4D printed tri-segment nerve conduit using zein gel as the ink for repair of rat sciatic nerve large defect.

Zein has enormous potential for application in biomedical field due to biodegradation and biocompatibility, we have recently prepared zein gel as a possible 3D printing ink. Our previous studies found that the pore structure in zein material can reduce early inflammation, promote the polarization of macrophages toward the M2 phenotype, and accelerate nerve regeneration. To further explore the role of zein in nerve repair, we used 4D printing technique to create nerve conduits with zein protein gel, and designed 2 types of tri-segment conduits with different degradation rates. Structural parts printed in support baths with higher water content show faster degradation rates than those printed in support baths with lower water content. The conduits that degraded quickly at both ends and slowly in the middle (CB75-CB40-CB75) and the conduits that degraded slowly at both ends and quickly in the middle (CB40-CB75-CB40) were 4D printed, respectively. Animal experiments suggest that the CB75-CB40-CB75 conduit is better for nerve repair, which may be because its degradation pattern can match to the pattern of nerve regeneration better. Our new strategy through 4D printing indicated that fine modulation in conduit degradation can affect efficacy of nerve repair significantly.

Upregulated lncARAT in Schwann cells promotes axonal regeneration by recruiting and activating proregenerative macrophages.

BACKGROUND: Axonal regeneration following peripheral nerve injury (PNI) depends on the complex interaction between Schwann cells (SCs) and macrophages, but the mechanisms underlying macrophage recruitment and activation in axonal regeneration remain unclear. METHODS: RNA sequencing (RNA-seq) was conducted to identify differentially expressed long noncoding RNAs (DElncRNAs) between crushed sciatic nerves and intact contralateral nerves. The putative role of lncRNAs in nerve regeneration was analyzed in vitro and in vivo. RESULTS: An lncRNA, called axon regeneration-associated transcript (lncARAT), was upregulated in SCs and SC-derived exosomes (SCs-Exo) after sciatic nerve injury. LncARAT contributed to axonal regeneration and improved motor function recovery. Mechanistically, lncARAT epigenetically activated C-C motif ligand 2 (CCL2) expression by recruiting KMT2A to CCL2 promoter, resulting in increased histone 3 lysine 4 trimethylation (H3K4me3) and CCL2 transcription in SCs. CCL2 facilitated the infiltration of macrophages into the injured nerves. Meanwhile, lncARAT-enriched exosomes were released from SCs and incorporated into macrophages. LncARAT functioned as an endogenous sponge to adsorb miRNA-329-5p in macrophages, resulting in increased suppressor of cytokine signaling (SOCS) 2 expression, which induced a proregenerative function of macrophages through a signal transducer and activator of transcription (STAT) 1/6-dependent pathway. CONCLUSIONS: LncARAT may represent a promising therapeutic avenue for peripheral nerve repair.

Zein-induced immune response and modulation by size, pore structure and drug-loading: Application for sciatic nerve regeneration.

Zein is a biodegradable material with great potential in biomedical applications. However, as a plant-derived protein material, body's immune response is the key factor to determine its clinical performance. Herein, for the first time, the zein-induced immune response is evaluated systemically and locally, comparing with typical materials including alginate (ALG), poly(lactic-co-glycolic) acid (PLGA) and polystyrene (PS). Zein triggers an early inflammatory response consistent with the non-degradable PS, but this response decreases to the same level of the biosafe ALG and PLGA with zein degradation. Changing sphere sizes, pore structure and encapsulating dexamethasone can effectively modulate the zein-induced immune response, especially the pore structure which also inhibits neutrophil recruitment and promotes macrophages polarizing towards M2 phenotype. Thus, porous zein conduits with high and low porosity are further fabricated for the 15 mm sciatic nerve defect repair in rats. The conduits with high porosity induce more M2 macrophages to accelerate nerve regeneration with shorter degradation period and better nerve repair efficacy. These findings suggest that the pore structure in zein materials can alleviate the zein-induced early inflammation and promote M2 macrophage polarization to accelerate nerve regeneration. STATEMENT OF SIGNIFICANCE: Zein is a biodegradable material with great potential in biomedical applications. However, as a plant protein, its possible immune response in vivo is always the key issue. Until now, the systemic study on the immune responses of zein in vivo is still very limited, especially as an implant. Herein, for the first time, the zein-induced immune response was evaluated systemically and locally, comparing with typical biomaterials including alginate, poly(lactic-co-glycolic) acid and polystyrene. Changing sphere sizes, pore structure and encapsulating dexamethasone could effectively modulate the zein-induced immune response, especially the pore structure which also inhibited neutrophil recruitment and promoted macrophages polarizing towards M2 phenotype. Furthermore, the pore structure in zein nerve conduits was proved to alleviate the early inflammation and promote M2 macrophage polarization to accelerate nerve regeneration.

