AIM: To investigate the changes of somatosensory evoked potential(SEP), nitric oxide (NO) levels both in serum and in brain tissue after subarachnoid hemorrhage(SAH) and the influence of Ginkgo biloba extract(GBE) on them. METHODS: Wistar rats were divided into sham-operated group, pure SAH group and GBE-treated group. Dynamic changes of regional cerebral blood flow( rCBF),SEP, and NO levels both in serum and in brain tissue were detected within 24 hours after operation. RESULTS: In pure SAH group, rCBF decreased immediately after operation, with no tendency to recover within 24 hours. Latency of SEP delayed progressively from 1 hour to 24 hours after SAH.NO levels in serum and in brain tissue decreased and increased respectively from 1 hour to 24 hours after SAH. GBE effectively antagonized the changes of above parameters. CONCLUSION: SEP is useful in the judgement of cerebral ischemic damage after SAH. Decrease of serum NO and increase of brain NO are important factors leading to cerebral vasospasm and neural damage respectively after SAH. GBE relieves cerebral ischemic damage by reversing the pathological alterations of NO.

Objective To explore the effect of protein kinase C inhibitor on the level of phosphralated extracellular regulated protein kinases in the spinal trigeminal nucleus of migraine model rats.Methods Healthy adult male SD rats were randomly divided into four groups:normal group (group C),sham operation group (group C),migraine model group(group M),and H-7group(H-7group),with 18 rats in each group.Dural blood flow and the extracellular discharge frequency in the spinal trigeminal nucleus was recorded.ERK1/2 phosphorylation was tested.Results (1) Dural blood flow:compared with group C((3.8± 1.0)％),the dural blood flow in M group ((78.0±4.2) ％)increased obviously(P＜0.01) ; compared with M group((78.0±4.2)％),the dural blood flow in H-7 group((-24.8±4.9) ％) decreased obviously(P＜0.01).(2) The percentage of extracellular discharge frequency change:two hours after treatment,the percentage of extracellula discharge frequency change in group M ((325.9 ±47.3)％)was higher than that in group C((107.3±16.4)％).The percentage of discharge frequency change in group H-7((136.0±26.5)％) was lower than that in group M((325.9±47.3)％).There was no significant difference in the percentage of discharge frequency change between group H-7((136.0±26.5) ％) and group C((107.3 ± 16.4)％).(2) ERK1/2 phosphorylation:the ERK1/2 phosphorylation in group M was higher than that in group C.The ERK1/2 phosphorylation in group H-7 was lower than than that in group C and group M.Conclusion ERK1/2 is a downstream PKC signal path and PKC may have indirect activation of ERK1/2.

AIM: To investigate the effect of Ginkgo biloba extrac (GBE) on cerebral ischemia during early stage of subarachnoid hemorrhage (SAH). METHODS: Noncraniotomy models of SAH in Wistar rats were used and animals were divided into sham-operated group, SAH group and SAH+GBE group. Dynamic change of regional cerebral blood flow (rCBF) was detected. Brain endothelin-1(ET-1) and calcium contents were also determined at different time point during 24 hours after the operation. Pathological change of neurons of hippocampus CA1 region was observed. RESULTS: In SAH group, rCBF decreased immediately and persistently after induction of SAH. Values of brain ET-1 content and calcium content at 1 hour, 6 hours and 24 hours were significantly higher than those in sham-operated group. Neurons of hippocampus CA1 region were damaged severely 3 days after onset of SAH. Above abnormal changes in SAH+GBE group were much slighter than those in SAH group. CONCLUSION: GBE may relieve cerebral ischemic damage after SAH.

AIM: To investigate the changes of somatosensory evoked potential(SEP), nitric oxide (NO) levels both in serum and in brain tissue after subarachnoid hemorrhage(SAH) and the influence of Ginkgo biloba extract(GBE) on them. METHODS: Wistar rats were divided into sham-operated group, pure SAH group and GBE-treated group. Dynamic changes of regional cerebral blood flow( rCBF),SEP, and NO levels both in serum and in brain tissue were detected within 24 hours after operation. RESULTS: In pure SAH group, rCBF decreased immediately after operation, with no tendency to recover within 24 hours. Latency of SEP delayed progressively from 1 hour to 24 hours after SAH.NO levels in serum and in brain tissue decreased and increased respectively from 1 hour to 24 hours after SAH. GBE effectively antagonized the changes of above parameters. CONCLUSION: SEP is useful in the judgement of cerebral ischemic damage after SAH. Decrease of serum NO and increase of brain NO are important factors leading to cerebral vasospasm and neural damage respectively after SAH. GBE relieves cerebral ischemic damage by reversing the pathological alterations of NO.

