The brain—type creatine kinase isoenzyme (CK—BB)in the cerebrospinal fluid(CSF) within 24 hs following trauma was measured in 62 cases with acute head injuries by chromatography and fluorometry, and the comparative analysis was made between CSF-CK-BB activities and the outcome of the patients in 6 months after trauma according to Glasgow Outcome Scale(GOS),The results showed that the determinations of CSF-CKBB activities,in combination with Glasgow coma Scale (GCS) can be used as an early objective diagnostic index for assessing the prognosis ,and there was a reference value for improving accuracy of evaluating recovery of neurological disfunction in the patients with acute head injuries.

Objective Angiotensin Ⅱ (Ang Ⅱ ) contributes to modulating blood pressure by stimulation of Ang Ⅱ AT1 receptors. We devised a rat transient middle cerebral artery occlusion (MCAO) model to assess whether oxidative damage is decreased after pretreatment with Angiotensin Ⅱ AT1 receptor blocker (ARB). Methods After 2 weeks pretreatment with ARB 0. 5 and 1 mg/kg, the male Wister rats were subjected to 2 h middle cerebral artery occlusion (MCAO). At 24 h, the lumen diameter of middle cerebral artery, the plasma level of 8-hydroxy-2'-deoxyguanosine (8-OHdG), and HIF-1 a levels were recorded and compared. Results After pretrcatment with ARB 0.5 and 1 mg/kg, blood pressure did not significantly change compared with that of controls. In the group of candesartan at 1 mg/(kg· day), the lumen diameter was significantly increased compared to that in control group [(86.0±5.0) μm vs. (69.0± 2.1) μm; P<0. 01, n = 6- 8]. The plasma 8-OHdG levels of ARB pretreatment groups were decreased. In immunohistochemical findings, 8-OHdG- and HIF-1α-containing cells in ARB pretreatment groups were decreased. Conclusion Brain ischemia and oxidative damage can be reversed by AT1 receptor blockade in normotensive rats after transient cerebral artery occlusion.

Objective Angiotensin Ⅱ (Ang-Ⅱ) increases NADPH oxidase activity and stimulates the production of reactive oxygen species (ROS) including superoxide anion through Ang Ⅱ AT1-receptor (AT1-R) activation. ROS is involved in various pathological processes in brain ischemia. We investigated whether the AT1-R blocker (ARB) candesartan can protect normotensive rats against brain ischemia. Methods After 2-week pretreatment with candesartan, rats were subjected to 2 hours middle cerebral artery occlusion-reperfusion (MCAO-R) and 24 hours later, the infarct volume, iNOS, and eNOS mRNA in the internal carotid artery was recorded and compared. Results Candesartan pretreatment reduced cerebral ischemia and oxidative brain damage after MCAO-R in normotensive rats, resulting in a decreased cortical infarct volume [0.5 mg/kg candesartan, (46.8±13.2)mm3; 1.0 mg/kg candesartan, (19.3±15.3)mm3 vs. control, (111.7±14.3)mm3; P<0.05, P<0.01, respectively]. Candesartan pretreatment increased the eNOS mRNA level in the internal carotid artery. Conclusion In normotensive rats exposed to MCAO-R, candesartan protectes against brain ischemia. This effect may represent a significant therapeutic advantage and may induce end-organ protection even at normal blood pressure.

Objective To investigate the Influence of Survivin and hTERT gene on cell proliferation and apoptosis in human colorectal carcinoma cell line SW 480 and to find experiment evidence for gene therapy of colorectal carci-noma .Methods Plasmids carrying shRNAs targeting survivin and hTERT were designed , constructed and trans-fected into SW480 cells.SW480 cells were then divided into blank group , blank Plasmid control group , survivin RNAi group , hTERT RNAi group and Survivin-hTERT RNAi group .The telomerase activity was examined by TRAP-PCR-ELISA analysis 48h after hTERT-shRNA transfection.Survivin and hTERT mRNA and protein expres-sion was analyzed by RT-PCR and Western blot .Cell apoptosis , proliferation were measured by flow cytometry , CCK-8 assay.Results Telomerase activity of SW480 cells in Survivin-hTERT RNAi groups were significantly decreased compared with the blank group ( P<0.01 ) .The expression of survivin and hTERT mRNA, proteins in the Survivin-hTERT RNAi group was reduced by 82.8%and 73.6%( P<0.01 ) ,79.2%and 66.7%( P<0.01 ) respectively .The inhibitory rate of cell proliferation of Survivin-hTERT RNAi group was 43.6% ±0.1%( P <0.01 ) .The apoptosis rate was 39.2%±2.3%( P<0.01 ) in the Survivin-hTERT RNAi group .Conclusions The Survivin-hTERT RNAi group could significantly reduces the protein expression of survivin and hTERT mRNA, in-hibit cell proliferation and induces cell apoptosis in human colorectal carcinoma cell line SW 480 .