Long Non-coding RNA MSTRG.24008.1 Regulates the Regeneration of the Sciatic Nerve via the miR-331-3p-NLRP3/MAL Axis.

Peripheral nerve injury (PNI) is a common clinical problem, which can cause severe disability and dramatically affect a patient's quality of life. Neural regeneration after PNI is a complex biological process that involves a variety of signaling pathways and genes. Emerging studies demonstrated that long non-coding RNAs (lncRNAs) were abnormally expressed after PNI and played pivotal roles in peripheral nerve regeneration. Based on the rat sciatic nerve injury model, we found that the expression levels of several lncRNAs were increased significantly in the sciatic nerve after injury. Software prediction prompted us to focus on one up-regulated lncRNA, MSTRG.24008.1. Dual-luciferase reporter assay, RNA pull-down assay and RNA interference approach verified that MSTRG.24008.1 regulated neuroregeneration via the miR-331-3p/nucleotide-binding oligomerization domain-like pyrin domain containing 3 (NLRP3)/myelin and lymphocyte protein (MAL) axis in vitro. Subsequently, we performed gastrocnemius muscle gravity and sciatic functional index experiments to evaluate the recovery of injured sciatic nerves after MSTRG.24008.1 siRNA interference in vivo. In conclusion, knockdown of MSTRG.24008.1 promotes the regeneration of the sciatic nerve via the miR-331-3p/NLRP3/MAL axis, which may provide a new strategy to evaluate and repair injured peripheral nerves clinically.

Exosomes produced by adipose-derived stem cells inhibit schwann cells autophagy and promote the regeneration of the myelin sheath.

Peripheral nerve injury (PNI) is encountered relatively commonly in the clinic and often results in long-term functional deficits. Research to develop methods to improve regeneration following nerve injury is ongoing. Numerous studies have shown that adipose-derived stem cells (ADSCs) promote the regeneration of peripheral nerve injury; however, the mechanism is unclear. Autophagy, a highly conserved intracellular process responsible for maintaining cellular homeostasis, and Schwann cells (SCs), play important roles in regeneration after PNI. In the present study, we explored the effect and mechanism of exosomes produced by adipose-derived stem cells (ADSC-Exos) on autophagy of SCs in PNI, as well as their effect on the regeneration of the nerve myelin sheath. The levels of autophagy and the expression of karyopherin subunit alpha 2 (Kpna2) in SCs increased markedly after the sciatic nerve was injured in SCs (SNI-SCs). The enhanced autophagy and the upregulated Kpna2 in SNI-SCs were inhibited after treatment with ADSC-Exos in vivo and in vitro. The effect of ADSC-Exos on inhibiting SC autophagy was blocked by overexpression of Kpna2 in SNI-SCs. Using quantitative real-time reverse transcription PCR, ADSC-Exos were demonstrated to contain a large amount of miRNA-26b, which was predicted to regulate Kpna2 on the TargetScan website. The effect of ADSC-Exos on inhibiting SCs autophagy was blocked after the silencing of miRNA-26b. Moreover, ADSC-Exos promoted the regeneration of the myelin sheath by inhibiting SC autophagy in rat SNI models. In conclusion, our results indicated that ADSC-Exos promote the regeneration of the myelin sheath by moderately reducing autophagy of injured SCs via miRNA-26b downregulation of Kpna2.

