Objective To investigate the altered glycotransferases and glycan profile in HCV-infected cells, the expression levels of 35 kinds of sialyltransferases and galactosyltransferases were evaluated in hepatitis C virus (HCV) infected Huh7.5.1 cell cultured model.Methods Western blot was used to measure the expressions of HCV-nonstructural protein NS3 and and quantitative real-time PCR (qRT-PCR) was performed to determine HCV-RNA levels and mRNA levels of 35 kinds of sialyltransferases and galactosyltransferases.Lectin microarray was used to compare the glycan profiles of sialyltransferases-and galactosyltransferases-associated glycan-profile in Huh7.5.1 cells and HCV infected Huh7.5.1 cells.Results Among the 35 kinds of sialyltransferases and galactosyltransferases, the mRNA levels of four sialyltransferases (including ST3Gal Ⅲ, ST6GalNAC Ⅲ, ST8Sia Ⅲ and ST8SiaⅣ) and five galactosyltransferases (including B3GALT1, B3GALT 2, B3GALT3, B3GALT4 and B4GALNT2) were found to be 11-45 fold higher in HCV-infected cells compared with naive Huh7.5.1 cells.The up-regulated level of B3GALT 1 was the most evident (45-fold change), followed by ST8Sia Ⅳ and ST8Sia Ⅲ, with 39-and 37-fold respectively.Consistently, lectin microarray showed that glycans recognized by EBL (binding terminal sialic acid, Neu5Acα6Gal, 1.97-fold change), ECA letin (binding terminal galactose, Galβ1,4GalNAc,4.3-fold change), and ACA(binding terminal galactose, Galβ1-3GalNAc, 3-fold change) were elevated in glycoprotein products of sialyltransferase and galactosyltransferase respectively.Conclusion HCV infection causes the increased expression levels of mutiple sialyltransferases and galactosyltransferases in Huh7.5.1 cell line and hence the upregulation of glycan profiles of the glycoproteins.These results provide potential therapeutic targets and diagnostic markers for HCV infection and insights on the molecular mechanism of HCV infection.

BACKGROUND:Oxidized low-density lipoprotein(ox-LDL) is recognized as the essential condition for atherosclerosis.As the ox-LDL-specific receptor,oxidized low-density lipoprotein receptor-1(LOX-1) involved in vascular inflammation and plaques develop-ment of the process.OBJECTIVE:To investigate the effect of fluvastatin on the expression of LOX-1 in human umbilical vein endothelial cells(HUVECs) induced by ox-LDL.DESIGN,TIME AND SETTING:The comparative observation was completed in the Medical Center of Second Xiangya Hospital,Central South University between August 2006 and May 2007.MATERIALS:Human umbilical vein endothelial cell line was purchased from the America ATCC Company.Fluvastatin original powder was supplied by Beijing Novartis Pharmaceutical Co.,Ltd.METHODS:HUVECs were incubated with:①Stimulation by ox-LDL with end concentration of 25,50,100 mg/L.②LOX-1 neutralizing antibody,and interfered with 50 mg/L ox-LDL.③Interfered with nuclear factor-?B(NF-?B) blockers pyrrolidine dithiocarbamate(PDTC),followed by 50 mg/L ox-LDL intervention.④Fluvastatin with concentration of 0.01,0.1,1 ?mol/L were used to interfere the cells,followed by 50 mg/L ox-LDL intervention.⑤There was an blank group as the control.MAIN OUTCOME MEASURES:The expression of LOX-1 mRNA level was detected by RT-PCR.RESULTS:The levels of mRNA of LOX-1 were increased after cells had beenwere incubated with different concertrations of ox-LDL,when compared with the control group.Within the dosage range of 25 to 50 mg/L,the above-mentioned indexes significantly changed in a concentration-dependent manner(P

Objective To investigate antiepileptic drug application and the influential factors in patients after epilepsy surgery.Methods During the period from 2002 through 2005,170 patients with intractable epilepsy who received epilepsy surgery in Xuanwu hospital were studied.They were divided into 3 groups according to the stages of operation.Patients in group A received epilepsy surgery during 2002 to October 2003,those in group B received initial phannacentieal care after epilepsy surgery during November 2003 to October 2004,and those in group C received integrated pharmaceutical care after epilepsy surgery during November 2004 to October 2005.The clinical outcomes,factors affecting antiepileptic drugs,safety and compliance with antiepileptic medication were analyzed.Results Patient's clinical outcomes(group B was 71%and group C was 81%),safety and compliance with antiepileptic medication in group B and group C are better than those in group A(46%)with a significance difference(X2=7.08,15.50,P<0.05).Conclusions The integrated pharmaceutical care rendered from the cooperation of neurosurgeon,neurologist and clinical pharmaceutist is a new and effective management mode for postsurgical epilepsy patients.

