Guided by the study theory of constructivism,we utilize informational environment and resource to design the teaching of “nursing of mental disorders who abuse psychoactive substances”,change teaching methods divorced from reality,advocate situation teaching,recommend student' s initiative and cooperative study and improve the students' practical and innovative ability.

For many reasons,the size of jointed implement for central oxygen supply are different in some small and mediate hospitals,and some equipments can not used in other departments.In order to share mutual resource and make good use of all important equipments,we design a fast jointed implement for respirator joint.In the application of 24 cases,it shows convenient and effective for saving patients.

Objective: To find out effects of the service unit floating payment system under the total budget control of the medical institution, and explore feasible solution. Methods: Through interpretation of the policy, compare with simple service unit floating payment system to find out the characteristics of the new policy. Results: Through the total budget pre-control system, the new policy can effectively restrain the trend of fast growing on medical expenses. Time of doctor and number of patient ratio indicators are introduced to effectively prevent the medical institution decompose the time of doctor. Conclusion: To achieve the steady growth of medical business, medical institutions need to enhance the management of expense control, develop new business to attract new patients , optimize the medical charge structure.

Objective: To clone the Fab gene of a monoclonal antibody (mAb) BDI against human bladder cancer and its expression in E. coli. Methods: Fd and K genes of mAb BDI were cloned by RT-PCR and inserted into an Fab expression vector. Phage displaying Fab and soluble Fab were expressed in E. coli. The N-terminal sequence of VH region was corrected by PCR mediated mutagenesis. The antigen-binding characteristics of the Fab were tested by ELISA and immu-nohistochemistry. Results: Fd and K genes were cloned into the expressing vector p3MH and the phage displaying antibody and soluble Fab were expressed in E. coli, which showed weak binding activity to bladder cancer cells. Correction of the N-terminal sequence of the VHimproved the biding activity dramatically. The feasibility of the application of the Fab in phage antibody library screening was confirmed by a simulated panning procedure. Conclusion: The Fab gene of an anti-human bladder cancer mAb was expressed in E. coli. The importance of the N-terminal sequence on antibody binding activity was suggested.

Objective To establish the assay method of hydroxyproline(HYP) level in rat lung tissue for evaluating the lung fi-brosis degree .Methods The impact of different acid hydrolyzable time ,oxidative time ,developing time on the assay results of the HYP level in rat lung tissue was studied .On this basis ,the assay method for determining the HYP level was established and prelim-inarily applied in the detection of the HYP level in rat lung tissue .Results The optimal condition for the HYP level detection in rat lung tissue was as follows :7 .50 mol HCL was hydrolyzed for 16 h at 110 ℃ ,oxidized for 10 min at room temperature and devel-oped for 25min at 60℃ .The sensitivity of this method was 0 .067μg/mL .The recovery rate and the average CV of this method were 88 .85% -110 .88% and 4 .70% -6 .60% ,respectively .In the study of bleomycin induced rat lung fibrosis ,the HYP level of the model group was obviously higher than that of the control group .Conclusion This method has high sensitivity ,high recovery rate and good reproducibility ,and may be used as a reliable quantitative method to judge the lung fibrosis level in clinic .

Objective:To explore the methods and condition in which calcium ionorphore(CI) induces the CML cells to differentiate into dendritic cells(DCs).Methods:Mononuclear cells were separated from peripheral blood or bone marrow of CML patients whose WBC counts were more than 30?30~9 L~ -1 when samples were collected,then lymphocytes and monocytes were discarded by pouring out supernatant twice at different culture time point. Slightly adherent cells were cultured in RPMI1640 containing 10%FCS(Fetal calf serum),with or without CI(375 ng/ml) and GM-CSF(200 ng/ml) at 37℃,5%CO_2 humidified atmosphere for 96 h. To evaluate the effect of CI on inducing CML cells to differentiate into DCs,the phenotype of these cells were analyzed by flow cytometry and the morphology change was observed under inverted microscope and electron microscope. Better condition was also explored for DCs differentiation from CML cells under the effect of CI.Results:After 96 h of culture with CI and GM-CSF,the CML cells acquired morphology of mature DCs and significantly up-regulation of CD80,CD86,CD40,CD86 and HLA-DR.Conclusion:CML cells might acquire typical morphology and immunological phenotype of mature DCs when being cultured with CI and GM-CSF.

