Objective:To construct the highly efficient expression recombinant adenovirus that expresses elafin protein for further study.Methods:The elafin gene was amplified by RT-PCR from lung tissues,then the fragment was inserted into the multiple clone site of pAdTrack-CMV. The linearized shuttle plasmid was homogenously recombined with pAdEasy-1 in BJ5183. The candidate clone was further analyzed by restriction endonuclease digestion, PCR, and sequence scanned. Then the recombined adenovirus plasmid was digested with Pac Ⅰ and transfected into 293 cells for packaging and amplifying. Infection titer and rate were monitored by green fluorescent protein(GFP) expression. The expression of elafin protein was detected by Western blot and ELISA to appraise the function of elafin protein for oppressing the activity of elastic protease.Results:Digestion with restriction endonuclease, PCR, Western blot and ELISA confirmed that elafin gene was cloned into the adenovirus vector successfully.Conclusion:The recombined adenovirus Ad-elafin is constructed successfully, which will be benefit for future study.

By analyzing the Military Health System winning "Heliang & Heli" Fund Award between 1994 and 2009,we found that the Military Health System possessed a good scientific basis and high level,but with an unbalanced scientific research development,which suggests the Military Health System should further strengthen its independent innovation ability.

Objective To construct the recombinant adenovirus for nature antibiotic elafin protein and observe its expression in primary airway epithelia.Methods The elafin gene was amplified by RT-PCR from lung tissues,then the fragment was inserted into the multiple clone site of pAdTrack-CMV.The linearized shuttle plasmid was homogenously recombined with AdEasy-1 in BJ5183.The candidate clone was further analyzed by restriction endonuclease digestion,PCR,and sequencing.Then the recombinant adenovirus plasmid was digested with PacⅠ and transfected into 293 cells for packaging and amplification.Infection titer and rate were monitored by green fluorescent protein(GFP)expression.Finally the primary airway epithelium was infected with Ad-elafin,and the GFP expression was observed by fluorescence microscope in epithelial cells 24 h after transfection.The elafin protein was detected by ELISA in the supernatant.Results Restriction endonuclease and PCR confirmed that elafin gene was cloned into the adenovirus vector successfully.Twenty-four hours after transfection,the GFP expression was seen by fluorescence microscopy.The concentration of elafin protein was(2.2?0.4)ng/ml in supernatant of transfected group,while that of control group was(1.9?0.3)ng/ml,P

Objective:To study the mechanism of protein kinase C regulating CD44 gene expression in vascular endothelial cells.Methods:Human umbilical vein endothelial cells (HUVECs)were taken as study model.The extract of Raf-1 kinase by immunoprecipitation was used for the Western blot analysis,and its activity was determined by enhanced chemiluminescene assay.CD44 gene expression was detected with RT-PCR,and phosphorylation was measurated by autoradiography.Results:CD44 phosphorylation in HUVECs was enhanced by 10ng/ml PMA treatment as compared with untreated cells, which reached the highest level at 30 minutes. Raf-1 kinase activity increased significantly after exposure to 10ng /ml PMA, and 0.05 ?mol/L Calphostinc could inhibit the role of Protein kinase C(PKC). CD44 gene expression level increased obviously after exposure to 10 ng /ml PMA (PKC activator) for only 1 minute(P

BACKGROUND: Peripheral blood stem cells (PBSCs) can be enhanced and differentiates into all kinds of neural cells and produce several kinds of neural growth factors in vitro, and have been used to treat several kinds of neural diseases and they showed satisfactory outcomes. It is now a research focus to optimize the culture conditions to induce the stem cell to differentiate into neural cells. On the other hand, it also becomes a focus of its amplification and differentiation. OBJECTIVE: To make a review on researches on treating brain lesion, genetic defection or degenerating diseases using PBSCs during recent years.METHODS: Researching Pubmed database(1999-01/2009-09), using "peripheral blood stem cell, neural, repair" as research words and these Chinese words in CNKI database(1999-01/2009-09). RESULTS AND CONCLUSION: 221 articles were collected altogether including 41 Chinese articles and 180 English articles. Totally 27 articles were adapted altogether after rejecting published early, repeat and similar articles. PBSCs can be enhanced and differentiates into all kinds of neural cells in vitro, and delivered into central neural system and improve the function of lesion neural area. It is not fully understood the differentiation mechanism of peripheral blood stem cells, but numerous studies have shown that surface marker antigen of peripheral blood stem cells is closely associated with its biological characteristics. There are many studies addressing surface marker of peripheral blood stem cells, but researchers cannot identify a reliable marker that can directly label peripheral blood stem cells. Function of CD34 remains unclear, so the principle of CD34 as a marker of hematopoietic stem cells receives challenge. It is necessary to do further researches on issues on how to enhance survival rate of PBSCs in vivo and the tendency to differentiate into neural cells.

OBJECTIVETo investigate the short-course and efficient way to treat mumps meningitis.

METHODSTotally 155 cases of mumps meningitis treated by intrathecal injection with dexamethasone only once were enrolled as experimental group and 55 similar cases treated with the common therapy as control.