Reducing Neuron Apoptosis in the Pontine Micturition Center by Nerve Root Transfer for Restoration of Micturition Function after Spinal Cord Injury.

OBJECTIVE: The rate of neuronal apoptosis increases after spinal cord injury (SCI). Anastomosing the normal nerve roots above the SCI level to the injured sacral nerve roots can enhance the functional recovery of neurons. Therefore, we evaluated the effect of sacral nerve root transfer after SCI on pontine neuronal survival. METHODS: Sprague-Dawley rats were randomly divided into three groups: Group A, reconstruction of afferent and efferent nerve pathways of the bladder after SCI; Group B, SCI only; and Group C, control group. We examined pontine neuronal morphology using hematoxylin and eosin (H&E) staining after SCI and nerve transfer. Bcl-2 and Bax protein expression changes in the pontine micturition center were quantified by immunohistochemistry. The number of apoptotic neurons was determined by TUNEL staining. We examined pontine neuronal apoptosis by transmission electron microscopy (TEM) at different time points. RESULTS: H&E staining demonstrated that the number of neurons had increased in Group A, but more cells in Group B displayed nuclear pyknosis, with the disappearance of the nucleus. Compared with Group B, Group A had significantly higher Bcl-2 expression, significantly lower Bax expression, and a significantly higher Bcl-2/Bax ratio. The number of apoptotic neurons and neuron bodies in Group A was significantly lower than that in Group B, as indicated by TUNEL staining and TEM. CONCLUSIONS: These findings demonstrate that lumbosacral nerve transfer can reduce neuronal apoptosis in the pontine micturition center and enhance functional recovery of neurons. This result further suggests that lumbosacral nerve transfer can be used as a new approach for reconstructing bladder function after spinal cord injury.

Syringic acid delivered via mPEG-PLGA-PLL nanoparticles enhances peripheral nerve regeneration effect.

Aim: To deliver syringic acid (SA) with a nanocarrier and enhance its function. Materials & methods: mPEG-PLGA-PLL (PEAL) nanoparticles were used to deliver SA. The characterization, storage stability, drug release, blood-compatibility and biocompatibility of SA-PEAL were detected by in vitro and in vivo assays. Cellular phenotypic experiments and rat sciatic nerve injury models were used to evaluate the function of SA-PEALs. Results: SA-PEAL had good storage stability, blood-compatibility and biocompatibility and could slowly release SA. SA-PEAL significantly enhanced the proliferation and migration ability of Schwann cells and function recovery of injured sciatic nerves. Conclusion: Our study provides an effective nano-delivery system for enhancing the neural repair function of SA and promoting further applications of SA.

Effect of exosomes from adipose-derived stem cells on the apoptosis of Schwann cells in peripheral nerve injury.

AIMS: Recovery after peripheral nerve injury (PNI) is often difficult, and there is no optimal treatment. Schwann cells (SCs) are important for peripheral nerve regeneration, so SC-targeting treatments have gained importance. Adipose-derived stem cells (ADSCs) and their exosomes can promote peripheral nerve repair, but their interactions with SCs are unclear. METHODS: Purified SCs from sciatic nerve injury sites were harvested, and apoptosis and proliferation of SCs at post-PNI 24 hours were analyzed. The effects of coculture with ADSCs and different concentrations of ADSC-derived exosomes (ADSC-Exo) were studied through in vitro experiments by flow cytometry, CCK8 assay, immunofluorescence staining, and histological analysis. The expression of the apoptosis-related genes Bcl-2 and Bax was also analyzed by qRT-PCR. RESULTS: ADSC-Exo reduced the apoptosis of SCs after PNI by upregulating the anti-apoptotic Bcl-2 mRNA expression and downregulating the pro-apoptotic Bax mRNA expression. Further, it also improved the proliferation rate of SCs. This effect was confirmed by the morphological and histological findings in PNI model rats. CONCLUSION: Our results present a novel exosome-mediated mechanism for ADSC-SC cross talk that reduces the apoptosis and promotes the proliferation of SCs and may have therapeutic potential in the future.

