Compare the effect of different chemical methods for preparation of acellular nerve scaffold and to provide an effective nerve scaffold for tissue engineering. Methods Fifteen male SD rats of 2 months old, weighing 200-250 g were selected; the bilateral sciatic nerves were harvested and divided into 3 groups according to preparation methods: group A (normal nerve), group B (Sondell method) and group C (optimal method by Triton X-200, SB-10 and SB-16). The morphology was compared by HE, immunohistochemistry and SEM after dispose; the degrees of decellularization, degrees of demyel ination and integrity of the nerve fiber tube were assessed by scoring system. Results HE staining: In group A, thecross section of nerve was roundness, the cell nuclei was dark blue and the myel in sheath was reticular structure. In group B, the axon and cell nuclei disappeared and the structure of endoneurium was destroyed. In group C, the axon and cell nuclei disappeared and the endoneurium become anomal istic round cavum. The immunohistochemistry staining of Laminin: In group A, the myel in sheath was surrounded by basement membrane with dark blue SC nuclei inside. In group B, the myel in sheath and SC nuclei disappeared and the structure of basement membrane destroyed. In group C, the myel in sheath and SC nuclei disappeared and basement membrane become anomal istic round cavum. The immunohistochemistry staining of S-100: In group A, the myel in sheath and SC were brown. In groups B and C, there were no apparent stained myel in sheath. SEM: In group A, the myel in sheath and axon were clear. In group B, the axon and myel in sheath disappeared and basement membrane became anomal istic. In group C, the basement membrane was more regular than that of group B. The degrees of acellularization and demyel ination of groups B and C were superior to that of group A (P lt; 0.05), and the degrees of demyel ination of group C were superior to that of group B (P lt; 0.05). The integrity of nerve fiber tube of group C was superior to that of group B (P lt; 0.05) and similar to that of group A (P gt; 0.05). The total score was the lowest in group C but the qual ity was the best. Conclusion The effect of decellularization of optimal method was similar to that of traditional Sondell method, but the effect of demyel ination and integrity of nerve fiber tube were better than that of traditional Sondell method. And this acellular nerve can be used as a new kind of nerve scaffold material.
Objective To determine the feasibility, safety, and efficacy of common pedicle screw placement under direct vision combined with dome shaped decompression via small incision for double segment thoracolumbar fracture with nerve injury. Methods A retrospective analysis was performed on the clinical data of 32 patients with double segment thoracolumbar fracture with nerve injury undergoing common pedicle screw placement under direct vision combined with dome shaped decompression via small incision between November 2011 and November 2015 (combined surgery group), and another 32 patients undergoing traditional open pedicle screw fixation surgery (traditional surgery group). There was no significant difference in gender, age, cause of injury, time of injury-to-surgery, injury segments and Frankel classification of neurological function between two groups (P>0.05). The length of soft tissue dissection, the operative time, the blood loss during surgery, the postoperative drainage, the visual analogue scale (VAS) of incision after surgery, and recovery of neurological function after surgery were evaluated. Results All cases were followed up 9 to 12 months (mean, 10.5 months) in combined surgery group, and 8 to 12 months (mean, 9.8 months) in traditional surgery group. The length of soft tissue dissection, the operative time, the blood loss during surgery, the postoperative drainage, and the postoperative VAS score in the combined surgery group were significantly better than those in the traditional surgery group (P<0.05). Dural rupture during surgery and pedicle screw pulling-out at 6 months after surgery occurred in 2 cases and 1 case of the combined surgery group; dural rupture during surgery occurred in 1 case of the traditional surgery group. The X-ray films showed good decompression, and fracture healing; A certain degree of neurological function recovery was achieved in two groups. Conclusion Common pedicle screw placement under direct vision combined with dome shaped decompression via small incision can significantly reduce iatrogenic trauma and provide good nerve decompression. Therefore, it is a safe, effective, and minimally invasive treatment method for double segment thoracolumbar fracture with neurological injury.
Objective To investigate the effects of exogenous pulmonary surfactant （PS） on ventilation weaning and respiratory mechanics in infants with acute lung injury（ALI） after cardiopulmonary bypass （CPB）.Methods Twelve infants underwent cardiopulmonary bypass cardiac surgery committed with ALI and difficulty in weaning from ventilation were included in this study.Exogenous PS was used in the treatment via intra-tracheal administration.The changes of blood gas，respiratory mechanics and the conditions of ventilation weaning were observed.Results After intra-tracheal PS administration，spontaneous breath remained steady；spontaneous respiratory rate significantly decreased from，tidal volume of spontaneous breath increased significantly.Three concave sign disappeared and koilosternia was alleviated.PaCO2 value decreased significantly and peak inspiratory pressure（PIP） decreased from （36.18±10.25）cm H2O to （25.11±5.14）cm H2O （Plt;0.01）.Static lung compliance （Cstat） increased from （1.49±0.65）mL·cm H2O-1·kg-1 before treatment to （1.95±0.50） mL·cm H2O-1·kg-1 6 h after treatment （Plt;0.01）；and airway resistance （Rstat）decreased from （128.17±26.34） cm H2O·L-1·s-1 before treatment to （78.56±18.22） cm H2O·L-1·s-1 6 h after treatment （Plt;0.01）.All 12 infants weaned from ventilator successfully.Conclusion Combined with PS intra-tracheal treatment，lung protective ventilation strategy can significantly improve parameters of respiratory mechanics，increase dynamic lung compliance，decrease airway resistance，which can decrease the breathing effort of the infants and make it easy to wean from ventilator.
Objective To study whether human amniotic fluid colony derived stem cells (hAFCSCs) are involved in regeneration of injured muscles in mice and to investigate the method and feasibil ity of hAFCSCs-based cytotherapy in the treatment of injured muscles. Methods Human second-trimester amniotic fluid was collected through ultrasound-guided amniocentesis, hAFCSCs were isolated from second-trimester amniotic fluid and cultured, and the cells at 6th-8th passages were spared. The mRNA was extracted to identify the stem cell related genes by RT-PCR. The muscular injury model of bilateral tibial is anterior muscle was establ ished by cardiotoxin and X-ray irradiation in 16 Nod/Scid mice (aged 6-8 weeks, and weighing 20-24 g). The hAFCSCs (3.3 × 107/mL, 30 μL) were injected into the right injured tibial is anterior muscles as the experimental group, while the same volume of complete medium (α-MEM containing 15%FBS, 18%Chang B, 2%Chang C, 1% penicill instreptomycin, and 1% L-glutamine) was injected into the left injured tibial is anterior muscles as the control group. At 2 and 4 weeks after cell transplantation, the immunofluorescence staining of tibial is anterior muscles was performed to detect hepatocyte growth factor receptor (c-Met), myogenic regulatory factor (Myf-5), Laminin, Desmin, and human specific nuclear mitotic apparatus protein (NuMa). Results The clone formation was observed at 5-7 days of primary hAFCSCs culture; after 8-10 days, the clones with homogeneous morphology were selected for subculture. Adequate stem cells were available after 6th-8th subculture. RT-PCR analysis showed that hAFCSCs expressed mRNA of the stem cell related genes. The immunofluorescence double-staining showed that NuMa expressed in tibial is anterior muscles of the experimental group and no myogenic phenotype expressed at 2 weeks after cell transplantation, and that single cell co-expressed NuMa and c-Met or Myf-5 at 4 weeks after cell transplantation. In some myofibers, NuMa and Laminin or Desmin were also co-expressed. No NuMa positive hAFCSCs were detected in the control group at 2 and 4 weeks after cell transplantation. Conclusion hAFCSCs can participate in the regeneration of injured mouse muscle.