Objective To discuss the indications and complications of primary closure of bile duct incision in laparoscopic bile duct exploration and balloon dilatation catheter dilatation to treat the papillary stenosis and the intrahepatic bile duct stenosis. Methods A pospective study of 42 ptients of bile duct incision closure primary in laparoscopic bile duct exploration and balloon dilatation catheter dilatation, laparoscopic bile duct exploration and extraction of bile duct stones with choledochotomy was first adopted in order to clear the stones, then followed by the balloon dilatation catheter(explosive pressure reached 2020 kPa, used 505kPa) to dilate the papillary stenosis and the intrahepatic bile duct stenosis (CT-7542～ CT-75104) until the stenosis was released. Whether the primary closure of duct incision was selected or not, it was based on the situation of intraoperative choledochoscopic exploration, if it had been selected, the closure of bile duct incision would accepted by using absorbable suture 4-0 or 5-0, without placing bile duct drainage.It was routinely to place the drainage tube in the oriffice of the lesser omentum. Results 41 out of 42 patients had obtained successful duct clearance, the dilatation of the stenosis to reach the expected expansion and without bile leakage. One patient had bile leakage about 30-150 ml daily persisted for 4 days through cured conservatively. Conclusion Eventually it was safe and effective for some patients who had completed successful duct clearance and the dilatation of the stenosis to reach the expected expansion with the balloon dilatation catheter. They were adopted to the primary closure of duct incision using absorbable suture and did not need to place bile duct drainage.

Objective To investigate the expression of receptor tyrosine kinase EphB2 and its relationship with peritumoral brain edema (PTBE) in meningiomas and to explore the possible mechanism of PTBE in meningiomas. Methods A total of 44 meningioma tissue samples from patients undergoing surgery and 14 normal meninges samples as the control were collected for the examination of the immunoreactive EphB2 expression. The expressions of EphB2 in different types of meningiomas were determined by immunohistochemical method. Peritumoral brain edema was assessed on a subjective 5 point(0～4) scale based on the data from preoperative magnetic resonance imaging scan. Results ① The expressions of EphB2 in 44 cases of meningiomas were: scale 4 in 7 cases, scale 3.5 in 5 cases, scale 3 in 6 cases, scale 2.5 in 2 cases, scale 2 in 8 cases, scale 1 in 13 cases, scale 0 in 3 cases, mean scale 2.19 . ② No expression of EphB2 was found in 10 out of the 14 normal meninges samples, but minimal (scale 1) expression of EphB2 was found in only 4 cases. ③ Preoperative MRI results showed different levels of PTBE in 33 out of 44 cases. Conclusion There is expression of EphB2 in the cytoplasm in most meningiomas. The expression of EphB2 in meningnioma is positively correlated with peritumoral brain edema, suggesting that EphB2 participates in PTBE of meningiomas.

