Objective To isolate the rat insulin gene enhancer binding protein 1(Islet-1)gene and construct plEGFP-C1-Islet-1 recombinant retroviral expression vector.Methods The cDNA encoding the rat Islet-1 gene was isolated by RT-PCR method,the cDNA was first cloned into PGET-1 TA vector to facilitate the sequence and then subcloned into the retroviral vector plEGFP-C1.plEGFP-C1-Islet-1 was transfected into PA317 packaging cells with lipofectamine 2000.Transformants were selected in medium containing G418.Results A 1 050 bp DNA fragment was obtained by RT-PCR;plEGFP-C1-Islet-1 recombinant retroviral expression vector was identified by restrictive enzymes digestion,PA317 cells transfected with recombinant vector expressed enhancer green fluorescent protein(EGFP).Conclusion The gene encoding the rat Islet-1 is obtained and plEGFP-C1-Islet-1 expression vector is constructed successfully.

Objective To evaluate the combined effects of vascular endothelial growth factor (VEGF) and placental growth factor (PLGF) on angiogenesis and cardiac function and compare with VEGF or PLGF only in acute myocardial infarction rats.Methods Seventy-five males Sprague-Dawley(SD) rats were randomly divided into five groups:sham group,NS group,VEGF group,PLGF group,and VEGF + PLGF group with 15 rats in each group.All the rats underwent LAD ligation and injection of NS,VEGF,PLGF,VEGF + PLGF,in the peri-infarct area,respectively,besides the sham group.Three weeks after coronary artery ligation and different agents injection,cardiac function,myocardial scar area,angiogenesis and arteriogenesis were studied.Cardiac structure and function,and infarct size were assessed by echocardiography.The number of new vessels and the number of new arterioles were evaluated by haematoxylin-eosin staining and immunohistochemistry staining.Results Three weeks after LAD ligation and different agents injection,the LVEDD and LVESD were significantly decreased (P ＜ 0.01)in NS group,VEGF group and PLGF group.While the LVEF and LVFS were higher in VEGF + PLGF group than that in other groups.Myocardial infarct size was reduced in VEGF group (P ＜ 0.05).Angiogenesis and arteriogenesis were higher in VEGF + PLGF group than that in VEGF group (P ＜ 0.01) and PLGF group (P ＜0.05).Angiogenesis and arteriogenesis were significantly higher in PLGF group than that in VEGF group (P＜0.01).The density of microvessels in VEGF group was higher than that in NS group (P ＜ 0.05),while arteriogenesis was of no statistical difference.Conclusion The combination of half VEGF and PLGF can increase angiogenesis and arteriogenesis in the ischemic marginal zone of myocardial infarction,decrease myocardial infarction area,and improve cardiac function.

ole on the development of septic AKI. It's useful to decrease the incidence of AKI by elevating the blood pressure and C1, but super-normal goal won't work better.

Objective To evaluate the clinical application of CT perfusion imaging (CTPI) and CT subtraction angiography (CTSA) in the diagnosis of acute ischemic cerebrovascular disease (AICVD). Methods 24 cases with AICVD onset within 24 hours were examined with regular CT, CTPI and CTSA. Some of them took CTPI, MRI, MRA, DSA, SPECT by follow up examinations. Results In 24 cases 11 had regular CT negative results after onset of stroke 3～6 hours in 6 cases,6～12 hours in 3 cases,12～24 hours in 2 cases. Ten cases of them were confirmed by CTPI as having ischemic lesions, 2 cases had middle cerebral artery occlusion (MCAO), and 1 had transient ischemic attack (TIA) with CTPI negative. In 24 cases 13 had regular CT positive rseults, 9 cases had ischemic lesions larger in CTPI than in regular CT,1 case had MCAO and 1 case had internal carotid artery occlusion(ICAO). There were 4 cases with ischemic lesions on regular CT almost having the same range as that of lacunar infarction in CTPI. The peak value of time(PT), mean transit time(MTT), relative flow (RF) in all 24 cases were found obviously changed. The side of ischemic lesion as compared with the opposite side, and the core of ischemic lesion as compared with peripheral zone were found changed significantly ( P

400?109/L and anemia was defined as Hb

Objective To assess whether end-tidal carbon dioxide partial pressure (PET CO2) can predict the fluid responsiveness in septic shock patients.Methods Septic shock patients under mechanical ventilation without spontaneous breathing and with the need of a fluid challenge test were included in this study.Heart rate,central venous pressure,pulse pressure,PErCO2,and CI before and after the fluid challenge test were conducted in all the patients.Results Of the 48 septic shock patients included,34 had preload responsiveness,14 had no responsiveness.△CI and △PET CO2 after the fluid challenge test involume responders were (0.85 ± 0.47) L · min-1 · m-2 and (3.5 ± 2.5) mmHg respectively,which were higher than those in no volume responders (P ＜ 0.05).The fluid-induced changes in PET CO2 and CI were correlated (r =0.072,P ＜ 0.05).The AUCRoc of fluid challenge-induced △PET CO2 as the predictor for volume responsiveness was 0.943,and its sensitivity was 87.9％ and specificity was 93.4％ with a critical value of 5％.The AUCRoc of △PP as the predictor for volume responsiveness was 0.801,and its sensitivity was 68.1％ and specificity was 73.2％ with a critical value of 10％.Conclusion The changes of PETCO2 induced by a fluid challenge test can predict fluid responsiveness with reliability,and have a better sensitivity and specificity than the changes of PP.

