Objective To investigate the feasibility of using PLGA loaded with SD rats' mesenchymal stem cells(rMSCs) transfected with green fluorescent protein (GFP) gene as scaffolds for combinations of molecular, cellular, and tissue-level treatments of spinal cord tissue engineering. Methods rMSCs infected with lentiviral vectors (lv-GFP) were seeded onto PLGA at 8000 cell/cm2, rMSCs-GFP grown under similar conditions on tissue culture plastic as control. The morphology of rMSC-GFP was examined by fluorescence microscopic. The activity of MSCs was detected by MTT assay everyday. Cell cycle analysis was performed after a 3-day culture on PLGA using flow cytometry. The rMSCs-GFP seeded on PLGA was identified by FITC-anti CD34,CD90 and PE-anti CD44, CD106,CD45,CDllb at the third day. Results Fluorescence microscopic examination revealed adherence of the cells to the PLGA surface within 24 h of initial plating. After 3 days, GFP cells were spindle shaped. The difference disappeared at 7 days when cells under both conditions had become confluent Cells proliferated at the same rate on the PLGA surface compared to tissue culture plastic. And cell's cycle was unaffected by the transduction process and seeded on PLGA. Cells maintained their stem cell phenotype as judged by expression of CD90, CD44, CD106 markers,and absence of the hematopoietic marker CD45, CD34. This demonstrated that the transduction and the PLGA surface were not adversely affecting the cells. Conclusions MSCs are a good candidate for spinal cord tissue engineering. Cells continued to express green fluorescent protein(GFP)on a long-term basis,and are compatible with polymer surfaces. Morphology,viability,and growth kinetics were maintained when cells were grown on a poly-lactic-co-glycolic acid(PLGA)polymer scaffold. Therefore,they could make further efforts for combinations of molecular, cellular,and tissue-level treatments of spinal cord tissue engineering.

The thyroid allografts in rats were cultured for 48h in the conditions of hyperbaric oxygen (a mixture of 95% O2,5% CO2 pressurized at 2 atmospheres)and low pH(5.5 and 6.9).Uncultured and air cultured allografts were taken as control groups.The thyroid slices were transplanted under the kidney eapsulse of thyroidectomized recipients.By examining histological changes,detecting the serum T3 and T4 levels and five weeks body weight gain,the graft survival was cvaluated.Though the thyroids cultured in hyperbaric O2 were contaminated with the passenger leukocytes,the grafts had prolonged survival,suggesting this method might effectively diminish MHC immunogenicity to thyroid grafts.

Objective To evaluate the value of lung ultrasound score (LUS) on assessing the severity and extubation opportunity in postoperative patients of general surgery, and to investigate the correlation between LUS and oxygenation index (PaO2/FiO2), acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ), sequential organ failure assessment (SOFA), stay length in ICU and stay length in hospital. Methods A prospective double- blind cohort study was conducted. Eighty- nine postoperative patients of general surgery with successful extubation were selected, and the patients were divided into 2 groups:group A ( admission ICU to extubation time less than 48 h, 52 cases) and group B(admission ICU to extubation time more than 48 h, 37 cases). Before extubation, the PaO2/FiO2 was recorded according the blood gas analysis, and APACHE Ⅱ, SOFA and LUS were examined, and the staying time in ICU and staying time in hospital were recorded. The correlation was analyzed. Results The LUS, APACHE Ⅱ, SOFA, staying time in ICU and staying time in hospital in group A were significantly lower than those in group B: (3.98 ± 2.31) scores vs. (13.41 ± 2.82) scores, (7.52 ± 1.96) scores vs. (14.92 ± 3.07) scores, (4.50 ± 2.24) scores vs. (9.70 ± 3.64) scores, (1.77 ± 1.41) d vs. (8.49 ± 4.35) d and (8.49 ± 2.28) d vs. (15.63 ± 6.10) d, and the PaO2/FiO2 was significantly higher than that in group B:(441.57 ± 45.31) mmHg (1 mmHg=0.133 kPa) vs. (305.78 ± 90.72) mmHg, and there were statistical differences (P<0.01). The LUS had negative correlation with the PaO2/FiO2 (r=-0.882, P<0.01), and it had positive correlation with APACHEⅡ, SOFA, staying time in ICU and staying time in hospital (r=0.711, 0.590, 0.930 and 0.709;P<0.01). Conclusions The LUS is simple and easily available. It can evaluate the changes of pulmonary ventilation, and also evaluate its degree of severity and prognosis. It is helpful in the prediction of the extubation time, staying time in ICU and staying time in hospital in patients with general surgery.

