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Neural Regeneration Research 2022年第2期17卷 418-426页

作者： yu-Song yuan Hai-Lin Xu Zhong-Di Liu yu-hui kou Bo Jin Pei-Xun Zhang Department of Trauma and Orthopedics Peking University People’s HospitalPeking UniversityBeijingChina Key Laboratory of Trauma and Neural Regeneration(Peking University) Ministry of EducationBeijingChina National Center for Trauma Medicine BeijingChina

Lower extremity nerve transposition repair has become an important treatment strategy for peripheral nerve injury;however, brain changes caused by this surgical procedure remain unclear. In this study, the distal stump of the right sciatic nerve in a rat model of sciatic nerve injury was connected to the proximal end of the left sciatic nerve using a chitin conduit. Neuroelectrophysiological test showed that the right lower limb displayed nerve conduction, and the structure of myelinated nerve fibers recovered greatly. Muscle wet weight of the anterior tibialis and gastrocnemius recovered as well. Multiple-model resting-state blood oxygenation level-dependent functional magnetic resonance imaging analysis revealed functional remodeling in multiple brain regions and the re-establishment of motor and sensory functions through a new reflex arc. These findings suggest that sciatic nerve transposition repair induces brain functional remodeling. The study was approved by the Ethics Committee of Peking University People's Hospital on December 9, 2015(approval No. 2015-50).

关键词： brain connectivity brain network fALFF MRI nerve transposition repair peripheral nerve injury ReHo remodeling

Chitin scaffold combined with autologous small nerve repairs sciatic nerve defects

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Neural Regeneration Research 2022年第5期17卷 1106-1114页

作者： Bo Wang Chang-Feng Lu Zhong-Yang Liu Shuai Han Pi Wei Dian-Ying Zhang yu-hui kou Bao-Guo Jiang Department of Orthopedics and Trauma Key Laboratory of Trauma and Neural Regeneration(Ministry of Education/Peking University)Peking University People’s HospitalBeijingChina Department of Orthopedics Chinese PLA General HospitalBeijingChina National Clinical Research Center for Orthopedics Sports Medicine&RehabilitationBeijingChina

Although autologous nerve transplantation is the gold standard for treating peripheral nerve defects,it has many clinical limitations.As an alternative,various tissue-engineered nerve grafts have been developed to substitute for autologous nerves.In this study,a novel nerve graft composed of chitin scaffolds and a small autologous nerve was used to repair sciatic nerve defects in rats.The novel nerve graft greatly facilitated regeneration of the sciatic nerve and myelin sheath,reduced atrophy of the target muscle,and effectively restored neurological function.When the epineurium of the small autogenous nerve was removed,the degree of nerve regeneration was similar to that which occurs after autogenous nerve transplantation.These findings suggest that our novel nerve graft might eventually be a new option for the construction of tissue-engineered nerve scaffolds.The study was approved by the Research Ethics Committee of Peking University People's Hospital(approval No.2019 PHE27)on October 18,2019.

关键词： autologous small nerve chitin scaffold nerve defect nervous system peripheral nerve injury peripheral nerve regeneration sciatic nerve trauma

Changes in proteins related to early nerve repair in a rat model of sciatic nerve injury

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Neural Regeneration Research 2021年第8期16卷 1622-1627页

作者： yu-Song yuan Fei yu Ya-Jun Zhang Su-Ping Niu Hai-Lin Xu yu-hui kou Department of Trauma and Orthopedics Peking University People’s HospitalBeijingChina Key Laboratory of Trauma and Neural Regeneration(Peking University) Ministry of EducationBeijingChina National&Local Joint Engineering Research Center of Orthopedic Biomaterials Department of Bone&Joint SurgeryPeking University Shenzhen HospitalShenzhenGuangdong ProvinceChina National Center for Trauma Medicine BeijingChina Office of Academic Research Peking University People’s HospitalBeijingChina Diabetic Foot Treatment Center Peking University People’s HospitalBeijingChina

