0.05).The correlation of reduction of intra-disc pressure and accumulation of ablation time was positive.Compared with intra-disc pressure in control group,the pressure change of all ablation groups was statistically significant(P

AIM:To study the direct reprogramming method of mouse embryonic fibroblasts (MEFs) conver-ted into induced neural stem cells (iNSCs).METHODS:Sox2-infected MEFs were cultured in NSCs culture medium for 10 d.Subsequently , repeated suspension and adherent culture were performed for 3 times for the purification of iNSCs .The iNSCs were cultured in suspension medium .Real-time PCR was used to detect the expression of neural stem cell marker genes and pluripotent marker gene .In vivo, iNSCs were microinjected into the mouse cerebral cortex .Immunofluorescence was performed to detect the expression of neural stem cell , neuron, oligodendrocyte and astrocyte markers in vitro and vivo. RESULTS:A variety of neural stem cell marker gene expression was significantly increased in iNSCs detected by real -time PCR.Immunofluorescence confirmed that iNSCs expressed nestin and differentiated into neurons , oligodendrocytes and as-trocytes in vitro and vivo.CONCLUSION:Sox2 is sufficient to trigger the direct reprogramming from MEFs to iNSCs .iN-SCs have the ability of self-renew and 3 differentiation potentials in vivo and vitro.iNSCs are the suitable seed cells of SCI .

BACKGROUND:Poly(L-lactic acid) (PLLA) scaffold is a kind of widely used biomaterial in tissue engineering. However, low hydrophilicity and lack of surface cell recognition site of PLLA hinder its further application. OBJECTIVE:To study the biocompatibility of multi-walled carbon nanotubes/PLLA (MWCNTs/PLLA) nanofiber scaffolds with mouse neural stem cells in vitro. METHODS:Mouse neural stem cells were isolated. Then we used electrospinning to fabricate PLLA nanofibers and modified them with multi-walled carbon nanotubes. We assessed their biocompatibility with passage 3 mouse neural stem cells in vitro. RESULTS AND CONCLUSION:Scanning electron microscope showed that the neural stem cells could survive on both scaffolds. No cytotoxic effects were detected on both scaffolds by Cell Counting Kit-8 detection. The adhesion and proliferation abilities of neural stem cells on the MWCNTs/PLLA scaffold were significantly greater than those on the PLLA scaffold. Neural stem cells were found grow well and have normal morphology on both scaffolds under scanning electron microscope and by Hoechst 33342 staining. Besides, immunofluorescence staining showed MWCNTs/PLLA could promote neural stem cells to differentiate into mature neurons and neurites grew along with the nanofiber scaffold. In conclusion, the MWCNTs/PLLA nanofiber scaffold has better properties than the PLLA for transplanted cells and provides a good growth carrier for neural stem cells to be induced to differentiate into neurons, which is expected to have a great potential of applications in nerve tissue engineering.

Objective To investigate the quantitative abnormality of CD~+_5B lymphocytes in the bone morrow of the patients with autoimmunic hemocytopenia and its clinical significance.Methods The patients were referred to the Institute of Hematology & Blood Diseases Hospital,Chinese Academy of Medical Sciences & Peking Union Medical College.Quantities of CD~+_5B lymphocytes in the bone morrow of 14 patients with autoimmune hemolytic anemia(AIHA)or Evans syndrome,22 immunorelated pancytopenia(IRP)patients and 10 normal controls were assayed by FACS.The correlation between their clinical and laboratory parameters with CD~+_5B lymphocytes was analyzed.Results The qutantity of CD~+_5B lymphocytes were significantly higher in AIHA and Evans and IRP patients than in normal controls;there was no significant difference between AIHA、Evans syndrome and IRP patients.The qutantity of CD~+_5B lymphocytes in the bone morrow of all cytopenic patients showed negative correlation with C_3 complement.In AIHA、Evans syndrome patients,the quantity of CD~+_5 B lymphocytes in their bone morrow showed positive correlation with IBIL.In Evans syndrome patients,the quantity of CD~+_5B lymphocytes in their bone morrow showed positive correlation with PAIgG and PAIgM.The qutantity of CD~+_5B lymphocytes in the bone marrow of all cytopenic patients showed negative correlation with treatment response,but no correlation with the yields of CFU-E、 BFU-E、CFU-F and CFU-GM cultured from their bone marrow mononuclear cells in vitro.Conclusion CD~+_5B lymphocytes in the bone marrow of the patients with autoimmunic hemocytopenia significantly increase and are related to the disease severity and clinical response.It is suggested that CD~+_5B lymphocytes might have important roles in the pathogenesis of autoimmunic cytopenia.

Objective To evaluate the clinical features and early management of combined thoracoabdominal injury (CTI).Methods A retrospective study was carried out in 29 cases with CTI treated surgically.There were 21 cases with penetrating injuries and 8 cases with blunt injuries.All cases were associated with penumothorax and/or hernothorax at various degrees and 13 cases complicated with shock.In this study thoracotomy was performed in 17 cases,laparotomy in 6 cases,thoracotomy plus laparotomy in 5 cases and combined thoraco-laparotomy in 1 case.Results The injury severity score in this series was (40.16±15.23) scores.Four cases were died from hemorrhagic shock (2 cases),pericardial tampenade(1 case) and multiple organ failure(1 case).Conclusions CTI has high mortality rate,and the most frequent cause of death is hypovolemic shock.The operative approach is based on individual injury condition.Early diagnosis and timely surgical intervention are key to improving the cure rate and reducing mortality in severe CTI.