We first study the photosensitization to bacteria and fungi with a definite structure, watersoluble porphyrin. We found that the porphyrin had strong sterilization on the Staphylococcus au- reus and Candida albicans. When the porphyrin concentrations increased, its sterilization strengthened. The sterilization was related to the photoradiating time. When photoradiating time was one hour, its sterilizing ability was the greatest. Its sterilizing ability was similar to the penicillin's.

Objective:To construct an eukaryotic expressing plasmid of IL-24,and investigate its inhibitory effects on the growth of tumor cells in vitro and in vivo.Methods:The eukaryotic expressing plasmid of IL-24(pEGFP-C1-IL-24) was constructed by DNA recombination technique.The recombination plasmid and empty vector were transfected into HIC cells,respectively with Lipofectamine 2000 Reagent and the expressions were determined by LSCM;the proliferation of HIC cells was measured by MTT assay;and apoptosis rate and cell-cycle distribution of HIC cells were measured with Flow Cytometry.Mice were inoculated with B16 cells,which were transfected with pEGFP-C1-IL-24 or empty vector,and the tumor size in mice was detected.The inhibitory effect of IL-24 gene transfection in mice solid tumor was observed and measured.Results:The expression of pEGFP-C1-IL-24 in HIC cells was determined by LSCM after the transfection of pEGFP-C1-IL-24.Comparing to the control group,the proliferation of HIC cells was inhibited by transfection with pEGFP-C1-IL-24 and the G2-M phase of the transfected cells was also increased.Moreover the percentage of mice with detectable tumor was significantly decreased after inoculated with B16 cells transfected with pEGFP-C1-IL-24.Growth rate of tumor in mice model was also significantly inhibited in IL-24 gene therapy group as compared with the control grouop(P

AIM: To construct full length hCD59 eukaryotic and extracellular domain of hCD59 (hsCD59) prokaryotic expression vectors and prepare polyclonal antibody of hCD59. METHODS: cDNA fragments encoding hCD59 and hsCD59 were amplified from human PBMCs by RT-PCR and cloned into the eukaryotic vector pVAX-1 and prokaryotic vector pGEX-KG, respectively. The recombinant fusion protein GST-hsCD59 was expressed in E. coil BL21 induced by IPTG. Then the fusion protein was purified and identified. Polyclonal antibody against hCD59 was prepared by immunizing rabbit with pVAX-1-hCD59 and boosting with GSThsCD59 fusion protein, and the titer was identified. RESULTS: The recombinant eukaryotic vector pVAX-1-hCD59 and prokaryotic vector pGEX-KG-hsCD59 were successfully constructed. The GST-hsCD59 fusion protein was over-expressed in E. coli BL21 and the relative molecular mass (M~r) of the expression product was identical with predicted size. The titer of the anti-hCD59 serum was 1:3 200. CONCLUSION: We got the recombinant eukaryotic vector pVAX-1-hCD59, prokaryotic vector pGEX-KG-hsCD59 and rabbit anti-hCD59 polyclonal antibody successfully. These work would be helpful for the further study of the biological function of human CD59.