Objective To explore the new methods to fill local temporal depression or augment superciliary with autotissue graft.Methods Applying upper eyebrow perdicled dermis -fat petal to fill the local temporal depression or augment superciliary, accompanied with eyebrow lifting.Results Augmentation superciliary arch plasty with upper eyebrow perdicled dermis-fat petal was performed in 5 patients. All 5 patients' superciliary arch showed good shape and accompanied with no complications. Filling of local depression of temporal with derma-fat flap was performed in 6 patients, with dermis-fat flap and prosthesis was performed in 8 patients. 2 cases had hematoma. Conclusion Upper eyebrow perdicled dermis-fat petal as a material being ultilized to augment superciliary or to fill local temporal depression has advantages of better localization and more natural shape.[

[Objective]To investigate the effect of cryopreservation on the antigenicity of vascularized bone allotransplantation,and to observe the patency of the pedicle and the osteosis of bone allografts,in order to find out the dose of immunosuppressive agent.[Method]Forty-eight New Zealand rabbits were involved in the study.Of them 16 rabbits served as donors,and 32 hosts.Allotransplantation of cryopreserved radius diaphysis with the vascular pedicles were implanted to 8 rabbits and immunosuppressive agent(CsA 10 mg/kg/d) was administrated 4 weeks after transplantation(group A),similar cryopreserved grafts with the vascular pedicles were implanted to 8 rabbits and immunosuppressive agent(CsA 5mg/kg/d) was administrated 4 weeks after transplantation(group B),similar cryopreserved grafts with the vascular pedicles were implanted to 8 rabbits and immunosuppressive agent(CsA 2 mg/kg/d) was administrated 4 weeks after transplantation(group C),allotransplantation of cryopreserved radius diaphysis with a vascular pedicle was performed in 8 rabbits(group D),and 16 donors were group E.Immunological analysis was performed at 1 weeks.Radiography was taken for each graft at 2,4,8,12 weeks.And 12 weeks after operation,the patency of the pedicle was observed through ink infusing and histological studies were performed.[Result]There was no difference of IFN-? between group A and group B.However,the levels of IFN-? in groups A and B were lower than that in group C.[Conclusion]Cryopreservation could reduce the antigenicity of vascularized allotransplant graft.After transplantation,the reject response is decreased,patency and osteosis of bone allografts are better,and the dose of immunosuppressive agent are decreased.

Objective To discuss the survivorship of rat mesenchymal stem cells (rMSCs) and the expression of human brain-derived neurotrophic factor (hBNDF) protein after transplantation of the hBDNF-modified rMSCs (hBDNF-rMSCs) to the adult rats with spinal cord injury (SCI) and discuss the effect of hBDNF-rMSCs on the apoptosis of rat neural cells. Methods A total of 240 adult male Sprague-Dawley rats were randomly divided into sham operation group,SCI group,hBDNF-rMSCs transplantation group and empty vector-rMSCs transplantation group,with60 rats in each group.The SCI model was established by using the modified Allen technique.At day 7 after modeling,an equal volume of hBDNFrMSCs suspension,empty vector-rMSCs suspension and phosphate buffered saline (PBS) were injected through the L4,5 subarachnoid space into the hBDNF-rMSCs group,empty vector-rMSCs group and SCI group,respectively.Then,the injured spinal cord tissues were obtained from each group at days 1,2,3,7 and 14 after transplantation to observe the viability and distribution of rMSCs with enhanced green fluorescent protein gene by fluorescent microscope,measure the expression of hBDNF protein by Western blot and detect the apoptosis of neural cells by TdT-mediated dUTP nick end labeling (TUNEL). Results Both hBDNF-rMSCs and empty vector-rMSCs groups showed green fluorescence expression of rMSCs.The hBDNF protein expression was observed in hBDNF-rMSCs group and changed with time,ie,the expression was detected at day 2 after transplantation,reached the highest level at day 7 and then decreased gradually.Among the hBDNF-rMSCs,empty vector-rMSCs and SCI groups,the number of TUNEL positive cells was the least in hBDNF-rMSCs group,followed by the empty vector-rMSCs group and the number was relatively more in SCI group at days 2,3,7 and 14 after transplantation,with significant differences among groups (P ＜ 0.05). Conclusions Transplantation of the hBDNF-modified rMSCs through subarachnoid approach are able to survive and assemble at the injured spinal cord area and express hBDNF protein.The hBDNF-modified rMSCs can inhibit the apoptosis of neural cells after SCI.