Objective To develop a vulnerable plaque targeting ultrasound contrast agent (UCA) and to evaluate its affinity and imaging performance in vitro.Methods E-selectin receptor-targeting UCA,which conjugated with monoclonal antibody of E-selectin,was prepared with filming-rehydration method and biotin-avidin linkage.The size and distribution of UCA were measured with particle size analyzer,the connectivity condition of microbubbles with E-selectin antibody was also detected with fluorescence analysis.The cytotoxicity from microbubble and ultrasound irradiation was evaluated through cell counting kit-8 (CCK8) assay.The adhesion effect of UCA was assessed after co-incubated with activated mouse endothelial cells (bEnd.3) and compared with that of free antibody intervention group and control group.The imaging performance of UCA at different time points was observed on an ultrasound equipment with a high-frequency transducer.Two-sample t test and one-way analysis of variance were performed to analyze the data.Results E-selectin receptor-targeting UCA was successfully prepared.The cytotoxicity result with CCK8 assay demonstrated the favorable biocompatibility of UCA.The connection amount of UCA on activated bEnd.3 cells ((6.23 ± 0.45) bubbles/cell) was significantly higher than that of the free antibody intervention group ((1.57±0.34) bubbles/cell) and control group ((0.07±0.03) bubbles/cell;F=291.43,P＜0.01).The performance of in vitro ultrasonography at the same time points showed no obvious difference between targeting UCA and control UCA (all t＜0.51,all P＞0.05).Conclusions The prepared E-selectin receptor-targeting UCA has favorable targeting and imaging capabilities.It might be a potentially ultrasound molecular imaging agent for early detection and prognosis evaluation of vulnerable plaque.

Objective To evaluate the growth inhibition and apoptosis of human monocytic leukemia THP-1 cell line by using 5,8-dimethyl-2-β-hydroxyisovalerylshikonin (SK36) and explore its preliminary mechanism. Methods CCK colorimetric assay and cell counting was used to examine the growth inhibition of shikonin on THP-1 cells. The apoptosis of THP-1 cells was detected by Annexin V/PI double labeling. The activation of Caspase-3 apoptosis pathway was determined by FCM. The apoptosis and the necrosis of THP-1 cells were detected by the laser scanning confocal microscopy. Results When the THP-1 cells were treated with SK36 at 1.02 μg/ml for 24 h and 48 h, the growth inhibition was dose-dependent. The cell apoptotic rate of THP-1 cells treated with 1.02 μg/ml evaluated by FCM with Annexin V/PI double labeling staining were (40.61 ±2.13) % and (67.40±9.15) % at 24 h and 48 h after treatment, respectively, which were significantly higher than that of the control group [(16.97±0.61) %] ([ = 18.444, t = 9.528, P <0.01). SK36 could induce THP-1 cells apoptosis involving the activation of Caspase-3 (F= 323.61, P<0.01). Conclusion SK36 can induce human THP-1 cells to undergo apoptosis, and its primary mechanism was to activate the Caspase-3.

AIM:To evaluate the effects of treatment with HP 1188-immunoglobulin yolk ( HP1188-IgY) on Helicobacter pylori ( H.pylori)-infected gastritis in BALB/c mice.METHODS:BALB/c mice were used to establish an animal model of H.pylori-infected gastritis, and the mice were divided into 8 groups (10 mice per group).Oral antibiotics were used in group 1, 1 mg HP1188-IgY in group 2, 1 mg HP1188-IgY plus 30%sucralfate in group 3, 5 mg HP1188-IgY in group 4, 5 mg HP1188-IgY plus 30%sucralfate in group 5, PBS in group 6, and 30% sucralfate in group 7 with the treatment once per day for 10 d;and 2.5 mg HP1188-IgY was injected hypodermically twice with a 48-h interval in group 8.Another 10 mice were used as normal control in group 9.The planting of bacteria in the stomach was assayed by bacteri-al culture, rapid urease test, PCR and pathological sectioning .RESULTS:Intragastric administration with 1 mg HP1188-IgY plus 30%sucralfate per day effectively cured the injury of gastric mucosa caused by H.pylori infection, and the effect has no significant difference compared with antibiotics (P>0.05).CONCLUSION:We establish a BALB/c mouse mod-el infected with H.pylori successfully.Sucralfate (30%) is an ideal protectant for HP1188-IgY, which might decrease H. pylori infection in the stomach of BALB/c mice by oral inoculation .

Objective The purpose of this study was to explore the risk factors of recurrence of atrial fibrillation (AF) after surgical radiofrequency ablation in patients undergoing cardiac valve surgery.Metbods Retrospective analysis was made in 255 cardiac valve patients undergoing concomitant radiofrequency modified Maze operation from April 2010 to December 2012 in Nanjing Drum Tower Hospital.Multivariate logistic regression analysis was used to screen independent risk factors for predicting recurrence of AF.Results Excluding 8 patients died after surgery,11 cases who were junctional rhythm after ablation or received pacemaker implantation and 60 lost case,176 patients were finished follow-up.107 cases maintained stable sinus rhythm (SR),whereas other 69 cases developed AF recurrence.Multivariate logistic regression analysis found larger left atrial size,higher B-type natriuretic peptide,lower preoperative heart rate and postoperative AF rhythm before discharge were independent risk factors for predicting AF recurrence.Conclusion Larger left atrial size,higher B-type natriuretic peptide and lower preoperative heart rate before surgery can impact outcome of surgical radiofrequency ablation in patients undergoing cardiac valve surgery after following 12(4-38)months.They can be used in clinical practice to choose more suitable cardiac valve patients with AF for surgical ablation.Postoperative appearance of AF rhythm before discharge indicated a poor prognosis,which should be interfered with drugs or electrical cardioversion.