Objective: To study how to activate transforming growth factor ? 1 (TGF ? 1) by coculturing bovine cerebral microvascular endothelial cells (BCMEC) and bovine cerebral microvascular smooth muscle cells (BCMSMC) in vitro , and to observe the effect of high concentration of glucose on active TGF ? 1 (aTGF ? 1) production by the cocultured cells. Methods: BCMEC and BCMSMC were cocultured, and compared with BCMEC or BCMSMC homotypically cultured. 3H TdR assays were used to measure the bioactivity of aTGF ? 1 in the conditioned media. Dot blots assays were used to evaluate the TGF ? 1 mRNA levels of the cultured cells. Then, the cocultured cells were cultured with 5.5 (normal level), 15 and 25(high level) mmol/L glucose for 24 h. The activity of aTGF ? 1 in the media was measured by the same way. Results: aTGF ? 1 was produced in the media of cocultured cells, but not in the media of homotypically cultured cells. Activity of aTGF ? 1 in the media with high concentration of glucose was significantly higher than that in the media with normal concentration of glucose. The dot blots assays implicated that both BCMEC and BCMSMC had TGF ? 1 mRNA expression. But there was no significant change after coculturing. Conclusion: TGF ? 1 can be activated by contacted coculture of BCMEC and BCMSMC in vitro , but the coculturing does not increase the TGF ? 1 mRNA expression. Cultured with high concentration of glucose can increase the aTGF ? 1 content in the media of the cocultured cells. [

pcDNA3.1/Hygro(-)+pCMV?. Conclusions Transactivation competence of genotype B HBx was higher than that of genotype C HBx, while the antiproliferative and apoptosis effects of genotype B HBx was lower than of genotype C HBx. B and C genotype-specific functional differences of HBx may closely co-related with the pathogenicity of HBV.

To investigate the inhibitory effects of silymarin on diabetic blood vessels chroniccomplication. Methods:Silymarin was given to streptozocin-induced diabetic rats. Rats were killed 9 weeksafter treatment. Plasma LPO, fructosamine and RBC-SOD were measured. LPO, fruct0samine, fluores-cence intensities of AGEs, pentosidine and liperperoxide adducts in aorta were also measured. Results:The early nonenzymatic glycation products-fructosamine were not inhibited, however, LPO and f1uores-cence intensities of AGEs, pentosidine,MDA and HNE adducts in aorta were more significantly reducedthan DM group. Conclusion: The results suggest that silymarin may inhibit nonenzymatic glycation andoxidation in aorta of streptozocin-induced diabetic rats and control the diabetic chronic complication.

Objective To study the correlation among acute cerebral infarction area ,serum high sensitivity C reactive protein(hs‐CRP)andhomocysteine(Hcy)concentrations.Methods 112acutecerebralinfarctionpatientsreceivedinneurologicaldepartment of the hospital were enrolled in the study .They were divided into small infarction size group(n=56) and large infarction size group (n=56) ,according to the cerebral infarction area determined by using MRI and MRA .Common information of patients such as age , sex ,smoking ,drinking ,hypertension ,hyperlipidemia and diabetes history were recorded ,serum hs‐CRP and Hcy concentrations were also determined .The comparison between the 2 groups were performed on those common information and test results .Results There was no statistically significant difference on age ,alcohol consumption ,smoking ,hypertension and high cholesterol between the two groups(P>0 .05) .Serum concentrations of hs‐CRP ,Hcy and blood glucose in small infarction size group were lower than those in large infarction size group(P<0 .05) ,and the infarct areas were positively correlated with hs‐CRP ,Hcy and blood glucose levels(r=0 .625 ,0 .833 ,0 .651 ,P<0 .05) .Conclusion Serum hs‐CRP ,Hcy level are high risk factor for atherosclerosis plaque for‐mation of acute cerebral infarction patients ,the serum levels help predict the infarction area ,and diabetes is an important cause of cerebral infarction .

Objective To investigate the clinical,MRI and pathological features of supratentorial primitive neuroectodermal tumor(PNET).Methods The clinical manifestations of 21 PNET patients were analyzed,the skull imaging examination were taken,including MRI with diffuse weighing imaging(DWI) and measured apparent diffusion coefficient(ADC) of tumor and its edema zone before surgery.After operation,the brain tumor tissues were routine and immunohistochemical staining.The relationship between the histopathologic changes and ADC were analyzed.Results In the group,the age of onset of 11cases(52%) were below 20 years old.Clinical manifestation include headache,dizziness and vomiting(16 cases),visual disorder(5 cases),dysosphresis or epilepsy(3 cases).MRI showed single PNET lesion in all the cases and which located at each brain region,the most of them were located at frontal,temporal,parietal lobes(18 cases),and could growing to cross a brain region.MRI T1WI showed that the lesions were iso-signal and lowiso-signal in 15 cases,interspersed high signal in 6 cases.T2WI showed that the lesions were iso and high mixed signal companing capsule change and necrosis,4 cases with lighter tumor edema,5 cases with vascular air flow sign.The imaging enhanced tumors showed uneven enhancement,and 4cases with meningeal tail sign.The pathological examination showed that PNET cell form was main differentiated to neuron(10 cases) and neuroglia(8 cases).There was no statistical significance between ADC and different cell differentiation.Immunity histochemistry showed that the positive of NSE,Syn and GFAP were more offen.Conclusions In the group,the age of onset is below 20 years old.Manifestations of supratentorial PNET are intracranial pressure incresing,visual disorder and dysosphresis.MRI features are mixed isgnal,vascular air flow sign and meningeal tail sign in the tumor.The tumor edema is lighter.The tumor is differentiation mainly toward nerurons and neuroglias in the pathology.There is no positive relationship between ADC and types of tumor differentiation.

