Objective To observe the effect of comprehensive rehabilitation in patients with peripheral nerve injuries after the Wenchuan earthquake. Methods A total of 24 cases of peripheral nerve injuries who were admitted to the Rehabilitation Center for Earthquake Victims of West China Hospital of Sichuan University were treated with comprehensive rehabilitation, including exercise therapy, acupuncture therapy, functional electrical stimulation, and occupational therapy (mainly sensory training and wearing orthosis). After 30 treatment sessions, patient motor and sensory function, upper limb functional activity, and electrodiagnostic parameters were evaluated. Meanwhile, concomitant injuries were also recorded. Results As for the recovery of motor and sensory functions, the effective rate was 41.66%. The difference in the scores of upper limb functional activities was statistically significant before and after treatment (Plt;0.01). As assessed by electromyogram and nerve conduction velocity, the response rate was 87.50%. Patients with more concomitant injuries were likely to have slower recovery. Conclusion Comprehensive rehabilitation is appropriate and effective for patients with peripheral nerve injuries after the Wenchuan earthquake.
Objective To observe the histomorphology and the biocompatibil ity of acellular nerve prepared by different methods, to provide the experimental evidence for the selection of preparation of acellular nerve scaffold. Methods Forty-eight adult Sprague Dawley rats, male or female, weighing 180-220 g, were selected. The sciatic nerves were obtained from 30 rats and were divided into groups A, B, and C (each group had 20 nerves). The acellular sciatic nerves were prepared by the chemical methods of Dumont (group A), Sondell (group B), and Haase (group C). The effect to remove cells was estimated by the degree of decellularization, degree of demyel ination, and intergrity of nerve fiber tube. The histocompatibil ity was observed by subcutaneous implant test in another 18 rats. Three points were selected along both sides of centre l ine on the back of rats, and the points were randomly divided into groups A1, B1, and C1; the acellular nerve of groups A, B, and C were implanted in the corresponding groups A1, B1, and C1. At 1, 2, and 4 weeks after operation, the rats were sacrificed to perform the general observation and histological observation. Results The histomorphology: apart of cells and the dissolved scraps of axon could be seen in acellular never in the group A, and part of Schwann cell basilar membrane was broken. In group B, the cells in the acellular never were not removed completely, the Schwann cell basilar membrane formed bigger irregular hollows, part of the Schwann cell basilar membrane was broken obviously. But in the group C, the cells were completely removed, the Schwann cell basilar membrane remained intactly. Group C was better than group A and group B in the degree of decellularization, degree of demyel ination, integrity of nerve fiber tube and total score, showing significant differences (P lt; 0.05). The subcutaneous implant test: there were neutrophils and lymphocytes around the acellular nerve in 3 groups at 1 week after implant. A few of lymphocytes were observed around the acellular nerve in 3 groups at 2 weeks after implant. The inflammation was less in groups A1, B1, and C1 at 4 weeks after implant, part of the cells grew into the acellular nerve and arranged along the Schwann cell basilar membrane. The reaction indexes of the inflammational cells in group A1 and group B1 were higher than that in group C1 at 1, 2, and 4 weeks after implant, showing significant differences (P lt; 0.01), but there was no significant difference between group A1 and group B1 (P gt; 0.05). Conclusion The acellular sciatic nerves prepared by Haase method has better acellular effect and the histocompatibil ity than those by the methods of Dumont and Sondell.
Objective To review researches of treatment of peripheral nerve injury with neuromuscular electrical stimulation (NMES) regarding mechanism, parameters, and cl inical appl ication at home and abroad. Methods The latest original l iterature concerning treatment of peri pheral nerve injury with NMES was extensively reviewed. Results NMES should be used under individual parameters and proper mode of stimulation at early stage of injury. It could promote nerve regeneration and prevent muscle atrophy. Conclusion NMES plays an important role in cl inical appl ication of treating peripheral nerve injury, and implantable stimulation will be the future.