Differential gene and protein expression in gastrocnemius and tibialis anterior muscle following tibial and peroneal nerve injury in rats.

Peripheral nerve injury is encountered quite commonly in the clinic, and treatment results are often not satisfactory. Therefore, promoting nerve regeneration and functional recovery is a primary goal of neuroscience research. Recovery of corresponding target muscle can differ following peripheral nerve injury, but the reasons are unknown. Herein, we investigated differential gene and protein expression in gastrocnemius and tibialis anterior muscle following tibial and common peroneal nerve injury using RNA sequencing and proteomics approaches, and analysed the results by bioinformatics. In total, 1794, 1765, 1656 and 2006 differential genes and 398, 400, 959 and 472 differential proteins were identified in gastrocnemius and tibialis anterior muscles at 1, 7, 14 and 21 days after surgery, related to activation of 51 signalling pathways. Differential expression of these genes and proteins may contribute to the degree of recovery of target organs following peripheral nerve injury. The findings provide a foundation for investigating regeneration mechanisms following peripheral nerve injury.

Syringic acid promotes proliferation and migration of Schwann cells via down-regulating miR-451-5p.

Schwann cells are the main force in spontaneous regeneration after peripheral nerve injury. The neurotrophic factors could promote the regeneration, but clinical applications of these factors are limited by some constraints. Hence, searching for new substances to elevate the function of Schwann cells and facilitate the regeneration of nerve is urgently needed. Syringic acid (SA) is a natural product with neuroprotective activity in vivo, but the role of SA on Schwann cells remains unclear. In this study, we for the first time found that SA was able to promote the proliferation and migration of Schwann cells, two important abilities in the process of regeneration. Then, microRNA (miRNA) microarray analysis was performed and 26 differentially expressed miRNAs (22 down-regulated and 4 up-regulated) were identified. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway analyses found that the target genes of these miRNAs were mainly enriched in cellular response to chemical stimulus and cancer-related pathways, respectively. Subsequently, the levels of top 6 down-regulated miRNAs were validated by RT-qPCR and miR-451-5p was shown to be the most down-regulated one. Further experiments demonstrated that inhibition of miR-451-5p significantly promoted the proliferation and migration of Schwann cells. These results suggested that SA promoted the proliferation and migration of Schwann cells via down-regulation of miR-451-5p, and SA could be developed into a promising nutritional supplement to assist peripheral nerve regeneration.

Differential gene and protein expression between rat tibial nerve and common peroneal nerve during Wallerian degeneration.

Wallerian degeneration and nerve regeneration after injury are complex processes involving many genes, proteins and cytokines. After different peripheral nerve injuries the regeneration rate can differ. Whether this is caused by differential expression of genes and proteins during Wallerian degeneration remains unclear. The right tibial nerve and the common peroneal nerve of the same rat were exposed and completely cut through and then sutured in the same horizontal plane. On days 1, 7, 14, and 21 after surgery, 1-2 cm of nerve tissue distal to the suture site was dissected out from the tibial and common peroneal nerves. The differences in gene and protein expression during Wallerian degeneration of the injured nerves were then studied by RNA sequencing and proteomic techniques. In the tibial and common peroneal nerves, there were 1718, 1374, 1187, and 2195 differentially expressed genes, and 477, 447, 619, and 495 differentially expressed proteins on days 1, 7, 14, and 21 after surgery, respectively. Forty-seven pathways were activated during Wallerian degeneration. Three genes showing significant differential expression by RNA sequencing (Hoxd4, Lpcat4 and Tbx1) were assayed by real-time quantitative polymerase chain reaction. RNA sequencing and real-time quantitative polymerase chain reaction results were consistent. Our findings showed that expression of genes and proteins in injured tibial and the common peroneal nerves were significantly different during Wallerian degeneration at different time points. This suggests that the biological processes during Wallerian degeneration are different in different peripheral nerves after injury. The procedure was approved by the Animal Experimental Ethics Committee of the Second Military Medical University, China (approval No. CZ20160218) on February 18, 2016.