Objective To investigate the diagnostic value of the craniocerebral magnetic resonance angiography(MRA) in adult moyamoya disease.Methods 20 adult patients with Moyamoya disease were examined by DSA,MRI and MRA.Results DSA showed that the ending of internal carotid artery system were stenosis or occlution,and the fog-like blood vessel networks at base of skull were observed in the 20 patients.By MRI,the cerebral ischemical changes were found in 11 cases,intracerebral hamrrahge was found in 9 cases.MRA showed that cerebral artery were stenosis or obstruction in all of the 20 patients,16 cases(80%) with the fog-like blood vessel net in the thalamus-basal ganglia region.The detectable rate of abnormal cerebral basal vascular network was no statistical difference between DSA and MRA.Conclusions MRA can provide more information of cerebral artery straitness or obstruction and fog-like blood vessel net in the adult patients with Moyamoya disease,and without hammerless.MRA has a very importent value of diagnosis in the adult moyamoya disease.

Silymarin 100mg?kg~(-1)/d was given to streptozotocin induced diabetic rats for 9 weeks. The results showed that LPO,fructosamine and the fluorescence intensities of AGEs,pentosidine,MDA adducts,HNE adducts in renal cortex of diabetic rats were significantly higher than in normal control rats. After being treated with Silymarin,LPO and the fluorescence intensities of AGEs,pentosidine,MDA and HNE adducts in renal cortex were significantly reduced than in untreated DM group.The albumin excretion in Silymarin group was significantly decreased than in untreated DM group.Silymarin may inhibit nonenzy- matic glycation and oxidation in kidneys of streptozotocin-induced diabetic rats and control the diabetic chronic complication.

Objective To screen the liver cell proteins interacting with the novel protein encoded by the 2.2 kb singly spliced variant of hepatitis B virus genome. Methods The splicing-specific gene TPss generated by the 2.2 kb singly spliced variant of HBV genome was amplified by PCR and cloned into the bait vector pGBKT7. After exclusion of self-activatian capacities of TPss protein, a two-hybrid library screening was performed using a pre-transformed human liver cDNA library to screen the liver cell proteins interacting with TPss. Mammalian two-hybrid assay was also done to further confirm the interactions between the bait and prey proteins in Huh7 and HepG2 hepatacytes. Results TPss gene with the size of 336 bp was successfully amplified and cloned, and the TPss protein expressed well in AHI09 yeast cells. Four liver cell proteins interacting with TPss, i. e. , cathepsin B, epoxide hydrolase 1, cathepsin D and fibrinogen gamma chain, were screened by yeast two-hybrid assay and further confirmed by mammalian two-hybrid assay. Conclusion TPss could interact with liver cell proteins.

Objective To assess the effects of hepatitis B virus(HBV) large envelope protein (LHB) on the apoptosis of HepG2 cells and explore the possible mechanism by proteomic approaches.Methods LHB gene was cloned into pShuttle-IRES-hrGFP-1,and the recombinant adenovirus either barboring LHB(Ad-LHB) or empty vector(Ad-GFP) were separately generated.Annexin V-FITC apoptosis detection kit,JC-1 mitochondrial membrane potential assay kit and propidium iodide(PI) staining kit were employed combined with flow cytometry to detect the apoptotic cells infected with Ad-LHB or control of Ad-GFP.The cellular proteins were collected after infection of HepG2 cells by Ad-LHBs or Ad-GFP,and a total of 600 μg proteins were submitted to two-dimensional gel electrophoresis(2-DE) and stained with R350.The gel images were captured by ImageScanner Ⅱ Imaging System,the differentially expressed proteins were identified by ImageMaster 2D Platinum analysis software and picked up by Ettan Spot Picker.After enzyme digestion,the protein samples were analyzed by MALDI-TOF-TOF MS.Results HepG2 cells infected with Ad-LHB were much more prone to apoptosis.There were thirty nine differentially expressed proteins were determined by 2-DE between HepG2 cells infected with Ad-LHB and Ad-GFP,and they were identified ultimately and categorized into thirty three kinds of proteins by MALDI-TOF-TOF MS.Among these proteins,nine were found to be closely related to cell apoptosis,in which CAPN2,eIF3K and PPP2CB were higher expressed in Ad-LHB infected HepG2 cells,and SERPINH1,LASP1,PRDX1,DHRS2,LDHA and PS-MA4 were lower expressed in Ad-LHB infected HepG2 cells.Conclusion LHB could induce apoptosis of HepG2 cells,and several apoptosis-related proteins participated in this process.

Objective To obtain protein of the major surface antigen P30 of Toxoplasma gondii by molecularcloning. Methods The gene of P30 (containing the whole P30 gene sequence, without the gene encoding signalpeptide) was obtained by polymerase chain reaction (PCR) using the primer designed according to the DNA sequence ofP30. The recombinant plasmid was constructed using EcoR Ⅰ, Xho Ⅰ and was then transformed into E. coli Top10. Thepositive clones were identified by restriction enzymes and DNA sequence analysis. The fusion protein was induced by IPTGand purified by affinity chromatography using ProBond~(TM) Resin (a kind of nickel-charged sepharose resin) and was identi-fied by SDS-PAGE and Western blotting. Results The products of PCR, cleavage and link reaction were same as ex-pected and the sequence of inserted fragment in the recombinant plasmid was same as reported except one synonymy muta-tion. A 58 kDa fusion protein was induced by IPTG and was purified by chromatography. Conclusion Fusion proteincontaining Toxoplasma gondii P30 was achieved and was provided as experiment material for further research.