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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.
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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.
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Objective To explore the modification of bionic silk fibroin nerve conduits (SF-NCs) by silk sencin.Methods The innovative SS/SF blended-NCs was fabricated by a vertical sequential cooling thermal induced phase separation (TIPS) processing with SF solution added sericin in proportion,its morphology was observed by Scanning electron microscopy (SEM),X-ray diffraction (XRD) and infrared spectroscopy (FTIR) were used to detect its internal molecular structure.MTT assay was used to quantitatively analyzed the PC12 cells viability co-cultured with the innovative SS/SF-NCs,SEM was used to observe the adhesion and morphology of PC12 cells seeded into the innovative SS/SF,PC12 cells were used to assess the NGF bioactivity released from the SS/SF.Results The SEM results showed that the new fabricated SS/SF-NCs had linearly oriented lamellar-like multiple-channel which distributed evenly,got great changes on the channel microstructure and their mechanical properties had been greatly improved,compared to SF-NCs.The XRD and FTIR results showed that the SS/SF-NCs had the similar internal molecular structure with natural silk.The spaces between parallel lamellar-like channels,porosities and compressive strengths of the SS/SF-NCs decreased with decreasing Sequential freezing temperature.MTT assay results showed that the viability of PC12 cell was better than the control group (P ＜ 0.05).The SEM observation indicated that PC12 cells showed good adhesion and differentiation with neuritis outgrowth during the period of co-culture with the SS/SF-NCs.NGF release from the innovative SS/SF-NCs was prolonged over 4 weeks,and remained bioactive.Conclusion The new fabricated SS/SF-NCs modified though silk sericin,which was highly bionic the structure of peripheral nerve fasciculus,had excellent mechanical properties and could be used as another alternative of artificial nerve conduits.
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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.
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Objective To evaluate the clinical effects of partial fascicle from the ulnar nerve to biceps branch of musculocutaneous nerve to treat brachial plexus injury. Methods Six cases of brachial plexus injury were involved in this group.3 cases were upper trunk injury and 3 cases were accompanied partial lower trunk injury.A partial fascicle of ulnar nerve transfered to repair biceps branch underwent in all cases,phrenic nerve or accessory nerve were transfered to repair suprascapular nerve.The mean time from injury to surgery was 2.8 months.Patients were evaluated with regard to elbow flexion and should abduction ansle,grip strengthen,morbidity of ulnar nerve function lose. Results Five cases out of six got follow up.The mean period of follow-up was 18 months(range from 9-30 months).The average reinnervation time for the biceps muscle was 3.3 months. All the patients' recovery of elbow flexion Was M_3~+-M_5; and the shoulder adduction was 90°-180°;the grip strength was not downgraded. No notable impairment of the donor site nerve function was observed in 4 cases,just 1 case with a little more fascicle been harvested had partial ulnar nerve impairments. Conclusion The use of ulnar nerve partial fascicle to biceps branch combined with phrenic nerve or accessory nerve to suprascapular nerve to reconstruct upper roots avulsion of the brachial plexus is a valid and convenient procedure.It can obtain good functional restoration in elbow flexion and shoulder adduction in a resonable time.The cases with partial lower trunk injury of brachial plexus,the partial fascicle of ulnar nerve can still be used for repair the musculocutaneous nerve.
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Objective To provide anatomical basis for one-stage full dynamic correction of late facial palsy with double latissimus dorsi segmental muscle flaps.MethodsForty sides of formaldehyde fixed latissimus dorsi in twenty cadavers were selected to microsurgery anatomy,three sides fresh latissimus dorsi specimens were blood vessel casted.The extramusclar and intramuscular neurovascular distribution were observed.Results①In 92.5%of the thoracodorsal nerve divides into a medial and a lateral branch be fore entering the muscle.In 7.5%of the thoracodorsal nerve divides into three major branches.The coordinate of the bifurcation point of the thoracodorsal Here was[(7.94±1.23)cm,(3.71±1.68)cm].In the district of this angle's middle line,the numbers of vessels and nerves is relative few.②The lateral muscle flaps of the latissimus dorsi can be divided into 3-5 independent segmental flaps,the medial muscle flaps of the latissimus dorsi Can be divided into 2-4 independent segmental flaps.③The segmental nerve usually locate the two sides of vessel,which are arranged(from medial to lateral)in a NVAV order in the reedial segmental branches(100%),in a VAVN order in the lateral segmental branches(85.0%)and in a NVAV order in the lateral segmental branches(15.0%).④The neural pedicle Was cut from the lateral bifurcation point,the mean length of the third medial muscle neural pedicle was 16 cm；the mean length of the third or the forth lateral muscle neural pedicle was 12 cm.Conclusion Late facial palsy can be cuwd with onestage cross-facial transplantation of neurovascularized free double latissimus dorsi segmental muscle flaps.
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Objective To evaluate the effect of microneurovascular muscle transplantation for long standing facial paralysis by employing phrenic nerve as recipient motor nerve source Method Three cases with long standing facial palsy were treated using this operative method The latissimus dorsi with thoracodorsal vessels and thoracordorsal nerve was used as donor The ipsilateral phrenic nerve was used as a recipient motor nerve source and the facial artery and vein as recipient vessels The transferred muscle was fixated on the zygomatic arch,the nasalabial fold and commissure of orbicular oris Results All of cases obtained function restoration of the transferred muscle during 12～16 weeks postoperatively Both static and dynamic facial appearance improved Among them,two cases showed a little over bulk in the operative side,among them,one case was improved by removing a part of subcutaneous fat and outer layer of transferred muscle in another operation Conclusion Comparing with the previous operative method,this new operation method could be completed in one stage,which had not to explore the buccal branch of facial nerve in the health side of face,therefore,there is no scar left in the health side In addition,phrenic nerve was easy to explore and longer enough for anastomosis to the nerve attached to the muscle,so the short length of nerve in the transferred muscle is more easy to regeneration after anastomosis to the recipient nerve All these suggests that the transplanted muscle with phrenic nerve as recipient motor source is a simple,short time consumption, less scar left and good effect operation method for facial paralysis
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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
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Objective To study the retrograde axonal transport of Schwann cell derived neurotrophic factor(SDNF)by motor neurons Methods SDNF was labeled with Chloramine T, 125 I SDNF and 100 fold excess of unlabelled SDNF was injected into the right hindlegs of neonatal rats separately The spinal cords was removed for counting in a counter Results The radioactive level of experimental side was more than contralateral side The transport was blocked by 100 fold excess of unlabelled SDNF Conclusion SDNF was transported retrogradely by spinal motor neurons
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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.