Objective To explore the feasibility of differentiation into neural cells from bone marrow stromal cells(BMSCs) in vitro by breviscapine,and to offer reference for the applying of BMSCs. Methods The bone marrow stromal cells of SD rat were separated and purified by passsage culture.After phenotype characterization,the 4th passage of BMSCs was exposed to breviscapine. Differentiation was observed under phase contrast microscope every 6 hours and the cells were stained immunocytochemically with neuronspecific enolase(NSE)and glial fibrillary acidic protein(GFAP).The induced cells activity was detected by MTT. NSE and GFAP were determined by flow cytometry and RT-PCR. Results The BMSCs were CD44~+,CD54~+,CD34~-. After 18 hours of induction, cytoplasm of BMSCs partly contracted with protruding;The immunocytochemical staining was performed on cells after induction for 24 hours,the rates of NSE and GFAP staining positive were(48.7±3.4)% and(56.8±4.2)%.By using flow cytometer, expressions of NSE and GFAP showed much higher in induced group than that in control group. By using RT-PCR, expressions of NSE and GFAP were positive in induced group. Conclusion Breviscapine could induce adult rat BMSCs to differentiate into neuron and glial cells.

BACKGROUND: Schwann calls are seed cells for constructing tissue engineered peripheral nerves. But their in vitro isolation, culture and purification are difficult. Acellular nerve allografts (ANA) have a great effect on repairing peripheral nerve defects, and it can induce marrow stromal cells (MSCs) into Schwann-like calls. Accordingly, MSCs can be used as seed calls theoretically in constructing tissue engineered peripheral nerves, instead of Schwann cells OBJECTIVE: To observe the repairing effect of tissue engineered peripheral nerves constructed with MSCs on sciatic nerve defects and to assess the feasibility of repairing peripheral nerve defects with MSCs as seed calls. DESIGN, TIME AND SE'I-rlNG: A randomized control animal experiment was conducted in the laboratory of Basic Medicine Collage of Dali Collage from July to December in 2008.MATERIALS: A total of 30 SD rats were divided randomly into 3 groups, with 10 in each group. In MSCs+ ANA group, end-to-end anastomosis were performed with 10/0 non traumatic suture to broken ends of rat sciatic nerves and tissue engineered nerves cultured using MSCs combined with ANA; In ANA group, ANA were bridged to broken ends of sciatic nerves; In METHODS: The 10ram sciatic nerve defects in rats were repaired using MSCs-constructed tissue engineered peripheral nerves, whose repairing effects were evaluated at week 12 post transplant through sciatic function index (SFI), gastrocnemius wet weight recovery rate, S-100 immunohistochemical staining and electron microscope observation, etc. MAIN OUTCOME MEASURES: Morphological changes of calls were observed during the culture of compounds; SFI and gastrocnemius wet weight recovery rates were detected after transplantation; New myelinization, axon growth and nerve fiber distdbuUon were observed with toluidine blue staining; Schwann calls growth and nerve fiber regeneration were observed with transmission electron microscope combined with S-100 protein immunohistochemical staining method.RESULTS: The detection results showed that SFI and gastrocnemius wet weight recovery rate were higher in the MSCs+ ANA group than in the ANA group (P < 0.05). Compared the ANA group, the MSCs+ ANA group had more S-100 proteins expressed in its compounds obviously, and the number, the diameter of its myelinated nerve fibers as well as its myelin sheath thickness were all greater than those of the ANA group (P < 0.05). The repairing effect of the MSCs+ ANA group was close to that of the autotransplantation group.CONCLUSION: MSCs-constructed tissue engineered nerves have a better effect on repairing peripheral nerve defects than ANA, and MSCs as seed cells have a high value in peripheral nerve tissue engineering.

OBJECTIVETo retrospectively analyze the experience of one-stage surgical correction for children with congenital heart diseases and tracheal stenosis and further to clear the principle of treatment for these patients.

METHODSFrom January 2006 to June 2013, 48 patients with congenital heart diseases and tracheal stenosis underwent surgical correction. There were 36 male and 12 female patients. The mean age at operation was (23 ± 27)months (range: 3-72 months) and the mean weight was (12 ± 8) kg (range: 3.4-46.0 kg). The underlying diagnoses were pulmonary sling in 33 patients, double aortic arch in 3, tetrology of Fallot in 6, ventricular septal defect in 4, double outlet right ventricle in 1, and pulmonary atresia in 1 patient. Among them, short tracheal stenosis was present in 15, long tracheal stenosis in 25 and bridging bronchus in 8 patients. Repairs with autogenous tracheal tissue were performed in 6, and end-to-end anastomosis in 11 and slide tracheoplasty in 31 patients. Two patients had granulation tissue growing in the airway postoperatively and were re-operated by autogenous rib tissue. All of patients were followed up after operation 1, 3, 6, 12 months and if the patient had symptoms that should be examined by bronchoscopy. The patients should be examined by CT post-operation one year.

RESULTSThere were 7 deaths in all 48 cases and the early mortality was 14.6%. Two deaths were not related to tracheal stenosis, and other 5 were associated with long segment tracheal stenosis. Forty-one patients were followed for 2 months to 6 years, and no patients required re-operations. Clinical symptoms of tracheal stenosis disappeared and the results of CT scan were satisfied.

CONCLUSIONSOne stage surgical repair of the patients with congenital heart diseases and tracheal stenosis have a good effect. Slide tracheoplasty is the effective surgical method for long segment tracheal stenosis. Postoperative granulation tissue growing in the airway is the leading cause of death.

Child ; Child, Preschool ; Female ; Heart Defects, Congenital ; complications ; surgery ; Humans ; Infant ; Male ; Retrospective Studies ; Tracheal Stenosis ; complications ; surgery