Study on the relationship of cytochrome C expression and cerebral edema in perihematomal brain tissue in patients with hypertensive cerebral hemorrhage

Objective To investigate the expression of cytochrome C in perihematomal brain tissue and its relationship with the histopathological change and formation of cerebral edema in patients with hypertensive cerebral hemorrhage.Methods Thirty four patients(23 male,11 female) of hypertensive cerebral hemorrhage in hospital from Sep.2001 to Sep.2002 were selected with a mean age 55.6± 10.2 years(from 35 to 75 years).The mean volume of hemorrhagic blood was 50.4±11.6 ml(from 25 to 85 ml).The perihematomal brain tissue was obtained from the minimally invasive surgery.Histopathological change and expressions of cytochrome C in perihematomal brain tissue was detected by histopathological and immunohistochemical techniques.The volume of perihematomal cerebral edema was determined by computed tomographic scan before operation.The results of staining and the volume of perihematomal cerebral edema were analyzed with double blind fashion.Results Perihematomal cerebral edema were found 12-72h after cerebral hemorrhage.Myelin sheath degeneration,condensation of nucleus and typical apopototic body were observed in perihematomal brain tissue.Expression of cytochrome C in perihematomal brain tissue was observed at 4 h and reached peak around 48-72 h after cerebral hemorrhage.Cytochrome C expressed higher positively in 16 patients and lower positively in 13 patients.Cytochrome C expression was not detected only in 5 patints.There were significant differences in volume of perihematomal cerebral edema with different expression of cytochrome C in perihematomal brain tissue(P<0.01).Conclusions Cytochrome C expression was upregulated in perihematomal brain tissue in patients with hypertensive cerebral hemorrhage.Cytochrome C might involve in the histopathological change and the formation of perihematomal cerebral edema     

Relationship between serum S-100 protein level and ischemic damage degree in patients with acute cerebral infarction

Objective： To investigate the time course of serum S-100 concentrations of patients with acute cerebral infarction,and their relation with the clinical data and the prognosis. Methods： Serum S-100 levels were serially determined in 36 patients with acute cerebral infarction within 12 h, at 24 h and day 2, 3, 4, 5, 7 and 10 after acute cerebral infarction and in 20 age- and sex-matched control subjects. An S-100 content assay was performed using a two-site radioimmunoassay technique. The clinical status was assessed using NIH Stroke Scale. The functional deficit at 4 weeks after acute cerebral infarction was scored using the modified Rankin scale. A cranial computed tomography was performed initially. Results： Elevated concentrations of S-100 （〉0.2μg/L） were observed in 29 of 36 patients with acute cerebral infarction,but none of the control subjects. The S-100 peak levels were at day 2 and 3 after acute cerebral infarction and were significantly high in those patients with severe neurological deficit at admission, with extensive infarction or with space-occupying effect of ischemic edema as compared with the rest of the populations. Conclusion： Serum S-100 level assay can be used as a peripheral marker of ischemic brain damage, and may be helpful for evaluation of therapeutic effects in acute ischemic stroke.     

Voxel-based statistical analysis of cerebral glucose metabolism in patients with permanent vegetative state after acquired brain injury

Background Permanent vegetative state is defined as the impaired level of consciousness longer than 12 months after traumatic causes and 3 months after non-traumatic causes of brain injury. Although many studies assessed the cerebral metabolism in patients with acute and persistent vegetative state after brain injury, few studies investigated the cerebral metabolism in patients with permanent vegetative state. In this study, we performed the voxel-based analysis of cerebral glucose metabolism and investigated the relationship between regional cerebral glucose metabolism and the severity of impaired consciousness in patients with permanent vegetative state after acquired brain injury.Methods We compared the regional cerebral glucose metabolism as demonstrated by F-18 fluorodeoxyglucose positron emission tomography from 12 patients with permanent vegetative state after acquired brain injury with those from 12 control subjects. Additionally, covariance analysis was performed to identify regions where decreased changes in regional cerebral glucose metabolism significantly correlated with a decrease of level of consciousness measured by JFK-coma recovery scare. Statistical analysis was performed using statistical parametric mapping.Results Compared with controls, patients with permanent vegetative state demonstrated decreased cerebral glucose metabolism in the left precuneus, both posterior cingulate cortices, the left superior parietal lobule （Pcorrected 〈0.001）, and increased cerebral glucose metabolism in the both cerebellum and the right supramarginal cortices （Pcorrected 〈0.001）. In the covariance analysis, a decrease in the level of consciousness was significantly correlated with decreased cerebral glucose metabolism in the both posterior cingulate cortices （Puncorrected 〈0.005）.Conclusion Our findings suggest that the posteromedial parietal cortex, which are part of neural network for consciousness, may be relevant structure for pathophysiological mechanism in patients with permanent vegetative state after acquired brain injury.     