Effect of exogenous spastin combined with polyethylene glycol on sciatic nerve injury.

Polyethylene glycol can connect the distal and proximal ends of an injured nerve at the cellular level through axonal fusion to avoid Wallerian degeneration of the injured distal nerve and promote peripheral nerve regeneration. However, this method can only prevent Wallerian degeneration in 10% of axons because the cytoskeleton is not repaired in a timely fashion. Reconstruction of the cytoskeletal trunk and microtubule network has been suggested to be the key for improving the efficiency of axonal fusion. As a microtubule-severing protein, spastin has been used to enhance cytoskeletal reconstruction. Therefore, we hypothesized that spastin combined with polyethylene glycol can more effectively promote peripheral nerve regeneration. A total of 120 male Sprague-Dawley rats were randomly divided into sham, suture, polyethylene glycol, and polyethylene glycol + spastin groups. In suture group rats, only traditional nerve anastomosis of the end-to-end suture was performed after transection of the sciatic nerve. In polyethylene glycol and polyethylene glycol + spastin groups, 50 µL of polyethylene glycol or 25 µL of polyethylene glycol + 25 µL of spastin, respectively, were injected immediately under the epineurium of the distal suture. Sensory fiber regeneration distance, which was used to assess early nerve regeneration at 1 week after surgery, was shortest in the suture group, followed by polyethylene glycol group and greatest in the polyethylene glycol + spastin group. Behavioral assessment of motor function recovery in rats showed that limb function was restored in polyethylene glycol and polyethylene glycol + spastin groups at 8 weeks after surgery. At 1, 2, 4 and 8 weeks after surgery, sciatic functional index values and percentages of gastrocnemius muscle wet weight were highest in the sham group, followed by polyethylene glycol + spastin and polyethylene glycol groups, and lowest in the suture group. Masson staining was utilized to assess the morphology of muscle tissue. Morphological changes in skeletal muscle were detectable in suture, polyethylene glycol, and polyethylene glycol + spastin groups at 1, 2, 4, and 8 weeks after surgery. Among them, muscular atrophy of the suture group was most serious, followed by polyethylene glycol and polyethylene glycol + spastin groups. Ultrastructure of distal sciatic nerve tissue, as detected by transmission electron microscopy, showed a pattern of initial destruction, subsequent disintegration, and gradual repair in suture, polyethylene glycol, and polyethylene glycol + spastin groups at 1, 2, 4, and 8 weeks after surgery. As time proceeded, axonal ultrastructure gradually recovered. Indeed, the polyethylene glycol + spastin group was similar to the sham group at 8 weeks after surgery. Our findings indicate that the combination of polyethylene glycol and spastin can promote peripheral nerve regeneration. Moreover, the effect of this combination was better than that of polyethylene glycol alone, and both were superior to the traditional neurorrhaphy. This study was approved by the Animal Ethics Committee of the Second Military Medical University, China (approval No. CZ20170216) on March 16, 2017.

Transferring of femoral nerve motor branches for high-level sciatic nerve injury: a cadaver feasibility study.