Objective To investigate a new technique to increase the survival rate of free autogenous pellet fat graft. Methods Many methods were used to graft free autogenous fat, and the results compared to look for the best method for it. Results After 3 and 6 months, the volume rate in the treatment group and control group were 70%, 60% and 40%, and 30%. The times of fat injection were 2 and 4 . Conclusion Some vasodilators, low negative pressure and plasma can enhance the survival rate of free autogenous pellet fat graft.

Objective To compare the effectiveness and safety of different concentrations of elastin-like polypeptides (ELP) as novel submucosal injection material for endoscopic submucosal dissection.Methods Forty healthy New Zealand white rabbits were randomly divided into two groups (n=20).The first group by submucosal injection of different drugs were randomly divide into five groups (n=4).Four concentrations of 50 ku ELP (0.05,0.025,0.012 5,0.005 g/ml) were used separately in each group,while glycerin fructose was used for control group.Each solution (2 ml) was injected into the submucosa through the resected margin,the increase of mucosal thickness and surface changes were observed and recorded at 0,5,10,and 30 min.The subgroup by submucosal injection of different drugs were randomly divide into five groups (n=4).The injection pressure of each solution (2 ml) with the 25-gauge needle was calculated by a manometer,which was connected between the needle and syringe.Results The submucosal uplift heights in groups using the 0.05 g/ml ELP and 0.025 g/ml ELP injection were significantly thicker than that of glycerin fructose (P＜0.05),the 0.012 5 g/ml ELP and glycerin fructose injection showed no significant difference (P＞0.05),whereas the uplift height in glycerin fructose group was thicker than that of the 0.005 g/ml ELP (P＜0.05).The injection pressure correlated with the ELP concentration.The injection pressures of 0.05,0.025,0.012 5,0.005 g/ml ELP solutions were (332±36) kPa,(223±24) kPa,(174±22) kPa and (142±19) kPa,respectively,and that of glycerin fructose was (269±17) kPa.The 0.025 g/ml ELP solution was easily injected into the porcine stomach to create submucosal uplift.The injection pressure of the 0.025 g/ml ELP solution showed significantly lower value compared with that of glycerin fructose (P＜0.05).Conclusions ELP might be a promising agent for submucosal injection for endoscopic submucosal dissection (ESD),and 0.025 g/ml ELP might be efficient concentration for maintaining mucosal elevation,injection pressure and safety.

Objective To investigate the effect of FK506 on the expression of axon guidance cue slit-2 after spinal cord injury(SCI)in rats.Methods A total of 75 adult Sprague-Dawley rats were randomly divided into three groups,ie,sham operation group,SCI group and FK506 treatment group.The SCI model was made by using the modified Allen's technique.Then,the rats were sacrificed and the spinal cord was removed at different time points(at days 1,3,7,14 and 28)for detection of the expression of slit-2 by means of reverse transcription polymerase chain reaction(RT-PCR)and immunohistochemistry.Results The expression of slit-2 changed with time.The expression of slit-2 could be detected at day 1 after SCI,reached the highest at day 7 and then decreased gradually,with higher expression level in the injury group compared with treatment group(P ＜ 0.05).Conclusion Following spinal cord injury,administration of FK506 can up-regulate the expression of slit-2 and may exert important effect on the guidance of the axon regeneration.

Objective To observe the effect of FTY720 on the expressions of Bel-2 and Bax protein and explore the neuroproteetive effect of FTY720 on neuron after acute spinal cord injury in the rats.Methods A total of 75 SD rats were divided randomly into three groups,ie,the test group(treated with FTY720),the injury group(treated with normal saline)and the control group.The test and injury groups were impacted to create T10 spinal cord injury(SCI)by Allen's method.Both the mRNA and protein expressions of Bcl-2 and Bax in the injured spinal cord section were studied respectively with hematoxylin and eosin(HE)staining,immunohistochemical examination and reverse transcription polymerase chain reaction(RT-PCR)technique at different time points(at6 h,12 h,24 h,3 d and 7 d). Results The expressions of Bcl-2 and Bax in the test group and the injury group were higher than that in the control group at all time points.Meanwhile,at 12 h,24 h,3 d and 7 d,the mRNA and protein expressions of Bcl-2 in the test group were significantly higher than that in the injury group(P<0.05),while the mRNA and protein expressions of Bax in the test group were obviously lower than that in the injury group (P<0.05). Condusion Early administration of FTY720(0.5 mg/kg)after spinal cord injmy Canincrease the mRNA and protein expressions of Bcl-2,decrease the expression of Bax and inhibit neuron apoptosis in the injured spinal cord.