The basic design plan of unified intelligence system with a demonstration effect was worked out using the system integration method, and its technical implementation was elaborated in aspects of enterprise service bus, database integration, platform service, and key business application from the operational point. Its extended appli-cation in clinical practice, scientific research, decision-making and hospital management was studied in order to further improve the hospital management level.

Objective To construct survivin-targeting siRNA-expressing plasmid.Methods DNA sequence correspond to siRNA targeting survivin was designed and synthesized,and cloned into plasmid pRNAT-U6.1/Neo to produce surviving-targeting plasmid.Two oligos in the template with cohesive BamHⅠ and HindⅢ sites were prepared and annealled to form the insert fragment for siRNA vector.The vector was cut with BamHⅠ and HindⅢ and ligated with the insert fragment using T4 ligase.The recombinant vector was confirmed by restriction digestion and DNA sequencing,and then was transfected into T24 cells with Lipofectamine TM2000 and the expression of survivin was detected by real-time quantitive PCR.Results DNA sequencing for the PCR product showed that the recombinant vector pRNAT-U6.1/Neo-survivin was successfully constructed without any base pair mutation.The plasmid pRNAT-U6.1/Neo-survivin could efficiently reduce the expression of survivin and confer G-418 resistance in T24 cells.Conclusion The siRNA-expressing plasmid which were successfully constructed and transfected into T24 cells in this study may facilitate the application of RNA interference technique,and lay foundation for further studies on the function of survivin.

Objective To investigate the elasticity reduction of liver tissue due to microwave ablation and the relevance to the histographic damages. Methods An experimental study using fresh porcine liver was designed. Elasto-ultrasonography scanning both in color display and strain ratio calculations was conducted before and 5 min after microwave ablation ( 2450 MHz) in manner of antenna insertion under ultrasound guidance to determine the alterations of the liver elasticity, in correspondence with the histopathologic assessment of each ROI. Results Elasto-ultrasonography showed a significant elasticity reduction and hardness augment of the targeted liver tissue and the corresponding histopathology revealed increases in the amount of massive coagulative necrosis and coking of liver cells after microwave irradiation,in proportional to the applied field power and working time. Conclusions Elasto-ultrasonography helps to demonstrate microwave-induced lesion in porcine liver got rapidly hardened. It is possible to estimate the tissue necrosis to the changing of tissue hardening.

AIM To investigate the effects of tranfection of IL 2R antisense RNA expression plasmids on mouse spleen cells' proliferation in vitro and its possible mechanism. METHODS Spleen cells were transfected with IL 2R antisense RNA eukaryotic expression plasmids using adhesion assisted lipofection method, and then the spleen cells were stimulated by mitogen. Cells' proliferation was tested by tetrazolium salt (MTT) method. IL 2R mRNA and protein expression level were measured by slot blot hybridization assay and flow cytometry method respectively. RESULTS The proliferation of spleen cells was inhibited obviously after transfecting with recombinant plasmids. The inhibitory rate of pcAnti mIL 2R?? and pciAnti mIL 2R?? transfected group was higher than that of pcAnti mIL 2R? and pcAnti mIL 2R? transfected group; the inhibitory rate of pcAnti mIL 2R? tranfected group was higher than that of pcAnti mIL 2R? tranfected group. No inhibitory effect on the growth of NIH3T3 cells was observed when they were transfected with recombinant plasmids. IL 2R mRNA and protein expression level were decreased in spleen cells after transfection of recombinant plasmids. CONCLUSION IL 2R antisense RNA can efficiently inhibit the proliferation of mouse spleen cells in vitro. IL 2R?? chimeric antisense RNA showed higher inhibitory rate than ? or ? antisense RNA. IL 2R? antisense RNA was more effective than ? antisense RNA. It can be concluded preliminarily that the inhibitory effect of IL 2R antisense RNA was exclusively on the growth of cells functionally expressing IL 2R. The inhibitory effect on the spleen cells proliferation was likely due to the blocking of IL 2R expression by antisense RNA.