Objective To investigate the effect of G-quadruplex (G4) RNA structure of core of hepatitis C virus (HCV) on the specific immune response. Methods Circular dichroism (CD) was usedto detect the G4 spatial structure of the G4 oligonucleotide chain RNA (named as G4R) and its mutant of G4 (named as G4RM) by G base site-specific mutation.The HCV wild-type core gene G4(DNA) sequence was mutated as G4M-core by PCR site-directed mutagenesis without changing the amino acid codon.Then wild type and mutated core genes were constructed into the eukaryotic expression plasmid pcDNA3.1-Myc, and produced as pcDNA3.1-core-G4-WT (named as pG4) and pcDNA3.1-core-G4-M (named as pG4M), the expression of core protein was examined by Western blot. The mice were immunized with the pG4 and pG4M plasmids DNA respectively, and their humoral and cellular responses were examined. Results CD results showed that the structure of G4RM was changed compared to Wild type G4R, and the melting curve analysis showed the melting temperature of GR4M was lower than that of G4R, which indicates that G4RM structure is unstable. Western blot analysis showed that pG4M had much higher protein expression level compared to pG4(P<0.05). Analysis of animal immunization showed that pG4M induced increased levels of total IgG and IFN-γ compared to pG4(P<0.05). The IgG level of the pG4M group was 1.61 times higher than that of the pG4 group. By enzyme-linked immunospot(ELISpot)assay, we found that the release IFN-γ level of pG4M was 1.39 times higher than those of pG4. Flow cytometry showed that the intracellular IFN-γ production in the splenic CD4+ T cells was 1.79 times than those of pG4. Conclusion The G-quadruplex structure of HCV core can inhibit its protein translation. The mutation of G-quadruplex of core led to increased Th1-type immune responses. This is the first report demonstrate that HCV core G-quadruplex mutation can enhance its immunogenicity and could be used as a new strategy ofexploring HCV vaccine with enhanced immunogenicity.

OBJECTIVE To promote the management of nosocomial infections continuous improvement in the clinical and medical technology department.METHODS According to the regulation of nosocomial infections management,a handbook of nosocomial infections management for clinical and medical technology department was designed,and the monitors of nosocomial infection could perform real time inspection and record according require of the handbook.The department of nosocomial infections management examined the monitoring work of clinical and medical technology department every month and summarized every year,and the results were internalized to the valuation of medical quality management.RESULTS After 3 years of the usage of the handbook,the capability of the monitor groups of nosocomial infection and the quality of the all monitoring items were significantly improved;the qualified rates were all above 96.30%.CONCLUSIONS The handbook of nosocomial infections management is useful to improve the quality of nosocomial infections management in the clinical and medical technology department.

Objective To study the effect of rhein on the process of tubular epithelial-mesenchymat transformation in kidney of diabetic rats. Methods Wistar male rats were randomly assigned to 3 groups: Control group (N group, n=12),diabetic group(D group, n=12), rhein treatment group(R group, n=12).The rats of rhein treatment group were treated with daily intragastric administration of periment. The excretion of urinary protein and serum creatine were measured. Histological changes of renal tissue were observed by HE and MASSON stain. Immunohistochemistry was performed to investigate the expression of E-cadherin, α-SMA,FN and TGF-β1 in kidney. Results Compared with the control group, the tubulointerstitial injury and the accumulation of extraeellular matrix protein in diabetic models were obvious(P＜0.01).Compared with the control group, the expression of E-cadherin was decreased significantly and the expression of α-SMA,FN and TGF-β1 was increased significantly in diabetic group. E-cadherin was negatively correlated with TGF-β1(rs=-0.60,P＜0.05),α-SMA and FN was positively correlated with TGF-β1(rs=0.88,P＜0.05;rs:0.91,P＜0.01).In comparison with diabetic group,rhein could up-regulate the expression of E-cad and down-regulate the expression of α-SMA and FN in renal tubular epithelial cells(P＜0.01).Conclusion Rhein could protect kidney by ameliorating interstitial fibrosis in diabetic rats. The mechanism may be depend on down-regulating the expression of TGF-β1 and suppressing tubular epithelial-mesenchymal transformation.

Objective To explore the demands of nursing staff on nursing repository and provide reference for the develop-ment of nursing repository. Methods In-depth interview was conducted on 21 nursing staff by qualitative research. The themes were formed by category analysis. Results There were four themes about the demands of nursing staff on nursing repository:necessity to develop nursing repository ,contents of the repository ,forms of the repository and prospect of the reposi-tory. Conclusions Nursing staff need a nursing repository. They hope that the repository can provide comprehensive,concrete and practical knowledge,and provide a good interface with digitization. The design of repository should be consistent with in-ternational standards.