Objective To analyse the molecular structure of Schwann cell derived neurotrophic protein (SDNP) by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI TOF MS) Methods The purity of SDNP was digested byTPCK trypsin and detected by MALDI TOF MS, mass mapping was determined and partial sequences of the protein have neen analyzed by the mass data of fragment ion peaks in the fragmentation analysis and structural TOF MS (FAST) spectra The primary partial structure of SDNP was identified by searching the protein databases Results The integrity SDNP in peak detected by MALDI TOF MS has 66?10 3 MW and higher purity, because no other peaks exists except double charge peak The mass mapping of many peptides was determined and 8 peptides of them have been retrieved in MS Fit database, there is not the same protein in database Hypothetical protein has 5 peptides homology with the sample (62%) FAST spectra of 2305 Da shows the primary partial structure of SDNP after searching the protein MS Tag database Conclusions The molecular weight of SDNP detected by MALDI TOF MS is 66?10 3, The sequence of partial amino acid is EPVKKVTNSRRAKRTKPNGHIAN

Objective To investigate the effect of perforator identification before DIEP flap dissection for the patient with abdominal scar.Methods Preoperative multidetector-row computed tomography angiography was used to identify that the dominant perforators of the abdominal wall were not damaged completely.During the second stage breast reconstruction operation,the located dominant perforator and the DIEP flap were dissected.Results The dominant perforator located by MDCTA was identified with the exploration in operation.Follow-up for half a year,the flap survived well and the patient was satisfied with the appearance.Conclusion Abdominal scar was not the definite contraindication for DIEP flap.MDCTA provided a good quality evaluation of the perforator vessels.The located dominant perforator was dissected to confirm the blood supply of the DIEP flap.Identification of perforator can be used as a routine preoperative evaluation for patients with scar on donor site.

Objective To study the neurotrophic activity of Schwann cells derived neurotrophic factor (SDNF) on developing motoneurons Methods The right facial nerve of 14 neonatal SD rats was transected neonatal SD rats was transected. The rats were randomly divided into two groups. SDNF (10?g/d) was applied for 7 days in experimental group and PBS(loul) was received in contral group. The mounts and morphology of motoneurons were studied. Results The number and morphometry of experimental group was better than that of controlled group significantly. Conclusion SDNF could save facial motoneurons fron axotomy induced cell death in neonatal rats.

Objective To probe the diagnostic value of MRI in wounded feet which were negative on X-ray film.Methods 77 cases with wounded feet displayed normally on X-ray film underwent MRI,using SE/T1WI,TSE/SPIR/T2WI and 3D-water sel.-FFE.Results The abnormalities on MRI were found in 54 cases,of them,47 cases were within 30 days including bone contusion in 20 cases,muscle injury in 12 cases,bone contusion and muscle injury in 6 cases,bone contusion and soft tissue space injury in 2 cases,simple soft tissue space injury in 3 cases,tiny bone fracture in 3 cases and bone fracture in combination with contusion 1 case.7 cases were 30 days after trauma including cancellous bone sclerosis and little ossature deformity.Conclusion MRI is of important value in the wounded feet which are negative on X-ray film and have obvious symptom.

Objective Comparison of induction time on the proliferation of induced adipose-derived stem cells (ADSC) to differentiate into Schwann-like cells (iSC).Methods From March,2017 to October,2018,ADSCs were isolated from inguinal adipose tissue of healthy adult female SD rats.Flow cytometry was performed to detect ADSC positive markers CD29,CD90 and negative marker CD45.iSC induction medium was used to culture ADSC.S-100 and GFAP were detected by immunofluorescence staining to confirm that ADSC had differentiated into iSC.Morphological changes of cells were observed by inverted microscope on day 1st,4th,7th,10th,13rd,16th and 19th after induction.MTS assay was used to evaluate cell proliferation ability.Tunel staining was applied to assess cell apoptosis.Results Both S100 and GFAP were expressed in iSC.On day 7th,the cell proliferation rate was significantly slower than that before induction (A value was 0.330±0.020 vs.0.400±0.004,P＜0.05).It was negatively correlated with induction time.On day 19th,the proliferation rate of iSC was lower than 50％ of the proliferation rate before induction (A value was 0.016±0.003 vs.0.400±0.004,P＜0.05).Apoptosis of iSC was more obvious than ADSC at the same time point.Conclusion The proliferation ability of ADSC-induced iSC is optimal within 7 days after induction.

Objective To establish a stable experimental model of vascularized composite allograft (VCA),which would facilitate us to study of the reaction and intervening measure regarding rejection reaction in the future.Methods From September,2016 to July,2017,30 healthy male New Zealand rabbits,weighted 2.5-3.0 kg each,were chosen.Their ears should be intact without defect or necrosis.All of them were randomly and eaqually divided into 2 groups:transverse amputated group and V-shaped amputated group.In situ ear replantation after the amputation was performed.Histology analysis of skin and cartilage were done through HE and TUNEL staining,in order to compare vital rate of these ears.Results Thirty rabbits underwent ear replantation,including 13 via transverse incision and 17 via V-shaped incision.In transverse group,no ear survived,and some of them encountered vein crisis gradually after operation.The survival time ranged from 1 day to 10 days.There were 2 ears survived in V-shaped group.From HE staining,it was found certain vacuolar degenerated cells within skin and cartilage in failure ears.The rates of cell necrosis and apoptosis were higher than the survived ears.Conclusion Rabbit ear replantation model is viable.However,the rabbit ear replantation model is not suitable to be used in large samples.

OBJECTIVETo investigate the effect of mammalian target of rapamycin(mTOR) inhibitor-rapamycin on cognitive function after chronic cerebral ischemia in mice and its molecular mechanism.

METHODSThe chronic cerebral ischemia model was induced by ligation of right common carotid artery (rUCCAO) in 6-week-old ICR mice. The expressions of mTOR, S6K, S6 and corresponding phosphorylated proteins were detected by Western blotting at different time interval (1 h, 3 h, 6 h, 24 h, 3 d, 7 d, 2 w, 4 w, 6 w) after rUCCAO to determine the changes of mTOR signaling pathway. Rapamycin was administrated i.p. at the dose of 3.0 mg/kg 24 h after rUCCAO. Fluoro Jade B staining was used to detect the apoptotic cells. The expressions of Beclin and LC3-Ⅱ were detected by Western blotting to determine the status of autophagy. Morris water maze test and Y maze test were performed to evaluate cognitive functions.

RESULTSThe mTOR signaling pathway was abnormally activated from 6 h to 6 w after rUCCAO in mouse cortex. The activation of mTOR signaling pathway induced by rUCCAO was reversed by administration of rapamycin, and the apoptotic cell number was significantly decreased (146.1±16.3 vs 84.5±9.6,<0.05). Meanwhile, the elevation of Beclin and LC3-Ⅱ protein induced by rUCCAO was reversed by rapamycin administration. Furthermore, compared with vehicle-treated mice, the latent period[(11.1±2.3) s vs (8.1±1.8) s,<0.05] and swimming distance[(672.8±128.5) cm vs (558.2±124.9) cm,<0.05] were significantly decreased and the number of crossing the platform quadrant in Morris water maze increased(2.8±0.9 vs 5.2±0.8,<0.05) in rapamycin-treated mice. Correct response rate in the Y maze was also increased significantly in rapamycin-treated mice[(38.5±9.2)% vs (64.9±7.9)%,<0.05].

CONCLUSIONSInhibiting mTOR pathway by rapamycin reverses the rUCCAO-induced cognitive impairment partly through the suppression of apoptosis and autophagy.