Objective To construct recobinant pmRNA IRES-hKDR and lay a primary foundation for further study of gene therapy for tumors.Methods pmRNA IRES-hKDR was obtained from pDC520 hkDR and pmRNA IRES-Luc which was reserved by our laboratory through molecular biology technology and was transfected into Hepall-6 via lipsome after identification of PCR,restriction enzyme digesting and DNA ELISA and Western blot.Results The recombiant plasmid had pmRNA IRES-hKD gene which was expressed in Hepall-6//G418 in vitro and the expressed protein was secreted out of the cells could response with specific hKDR antibody,which was identified by Western blot.Conclusion An eukaryotic expression recombiant plasmid pmRNA IRES-hKDR can be constructed successfully which provides the possibility of further researches on its anti-tumor effect.

BACKGROUND: Surface roughness of implants can directly influence cellular proliferation, differentiation, and gene expression. OBJECTIVE: To observe the early attachment of periodontal ligament cells (PDLCs) to pure titanium with different surface roughness levels, and to study the effect of surface performance on cell differentiation. DESIGN, TIME AND SETTING: Randomized controlled observation/multi-sample comparison study, which was performed at Center of Science and Technology, Gannan Medical College between January 2005 and July 2006. MATERIALS: Pure titanium stick was cut into pieces, of 10 mm diameter and 2 mm thickness, using cutting-off machine, and there were 24 sections in total. Then, the titanium sections were randomly divided into four groups: simple mechanical processing, nitric acid processing, sand blasting processing, and combination group, with 6 sections per group. METHODS: TR240 portable-type surface roughness meter was used in this study. In the simple mechanical processing group, sections were scoured by sand paper alone; in the nitric acid processing group, sections were etched with 65% HNO3 for 1 hour at 100 ℃ after scoured by sand paper; in the sand blasting processing group, sections were sandblasted by 100 μ m AI203 after scoured by sand paper; in the combination group, sections were etched with 65% HNO3 for I hour at 100～C after scoured by sand paper and sandblasted by 100 μ m Al2O3. MAIN OUTCOME MEASURES: Samples were maintained in DMEM for 30 minutes, and the third-passage cells were inoculated. Then, titanium sections were taken out at different time points of 30, 60, 120, 240 minutes, 1, 3, and 7 days. Surface roughness and early attachment of PDLCs were detected under fluorescent microscope. RESULTS: O Quantitative analysis: Surface roughness was (599.5±8.3) nm in the simple mechanical processing group, (406.5 +4.6) nm in the nitric acid processing group, (358.8±11.8) nm in the sand blasting processing group, and (8.7±2.0) nm in the combination group. On the other hand, surface roughness in the simple mechanical processing group was significantly higher Fluorescence observation exhibited that number of PDLCs attaching to pure titanium surface was increased, and the proliferation was greater with the time passing by. in addition, surface roughness of pure titanium was positively associated with number of PDLCs. CONCLUSION: The lighter the surface roughness is, the more the early attachment of PDLCs is, benefiting for cell adhesion and proliferation.

Aim To study the effects of propofol on apoptosis of cortical astrocytes isolated from neonatal rats.Methods Pure astrocytes(AST)were obtained from the cultured cerebral cortical cells and identified by the GFAP stain technology in neonatal rats.AST cells were treated with different concentrations of propofol(0,10,30,90 μmol·L-1)for 8 hours.Cell viability was measured by MTT method,and the apoptosis was detected by Annexin V/PI double staining.Cytochrome C(cyt-C)leakage was detected by Western blot.Caspase-3 and caspase-9 activities were measured by spectrophotometry.Results AST cells were administered with different concentrations of propofol(0,10,30,90 μmol·L-1)for 8 hours.It decreased the survival rate in a concentration-dependent manner,induced the leakage of cyt-C in mitochondria,up-regulated the expression of pro-apoptotic protein caspase-3 and caspase-9,and induced AST apoptosis.Conclusion Propofol induces the apoptosis of astrocytes in neonatal rat cortex in vitro,which may be related to the activation of mitochondria apoptosis pathway induced by mitochondrial cyt-C release.