The application of simulated therapy in surgical clinical teaching can stimulate the students' learning enthusiasm and improve their unified qualities in all-rounded way,improve the teacher's teaching level to acquire satisfactory results.

Objective:To investigate the methods of the anesthetization of the foreign body extraction in respiratory tract of infants.Methods:This clinical study compared two anesthetic methods in foreign body extraction of respiratory tract of infants:Ketamine+Sodium r-hydroxybutyrate intravenous (group A) and Ketamine+propofol intravenous (group B) and analyzed the advantages and disadvantages of both methods.Results:Compared with group B,the extent of the changes of heart rate and blood oxygen saturation in patients of group A is littler,and the incidence rate of breath holding and asphyxiation is littler too.Conclusions:The anesthetic effect of Ketamine+Sodium r-hydroxybutyrate intravenous in better than that of Ketmaine+propcfol intravenous.

Objective To construct contains human interleukin-10 gene recombinant lentiviral-vector(LV-IL-10)and to form a basis to further explore the therapy of chronic pain.Methods hIL-10 gene fragment was isolated and amplified from pCYIL-10 plasmid by PCR,and was cloned into pWPXL-GFP.The inserted hIL-10 fragment was verified by Pme I digestion and DNA sequencing.The recombinant plasmid pWPXL-IL-10-GFP,envelope plasmid pMD2.G and packaging plasmid psPAX2 were cotransfected into 293T cells,to pack out lentivirus particle that has the ability of duplicated-deficiency,then virus titer determination was undertaken.Results The 530bp IL-10 gene fragment was amplified from pCYIL-10 plasmid by PCR,and was recombinated into pWPXL-GFP plasmid.DNA sequencing confirmed that the cloned gene segment was 100% homologous to the published hIL-10 sequence in genebank.High titer(2?1010)and highly purified lentiviral particles was obtained.Conclusions The lentivirus vector LV-hIL-10 was constructed successfully,which form a basis of research of chronic pain therapy.

Objective To investigate the intervention effect of lentivirus expressing human IL-10 (LV-hIL-10) on activated astrocytes.Methods DI TNC1 cell line was treated with different concentrations of lipopolysaccharide (LPS) and time points.The expression of proinflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukinb-1β (IL-1β) was detected by RT-PCR and ELISA.Moreover,the effect on the expression of proinflammatory cytokines TNF-α and IL-1β was analyzed in DI TNC1 cell lines infected with and without LV-hIL-10.Results The expression of TNF-α and IL-1β was increased in DI TNC1 induced by LPS.The expression of IL-10 was upregulated in DI TNC1 infected with LV-hIL-10.TNF-α and IL-1β were inhibited by IL-10 overexpression in DI TNC1 actived by LPS.Conclusion DI TNC1 is activated by LPS and secretes proinflammatory cytokines TNF-α and IL-1β as immune-like cells,and these activation is inhibited by hIL-10 overexpression.

Objectives To analyze pathogen distribution and antimicrobial resistance of bacilli among children with otitis media. Methods Pathogenic bacteria was isolated from children with suppurative otitis media. The VITEK32 was used for iden-tification. The bacterial susceptibility testing was done by Kirby-Bauer method. According to CLSI standard the antimicrobial susceptibility was determined. Results From Jan 2010 to Dec 2012, 425 children with suppurative otitis media were examined. 347 strains were isolated, of which the detectable rate was 81.65%. The detectable rate of bacteria and fungus was 93.37%(324/347) and 6.63%(23/347), respectively. Among bacteria, the detectable rate of streptococcus pneumoniae was 40.92%(142/347) and staphylococcus aureus was 33.43%(116/347). The detectable rate of haemophilus influenza was 7.78%(27/347). The preva-lence of streptococcus pneumoniae is high in children aged 1-3years, with detectable rate at 47.09%. There was no statistical dif-ference among different age groups. The prevalence of methicillin-resistant staphylococcus aureus (MRSA) in middle ear secre-tion was 1.11%(5/45), 18.75%(9/48)and 30.43%(7/23)in 2010, 2011 and 2012 respectively, with no statistical difference (χ2=3.86, P=0.145). The prevalence of penicillin-resistant streptococcus pneumoniae (PRSP) in middle ear secretion was 9.26%, 3.92%and 27.03%in 2010, 2011 and 2012 respectively, with statistical difference (χ2=11.47, P=0.003). Conclusions Choosing correct therapy according to the result of middle ear secretion culture and antibiotics sensitive test can increase the recovery rate of otitismedia.

Objective: To prepare an immunoconjugate of anti-EGFR monoclonal antibody (ETS) and adriamycin(ADR) and to investigute it's inhibitory effect on human epidermoid carcinoma (A431) in vitro and in vivo and it's side effect on C57BL/ 6J mice.Methods:The immunoconjugate(ETS-ADR) was prepared with an improved glutaraldehyde conjugate method and the cytotoxicity effects of EQ75-ADR on A431 cells and Wish cells were measured by MTT assay. A431 tumor-bearing nude mice and C57BL/6J experimental model were established and were used for testing the inhibitory edicts of EQ75-ADR. The side effect of EQ75-ADR was compared with ADR. Results:The EQ75-ADR showed specific cytotoxicity on A431cells,the ytotoxicity of EQ75-ADR was 50-fold, 14-fold and 10-fold more efficient than that of EQ75, ADR and mixture of EQ75 and ADR (EQ75+ ADR) respectively.The effect of EQ75-ADR on with cells was 10-fold less efficient than that of ADR and EQ75+ADR. As compared with the cytoxicity effect of EQ75-ADR on Wish cells with A431 cells, it showed 300-fold less efficient.Similar cytotoxicity results were shown in 2 h treatment experimental group. The EQ75-ADR exhibited significant anti-tumor activity on tumor-bearing nude mice,and the inhibition late is 92.2% (P

