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.

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.

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

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.

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.

Objective To study the feasibility of engineering cartilage tissue in a tube lined with epithelium and implanting allogenic chondrocytes into a novel scaffold consisting of chitosan nonwoven cloth coated with poly (DL-lactide-co-glycolide)(PLGA).The focus of the present study is to explore a new way of repairing laryngeal and tracheal defects.Methods Allogenic chondrocytes were obtained from the auricles of 1-month-old rabbits.After being cultured in vitro for three to four passages,the cells were implanted into the scaffolds to form composite grafts and then transplanted into the rabbits.After 6,12,and 18 weeks,the general,histological characteristics were investigated.Results The cobweb-like matrix was observed approximately 1 week after the chondrocytes had been implanted into the scaffolds.At 6 weeks,the matrix was secreted,and there were immature chondrocytes in the grafts.At 12 weeks,the allogenic cartilage in the tube lined with epithelium had been created.Chondrocytes were almost mature and the lacunae had formed.At 18 weeks,the neocartilage was similar to native cartilage.Conclusion It is feasible to fabricate allogenic cartilage in a tube lined with epithelium by implanting allogenic chondrocytes into a novel scaffold made of chitosan nonwoven cloth coated with PLGA.

Objective To assess the incidence,risk factors and mortality of acute kidney injury(AKI)in patients with chronic myelogeneous leukemia(CML)after myeloablative allogenetic hematopoietic stem cell transplantation(HSCT). Methods Renal function in 93 CML patients undergone myeloablative allo-HSCT was retrospectively analyzed by the RIFLE criteria. Results Thirty-nine patients (41.9%) developed AKI at a median of 40 days after allo-HSCT, including 24 AKI-R patients(25.8%), 10 AKI-I patients(10.8%) and 5 AKI-F patients (5.4%). The morbidity of AKI in patients with ≥Ⅲ acute graft-versus-host disease (aGVHD) and without ＜Ⅲ GVHD was (81.82±11.63)% and (36.59±5.32)% (P=0.0037)rospectively. The morbidity of AKI in patients with increased total bilirubin and without increased total bilirubin was (72.73±13.43)% and (37.04±5.37)%(P=0.0192) respectively. ≥Ⅲ aGVHD was peor-prognostic factor of AKI and RR was 2.773 [95%CI (1.073-7.167), P=0.035]. RR of AKI-I and AKI-F in patients with ≥Ⅲ aGVHD was 6.320195%CI (1.464-27.291), P=0.013]. The mortality within 100 days after allo-HSCT of patients with AKI was significantly different as compared to patients without AKI (P=0.001). Six-mouth survival rates of different class AKI patients after myeloablative allo-HSCT were (86.96±7.02)% (AKI-R), (70.00±14.49)% (AKI-I), 0 (AKI-F) (P=0.000)respectively. Conclusions AKI is one of the main complications in CML patients after myeloablative allo-HSCT. ≥Ⅲ aGVHD and increased total bilimbin are poor-prognostic factors of AKI, and higher morbidity of AKI-I and AKI-F can be found in patients with ≥Ⅲ aGVHD. With the deteriorated AKI, 6-month survival is decreased. RIFLE criteria is sensitive to the early diagnosis of renal function. Moreover RIFLE can monitor the progression of AKI and predict the clinical outcome.