BACKGROUND: It is a new tendency to treat central nervous system injury or tumor therapy using the combination of seed cells and gene therapy.OBJECTIVE: To observe the dose-relationship between transfection and expression of rabbit bone marrow mesenchymal stem cell (BMSCs) with adenovirus vector carrying green fluorescent protein (Ad-GFP), and to study its effects on cell biological properties, in addition, to explore the feasibility of using Ad-GFP vector to construct gene modified BMSCs.DESIGN, TIME AND SETTING: A randomized grouping, contrast observation. The experiment was performed at the Southern Medical University between August 2008 and March 2009.MATERIALS: New Zealand white rabbits, irrespective of genders, weighing 2.0-3.0 kg, were selected.METHODS: BMSCs were separated and cultured in vitro, and then the cell immunophenotypes were detected by flow cytometry.The adenovirus was obtained by packaging 293 cells and was used to transfect BMSCs with various liters (1 ×-10~3-1×10~(10) PFU/mL).Cytometry was used to analyze the transfection efficiency.MAIN OUTCOME MEASURES: Cell morphological changes were detected under an invert microscope. The cell proliferation was detected by CCK8 kits. BMSCs transfected with Ad-GFP were induced differentiating into neuron-like cells by adding of β-mercaptoethanol.RESULTS: The surface markers of 3-6-generation BMSCs were negative to CD34 and CD45, but positive for CD29 and CD44.When the virus titers were 1 ×10~7 PFU/mL, the transfection rate was 55%, which were 85% when the virus titers were 1 ×10~9 and1×10~(10) PFU/mL. However, cell pathology phenomenon occurred when the virus titer was 1 ×10~(10) PFU/mL. The fluorescence was strongest expressed at day 7, and it still can be seen at day 28. The BMSCs trasfected with Ad-GFP could differentiate into neuron-like cells under induction of p-mercaptoethanol, with positive neuron-specific enolase.CONCLUSION: Ad-GFP with suitable titers can infect BMSCs effectively with little influence on the biology property or differentiation function. BMSCs can serve as seeds cell in gene therapy field when utilizing ad-GFP vector system.

0.05);in treating gradeⅢto IV aGVHD,the effectiveness of FK506 with MP was significantly better than that of CsA with MP(P

Animals ; Arrhythmias, Cardiac ; etiology ; therapy ; Captopril ; therapeutic use ; Disease Models, Animal ; Electric Stimulation Therapy ; methods ; Myocardial Ischemia ; physiopathology ; therapy ; Rabbits ; Ventricular Fibrillation

Objective To explore the value of Gd-EOB-DTPA enhanced magnetic resonance imaging (MRI) in the quantitative evaluation of liver function.Methods Between June 2012 and June 2013,we prospectively included 8 patients with cirrhosis and 6 patients with primary hepatic carcinoma (PHC).The Child-Pugh classification,indocyanine green retention rate at 15 minutes (ICGR15),and Primovist enhanced MR imaging were performed in all the patients.Using the deconvolution method to calculate hepatic extraction fraction (HEF),the correlations between HEF and Child-Pugh and ICGR15 were analyzed.Results For cirrhosis,there was a significant correlation between the HEF and the ICGR15 by the Pearson correlation coefficient (r =-0.971,P ＜ 0.05).For hepatocarcinoma,there was also a significant correlation (r =-0.855,P ＜ 0.05).The average HEF of PHC was 21.80％ (14.89％ ～ 35.90％).Conclusions The Gd-EOB-DTPA enhanced MRI accurately evaluated liver function quantitatively.It may be used for patients with PHC for preoperative examination.

Objective To explore and evaluate the implementation process and prelimnary achievements of 4+4 medical education program in Shanghai Jaiotong University School of Medicine. Methods The data of student background,curriculum,exam grades and graduate employment of class 2002 and 2003 were collected and analysed. Results The data showed that this program has enrolled high quality students,teaching hours of key courses of medicine has been reduce to 70%,while the average grades of the comprehensive examination were higher than those of the 5 year students,and the employment prospects for 4+4 program has been optimistic. Conclusion The 4+4 program has achieved preliminary success,and the experiences is beneficial for the further reform of medical educational system.

Objective:To study the effects of recombinant human platelet-derived growth factor-BB(rhPDGF-BB)combined with recombinant human transforming growth factor-β1 (rhTGF-β1 )on the expression of FAK mRNA of osteoclasts in the alveolar bone on the pressure side of orthodontic teeth in rats.Methods:Orthodontic tooth movement model was established in 160 male SD rats.The rats in experimental group were treated by injection of 10 ng rhPDGF-BB and 5 ng rhTGF-β1 in the buccal submucosal area of the mo-lar every other day from day 1 afterburner,while those in control group received equivalent volumes of PBS.The rats were sacrificed at 1,4,7,10 and 14 days(n=16)after appliance placement.Specimens were collected.Osteoclasts in the alveolar bone on the pres-sure side of the orthodontic teeth were observed by TRAP staining,the FAK mRNA expression was quantified by quantitative RT-PCR.Results:rhPDGF-BB combined with rhTGF-β1 significantly promoted an increasing number of osteoclasts on the compressing side(P<0.05),increased the expression of FAK mRNA at day 4 and 7(P<0.05),then decresed it to the control level(P>0.05) at day 14.Conclusion:Combination of rhPDGF-BB and rhTGF-β1 can increase the number of osteoclasts in the alveolar bone on compressing side,and promote FAK mRNA expression in osteoclasts.