Changes of c-fos,malondialdehyde and lactate in brain tissue after global cerebral ischemia under different brain temperatures

Summary： Under global cerebral ischemia, the effect of different brain temperature on cerebral ischemic injury was studied. Male Sprague-Dawley rats were divided into normothermic （37-38℃） ischemia, mild hypothermic （31 32℃） ischemia, hyperthermic （41-42℃） ischemia and sham-operated groups. Global cerebral ischemia was established using the Pulsinelli four-vessel occlusion model and brain temperature was maintained at defined level for 60 min after 20omin ischemia. The expression of c-fos protein and the levels of malondialdehyde （MDA） and lactate in brain regions were detected by immunochemistry and spectrophotometrical methods, respectively. C-fos positive neurons were found in the hippocampus and cerebral cortex after cerebral ischemia reperfusion. Mild hypothermia increased the expression of c-fos protein in both areas, whereas hyperthermia decreased the expression of c-los protein in the hippocampus at 24 h reperfusion, and the cerebral cortex at 48 h reperfusion when compared to normothermic conditions. In normothermic, mild hypothermic and hyperthermic ischemia groups, the levels of MDA and lactate in brain tissue were increased at 24, 48 and 72 h reperfusion fol- lowing 20-min ischemia as compared with the sham-operated group （P〈0.01）. The levels of MDA and lactate in mild hypothermic group were significantly lower than those in normothermic group （P〈0.01）. It is suggested that brain temperature influences the translation of the immunoreactive protein product of c-fos after global cerebral ischemia, and MDA and lactate are also affected by hypothermia and hyperthermia.     

AQP4 expression and its correlation with the Lac and NAA using proton magnetic resonance spectroscopy after rat cerebral ischemia

Objective： To determine whether AQP4 expression is associated with lactate （Lac） and Nacetyl aspartate （NAA） and with apparent diffusion coefficient （ADC） abnormality after rat cerebral ischemia. Methods： The time courses of ADC and lactate and NAA assessed by proton magnetic resonance spectroscopy （^1HMRS） were investigated at the time point of 6 h, and 1, 3, 7 d after rat cerebral ischernia induced by middle cerebral artery occlusion. Expression of AQP4 mRNA and protein were measured using RT-PCR and Western blot analysis respectively. Results： Significant reductions of NAA concentration and increases of lactate concentration were found after rat cerebral ischemia. The expressions of AQP4 mRNA t and protein were increased at 6 h, and reached the peak at 1-3 d, then began to decrease at 7 d after rat cerebral ischemia. The expression of AQP4 was significantly correlated with NAA （rRT = -0. 856, rw = -0. 927, P〈0.01）, and with lactate （rw=0. 473, rRT=0. 413, P〈0.05）, and with ADC values during the period of 1-7d after rat cerebralischemia （rw=0.984, rRT=0.925, P〈0.05）. In addition, correlations between Lac and the ADC values（r=-0. 677, P〈0.05）and between NAA and ADC values during the period of 1-7 d after rat cerebralischemia （r=0.909, P（0.05） were also observed. Conclusion： The data suggest that AQP4 is involved in the transport of water when brain edema is formed and cell membrane integrity is lost.     

EFFECT OF DL-3-N-BUTYLPHTHALIDE ON BRAIN EDEMA IN RATS SUBJECTED TO FOCAL CEREBRAL ISCHEMIA

The present study evaluated the effect of dl-3-n-butylphthalide(NBP) ,a novel brain protective agent, on brain edema in rats following focal ischemia. Edema was induced by occluding the right middle cerebral artery (MCAO).producing permanent focal ischemia in the right cerebral hemisphere,which developed ip-silateral brain edema reproducibly. Edema was assessed 24 h after MCA occlusion by determining the brain water content from wet and dry weight measurements,and the sodium,potassium concentrations with ion-selective electrodes. In this model,NBP at the dose of 80,160 and 240 mg/kg po 15 min after MCAO prevented from brain edema in a dose-dependent manner. A significant reduction of sodium content and an increase in potassium level were observed in all drug-treated groups. It showed that NBP strongly attenuated brain water entry,sodium accumulation and potassium loss. Nimodipine treatment(5mg/kg sc) also reduced brain edema (P<0. 05). The results suggest that a strong anti-edema activity of NBP may pla     

Effect of Rosiglitazone Maleate on Inflammation Following Cerebral Ischemia/Reperfusion in Rats