Objective To observe the delaying effect of neural stem cell (NSC) transplantation on denervated muscle atrophy after peri pheral nerve injury, and to investigate its mechanism. Methods NSCs were separated from the spinal cords of green fluorescent protein (GFP) transgenic rats aged 12-14 days mechanically and were cultured and induced to differentiate in vitro. Thirty-two F344 rats, aged 2 months and weighed (180 ± 20) g, were randomized into two groups (n=16 per group). The animal models of denervated musculus triceps surae were establ ished by transecting right tibial nerve and commom peroneal nerve 1.5 cm above the knee joints. In the experimental and the control group, 5 μL of GFP-NSCsuspension and 5 μL of culture supernatant were injected into the distal stump of the tibial nerve, respectivel. The generalcondition of rats after operation was observed. At 4 and 12 weeks postoperatively, the wet weight of right musculus tricepssurae was measured, the HE staining, the Mallory trichrome staining and the postsynaptic membrane staining were adopted for the histological observation. Meanwhile, the section area of gastrocnemius fiber and the area of postsynaptic membrane were detected by image analysis software and statistical analysis. Results The wounds in both groups of animals healed by first intension, no ulcer occurred in the right hind l imbs. At 4 and 12 weeks postoperatively, the wet weight of right musculus triceps surae was (0.849 ± 0.064) g and (0.596 ± 0.047) g in the experimental group, respectively, and was (0.651 ± 0.040) g and (0.298 ± 0.016) g in the control group, respectively, showing a significant difference (P lt; 0.05). The fiber section area of the gastrocnemius was 72.55% ± 8.12% and 58.96% ± 6.07% in the experimental group, respectively, and was 50.23% ± 4.76% and 33.63% ± 4.41% in the control group, respectively. There were significant differences between them (P lt; 0.05). Mallory trichrome staining of muscle notified that there was more collagen fiber hyperplasia of denervated gastrocnemius in the control group than that in the experimental group at 4 and 12 weeks postoperatively. After 12 weeks of operation, the area of postsynaptic membrane in the experimental group was (137.29 ± 29.14) μm2, which doubled that in the control group as (61.03 ± 11.38) μm2 and was closer to that in normal postsynaptic membrane as (198.63 ± 23.11) μm2, showing significant differences (P lt; 0.05). Conclusion The transplantation in vivo of allogenic embryonic spinal cord NSCs is capable of delaying denervated muscle atrophy and maintaining the normal appearance of postsynaptic membrane, providing a new approach to prevent and treat the denervated muscle atrophy cl inically.
Objective To study the functional change of nerve trunk after removing the partial bundles of ulnar nerve, to propose the concept of functional reserve of peripheral nerves and to investigate the functional reserve quantity of peripheral nerves. Methods Two hundred and twenty SD rats (male or female), aging 3 months and weighing 300-350 g, were randomized into the experimental group and the control group (n=110 per group). And the experimental group wassubdivided into group 1/8, group 1/4, group 1/3, group 1/2 and group 2/3 according to the resection portion (n=22 per group). In the experimental group, the section of the lowest level on ulnar nerve trunks was exposed, and a certain portion of its bundles was separated and cut, while in the control group the bundles were only separated without resection. The general condition of all rats was observed, and the motoneurons in cornu anterius medullae spinal is were detected at 1 week, 2 weeks and 2 months after operation. The neuro-electrophysiology and the function of dominated muscles were detected at 2 weeks, 2 months, 3 months, and 4 months after operation. Results All the rats survived without infection and obvious ulcer in the l imbs. The number of motoneurons in cornu anterius medullae spinal is in various experimental subgroups witnessed no obvious changes (P gt; 0.05). The superstructure changed obviously at the early postoperative stage in group 1/2 and group 2/3, but restored well at 2 months after operation. For the latent period of evoked potential, there was no significant difference between the various experimental subgroups and the control group at each time point (P gt; 0.05), but there was a significant difference among the various experimental subgroups when compared the time points of 2, 3 and 4 months to that of 2 weeks (P lt; 0.05) and no statistically significant difference at other time points (P gt; 0.05). For the wave ampl itude of evoked potential of motor nerves, the maximum wave ampl itude and the persistence time of the dominate muscle, there were significant differences between the various experimental subgroups and the control group at each time point (P lt; 0.05), and there were significant differences among the various experimental subgroups when comparing the time points of 2, 3 and 4 months to that of 2 weeks (P lt; 0.05) and no statistical significance at other time points (Pgt; 0.05). Conclusion The functional reserve of the ulnar nerve withoutcompromise accounts the 1/3 of the whole trunk diameter.
Objective To explore effects of several immunosuppressants on cytokine expressions after repair for a sciatic nerve injury in a rat model. Methods The sciatic nerves of 42 rats were cut and suturedend to end. After operation, the rats were divided into 6 groups. Group A(n=9) was served as a control with no medicines given. Group B (n=9) was given methylprednisolone 20 mg/(kg·d) for 2 days. Groups C(n=9) and D(n=3) were given FK506 1 mg/(kg·d) for 2 weeks and 4 weeks respectively, and were given the same doses of methylprednisolone as Group B. Groups E and F were given CsA 2 mg/(kg·d) for 2 weeks and 4 weeks respectively, and were given the same doses of methylprednisolone as Group B. The sciaticnerves were sampled at 1, 2 and 4 weeks postoperatively. And immuneohistochemistry stainings of interleukin 1β(IL-1β)， tumor necrosis factor α(TNF-α), interferon γ(IFN-γ) and macrophage migration inhibitory factor(MIF) were performed. The staining results were compared and analyzed. Results The expression peaks of IL-1β and IFN-γ were found at the 1st week postoperatively in Group A. Then, the expression decreased rapidly at the 2nd week and disappeared at the 4th week. As for TNF-α and MIF, they were only found to have a low expression until the 1st week in Group A. In groups C-F, the expression peaks of IL-1β, TNF-α and IFN-γ were found at the 2nd week, while the expression peak of MIF was still at the 1st week, and the expression of all the cytokines extended to the 4th week. The expressions of these cytokines in Group B were just between the expression levels of Group A and Groups C-F. Conclusion Immunosuppressants can delay the expression peaks and significantly extend the expression time of IL-1β, TNF-α, IFN-γ and MIF after repair for a sciatic nerve injury in a rat model.