AIM: To investigate the effect of autologous bone marrow mononuclear cells (BM-MNCs) local transplantation (BM-LT) on a rabbit model of hindlimb ischemia. METHODS: After unilateral femoral artery and its branches were surgically ligated and excised in rabbits (n=14) 1 hour or so, autologous BM-MNCs were injected into the ischemic skeletal muscles in 7 rabbits, and phosphate buffered saline for another 7 ones as control. Hemodynamics parameters were measured at day 0, day 7 and day 28, respectively. On day 28, the animals were sacrificed in groups for pathological analysis. RESULTS: Compared with control group, color Doppler flow image in BM-LT group was recovered significantly, and its' systolic peak velocity and end-diastolic velocity were also improved (P

Objective To establish a flow cytometric measurement of detecting minimal residual disease(MRD) according to the leukemia-associated immunophenotypes in children with acute lymphoblastic leukemia(ALL) and to explore the significance of MRD detection in ALL children for a individualized treatment. Methods A variety of four-color fluorescent antibody combinations were used to investigate the children's normal bone marrow. The normal bone marrow pattern at two-parameter plots was established to identify the residual tumor cells, seventy-five bone marrow samples from newly diagnosed ALL children were analyzed with four-color cytometry to determined the optimal combinations which can clearly distinguish the tumor cells from normal cells. The bone marrow samples were monitored with the combination panel in 60 patients at the end of induction therapy and follow-up treatment. Cytomorphology test, PCR amplification of 29 fusion genes as well as IgG and TCR gene rearrangements were performed simultaneously. Results Sixty-nine cases (92.0%) could be identified for effective antibody combinations to monitor MRD by four-color cytometry. Fusion genes or IgG and T cell receptor (TCR) gene rearrangements can be detected in 21 cases (28.0%) to monitor MRD by PCR. No MRD can be detected in 25 bone marrow samples at the end of induction therapy and follow-up treatment. Four-color cytometry could detect as low as 0.021%-4.130% residual leukemia cells. Conclusion MRD can be monitored by flow cytometry which is faster than PCR, and the sensitivity is superior to morphology method.

Objective To investigate characteristics of cage migration after transforaminal lumbar interbody fusion (TLIF) and related risk factors.Methods A retrospective study was conducted to review cage migration in 512 patients who had undergone TLIF procedure from January 2010 to June 2011 in 5 spinal research centers.There were 255 males and 257 females,aged from 37 to 77 years (average,54.7 years).All patients were followed up at 3,6,12 months after operation.The clinical outcomes were evaluated using the visual analogue scores (VAS) and Oswestry disability index (ODI).X-rays and 3D CT scans were used to analyze the incidence and related risks factors of cage migration in these patients.Results Cage migration was found in 6 of 512 patients,the total incidence was 1.17％.Significant difference was found between each center.Cages with different shapes had different incidence.The analysis showed that the incidence of migration of rectangular-shaped cage (3.11％,5/161) was significantly higher than that of kidney-shaped cage (0.28％,1/351).The cage in double-segment TLIF (5.75％,5/87) was easier to migrate than that in monosegment TLIF (0.24％,1/425); furthermore,linear type endplate(3.50％,5/143) was remarkably easier to migrate than concave-concave one (0.27％,1/369).Conclusion Difference in operative skills,cage shape,number of fused segments,adjacent endplate shape,and lumbar spondylolisthesis might be risk factors for cage migration after TLIF.

Objective To establish an animal model of rapid eye movement (REM) sleep deprivation (SD) and an animal model for perifornical nucleus microdialysis and investigate the change of cognition, hypocretinergic system and GABAergic system in rats' hypothalamus after various degrees of REM sleep deprivation and sleep revival and two GABAergic drugs intervention. Methods The modified multiple platform method (MMPM)was used to establish sleep deprivation model and the cognitive function was assessed by Morris' water maze. Immunofluorescence technique was used to analyze the number of Hypocretin (Hcrt) immunoreactive neurons, total Fos immunoreactive neurons, Hcrt and Fos colabeled neurons, and the integrated optical density ( IA ) of GABAA Rαl immunoreactive area in rats' hypothalamus.High performance liquid chromatograph (HPLC) was used to quantitatively analyze the level of GABA and Gluin in the rats' hypothalamus. Two GABAergic drugs, a selective GABAA R antagonist, SR-95531, and a selective blocker of type 1 GABA transporter (uptake blocker), NO-711, were used for perifornical nucleus microdialysis. Results There was no statistically significant difference in tests between CC and TC ( Define CC and TC). There was a significant decrease (P ＜ 0. 05 ) of cognitive function measured by Morris maze test in SD 3 d, SD 5 d and RS 6 h of SD groups compared with CC and TC groups. Number of Fos immunoreactive, F+ &H+ immunoreactive neuronsand IA of GABAA Rαl immunoreactive area were all significantly increased ( P ＜ 0. 05 ). Content of GABA measured by HPLC was also increased ( P ＜ 0. 05 ). However, all these changes were partly reversed by sleep revival SR-95531 and NO-711 had different effect on these changes. Conclusions Sleep deprivation, no matter mild or severe, has adverse effects on cognitive function. Activities of both GABAergic and Hcrtergic system are increased during REMSD. These two neurons system could be regulated by each other and the relationship between them is positive correlation. GABAergic system also had self-regulation during REMSD, but microdialysision of either SR-95531 or NO-711 acquired adverse effects on cognitive function of rats. So GABAergic system is not an optimal therapeutic target. Because GABAergic and Hcrtergic system has inhibitory effect on each other,suppressing activity of Hcrtergic system might be a promising therapeutic target.