Rhubarb is a traditional Chinese medicines which possess laxative, lipid-lowering, and weight-loss activities, but the active compounds of lipid-lowering and underlying molecular mechanisms are not yet clear. This study aims to explore the effects of chrysophanol on the mRNA expressions of sterol regulatory element-binding proteins (SREBPs) and lipid metabolism in human liver carcinoma Huh-7 cells, which is one of the active compounds obtained from Rhubarb. A reporter gene assay was used to test the transcription of SREBP. The intracellular triglyceride and total cholesterol contents were measured by using commercially available test kits. The SREBPs target genes expressions were measured by Quantitative Real-Time PCR. Cell viability was evaluated by Cell Counting Kit-8. As the results shown, chrysophanol (40 μmol · L(-1), 16 h) could notably inhibited human SRE promoter activity in a dose-dependent manner and decrease intracellular cholesterol and triglyceride levels. Furthermore, the mRNA expressions of SREBPs target genes were significantly downregulated by chrysophanol treatment. However there are no significant differences on cell viability when compared with the control group. These results suggested that chrysophanol might improve lipid metabolism through suppressing the mRNA expressions of SREBPs target genes to attenuate intracellular lipid accumulation.

Objective To study the promotive effect of neovascularization on rats with cerebral infarction by nasal administration of granulocyte colony-stimulating factor.Methods A blinded,vehicle-controlled study of ING-CSF and IHG-CSF administration was performed by intraluminal middle cerebral artery occlusion (MCAO) model.All Sprague-Dawley rats were randomly divided into sham-operation group,model group,INNS group,IHGCSF group and ING-CSF group.The neurologic behavioral tests were assessed after reperfusion 72 h.Mter 72 h of MCAO,the brains of rats were stainned with TTC and the infarcted volume was calculated by computer image analysis.The expression of vascular endothelial growth factor (VEGF) in the brain was determined by immune-histochemistry.The density of angiogenesis in the brain was counted under fluorescence microscope.Results The score of neurological function of ING-CSF group(3.90± 1.65)was improved significantly compared with the IHG-CSF group (10.55±2.19) at the point of 72 h after cerebral infarction (P＜0.01).The cerebral infarct volume of ING-CSF group((20.01±3.29) ％) was reduced evidently compared with the IHG-CSF group((33.48±4.49) ％) at 72 h (P＜ 0.01);while the cerebral infarct volume of INNS group ((60.20±7.72) ％)was not markedly different compared with the model group((61.49±6.41)％) at 72 h (P＞0.05).The expression of VEGF in the brains of ING-CSF group was significantly higher than other groups at 72 h.Conclusion Intranasal administration G-CSF can improve neurological function and vascular angiogenesis in rats following MCAO.

Objective To observe the effects of CoC12 treatment on the expression of Hypoxia-inducible factor-1(HIF-1α) in mice hippocampus at different time point.Methods Balb/c mice were injected with CoCl2 and the change of HIF-1 α was detected by western blot and immunofluorescence and confocal laser scanning microscope at different time point(0h,1h,2h,3h,4h,5h and 6h) after injection.Results The relative protein level of HIF-1α was 0.135 ±0.01,0.572 ±0.01,0.595 ±0.03,1.09 ±0.03,1.30 +0.04,1.275 ±0.03,0.947 ±0.03respectively at different time point after the injection.The HIF-1α protein level reached its peak value at 4 h and decreased at 5h and 6h.Fluorescence intensity of HIF-1α was 13.33 ± 3.42,30.95 ± 7.86,46.50 ± 9.65,61.50± 10.02,88.30 + 15.69,71.39 ± 11.28,67.41 ± 10.78 respectively at different time point after the injection.The HIF-1α fluorescence intensity also reached its peak value at 4 h and decreased at 5h and 6h.Conclusion Time dependent HIF-1α accumulation was in close correlation with the CoCl2.

Thirty patients with confirmed primary sjogren syndrome and thrombocytopenia during January 2002 and December 2007 were investigated.All the participants received 0.5 mg·kg-1·d-1 Prednisone and 100 ms/d CosA for 1 year.Clinical symptoms and the levels of white blood eell.platelet count,erythrocyte sedimentation rate,liver function,immunoglobin,and rheumatoid factors were recored before and after the treatment.After one-year follow.up,platelet count was elevated(t=11.4179,P< 0.01).There was no significant change in erythrocyte sedimentation rate.The concentrations of immunoglobin and rheumatoid factors were significantly decreased at 1 year(t=5.4222,P<0.01;t= 9.2857.P<0.01).Prednisone plus CosA could effectively treat primary sjOgren syndrome combined with thrombocytopenia and result in fewer side effects.