Objective To study the effect of olfactory ensheathingcells(OECs) transplantation on protecting spinal cord and neurons after peripheral nerve injury. Methods Fifty-five SD rats were randomly divided into blank group (n=5), experimental group (n=25) and control group (n=25). The right sciatic nerves of all the rats were transected. The proximal end was embedded in muscle and treated with OECs (experimental group) and DMEM (control group). No treatment was given to the blank group. The rats were sacrificed 1, 2, 3, 7, and 14 days after the transplantation, the related neurons were observed with histological and TUNEL methods. Results After sciatic nerves were transected, death of neurons occurred in spinal cord and ganglion. One, 2, 3 days after treatment, the neuron survival rate in experimental group was 98.4％±6.5%，97.6％±6.5%，95.2％±6.7% respectively. The neuron survival rate in control group was 97.8％±6.7%，97.4％±6.4%，94.3％±6.8% 1, 2, and 3 days after treatment respectively. There was no significant difference between experimental group and control group. Seven and 14 days after treatment, the neuron survival rate in experimental group was 92.4％±8.9%，87.7％±9.4% respectively. The neuron survival rate in control group was 87.4％±8.6%，83.4％±8.5% 7 and 14 days after treatment respectively. There was significant difference between experimental group and control group. On 1st and 2nd day, no apoptosis was seen in spinal cord anterior horn of the rats in both experimental group and control group. On 3rd, 7th, and 14th day, the apoptosis index of spinal cord anterior horn motoneuron in experimental rats were lower（1.2±0.8，1.4±0.6，4.1±1.3） than that in the control group（2.1±1.1，3.1±1.1，6.1±1.8）（Plt;0.05）. One, 2, and 3 days after the operation, no ganglion neurons apoptosis was observed in all rats. On 7th day the apoptosis index of ganglion neurons in experimental group（2.10±0.32）were lower than thatin control group （4.40±0.56）（Plt;0.05）. On 14th day there was no significant difference in the apoptosis index of ganglion neurons between experimental group （4.30±1.80）and control group（6.70±2.50）(P＜0.05). Conclusion Apoptosis of neurons occur after peripheral nerve injury in spinal cord and ganglion. OECs transplantation is effective in preventing apoptosis.
Objective Targeted adenoviral gene delivery from peripheral nerves was used to integrally analyse the characterization and time course of LacZ gene (AdLacZ) retrograde transfer to spinal cord and transgene product anterograde labeling ofperipheral nerve. Methods Recombinant replication-defective adenovirus containing AdLacZ was administrated to the cut proximal stumps of median and tibial nerves in Wister rats. Then the transected nerve was repaired with 10-0 nylon sutures. At different time point postinfection the spinal cords of C5 to T1 attached with DRGs and brachial plexuses, or L2 to L6 attached with DRGs and lumbosacralplexuses were removed. The removed spinal cord and DRGs were cut into 50 μm serialcoronal sections and processed for X-gal staining and immunohistochemical staining. The whole specimens of brachial or lumbosacral plexuses attaching with theirperipheral nerves were processed for X-gal staining. The number of X-gal stained neurons was counted and the initial detected time of retrograde labeling, peaktime and persisting period of gene expression in DRG sensory neurons, spinal cord motor neurons and peripheral nerves were studied. Results The gene transfer was specifically targeted to the particular segments of spinal cord andDRGs, and transgene expression was strictly unilaterally corresponding to the infected nerves. Within the same nerve models, the initial detected time of gene expression was earliest in DRG neurons, then in the motor neurons and latest in peripheral nerves. The persisting duration of β-gal staining was shortest in motor neurons, then in sensory neurons and longest in peripheral nerves. The initial detected time of β-gal staining in median nerve models was earlier in mediannerve models compared with that in the tibial nerve models. Although the initial detected time and the beginning of peak duration of β-gal staining were not same, the decreasing time of β-gal staining in motor and sensory neurons of thetwo nerve models were started at about the same day 8 post-infection. The labeled neurons were more in tibial nerve-models than that in median nerve models. Within the same models, the labeled sensory neurons of DRGs were morethan labeled motor neurons of ventral horn. The β-gal staining was tenser in median nerves than that in tibial nerves. However the persisting time of β-gal staining was longer in tibial nerve models. Conclusion The b gene expression in neurons and PNS renders this system particularly attractive for neuroanatomical tracing studies. Furthermore this gene delivery method allowing specific targeting of motor and sensory neurons without damaging the spinal cord might offer potentialities for the gene therapy of peripheral nerve injury.