BACKGROUND: Sciatic nerve injuries cause significant disability. We propose here a novel reconstructive procedure of transferring the motor branches of the femoral nerve as donor nerves to reconstruct both the peroneal and tibial nerve function as a novel approach to treat high sciatic nerve injury. METHODS: The autopsies of donor nerves (vastus lateralis nerve branch (VLN), vastus medialis nerve branch (VMN), saphenous nerve (SAN)) and respective recipient nerves (deep peroneal nerve branch (DPN), medial gastrocnemius nerve branch (MGN), sural nerve (SN)) were conducted in six fresh-frozen lower limbs. The distance between the origin or bifurcation points of the nerves to the head of fibula and the diameter of the end at the coaptation site were measured. The feasibility of tensionless direct suturing or grafting between the donor nerves and the recipient was evaluated. Finally, the nerve end at the coaptation site was harvested for observation with toluidine blue staining and nerve fiber count. RESULTS: The mean diameter of the VMN, VLN, MGN, DPN, SAN, and SN nerves were 1.5 ± 0.1, 1.4 ± 0.1, 1.3 ± 0.1, 2.3 ± 0.1, 2.1 ± 0.3, and 1.3 ± 0.2 mm, respectively. Histological observation showed that the abovementioned six nerve bundles had a respective nerve fiber number of 392 ± 27, 205 ± 520, 219 ± 67, 394 ± 50, 308 ± 77, and 335 ± 49. A total of 5/6 specimens needed grafting for a length ranging from 5 to 15 cm to bridge the VMN-MGN, 6/6 needed a graft length of 10-20 cm for VLN-DPN bridging, and 2/6 needed a graft length of 0-4 cm for SAN-SN bridging. CONCLUSION: The study demonstrated the feasibility of the transferring femoral nerve branches to sciatic nerve branches to restore the function for sciatic injury.

[Effect of exosomes from adipose-derived stem cells on peripheral nerve regeneration].

Objective: To investigate the effects of exosomes from adipose-derived stem cells (ADSCs) on peripheral nerve regeneration, and to find a new treatment for peripheral nerve injury. Methods: Thirty-six adult Sprague Dawley (SD) rats (male or female, weighing 220-240 g) were randomly divided into 3 groups ( n=12). Group A was the control group; group B was sciatic nerve injury group; group C was sciatic nerve injury combined with exosomes from ADSCs treatment group. The sciatic nerve was only exposed without injury in group A, and the sciatic nerve crush injury model was prepared in groups B and C. The SD rats in groups A and B were injected with PBS solution of 200 µL via tail veins; the SD rats in group C were injected with pure PBS solution of 200 µL containing 100 µg exosomes from ADSCs, once a week and injected for 12 weeks. At 1 week after the end of the injection, the rats were killed and the sciatic nerves were taken at the part of injury. The sciatic nerve fiber bundles were observed by HE staining; the SCs apoptosis of the sciatic nerve tissue were detected by TUNEL staining; the ultrastructure and SCs autophagy of the sciatic nerve were observed by transmission electron microscope. Results: Gross observation showed that there was no obvious abnormality in the injured limbs of group A, but there were the injured limbs paralysis and muscle atrophy in groups B and C, and the degree of paralysis and muscle atrophy in group C were lighter than those in group B. HE staining showed that the perineurium of group A was regular; the perineurium of group B was irregular, and there were a lot of cell-free structures and tissue fragments in group B; the perineurium of group C was more complete, and significantly well than that of group B. TUNEL staining showed that the SCs apoptosis was significantly increased in groups B and C than in group A, in group B than in group C ( P<0.01). Transmission electron microscope observation showed that the SCs autophagosomes in groups B and C were significantly increased than those in group A, but the autophagosomes in group C were significantly lower than those in group B. Conclusion: The exosomes from ADSCs can promote the peripheral nerve regeneration. The mechanism may be related to reducing SCs apoptosis, inhibiting SCs autophagy, and reducing nerve Wallerian degeneration.

Transfer of obturator nerve for femoral nerve injury: an experiment study in rats.