Objective To investigate the influence of cerebral lymphatic blockade (CLB) on apoptosis of hippocampal neurons after subarachnoid hemorrhage (SAH) in rats. Methods Healthy adult Wistar rats were randomly assigned to normal control group,SAH group and SAH + CLB group. SAH model was induced by double injection of autologous blood into the cistema magna. On day 3 after second injection, hippocampal cell shape structure of each group were determined by hematoxylin-eosin staining (HE) and propidium iodide (PI) staining. Terminal-deoxynucleotidy transferase mediated nick end labeling (TUNEL) fluorescent was used to determine the situ apoptosis. Immunohistochemistry was conducted to study the expression of caspase-3 and Bcl-2 in hippocampal neurons. Results (1) HE staining and PI staining showed the hippocampal neurons of SAH rats were partly shrink,and nuclei showed wavy or folded seam-like,some crescent-shaped; the hippocampal neurons in SAH + CLB group distributed sparsely,nuclear fragmentation,apoptotic bodies could be seen,surrounded by vacuole formation, Compared with the SAH group, the number of apoptotic cells in SAH + CLB group was significantly increased(the number of apoptotic cells: 0.71 ±0.05,25.36 ±4. 02,37. 82 ±5.93, P<0.01). (2) The fluorescence intensity of positive cells by TUNEL stain in SAH group and SAH + CLB group was higher than in normal control group,while the SAH + CLB group was significantly higher than the SAH group (the fluorescence intensity: 0.19 ±0.03,1.70 ±0.37,2.54±0.53, P<0.01). (3) The fluorescence intensity of caspase-3 in SAH group and SAH + CLB group was higher than the normal control group, while the SAH + CLB group was significantly higher than the SAH group (the fluorescence intensity: 0.14 ±0.03,2.45 ±0.49,2.96 ±0.44, P<0.01). (4) The fluorescence intensity of Bcl-2 in SAH group and SAH + CLB group was higher than the normal control group, while the SAH + CLB group was significantly lower than the SAH group(the fluorescence intensity: 0.58 ±0.08, 3.40 ±0.61,2.67 ±0.44, P<0.01). Conclusion Cerebral lymphatic blockade induce the apoptosis of hipp-ocampal neurons in rats after SAH,which mechanism may be related to high expression of caspase-3 and low ex-pression of Bcl-2.

Objective To investigate the pathway of lymphatic drainage of proteins from cerebral parenchyma in subarachnoid hemorrhage rat models. Methods Healthy adult male Wistar rats were divided into Saline group, Evans blue-labeled albumin (EBA) group, and SAH + EBA group. SAH models were produced by double injection of autologous arterial blood into cisterna magna. Using a modified microinjection method, EBA was injected into left candate-putamen of the EBA group and EBA + SAH group rats. In Saline control group, saline was injected. After injection, at 12 hours, 1 day, 2 days, 3 days and 5 days, the animals were sacrificed and the fluorescence signals of EBA were imagined and analyzed along the possible lymphatic drainage pathway, e.g. the brain tissue, the wall of common carotid artery, and cervical lymphatic nodes. Results One day after injection, in EBA group, the fluorescence of EBA initially appeared on the left of the brain, the wall of common carotid artery, left lateral cerebral ventricle, and the perivascular spaces of cerebral vessels. The fluorescence signals gradually expanded to the opposite side.Large amount of fluorescence granules accumulated in the outer layer of common carotid artery. Fluorescence was also found in cervical lymphatic nodes. Two days after injection in this group, the density of fluorescencein the brain became weaker while the density of fluorescence in rhinencephalon became stronger. The fluorescence of EBA was found in lymphatic nodes adjacent to abdominal aorta. In SAH + EBA group,reduced amount and velocity of the drainage of EBA from left caudate-putamen to rhinencephalon, cervical lymphatic nodes, and lymphatic nodes adjacent to abdominal aorta were observed. From 12 hours to 5 days after injection, fluorescence intensity of EBA in deep cervical lymphatic nodes in SAH + EBA group(8.9 ±2. 0, 11.9 ± 2. 5, 17.4 ± 3.7, 26.7 ± 4. 5 and 59.0 ± 8. 1 ) were lower than those in EBA group ( 14. 5 ±3.2, 27.5 ±7.4, 60.3 ±12.3, 138.0±12.0 and 108. 1 ±13.4, F=13. 17, 24.04, 66.81, 302.77 and 59.36, P ＜ 0. 01 ). From 2 to 5 days, fluorescence intensity of EBA in lymphatic nodes adjacent to abdominal aorta was also lower in SAH + EBA group( 11.0 ± 1.5, 12. 5 ±2. 8, 23.6 ±3. 2) than those in EBA group(26. 3 ±5.9, 47.5 ±9.6, 41.0 ±9.3; F =38. 17, 72.52, 19.01, P ＜0.01). Conclusion SAH can result in reduced drainage of macromolecular substances, e.g. protein, from the brain via lymphatic pathway.