Purpose:To study the effect of early restrictive fluid resuscitation (EFR) on inflammatory and immune factors in patients with severe pelvic fracture (SPF).Methods:A total of 174 SPF patients in the Department of Orthopaedics,the First Affiliated Hospital of Chengdu Medical College from July 2015 to June 2018 were involved in this study and divided into EFR group (n =87) and control group (n =87) using the random number table method.Conventional fluid resuscitation (CFR) was performed in control group,and EFR was performed in EFR group.The incidences of acute respiratory distress syndrome (ARDS) and multiple organ dysfunction syndrome (MODS) during rescue,successful rescue rate,blood transfusion volume,fluid input,and resuscitation time were compared between the two groups.The parameters including prothrombin time (PT),hematocrit (HCT),platelet (PLT) and blood lactate (BL) at the 4th hour after fluid resuscitation were recorded.The levels of inflammatory factors (TNF-α,IL-6,CRP) and immune factors (CD3+,CD4+,CD8+,CD4+/CD8+) were compared between the two groups before treatment and 7 days after treatment.The revised acute physiologic and chronic health evaluation system and the sequential organ failure assessment scores were adopted for evaluation before treatment and 7 days after treatment.Results:The incidences of ARDS and MODS during rescue in EFR group were significantly lower than those in control group (p=0.015 and 0.010 respectively),and the successful rescue rate in EFR group was significantly higher than that in control group (p =0.011).The blood transfusion volume,fluid input,resuscitation time in EFR group were significantly lower than those in control group (p =0.016,0.002 and 0.001 respectively).At the 4th hour after fluid resuscitation,PTand BL in EFR group were significantly lower than those in control group (p =0.021 and 0.003 respectively),while HCT and PLT in EFR group were significantly higher than those in control group (p =0.016 and 0.021 respectively).On day 7 after treatment,TNF-α,IL-6,CRP and CD8+ in EFR group were significantly lower than those in control group (p =0.003,0.004,0.007 and 0.003 respectively),while CD3+,CD4+ and CD4+/CD8+ in EFR group were significantly higher than those in control group (p =0.004,0.000,0.007 respectively).On day 7 after treatment,the revised acute physiologic and chronic health evaluation (APACHE) system and the sequential organ failure assessment (SOFA) scores in EFR group were significantly lower than those in control group.Conclusion:EFR can effectively eliminate inflammatory factors,improve immune function,maintain the stability of blood components,reduce the incidences of ARDS and MODS,and elevate the successful rescue rate in patients with SPF.

Antipsychotic Agents ; poisoning ; Clozapine ; blood ; poisoning ; Exchange Transfusion, Whole Blood ; methods ; Humans ; Infant, Newborn ; Male ; Treatment Outcome

Objective To investigate the effect of parecoxib on postoperative hyperalgesia induced by remifentanil-based anesthesia.Methods One hundred ASA Ⅰ or Ⅱ patients,aged 21-64 yr,weighing 50-80 kg,undergoing elective laparoscopic operation,were randomly divided into 5 groups ( n =20 each):parecoxib group (group P),small dose remifentanil group (group S),large dose remifentanil group (group L),small dose remifentanil + parecoxib group (group SP) and large dose remifentanil + parecoxib group (group LP).Parecoxib 40 mg was injected intravenously at 30 min before anesthesia in groups P,SP and LP.Anesthesia was induced with midazolam0.05 mg/kg,etomidate 0.2 mg/kg,cisatracurium 0.15 mg/kg and remifentanil 1 μg/kg (fentanyl 4 μg/kg in group P).The patients was tracheal intubated and mechanically ventilated.PETCO2 was maintained at 35-45 mm Hg.Anesthesia was maintained with infusion of remifentanil at 0.05 μg·kg-1 ·min-1 (in groups S and SP) or at 0.3 μg· kg- 1· min- 1 (in groups L and LP) combined with inhalation of sevoflurane and infusion of cisatracurium at 0.12 mg·kg-1·h-1.At 30 min after operation,numeric rating scale (NRS) was used to assess the degree of pain at rest and during activity.Tramadol 1.5 mg/kg was injected intravenously after operation if needed.NRS scores were maintained ≤5.The use of tramadol and adverse effects during 24 h after operation were recorded.Results Compared with group P,NRS scores at rest and during activity were significantly increased at 30 min after operation in groups S and L,the incidence of shivering and the number of patients who needed tramadol were significantly increased in group L,and no change was found in NPS scores at rest and during activity at 30 min after operation,the incidence of adverse effects and the number of patients who needed tramadol in groups SP and LP.Compared with group S,NRS scores at rest and during activity at 30 min after operation,the incidence of shivering and the number of patients who needed tramadol were significantly increased in group L,NRS scores at rest and during activity at 30 min after operation were significantly decreased and no change was found in the incidence of adverse effects and the number of patients who needed tramadol in group SP.Compared with group L,NRS scores at rest and during activity at 30 min after operation,the incidence of shivering and the number of patients who needed tramadol were significantly decreased in group LP.Conclusion Intravenous injection of parecoxib 40 mg at 30 min before anesthesia can attenuate postoperative hyperalgesia induced by remifentanil-based anesthesia.

Adult ; China ; Humans ; Male ; Neisseria meningitidis, Serogroup C ; drug effects ; genetics ; isolation & purification