RESULTSThe time for recoveries of temperature, headache, the pathologic reflexes and the total time of treatment were 32 hours, 15 hours, 12 hours and 3.1 days, respectively, while those of the control group were 58 hours, 24 hours, 32 hours, 6.5 days respectively. There was statistical difference between the two groups (P<0.01 or 0.05).

CONCLUSIONSIntrathecal injection with dexamethasone only once is efficient and safe in the treatment of mumps meningitis.

Anti-Inflammatory Agents ; therapeutic use ; Child ; Child, Preschool ; Dexamethasone ; therapeutic use ; Female ; Humans ; Infant ; Injections, Spinal ; Male ; Meningitis, Viral ; complications ; drug therapy ; Mumps ; complications ; drug therapy ; Treatment Outcome

Objective:To analyze the features and clinical significance of damages to non pulmonary organs like the heart,kidney and peripheral blood in SARS patients.Methods:SPSS 11.0 and SDAS software packages were used to retrospectively analyze the damages to the heart,kidney and peripheral blood in 330 clinically confirmed SARS cases.Results:(1)The heart damage occurred within 1 week after SARS onset in mild cases; the rates of LDH,HBDH,CK and CK MB returning to normal were 74.2%,80.6%,100% and 100%,respectively.In severe cases,the heart damage usually occurred 1 week after SARS onset; the rates of LDH,HBDH,CK and CK MB returning to normal were 32.2%,29.5%,55.2% and 44.4%,respectively.Ten patients whose CK MB didnot return to normal died.(2) The changes of RBC count appeared after an average of 1 week (1 10 d) in 54.2% of the patients.It returned to normal in all the mild SARS cases after an average of 12.5(4 21) d but didnot in 18.4% of the severe cases ( P

Objective To explore the differences between ABI 7500 and TL 988 fluorescent quantitative instrument for polymerase chain reaction. Methods Based on EP9-A2 Files, quantitative influenza A virus positive controls of five kinds of concentrations were detected by two kinds of fluorescence quantitative PCR instruments at the same time, and each sample was detected 20 times. The precision and sensitivity between the two kinds of instruments were compared. Results Inner group CV of TL 988 ranged from 0.68% to 1.52%. The CV for ABI 7500 ranged from 1.93% to 2.69%. The high (107, 106, 105 copies/mL) levels of IFVA positive control were detected by ABI 7500 and TL 988 instruments, and there were no significant difference between the results(P>0.05) while the low(104, 103 copies /mL)levels were de-tected by ABI 7500 and TL 988 instruments, and there was a significant difference between the results(P<0.05). When samples were detected at the level of 103 copies/mL, the positive detection rate was 100% with TL 988 instrument, but the positive detection rate was 0 with ABI 7500 instrument. The related index of ABI 7500 and TL 988 was better (r=0.998). Conclusion The performances in precision and sensitivity of TL 988 PCR instrument are better than ABI 7500 PCR.

Objective:To investigate the effect of peripheral blood neutrophil to lymphocyte ratio (NLR) and platelet distribution width (PDW) on the clinical prognosis of childhood medulloblastomaMethods:Clinical data and survival data of 76 pediatric patients who were diagnosed as medulloblastoma by post-operative pathology in the First Affiliated Hospital of Zhengzhou University from January 2009 to December 2016 were collected. Kaplan- Meier method was used to calculate the overall survival(OS) and progression free survival(EFS) rates, Log- rank test was employed to compare the survival rates of different groups, and Cox proportional hazards regression model was used for multivariate analysis. Results:The log- rank test revealed that 5-year PFS rate and OS rate (22.2%, 22.2%) in the high NLR group (NLR>4.94) were significantly lower than those in the low NLR group (NLR≤4.94) (45.6%, 55.7%), and the differences were statistically significant(PFS: P=0.009, OS: P=0.001), and the 5-year PFS and OS (52.3%, 66.5%) of the high PDW group (PDW>15.90) were significantly higher than those in the low PDW group (PDW ≤ 15.90) (27.1%, 32.5%), and the differences were statistically significant(PFS: P=0.032, OS: P=0.039). Univa-riate analysis showed that the extent of resection (PFS: P=0.006, OS: P=0.009), and postoperative radiotherapy (PFS: P=0.011, OS: P=0.001) and postoperative radiotherapy(PFS: P=0.011, OS: P=0.001) were the factors influencing the prognosis of children with medulloblastoma.Multivariate Cox proportional hazards regression model analysis suggested that no postoperative radiotherapy (PFS: P=0.048, OS: P=0.008), NLR>4.94 (PFS: P=0.023, OS: P=0.003) and PDW≤15.90 (PFS: P=0.028, OS: P=0.006) were the independent risk factors for the prognosis of childhood medulloblastoma. Conclusions:Increased NLR and decreased PDW indicate unfavorable prognosis of the childhood medulloblastoma.Therefore, preoperative NLR and PDW may be the potential prognostic markers for childhood medulloblastoma.