Objective To analyze the effects of overexpressed iASPP on breast cancer cell line MCF-7. Methods The iASPP cDNA was obtained from breast carcinoma tissue by RT- PCR. The breast cancer cell line MCF-7 that overexpressed iASPP was established by stable transfection and G418 selection. The proliferation and apoptosis of the MCF-7 cells that overexpressed iASPP after exposure to cisplatin were detected by MTT and flow cytometry. Results The MCF-7 cells that overexpressed iASPP were successfully constructed. Overexpressed iASPP could significantly decrease the apoptosis rate of MCF-7 cells exposed to cisplatin and weaken the inhibitory effects of DDP on the proliferation of MCF-7 cells. Conclusion Overexpressed iASPP attenuates the sensitivity of human breast cancer cell line MCF-7 to cisplatin.

Objective To construct and identify the recombinant adenovirus vector containing the Toll-like receptor 2(TLR2) extracellular domain gene of human.Methods Human TLR2 extracellular domain cDNA was amplified by RT-PCR from peripheral blood mononuclear cells(PBMCs) and inserted into pMD18-T vector.After being confirmed by enzyme digestion and sequencing,the DNA fragment,digested with Kpn Ⅰ and Hind Ⅲ,was directionally cloned into adenovirus shuttle plasmid pAdTrack-CMV.After linearization by Pme Ⅰ digestion,the recombinant plasmid pAdTrack-CMV-TLR2 was transformed into competent AdEasier-1 germs and then homologically recombined with an adenoviral backbone plasmid pAdEasy-1 in bacteria BJ5183 to obtain the recombinant adenovirus plasmid.After confirmation,the recombinant adenovirus plasmid pAd-TLR2 was linearized with Pac Ⅰ digestion and transfected into 293 cells via liposome,and then package and adenovirus amplification were performed.The expression of green fluorescent protein(GFP) was observed,the virus titer was determined and the recombinant adenovirus was identified by PCR.ResultsThe gene fragment obtained by RT-PCR was of the same sequence as in GenBank.It was certified by restricted endonuclease digestion and PCR analysis that the recombinant adenovirus containing the TLR2 extracellular domain gene of human had been successfully constructed with a satisfactory high titer of 3?109pfu/ml.Conclusion The recombinant adenovirus containing TLR2 extracellular domain gene of human has been successfully constructed,which lays a foundation for further study on the structure and biological activity of TLR2.

Objective To explore the role and significance of matrix metalloproteinase-9 (MMP-9) in angiogenesis through observing the relationship between the expression of MMP-9 and microvessel density (MVD) in glioma. Methods The expressions of MMP-9 and CD34 in 10 cases of normal brain tissues and 58 cases of glioma (14 cases of grade Ⅰ , 20 cases of grade Ⅱ , 15 cases of grade Ⅲ, and 9 cases of grade Ⅳ ) were detected by immunohistochemical streptavidin-peroxidase technique. The positive cells of MMP-9 and the positive microvessels were examined under binocular light microscope. Results The positive expression of MMP-9 in glioma was located in the tumor-cell cytoplast and endothelial cells. The positive rate of MMP-9 in glioma of grade Ⅰ, Ⅱ, Ⅲ and Ⅳ was 42.9%, 65.0%, 86.7% and 88. 9%, respectively. The expression of MMP-9 was obviously higher than that of normal brain tissues (P<0.01) and positively correlated with glioma malignancy (rz =0. 597, P<0.05). MVD was correlated with glioma malignancy (H=47. 865, P<0. 05). The expression of MMP-9 was significantly correlated with MVD (rz =0.897, P<0.01). Conclusion The expressions of MMP-9 and MVD are correlated with glioma malignancy, which may be helpful in judging the malignancy, invasion and prognosis. MMP-9 plays an important role in angiogenesis of glioma and accelerates glioma malignancy development by promoting angiogenesis.