BACKGROUND: Quadriceps palsy is mainly caused by proximal lesions in the femoral nerve. The obturator nerve has been previously used to repair the femoral nerve, although only a few reports have described the procedure, and the outcomes have varied. In the present study, we aimed to confirm the feasibility and effectiveness of this treatment in a rodent model using the randomized control method. METHODS: Sixty Sprague-Dawley rats were randomized into two groups: the experimental group, wherein rats underwent femoral neurectomy and obturator nerve transfer to the femoral nerve motor branch; and the control group, wherein rats underwent femoral neurectomy without nerve transfer. Functional outcomes were measured using the BBB score, muscle mass, and histological assessment. RESULTS: At 12 and 16 weeks postoperatively, the rats in the experimental group exhibited recovery to a stronger stretch force of the knee and higher BBB score, as compared to the control group (p < 0.05). The muscle mass and myofiber cross-sectional area of the quadriceps were heavier and larger than those in the control group (p < 0.05). A regenerated nerve with myelinated and unmyelinated fibers was observed in the experimental group. No significant differences were observed between groups at 8 weeks postoperatively (p > 0.05). CONCLUSIONS: Obturator nerve transfer for repairing femoral nerve injury was feasible and effective in a rat model, and can hence be considered as an option for the treatment of femoral nerve injury.

[Biomechanics test of fixation of star-shaped six-part patellar fractures with petal-shaped poly-axial locking plate].

Objective: To compare the biomechanical difference between petal-shaped poly-axial locking plate and tension band wire cerclage in fixing star-shaped 6-part patellar fractures in cadaver model, and provide the experimental data for clinical use. Methods: The paired 12 knee specimens from 6 human cadavers were randomly divided into 2 groups (the control group and the test group) after a star-shaped 6-part patellar fracture model was established. The specimens were weighted, and the control group was fixed with tension band wire cerclage and the test group was fixed with petal-shaped poly-axial locking plate. The specimens were connected to CMT5105 biomechanics test machine by a customized fixture, the total fracture gap of patellar fracture blocks was measured before testing. The knee extensor load test was performed to record the extensor load of knees at 90° flexion to extension. Then the anti gravity physiological knee extension process at 90° flexion was stimulated according to the knee extensor load. The cyclic times until failure and the total fracture gap of patellar fracture blocks after failure were recorded. Results: The specimens weight and the total fracture gap of patellar fracture blocks before testing between 2 groups had no significant difference ( t=0.410, P=0.690; t=0.650, P=0.530). In the biomechanical test, there was no significant difference of knee extension load between 2 groups ( t=0.490, P=0.638). The total fracture gap after failure in test group was significantly smaller than that in control group ( t=3.026, P=0.013), and the cyclic times until failure in test group was significantly more than that in control group ( t=2.277, P=0.046). The failure reasons in control group were all the wires slipped off the Kirschner wires, while the failure reasons in test group were the screws pulled out from the upper pole in 5 cases (83.3%) and from the lower pole in 1 case (16.7%). Conclusion: The petal-shaped poly-axial locking plate has better biomechanical stiffness to fix the star-shaped 6-part patellar fractures when compared with tension band wire cerclage method. However, this type of fracture is a serious comminuted type, and the early excessive activity still carries the risk of displacement.

[Experimental study of urinary center change in pons after conus medullaris injury in rats].

Objective: To observe the structural changes of urinary center and the expression of Bcl-2 after conus medullaris injury in rats brain so as to explore the possible influence factors of degeneration in brain. Methods: Thirty-six adult Sprague-Dawley rats were randomly divided into experimental group ( n=30) and control group ( n=6). In the experimental group, the conus medullaris injury model was established by cutting off the spinal nerve below L 4, and no treatment was done in the control group. The modeling operations in the experimental group were successful, and 2 rats died at 3 months and 5 months after modeling operation respectively, which may be caused by renal failure or urinary tract infection. In the experimental group, 6, 6, 6, 5, and 5 rats were killed at 1 day, 1 week, and 1, 3, 6 months after operation respectively, and 1 rat was killed at each time point in the control group. The dorsolateral tissue of the pontine tegmentum was harvested to perform HE staining and Bcl-2 immunohistochemical SP staining. Results: HE staining showed that there was no obvious difference between the experimental group and the control group at 1 day after operation, the neurons were densely packed, arranged neatly, and the nucleoli were clear; at 1 week, the space between the neurons in the experimental group were slightly widened; at 1 month, nucleus retraction in some neurons happened in the experimental group; at 3 and 6 months, the nuclei in the experimental group were more and more condensed, and even some cells disappeared. Bcl-2 immunohistochemical SP staining showed that the expression of Bcl-2 in the control group was weakly positive. The positive expression of Bcl-2 was found at 1 day after operation in the experimental group; the positive expression of Bcl-2 at 7 days after operation was significantly higher than that in the control group, and reached the peak; the positive expression of Bcl-2 decreased gradually at 1, 3, and 6 months after modeling operation, but it was still higher than that of the control group. Conclusion: The urinary center appears structure degeneration and necrocytosis after conus medullaris injury in rats brain. The elevated expression of Bcl-2 may be associated with brain tissue repair and function remodeling.