BACKGROUND: Poly-3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV) is a novel scaffold made by solvent casting/particulate leaching procedure, composed of polyhydroxybutyrate and polyhyclroxyvalerate at certain ratio, which has good biocompatibility as well as high intensity and modulus. It has three-dimensional porous net structure and good biodegradation. OBJECTIVE: To evaluate the biocompatibility between copolymers of PHBV and canine bone marrow mesenchymal stem cells(BMSCs).DESIGN, TIME AND SETTING: In vitro comparative observation. The study was performed at the Laboratory of Histology and Embryology, Sun Yat-sen University between June 2003 and March 2004.MATERIALS: PHBV scaffold, film porosity > 85% and 100-350 μ m aperture size.METHODS: Canine BMSCs were isolated and cultured. The 3-4 passage cells were seeded onto the PHBV films and three-dimensional foam scaffold. Cells cultured alone served as control.MAIN OUTCOME MEASURES: The seeded cells were observed under inverted microscope; at 1, 2, 3 weeks after seeding, the BMSCs were treated with 4% paraformaldehyde and stained with hematoxylin-eosin (HE); The protein content in seeded cells was determined by bicinchoninic acid assay (BCA), and the content of DNA was quantified using Hoechst33258 assay at 5, 10, 14 days after culture.RESULTS: Inverted microscopic observation showed that the PHBV fibers were fairly thick with weak lucency, and the fibers were hardly detectable under contrast phase microscope. Majority of cells attached onto the PHBV films 2 hours after seeding, and extended well in a spindle shape at 3 days. One week after culture, 2 PHBV were fixed, and BMSCs proliferation was observed after HE staining. At two weeks, cells continued to proliferate and densely covered the PHBV film. The cells grew in the three-dimensional pores, connected at 1 week, extended at 3 weeks, secreting a large amount of material around cells. Cell proliferation did not change much at 3 weeks compared with 2 weeks, and there was no significant difference in DNA and protein contents between control and PHBV groups (P > 0.05).CONCLUSION: As a kind of tissue-engineered scaffold material for BMSCs, PHBV displays good biocompatibility.

Objective To explore the roles of insulin in the early treatment of acute brain injury and its administration methods.Methods 253 patients were randomly divided into the intensive insulin therapy group and the conventional therapy group.Infection rates,the short-term effect (APACHE Ⅱ assessment),and long-term efficacy (GOS prognosis) was compared between two groups.Results The results of the strengthen treatment group in the rate of infection (25.95％ vs 39.34％,x2 =5.17,P ＜0.05),the short-term effect (11.33 ± 7.66 vs 16.49 ± 14.97,u =3.42,P ＜ 0.05) and the long-term efficacy (40.46％,55.73％,3.82％ vs 25.41％,68.85％,5.74％,x2 =7.62,P ＜0.05) were significantly better than the conventional therapy group with the statistically significant differences (P ＜ 0.05).Conclusions The hypoglycemic effect,neuroprotective effect,regulatory role,and nutrition role of insulin occurred in the early treatment of acute brain injury.After acute brain injury,patients with hyperglycemia should be treated early with an enough volume,continuous,and uniform insulin.

Objective To discuss the application of scenario simulation technique in emergency rescue training of hospital grave pubic health events.Methods Based on the simulate function in the scenario simulation technique,we designed the emergency drill script of hospital grave pubic health events,set the simulate scene,communication system and indicators measuring emergency rescue,drilled and trained the hospital rescue crews,compared with the rescue crews trained in routine method.40 nurses were selected before or after the application of scenario simulation technique separately.Results The qualified rate of indicators such as communication and medical rescue system that were trained by scenario simulation method was higher than the rescue crews trained in routine method.Conclusions Scenario simulation technique would improve the training effect of emergency rescue in hospital grave pubic health events.

Objective To investigate the effect of galectin-3 (Gal-3) on the pathogenesis of primary biliary cirrhosis (PBC).Methods The clinic data of 72 PBC patients at different stages were analyzed and the serum levels of Gal-3 were detected in 72 PBC patients and 20 controls by enzyme linked immunosorbent assay (ELISA).Independent t-test,variance analysis,LSD-t and Pearson correlation analysis were adopted for data analysis.Results The serum Gal-3 levels were significantly higher in PBC patients than those of healthy controls [(855±634) pg/ml,(463±446) pg/ml,P＜0.05].With the progression of disease,the levels of Gal-3,platelets,hemoglobin,albumin,IgM and complement C3 gradually declined,but the level of total bilirubin gradually elevated(P＜0.05).There was positive correlation between Gal-3 and immunoglobin IgM,complement C3 levels (r=0.330,P=0.005; r=0.357,P=0.002).There was negative correlation between Gal-3 and total bilirubin levels (r=0.350,P=0.003).Conclusion The Gal-3 can participate in the immune-mediated inflammation of PBC and the formation of liver fibrosis.The levels of Gal-3,platelet count,hemoglobin,albumin,IgM,complement C3 and total bilirubin could be regarded as laboratory parameters for the evaluation of the disease severity and prognosis.

The case design,standardized patients(SPs)training and training communication skills for medical students with SPs were illustrated in this paper.And our experiences of SPs using Were summarized.