Objective To explore the application of a three-dimensional simulation system in extended liver resections.Methods A three-dimensional surgical simulation system was used for preoperative assessment and for computer simulation to estimate the resected liver volume,the residual liver volume,and the surgical resection margins.The software virtual liver resection was used to choose the most optimal surgical option.The actual resection,the postoperative liver volume and the actual surgical resection margins were compared.Results 1 patient was diagnosed by ultrasound to develop ascites on post-operative day 2.Another patient was diagnosed by transthoracic ultrasound to have moderate pleural effusion on postoperative day 1.Postoperative liver failure developed in 1 patient.There was no patient who developed bile leakage.The average length of stay was 6 ～ 85 days (average 22 d).There was no recurrence within 1 month after surgery and there was no postoperative deaths.Condusions The three-dimensional surgery simulation system was effective.It gave an accurate evaluation and simulation of the liver surgery.In extended liver resection,it gave good guidance and help.

Objective CD8 +T cells increased in the airway of patients with chronic obstructive pulmonary disease and exis -ted constantly .The aim was to investigate the role of CD 8 +T-cells in rats with chronic bronchitis ( CB) which was induced by cigarette smoking and intratracheal injection with lipopolysaccharide ( LPS) . Methods 18 health Wistar rats were radomly divided into sham smoking group(group A), CB group(group B) and N-acetylcysteine prevention group (group C).The rats in group B and group C re-ceived intratracheal injection with LPS twice and exposed to cigarette smoking for 4 weeks to induce CB model .The rats in Group C re-ceiving intragastric administration with N-acetylcysteine (NAC)(200mg/kg) before received LPS and smoking.Group A was the sham smoking group.The lung tissue of all rats were stained by HE then evaluated about pathological scores .The expression of nuclear fac-tor-κB (NF-κB), major histocompatibility complex class I (MHCI), CD8 +T cell and Vascular endothelial growth factor (VEGF) in airway were detected by immunohistochemisty which was stained by labeled streptavidin biotin method . Results The pathological scores of airway ( 10 .83 ±3 .31 ) in group B were higher than (1.17 ±2.40) in group A(P <0.05).The pathological scores of airway(4.66 ±2.25) in group C were less than (10.83 ±3.31) in group B(P <0.05).The expression of NF-κB(4.84), MHC I (2.48),CD8 +T cell(5.35)and VEGF(5.02) in airway increased in group B when compared with (1.18, 1.25, 1.33) and (1.18) in group A respectively(P <0.05).The expression of NF-κB (2.18), MHC I(1.46),CD8 +(2.35)and VEGF(2.02) in airway decreased in group C when compared with (4.84), MHC I(2.48),CD8 +T cell(5.35)and VEGF(5.02) in group B respectively (P<0.05 ). There were positive correlations between the expression of NF-κB, MHC I and CD8 +T cells in airways(r=0.670, r=0.701, respec-tively, all P<0.01).There were positive correlations between the expression of CD 8 +T cells and VEGF the pathological scores of air-ways(r=0.689, r=0.782, respectively, all P<0.01). Conclusion NAC can inhibit airway inflammation which is regulated by CD8 +T-cells and VEGF through suppressing the expression of NF -κB and MHC I.

Objective To evaluate the value of ATP content of CD4+ T lymphocytes in the diagnosis of infection and its correlation with drug concentrations in renal transplant recipients.Methods 45 renal transplant recipients were reviewed from May 2010 to October 2011.There were 33males and 12 females,aged from 21 to 58 years old.The recipients were divided into non-infection group (n =34) and infection group (n =11) according to their clinical manifestation.11 cases of infection were diagnosed by the chest X-ray,CT imaging manifestations and etiological examination,among them 5 cases were pulmonary infection,4 cases were upper respiratory infection,1 case was urinary tract infection and 1 case was perineal abscess.23 healthy volunteers were enrolled as the control group.They were detected ATP content of CD4+T lymphocytes by Immuknow method.Thetrough concentrations of the FK506 and CsA were detected by microparticle enzyme immunoassay and fluorescence polarization immunoassay,respectively.The hs-CRP concentration was detected by immunoturbidimetry.Results The ATP content of CD4+ T lymphocytes of the control group,non infection group and the infection group were (295±74) μg/L,(35± 189) μg/L and (212± 155) μg/L respectively.The levels of ATP of infection group were obviously lower than the control group and non-infection group.There were statistically differences (P ＜0.05).24 recipients were followed up dynamicly.There were 4 cases whose ATP value was lower than the postoperative average levels in 5 infection recipients.The hs-CRP concentration of infection group were (12.4±4.8) mg/L,obviously higher than the non infection group's (3.3 ± 4.7) mg/L and the control group' s (0.5 ± 0.5) mg/L.There were statistically differences (P＜0.05).The ATP content of CD4+ T lymphocytes were no significant associated with drug trough concentrations (P＞0.05).Conclusions Low ATP level after renal transplantation is a risk factor for infection recipients.Immuknow cell function assay can make up for the inadequacy of the drug concentration monitoring,reduce the risk of infection,and guide clinical immunosuppressive adjustment.