In order to evaluate the neuroprotective effect of Rosiglitazone Maleate (RSG) against brain ischemic injury, the effects of Rosiglitazone Maleate on the inflammation following cerebral ischemia/reperfusion were investigated. Focal cerebral ischemia was induced by the intraluminal thread for cerebral middle artery (MCA) occlusion. Rosiglitazone Maleate at concentrations of 0.5, 2 and 5 mg/kg was infused by intragastric gavage twice immediately and 2 h after MCA occlusion, respectively. The effects of Rosiglitazone Maleate on brain swelling, myeloperoxidase and inter- leukin-6 mRNA level in brain tissue after MCA occlusion and reperfusion were evaluated. The results showed that as compared with the model control group, RSG (0.5 mg/kg) had no significant influence on brain swelling (P>0.05), but 2 mg/kg and 5 mg/kg RSG could significantly alleviate brain swell- ing (P<0.05). All different doses of RSG could obviously reduce MPO activity in brain tissue after MCA occlusion and reperfusion in a dose-dependent manner. RSG (0.5 and 2 mg/kg) could decrease the expression levels of IL-6 mRNA in brain tissue after MCA occlusion and reperfusion to varying degrees (P<0.05) with the difference being significant between them. It was concluded that RSG could effectively ameliorate brain ischemic injury after 24 h MCA occlusion and inhibit the inflam- matory response after ischemia-reperfusion in this model.     

Change profiles in matrix metalloproteinase-2 and -9 in induced endometriosis in mice

To examine the changes in matrix metalloproteinase-2 （MMP-2） and -9 （MMP-9） in the development and progression of endometriosis, real time quantitative polymerase chain reaction, enzyme-linked immunoabsorbent assay and gelatin zymography were employed to determine the mRNA and protein levels and activities of MMP-2 and MMP-9 from the first day to the 21^st day after the induction in mice with induced endometriosis （experimental group） and sham-operated animals （controls）. The results showed that the mRNA and protein levels and activities of the MMP-2 and MMP-9 were significantly increased on the first day after the induction and the level of MMP-2 stayed at a level higher than that in the control group. MMP-9 had two or three peaks during the 21 days, taking place at day 1, 4 and 15. It is concluded that the changes in the MMP-2 and MMP-9 might be involved in pathogenesis of endometriosis.     

The relationship between the expression of tumor matrix-metalloproteinase and the characteristics of magnetic resonance imaging of human gliomas

Objective： To investigate the relationship between the expression level of matrix-metalloproteinases （MMPs） with the pathological grades and MRI characteristics of human gliomas. Methods： Prior pre- and post-contrast enhancement MRI was performed on 31 patients with gliomas, which were confirmed by post-operational pathology. The expression of MMP-2 and MMP-9 were determined by immunohistochemical staining in both a low grading group （grades I and II, n = 20） and high grading group （grades III and IV, n = 11）. Results： Compared to the low grading group, the expression levels of MMP-2, MMP-9, as well as the tumor edema index （EI）, enhanced percentage （EP） and maximum diameters were significantly greater in the high grading group. MMP-2 and MMP-9 expression were correlated with the tumor EI, EP and the maximum diameters. There were no differences in MMP-2 and MMP-9 expression between the unclear border definition group and the clear border definition group, whereas the MMPs expression levels were greater in the heterogeneous signal group than in the homogeneous signal group. Conclusion： The expression level of MMPs is correlated with the invasion ability of human gliomas. The MRI parameters, such as tumor El, EP, maximum diameter, and signal heterogeneity technically reflect the expression level of MMPs, and can be used to estimate the tumor＇s malignant behavior, thus providing the guidance for clinical therapies.     

Attenuation of Brain Inflammatory Response after Focal Cerebral Ischemia/Reperfusion with Xuesaitong Injection(血塞通注射液) in Rats

Objective： To investigate the neuro-protective effect of Xuesaitong Injection （血塞通注射液, XST） on brain inflammatory response after transient focal cerebral ischemia/reperfusion in rats. Methods： Focal cerebral ischemia/reperfusion models of male rats were induced by transient occlusion for 2 h of middle cerebral artery （MCA） which was followed by 24 h reperfusion. XST was administered through intraperitoneal injection of 25 mg/kg or 50 mg/kg at 4 h after the onset of ischemia. After reperfusion for 24 h, the neurological function score was evaluated, the brain edema was detected with dry-wet weight method, the myeloperoxidase （MPO） activity and the expression of intercellular adhesion molecule-1 （IOAM-l） of ischemic cerebral cortex and caudate putamen was determined by spectrophotometry and immunohistochemistry respectively. Results： XST not only lowered neurological function score at the dose of 50 mg/kg, but reduced brain edema and inhibited MPO activity and IOAM-1 expression as compared with the ischemia/reperfusion model group （P〈0.01）. Conclusion： XST has a definite effect on inhibiting the expression of IOAM-1 and neutrophil infiltration in rats with cerebral ischemia/reperfusion when treatment started at 4 h after ischemia onset, and also attenuates inflammation in the infarcted cerebral area.