Objective To evaluate if 18F-FLT PET imaging could be used as a new clinical method to predict tumor radiosensitivity.Methods MDA-MB-231 and LN229 cells were irradiated with doses of 0,8 and 16 Gy of 6 MV photon energy,then soft agar assay and cellular uptake of 18F-FLT were performed on the 2 cell lines.The t test and one-way analysis of variance were used for the two groups and data before and after irradiation.The MDA-MB-231 and LN229 tumor xenografts were prepared by injecting the tumor cells into the right limbs of female BALB/c nu/nu mice.Once tumors reached a diameter of 10 mm,the two types of mice were divided randomly into 3 groups (20 mice per group) according to the irradiation doses (0,8 and 16 Gy).After irradiation,18F-FLT PET imaging and immunohistochemical staining were conducted.Then correlations between 18F-FLT SUVtumor/SUVmuscle ratio (T/M ratio) and TK1 labeling index percentages (LITK1) were tested using linear correlation analysis.Results The survival fraction of MDA-MB-231 and LN229 cells after irradiated with 8 Gy were (59.73 ± 4.3) ％ and (93.41 ± 3.75) ％,respectively (t =-13.20,P ＜ 0.001).When the dose increased to 16 Gy,the survival fraction decreased to (43.57 ±4.06) ％ and (81.77 ± 4.42) ％,respectively(t =-14.24,P ＜ 0.001).In MDA-MB-231 cells,the cellular uptake of 18F-FLT after irradiation with 8 Gy declined rapidly to (18.32 ± 1.38) kBq/105 cells ((128.22 ± 8.24) kBq/105 cells with the dose of 0 Gy,F =266.41,P ＜ 0.01),and maintained this low level till 72 h.For the LN229 cells,the cellular uptake decreased to (9.87 ± 1.30) kBq/105 cells after 8 Gy irradiation ((134.88 ± 6.59) kBq/105 cells with the dose of 0 Gy,F =346.06,P ＜ 0.01),then increased gradually to (127.17 ± 9.08) kBq/105 cells at 72 h (F =346.06,P ＞ 0.05).The dynamic changes of 18F-FLT cellular uptake in the two cells had the same pattern after being treated with 16 Gy irradiation.In the 18F-FLT PET image of MDA-MB-231 tumor mice after 8 Gy radiotherapy,the T/M ratio decreased to 0.78 ± 0.39 at the first day,but it was 2.84 ± 0.29 before radiotherapy (F =39.78,P ＜0.01).Then the ratio increased slowly,and it was still lower than the baseline at 7 d after radiation (F =39.78,P ＜0.01).The same pattern could be seen in the group of 16 Gy irradiation.In LN229 tumor mice treatment with 8 Gy irradiation,the T/M ratio increased to 2.41 ±0.47 at the first day,and it was 1.58 ±0.29 before radiotherapy (F =34.01,P ＜ 0.05).The ratio decreased steadily to 0.66 ± 0.32 (F =34.01,P＜0.05) at 7 d after radiotherapy.However,in the treatment group with 16 Gy,the T/M ratio decreased gradually and reached 0.44 ± 0.22 at 7 d (F =41.85,P ＜ 0.01).A correlation was found between 18F-FLT T/M ratio and LITK1 (8 Gy:r=0.67,0.73; 16 Gy:r=0.73,0.69; all P＜0.01) in both tumor models.Conclusion 18F-FLT PET imaging may be used as a new assay to predict tumor radiosensitivity,but further investigation is needed before clinical application.

Objective To investigate the risk factors for postoperative nausea and vomiting (PONV) in spinal anesthesia patients.Methods A total of 841 patients received spinal anesthesia were visited after operation.Data were analyzed using univariate analysis and multivariate Logistic regression to identify risk factors related to PONV.Results PONV occurred in 94 patients (11.2％,94/841).Univariate analysis showed that PONV was unrelated with gender,age,ASA classification,anesthesia mode (P ＞ 0.05),related with operation department (P =0.026),body mass index (P =0.020),education level (P =0.000),history of previous surgery anesthesia (P =0.005),history of PONV (P =0.000),history of kinesia (P =0.002),smoke (P =0.019),intraoperative using of tramadol (P =0.018).Multivariate analysis showed that operation department (OR =4.039,95％ CI 1.331-12.259,P=0.048),education level (OR =3.504,95％ CI 1486-8.260,P=0.015),history of PONV (OR =5.113,95％ CI 1.790-14.606,P =0.002),intraoperative using of tramadol (OR =5.316,95％ CI 1.091-25.908,P =0.039) were identified as independent risk factors for PONV.Conclusions The independent factors associated with PONV following spinal anesthesia include operation department,education level,history of PONV,intraoperative using of tramadol.Identifying patients who are at high risk for PONV will enable the formation of more timely management project.

