Purpose To construct a lentiviral ventor-mediated RNA interference of human midkine ( MK) gene and to provide the ba-sis for further experiment in vivo and in vitro. Methods Four RNAi sequences targeting human MK gene were designed, and cloned into the lentiviral vector to construct lentiviral vectors:GV115-MK-1, GV115-MK-2, GV115-MK-3 and GV115-MK-4. After transfec-tion into competent E. Coli bacteria, the candidate clones were identified by PCR and DNA sequencing. The titer of lentiviruses was determined after 293 T cells were co-transfected with GV115-MK, pHelper 1. 0 and pHelper 2. 0. The four kinds of recombinant lenti-viruses were used to infect human breast cancer cell MDA-MB-231, and the expression levels of MK mRNA were detected by real-time PCR. Results PCR analysis and DNA sequencing confirmed that the four inserted MK RNAi sequences were corrected. Strong green fluorescence was observed in the 293 T cells under the fluorescent microscope after co-transfection. The titer of the concentrated virus was 6 × 108 , 5 × 108 , 5 × 108 and 6 × 108 TU/ml, respectively. The MK expression in MDA-MB-231 cells infected with lentiviral vec-tors GV115-MK-1 was decreased by 87. 2% (P<0. 05). Conclusion Lentiviral RNAi vector for MK gene is successfully construc-ted, and it could effectively inhibit the expression of MK gene in MDA-MB-231 cells in vitro.

Since the early symptoms are very hidden,most of patients with colorectal cancer (CRC)have occurred metastasis when diagnosed.Recently,chemotherapy combined with targeted therapy is the major therapeutic strategy for metastatic CRC and the high frequency activation of phosphatidylinositol 3-kinase (PI3K) signaling pathway has been validated sharing a close relationship with the development of CRC,contributing important values for the diagnosis and therapy of CRC.

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. [

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.

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 research and develop the mobile platform-oriented echocardiogram data visualization technology for the evaluation of left ventricular diastolic function,and investigate its application value.Methods First,diagnostic algorithms for the evaluation of left ventricular diastolic function (based on recommendations from ASE & EAE guidelines) were programmed into a mobile application software package,using Adobe Flex builder 4.0 as a basic tool for data visualization.Second,the program was installed in the Apple iPad4 tablet computer,then debugged and improved according to feedback information from practical applications.Results The echocardiogram data visualization for the estimation of left ventricular filling pressures in patients with depressed ejection fraction(EF) or normal EF and grading diastolic function can be implemented in mobile platform device,and simultaneously with emergency bedside echocardiography.The evaluation process can be visually presented by using the graphics to show the algorithm framework,the animation to express the analysis step,the axis to indicate the direction of information,and the color to differentiate the degree of the diastolic dysfunction.Conclusions The mobile platform-oriented echocardiogram data visualization technology makes the evaluation of left ventricular diastolic function more convenient,efficient and fast,and shows its potential to become an important adjunct to evaluation of cardiac function in the context of mobile health.

Objective To study the mechanisms and expressions of Fas/FasL in the lungs,spleens and lymph nodes of SARS patients.Methods The lungs,spleens and lymph nodes specimens of four autopsy cases and one control subject were studied with immunohistochemical mehod.Results Compared with normal tissues,apoptotic cells increased significantly in the spleens,lungs and lymph nodes of SARS patients.The apoptotic cells of lungs included alveolar epithelial ones,bronchial epithelial ones,monocytes/macrophages and lymphocytes.Apoptosis in the spleens and the lymph nodes was observed primarily in monocytes/macrophages and lymphocytes.Conclusion Over-expression of Fas/FasL is a general phenomenon in SARS,suggesting that Fas/FasL may play a key role in the pathogenesis of SARS.

Objective: To evaluate the long-term result after bioprosthesis replacement, and analyse the risk factors of the late deaths. Methods: From September 1984 to March 1988, 90 patients underwent bioprosthesis replacement in our hospital. There were 39 males (43%) and 51 females (57%) with an average age of 36.1 years. 78 patients had rheumatic disease, 8 degenerative disease, 3 congenital mitral valve anomaly and 1 Ebstein's anomaly. The preoperatively heart function (NYHA) was grade II in 15, grade III in 54, and grade IV in 21. C/T was 0.51～0.90 (average 0.65?0.10). Results: The operative mortality rate was 7。8%. All survived patients were followed up. The mean follow-up time was 7.8 years (range from 3 months to 15.7 years. 35 patients died during the follow-up period. The causes of death cardiac related deaths in 33 patients (88.6%), noncardiac related death in 3 (8.6%), and unknown in 1 (2.8%). Reoperation was performed in 20patients. At 1, 5 and 10 years after operation, the actuarial survival rate of patients were 92.7%, 80.7% and 57.8%, respectively. Multiple regression analysis showed that preoperatively cardiac function, C/T ratio and EF were risk factors for late deaths. Conclusion: The long-term results with bioprosthesis replacement appear satisfactory.

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.

BACKGROUND: Magnesium alloy studies in orthopedic field have been carried out,and good biocompatibility has been reported.However magnesium alloys have not yet been researched in the intestine.OBJECTIVE: The biodegradable magnesium-zinc alloy samples are implanted around the rat cecum to investigate the biocompatibility in rat.METHODS: Sprague Dawley rats were randomly divided into magnesium alloy group,medical titanium group and the sham-operated group.Then magnesium-zinc alloy samples with the dimension of 5 mm × 1 mm× 1 mm were embedded in the cecum incision in the magnesium alloy group.The medical titanium was embedded in the medical titanium group,and just suture in the sham-operated group.Prior to surgery and at 7,14,21 and 28 days after operation,the serum glutamic-oxaloacetic transaminase,creatinine and magnesium ion concentration were examined in each group.X-ray film on implanted region.The pathological changes in liver,kidney and cecum were examined.RESULTS AND CONCLUSION: There were no significant differences in serum glutamic-oxaloacetic transaminase,creatinine and magnesium ion concentrations among each group(P > 0.05).Magnesium-zinc alloy degraded gradually during 28 days.The pathology of liver,kidney and cecum was normal.Results suggested that magnesium-zinc alloy had no obvious effect on the cecum.The degradation time to play a fixed function of time was longer than the intestinal healing time,with good biocompatibility.Magnesium-zinc alloy can be used as anastomotic nail for stomach intestine anastomat.