Analysis Of Factors Related Hemorrhagic In Intracranial Cavernous Malformation

PrefaceCerebral cavernous malformations were about 9%in Central Nervous System Vascular Malformations Population incidence of 0.45%-0.9%,As a common neurosurgical vascular diseases,Some scholars had carried out further explore in clinical and basic research,and had great progress.but to the recurrent hemorrhagic, studies were less,on hemodynamic,CCMs were a low-voltage,low-flow vascular malformation,and no main artery,It was not known why they bleed and had such a high rate of recurrent hemorrhage.Although most bleeding type belong to sub-clinical micro-hemorrhage and obvious clinical symptoms of bleeding relatively small,about 8%-37%,However,they had a relatively high rate of bleeding in bleeding reported in the literature past history.In particular,the brain stem and located in important functional areas of bleeding can caused neurological deficits And even had life-threatening.To further explore the pathogenesis of recurrent hemorrhagic and the potential for the optimal treatments for the disease,the research had been carried out through the study of Ultrastructure and Immunohistochemistry of the CCMMaterials and Methods1.General InformationBetween 2002-2008,35 patients who received a diagnosis of cerebral cavernous malformation were treated microsurgically and Pathologically confirmed at the department of neurosurgery sheng jing hospital china medical university Including frontal lobe 8 cases,occipital 6 cases,temporal lobe 4 cases,Basal ganglia 2 cases, Pons 7 cases,Cerebellar hemisphere 6 cases,Lateral sphenoid ridge 2 cases,male 19 cases,Female 16 cases,age from 1to78,The average age was 34 years,the Course from 6 hours to 20 years;epilepsy 12 cases hemorrhage 17 cases Asymptomatic 17 cases(routine physical 2 cases films by chance after traumatic brain injury 4 cases) Choose cavernous hemangioma of the liver 35 cases and normal brain tissue 4 cases as control,Specimens from severe traumatic brain injury patients with internal and external decompression.2.Methods(1)H.E and immunohistochemical stainingSpecimens were Conventional paraffin-embedded,sectioned at 10 mmon a cryostat,and processed for HE and immunohistochemical staining.Experiment were performed in strict accordance with the immunohistochemical SABC method steps of conventional staining.Ang-1 and CD68 I-anti-concentration were 1:200.MMP-9 I-anti-concentration was 1:160 specimens were Water-bath heated 25 Minutes.SABC kit and other experimental reagents were purchased in Wuhan bo shi de Company Each experiment are equipped with negative and positive control(2)Transmission electron microscopy studyFresh specimens which were cutted the size of a grain of rice and were fixed in 2.5%glutaraldehyde and were Rinsed with 0.1MPBS and post fixed in 1%Osmium tetroxide the specimens were dehydration in Gradient alcohol fixed by EPON812 and Were sliced 70nm,the specimens were stained with Uranyl acetate and AlCit and were Observated,filmed under JEM-1200EX Transmission electron microscopy3.ObservateControl the corresponding general pathology biopsy,we Observated Endothelial cell layer,under-Endothelial cell and the Matrix between Sponge gap and Vascular histology characteristics of the surrounding in Immunohistochemical staining of biopsy.Number of the total endothelial cells(ECs) and ECswith postitive immunohistochemical staining of eachmarker were microscopically counted,and the percentage of ECs with positive expression of CD 68 and MMP-9 were calculated.The amount of the immunohistochemical expression was graded to four groups asshown, that is,negative expression or positive in endothelial cells(ECs) less than 10%among all ECs as(－),strong expression in ECs from 10 to 33%among all ECs(+),strong expression in ECs from 34 to 66%among all ECs as(++),and strong expression in more than 67%of all ECs as(+++).Statistics deal with the use of SPSS 10.0 Package, Inter-group comparison used rank sum test. Results1,Under Light microscopy,lesions were primarily composed of outstretched vascular wall,the sizes of Lumens varied and the lumens were extremely thin and only had thin monolayer of endothelial cells and adventitial without basal lamina and Elastic layer.Erythrocytes were present in the lesions but no Brain tissue.2,Positive immunohistochemical expression of Ang-1 had Yellow to brown granules and Mainly distributed in vascular endothelial cells,plasma packets.CCM expressed Less Ang-1 than control and the difference had no Statisticalsignificance (P>0.05)Positiveimmunohistochemical expression of CD68 and MMP-9 also had Yellow to brown granules and Mainly distributed in Adventitial cell plasma packets. CCM expressed more CD68 and MMP-9 than control and the difference had Statistical significance,we had the expression of CD68 and MMP-9 in Rank Correlation Analysis,the result was 0.870 and had A high degree of positive correlation.3,With the aid of Electron microscopy,The CMs demonstrated evidence of incompetent Vascular structure,The endothelial cells distributed Single-layer and nuclear ruled mostly,The Heterochromatin in nuclear Gathered at the edge of cells.Gap junctions were Obviously observed between endothelial cells Basement membrane showed unintelligible,and The mitochondrial Increased,a large number of high electron density of sediment was present surrounding Blood vessels.In contrast, The junctions of Cavernous hemangioma of the liver were uniformly tight and The Basement membrane showed intelligible but The nuclear were not ruled.conclusionThe defection and instability in the vascular structure and function of CCM Provide favorable conditions for the recurret hemorrhage,CD68 and MMP-9 play an important role in promoted recurrent hemorrhage.