Objective To investigate efficiency of centipede extracts on apoptosis induction, proliferation inhibition to Human A549 cell line and growth suppression of subcutaneous transplanted sarcoma in nude mice. Methods Centipede extracts prepared by enzymolysis and acetone precipitation methods were used to treat human lung cancer A549 cell line. Proliferation inhibition was evaluated by MTT assay and half inhibit concentration (IC50) was calculated. Cell morphological change and apoptosis were detected by flow cytometry and Hoechst stain. The subcutaneous transplanted sarcoma models were prepared with nude mice and randomly divided into model group, control group and centipede extracts group, with 10 mice in each group. Changes of tumor volume, quality and anti-tumor rate were observed.Results In vitro experiment, proliferation of A549 cells was inhibited with dose-dependency and IC50 value was 0.603 mg/mL. The G0/G1 phase of cells was down regulated and G2/M and S phase cells were up-regulated. The apoptotic character cells were been found by Hoechst stain. In vivo experiment, the tumor weight and volume decreased significantly compared with model control group, with statistical significance (P<0.01).Conclusion The centipede extracts shows dose-dependent anti-proliferative effect on A549 cells, which can induce apoptosis by arresting A549 cells at G2/M phase and suppressing growth of subcutaneous transplanted sarcoma of lung cancer in nude mice.

BACKGROUND:Schwann cell is one of the major seed cells In peripheral nervous system and plays an important role in neural injury and neural disease.However,the source of Schwann cells is limited.And the purity of Schwann cells is affected due to the pollution of fibroblasts.Many purified methods have been proposed,but every one has its defect to satisfy the clinical demand.OBJECTIVE:To compare the differences among differential adhesion purified method,cold jet purified method,immunomagnetic beads selection purified method and G418 selection purified method to purify Schwann cells of neonatal rat in vitro.METHODS:Bilateral sciatic nerves of SD rats were harvested under sterile condition.Schwann cells were purified respectively using differential adhesion purified method,cold jet purified method,immunomagnetic beads selection purified method and G418 selection purified method.Cell viability was compared,and cell purity was determined by immunohistochemistry.RESULTS AND CONCLUSION:The purity of Schwann cells separated by differential adhesion method was low,but the viability was fair.The purity and viability of cells following cold jet method immunomagnetic beads selection method was high.The purity of cells separated by immunomagnetic beads selection methods was similar to that of cold jet method immunomagnetic beads selection method,but the cell viability was worse.The cell viability following G418 selection method was bad,but the purity was high.

BACKGROUND:Pore diameter,size and porosity of biomaterials are important for cell attachment,infiltration and growth.OBJECTIVE:To primarily evaluate the biocompatibility of novel bionic scaffold of poly(3-hydroxybutyrate-co-3-hydroxyvalerate)/sol gel bioactive glass (PHBV/SGBG) with nanostructure in vitro.DESIGN,TIME AND SETTING:Cell-material experiment was performed at the Central Laboratory of Third Hospital of Sun Yat-sen University from March to September 2006.MATERIALS:PHBV/SGBG was provided by the Institute of Materials Science and Engineering,South China University of Technology;bone marrow stroma cells (MSCs) were prepared by our team.METHODS:The standard concentration of PHBV/SGBG extracting solution should be 10 mL/ cm2 of the ratio of culture solution to material surface.PHBV/SGBG was immersed into the complete culture solution and incubated in 5% CO2 at 37 ℃ for 2-3 days.The extracting solution was drawn and stored under sterile condition.In addition,PHBV/SGBG extracting solution of 8,4,2,1,0.5,0.25 and 0.125 times of standard concentration was prepared.MAIN OUTCOME MEASURES:PHBV/SGBG ultrastructure was observed by scanning electron microscopy;PHBV/SGBG porosity was measured by routine measurement.MTT methods were used as a quantitative assessment for cytotoxicity of the biomaterials.Adherence and spreading of MSCs on the surface of specimen was observed using direct contact cultivation.RESULTS:The PHBV/SGBG was porous,with connecting pores under electron microscopy.Nanometer SGBG particles were imbedded or encapsulated in pore wall of PHBV,with the porosity ＞90%.The toxicity gradation of the novel bionic scaffold ranked from grade 0 to 1 at 1,3,5 days of culture.MSCs slightly attached and grew on the surface of the biomaterials,and proliferated rapidly.Obvious cell processes stretched into the micro-pores structure.CONCLUSION:The novel bionic scaffold of PHBV/SGBG has excellent cellular affinity,possibly due to the porous structure,with no cytotoxicity to MSCs.

