samples were collected from the mice for determining brain edema , measuring the expressions of cytochrome c , Bcl-2-associated X protein ( Bax ) , and caspase-3 in the mitochondria and cytosol of the brain tissue by Western blot and immunohistochemistry respectively , and detecting the survival of the neurons and apoptosis of neural cells in the cortical area by Nissl staining and TUNEL , re-spectively. Results The brain water volume , caspase-3 expres-sion, and neural cell apoptosis were markedly higher while the neuron survival remarkably lower in the trauma +vehicle group than in the sham operation +vehicle and sham operation +α-LA groups ( P<0.01).Compared with the mice in the trauma +vehicle group, those in the trauma +α-LA group showed significantly reduced proportion of water in the brain tissue ([79.89 ±0.55] vs [81.71 ± 0.66]%, P<0.05), expression of caspase-3 ([58.40 ±7.31] vs [47.42 ±7.74]%, P<0.05), and apoptosis of neural cells ([59.63 ±8.61] vs [44.86 ±7.32]%, P <0.05), but increased survival rate of neurons ([44.45 ±10.56] vs [57.46 ± 11.01]%, P<0.05).The expression of cytochrome c in the mitochondria was remarkably decreased and that of Bax markedly in -creased in the trauma +vehicle than in the sham operation +vehicle and sham operation +α-LA groups (P<0.01). Conclusion Alpha lipoic acid has a protective effect against traumatic brain injury in mice , probably by inhibiting the apoptosis of neural cells through the mitochondrial pathway .

Objective:The aim of the current study was to explore the alterations of TLR4 after traumatic brain injury. Methods: 20 Male ICR rats were randomly divided into four groups(5 rats each group)including control group without brain injury and traumatic brain injury groups(4,24 hours and 7 days).All rats were decapitated at corresponding time point and mid-jejunum samples were taken.The intestinal mucosa structure was detected by histopathological examination.Immunohistochemistry and realtime RNA were used for detection of TLR4 expression in ileum samples.Results: After traumatic brain injury,the histopatholocal alterations of gut mucosa occurred at 4 hours and progressed to a serious state.Compared to control group,TLR4 was significantly up-regulated on the endothelia of microvessels in villous interstitium and lamina propria by 4 h following TBI and maximally expressed at 24 h post-injury.The endothelial TLR4 immunoreactivity in ileum mucosa still remained strong on 7 d post-injury.There was a very low mRNA expression of TLR4 in the gut of control rats and significantly increased at 4 h following TBI(P

Objective To evaluate the potential involvement of melatonin in the activation of nuclear factor-E2-related factor 2-antioxidant response element (Nrf2-ARE) signaling pathway and anti-oxidative stress in an experimental model of traumatic brain injury (TBI).Methods One hundred and sixteen ICR male mice,aged 6-8 weeks,were randomly divided into sham-operated group,model group,vehicle group and melatonin treatment group (n=29);mice in the later three groups were established TBI models by Flierl improved free-fall method;0,1,2,3 and 4 h after that,mice in the melatonin treatment group were performed intraperitoneal injection of melatonin (10 mg/kg,3 mg/mL diluted in normal saline),and those in the vehicle group were given the same volume of normal saline.Twenty-four h after TBI,the neuronal degeneration was investigated by Fluoro-Jade C (FJC) staining and brain water content was measured by wet-to-dry weight ratio method;colorimetric method was employed to detect the malondialdehyde (MDA),superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels;Western blotting was engaged to analyze the protein content of nuclear Nrf2,cytoplasmic Nrf2,NADPH quinine oxidoreductase-1 (NQO1) and heme oxidase-1 (HO-1);real-time quantitative-PCR was employed to evaluate the mRNA content of HO-1 and NQO-1;electrophoresis mobility shift assay (EMSA) was performed to measure the Nrf2 DNA binding activity.Results As compared with the sham-operated group,the other three groups had significantly increased number of cortical degenerative neurons,water content,MDA content,nuclear Nrf2 expression,and nuclear HO-1 and NQO-1 protein and mRNA expressions,and had significantly lower levels of cytoplasmic Nrf2 and GPx.As compared with the model group and vehicle group,the melatonin treatment group had significantly decreased number of cortical degenerative neurons,water content,MDA content and nuclear Nrf2 expression,and had significantly increased cytoplasmic Nrf2 level,GPx content and nuclear HO-1 and NQO-1 protein and mRNA expressions (P＜0.05).The SOD content in the sham-operated group and melatonin treatment group was significantly higher than that in the model group and vehicle group (P＜0.05).EMSA showed that the Nrf2 DNA binding activity increased accordingly in the order of sham-operated group,model group,vehicle group and melatonin treatment group.Conclusion The melatonin can improve the anti-oxidative stress neuroprotection following TBI,which may be associated with the activation of Nrf2-ARE signaling pathway.