[Changes of endogenous Spastin expression after sciatic nerve injury in rats].

Objective: To investigate the expression change of endogenous Spastin after sciatic nerve injury in rats, and to discuss the role and significance in the peripheral nerve regeneration. Methods: Thirty-six adult male Sprague Dawley rats weighing 180-220 g were randomly divided into the experimental group ( n=30) and the control group ( n=6). Sciatic nerve compression damage model was established in the experimental group, and the sciatic nerve was only exposed in the control group. The L 4-6 spinal cord tissue was obtained to detect Spastin mRNA and protein levels by real-time fluorescence quantitative PCR and Western blot at 1, 3, 7, 14, and 28 days after operation in the experimental group ( n=6) and at 7 days in the control group. Meanwhile, the sciatic nerve at 5 mm distal to the injured site was obtained to observe the ultrastructure of the distal axon by transmission electron microscope (TEM). Results: The expression trends of Spastin gene and Spastin protein in L 4-6 spinal cord tissue of 2 groups were basically identical. In the experimental group, the expressions of Spastin gene and protein decreased at the beginning, and then increased; the expressions reduced to the minimum at 7 days after operation, and came back to the initial level at 28 days. The expression levels of Spastin mRNA and protein at 3, 7, and 14 days were significantly lower in the experimental group than the control group ( P<0.05), but no significant difference was noted between 2 groups at 1 and 28 days ( P>0.05). The expression levels of Spastin mRNA and protein at 3, 7, and 14 days were significantly lower than those at 1 and 28 days in the experimental group ( P<0.05), but no significant difference was noted between at 1 day and 28 days ( P>0.05). At 1, 3, and 7 days after operation, the myelin damage was observed by TEM; at 14 days, there were regenerating Schwann cells; at 28 days, a large number of myelinated nerve fibers were seen, which were closed to normal form. Conclusion: In the process of sciatic nerve regeneration after injury, a complex succession of changes take place in the expression of endogenous Spastin protein in rats, indicating that Spastin protein plays an important role in the process.

[Anatomical study on the repair of femoral nerve injury with anterior branch of obturator nerve].

Objective: To investigate the feasibility of the anastomosis of the anterior branch of obturator nerve and the muscular branch of femoral nerve. Methods: Five fresh frozen cadavers, including 3 males and 2 females, were included. Both of the obturator nerve, femoral nerve and their branches were dissected, then their routes and anatomical positions were observed. The diameter and the number of myelinated nerve fiber of the anterior branch of obturator nerve and femoral nerve muscular branches were measured, as well as the overlap distance between them. Results: The diameter of myelinated nerve fiber of the anterior branch of obturator nerve was (3.80±1.22) mm; the number of myelinated nerve fiber was 11 358±800. The diameters of the rectus femoris branch and the medial femoral branch were (1.60±0.54) mm and (2.20±0.66) mm, respectively; the number of myelinated nerve fiber were 4 961±655 and 6 666±466. Both the diameter and number of myelinated nerve fiber were close to the anterior branch of obturator nerve. The anterior branch of obturator nerve could be directly anastomosed with each nerve branch of femoral nerve in nontension, and the overlap distance was about 30 mm. Conclusion: It is feasible to repair the femoral nerve by transposed the anterior branch of obturator nerve and anastomosed with the femoral nerve muscular branches. And the rectus femoris branch and the medial femoral branch should be taken as the recipient nerve.