Objective To detect the changes of the brain white matter microstructure at the acute stage of posttraumatic stress disorder(PTSD)resulting from a single,extreme and long-lasting trauma.Methods DTI scans were performed on 1 7 survivors of coal mine disaster(PTSD group)and 1 7 cases of normal control(control group).The differences of the mean diffusivity(MD)values measured from the whole brain DTI between the two groups were analyzed based on tract based spatial statistics (TBSS).MD data were statistically compared between the two groups based on nonparametric random permutation test(RPT),and the brain areas of significant differences between the two groups were obtained.Results Compared with the control group,MD values were increased in the bilateral rostral corpus callosum body and left precorona radiata,and decreased in the bilateral superior and posterior corona ra-diate,posterior limb of the left internal capsule,left cerebral peduncle and left thalamic.The differences were statistically significant (P <0.01 TFCE-corrected).Conclusion TBSS is a comprehensive and accurate method for evaluate the changes of brain white mat-ter in PTSD cases.TBSS can provide an objective basis of the pathological brain neural structures imaging for early diagnosis and in-tervention of PTSD.

Objective To investigate the protective effects of the 14-3-3 protein overexpression on the injury of PC12 cell induced by MPP~+ and its mechanisms.Methods For expression in mammalial cells, pcDNA3.1(+)-14-3-3 plasmid was constructed and transfeeted into PC12 cell with Lipofectamine~(TM)2000. The overexpression of transfected 14-3-3 gene in PC12 cell was determined by immunofluorescence and Western blotting.The effects of 14-3-3 overexpressing on the cells viability,apoptotie ratio and the activity of superoxide dismutase(SOD)as well as glutathione peroxidase(GSH-Px)of PC 12 cell treated with MPP~+ were measured by MTT assay,flow cytometry analysis and microplate reader respectively.Results The expression of 14-3-3 protein in transfection group(1.19?0.06)increased evidently compared with control group(0.75?0.05).And the antioxidant enzyme activity assession,MTT assay and flow cytometry analysis shows that the overexpression of 14-3-3 protein elevates the activity of SOD(transfection group:(9.13? 0.41)U/mg protein,MPP~+ group:(6.45?0.52)U/mg protein)and GSH-Px(transfection group: (89.66?3.42)?mol/mg,protein MPP~+ group:(82.73?4.15)?mol/mg protein),increases the cell viability(transfection group:0.78?0.06,MPP~+ group:0.54?0.07),and inhibits cell apoptosis (transfeetion group:11.87%?3.26%,MPP~+ group:36.30%?2.39%)of PC12 induced by MPP~. Conclusion The overexpression of 14-3-3 protein could elevate the activity of antioxidant enzymes SOD and GSH-Px,reduce oxidant stress,alleviate MPP~+ toxicity,and thus inhibit the apoptosis of PC12 cell induced by MPP~+.

Objective To evaluate the clinical effect of itraconazole in prevention of invasive fungal infections in allogeneic hema-topoietic stem cell transplantion .Methods In this retrospective study ,110 patients receiving allogeneic hematopoietic stem cell transplants were administed itraconazole or fluconazole for prevention of fungal infection .The occurrence and prognosis of invasive fungal infection ,and the side effect of both pyrroles were observed .Results Proven and probable invasive fungal infections occurred in 5 of 69 itraconazole recipients(7 .2% ) and in 8 of 41 fluconazole recipients(19 .5% ) during the first 180 days after transplanta-tion ,the difference had statistical significance(P<0 .05) .The fatality rate related to fungal infection had no statistical difference be-tween the two groups(2 .9% vs .7 .3% ) .The occurrence of itraconazole adverse reactions were more than fluconazole (26 .9% vs . 7 .0% ) ,and both itraconazole and fluconazole were well tolerated .Conclusion Itraconazole significantly reduces the incidence of inva-sive fungal infection in the patients receiving allogeneic hematopoietic stem cell transplants ,and it is a effective and safe prophylaxis .