BACKGROUND: Studies have demonstrated that incidence rate of acute rejection in renal transplant recipients with pre-production of major histocompatibility complex class I chain-related gene A (MICA), including parts of autoantibody, before transplantation in body, is obviously greater than that of recipients with negative antibody. OBJECTIVE: To investigate effects of exogenous antigen on MICA expression in endothelial cells. METHODS: The endothelial cells were cultured with exogenous recombinant MICA protein (group M5, M10 and M25) and heat shock protein-70 (group H5, H10 and H25) with dosages of 5, 10 and 25 μg/L, respectively, for 48 hours. Same volume of phosphate buffer saline was added into the control groups. RESULTS AND CONCLUSION: At 48 hours after induction, the expressions of MICA mRNA and protein were increased significantly in each experimental group (M5, M10 and M25) than that of the control group with significant (P < 0.05). The expression of MICA mRNA and MICA protein of group M5 and group M10 were remarkably higher than group M25 (P < 0.05); however, there was no significant difference between group M5 and M10 (P > 0.05). The expression of MICA membrane protein in the group M10 was obviously greater than that of the group M5 and M25 (P < 0.05). The level of soluble MICA (sMICA) in experimental groups (M5, M10 and M25) was decreased obviously comparing with that of the control group. These differences had statistical significances (P < 0.05). But there was no significant difference among the experimental groups (P > 0.05). However, the expression of MICA gene and sMICA level did not change after heat shock protein-70 stimulation. The exogenous MICA antigen up-regulates the expression of MICA mRNA and protein, especially increases the expression of membrane protein on the cell surface significantly, but sMICA in supernatant was dramatically decreased.

Objective To detect the gene expression of PCA3 and PSA in peripheral blood and urine simultaneously to investigate whether PCA3 combining PSA gene could become new markers for diagnosis of Pca. Methods From June 2009 to December 2009,the initial urine after prostatic massage and the peripheral blood specimens were collected from 37 patients with PCa and 68 patients with BPH that were pathologically confirmed,g patients with urinary stone were used as normal control,the expression of PCA3 and PSA mRNA of mononuclear cells in urine sediments and peripheral blood were detected by fluorescence real-time quantitative PCR,with β-actin mRNA as internal control. Results The sensitivity and specificity of the expression of PCA3 mRNA in peripheral blood for diagnosis of prostate cancer were 48.6％ and 100％ respectively.ROC curve analysis was performed for the PCA3 score and the area under the ROC curve was 0.908.Using 64.6 as the cutoff,the sensitivity was 81.1％ and the specificity was 86.8％.In group with serum tPSA value ＜4 pg/L,the positive rate and negative rate of urinary PCA3 score for diagnosing prostate cancer were 80％ (4/5) and 89.4％ (20/22) respectively.In group with serum tPSA value 4 - 10 μg/L,the positive rate and negative rate of urinary PCA3 score were 66.7％ ( 2/3 ) and 84.2％(16/19) respectively.In group with serum tPSA value ＞ 10 μg/L,the positive rate and negative rate of urinary PCA3 score were 82.8％ (24/27) and 81.5％ (22/27) respectively.The sensitivity of simultaneous detection of PCA3 mRNA in peripheral blood and urinary PCA3 score was 86.5％. Conclusions The expression of PCA3 mRNA in peripheral blood was a specific marker for the diagnosis of PCa.The simultaneous detection of PCA3 mRNA in peripheral blood and urinary PCA3 score could increase the sensitivity for the diagnosis of PCa.

Through the construction practice of the optic nerve disease picture database,the paper discusses the system architecture,database field,data content,picture processing,organization and implementation,and other issues about the construction of the clinical case picture database,states and analyzes the operation effect,points out deficiencies,and provides reference for the construction of relevant picture databases.