Peripheral nerves have a limited capacity for self-repair and those that are severely damaged or have significant defects are challenging to repair. Investigating the pathophysiology of peripheral nerve repair is important for the clinical treatment of peripheral nerve repair and regeneration. In this study, rat models of right sciatic nerve injury were established by a clamping method. Protein chip assay was performed to quantify the levels of neurotrophic, inflammation-related, chemotaxis-related and cell generation-related factors in the sciatic nerve within 7 days after injury. The results revealed that the expression levels of neurotrophic factors(ciliary neurotrophic factor) and inflammationrelated factors(intercellular cell adhesion molecule-1, interferon γ, interleukin-1α, interleukin-2, interleukin-4, interleukin-6, monocyte chemoattractant protein-1, prolactin R, receptor of advanced glycation end products and tumor necrosis factor-α), chemotaxis-related factors(cytokine-induced neutrophil chemoattractant-1, L-selectin and platelet-derived growth factor-AA) and cell generation-related factors(granulocyte-macrophage colony-stimulating factor) followed different trajectories. These findings will help clarify the pathophysiology of sciatic nerve injury repair and develop clinical treatments of peripheral nerve injury. This study was approved by the Ethics Committee of Peking University People's Hospital of China(approval No. 2015-50) on December 9, 2015.

关键词： animal model cell generation chemotaxis clamp injury inflammation injury neurotrophic factor peripheral nerve protein array repair sciatic nerve Wallerian degeneration

Combining CUBIC Optical Clearing and Thy1-YFP-16 Mice to Observe Morphological Axon Changes During Wallerian Degeneration

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Current Medical Science 2021年第5期41卷 944-952页

作者： yu-song yuAN Fei yu Su-ping NIU Hao LU yu-hui kou Hai-lin XU Department of Trauma and Orthopaedics Peking University People's HospitalPeking University'Beijing 100044China National and Local Joint Engineering Research Center of Orthopaedic Biomaterials Department of Bone&Joint SurgeryPeking University Shenzhen HospitalShenzhen 518036China Key Laboratory of Trauma and Neural Regeneration(Peking University) Ministry of EducationBeijing 100044China Office of Academic Research Peking University People's HospitalPeking UniversityBeijing 100044China Diabetic Foot Treatment Center Peking University People's HospitalPeking University'Beijing 100044China

Objective:Wallerian degeneration is a pathological process closely related to peripheral nerve regeneration following injury,and includes the disintegration and phagocytosis of peripheral nervous system cells.Traditionally,morphological changes are observed by performing immunofluorescence staining after sectioning,which results in the loss of some histological information.The purpose of this study was to explore a new,nondestmetive,and systematic method for observing axonal histological changes during Wallerian degeneration.Methods:Thirty male Thy1-YFP-16 mice(SPF grade,6 weeks old,20±5 g)were randomly selected and divided into clear,unobstructed brain imaging cocktails and computational analysis(CUBIC)optical clearing(n=15)and traditional method groups(n=15).Five mice in each group were sacrificed at 1st,3rd,and 5th day following a crush operation.The histological axon changes were observed by CUBIC light optical clearing treatment,direct tissue section imaging,and HE staining.Results:The results revealed that,compared with traditional imaging methods,there was no physical damage to the samples,which allowed for three-dimensional and deep-seated tissue imaging through CUBIC.Local image information could be nicely obtained by direct fluorescence imaging and HE staining,but it was difficult to obtain image information of the entire sample.At the same time,the image information obtained by fluorescence imaging and HE staining was partially lost.Conclusion:The combining of CUBIC and Thy1-YFP transgenic mice allowed for a clear and comprehensive observation of histological changes of axons in Wallerian degeneration.

关键词： Wallerian degeneration axon optical clearing clear,unobstructed brain imaging cocktails and computational analysis Thy1-YFP transgenic mice

Identification of four differentially expressed genes associated with acute and chronic spinal cord injury based on bioinformatics data

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Neural Regeneration Research 2021年第5期16卷 865-870页

作者： Su-Ping Niu Ya-Jun Zhang Na Han Xiao-Feng Yin Dian-Ying Zhang yu-hui kou Key Laboratory of Trauma and Neural Regeneration(Peking University) Ministry of EducationBeijingChina Department of Trauma and Orthopedics Peking University People’s HospitalBeijingChina Trauma Medicine Center Peking University People’s HospitalBeijingChina Office of Academic Research Peking University People’s HospitalBeijingChina

Complex pathological changes occur during the development of spinal cord injury(SCI),and determining the underlying molecular events that occur during SCI is necessary for the development of promising molecular targets and therapeutic strategies.This study was designed to explore differentially expressed genes(DEGs)associated with the acute and chronic stages of SCI using bioinformatics analysis.Gene expression profiles(GSE45006,GSE93249,and GSE45550)were downloaded from the Gene Expression Omnibus database.SCI-associated DEGs from rat samples were identified,and Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed.In addition,a protein-protein interaction network was constructed.Approximately 66 DEGs were identified in GSE45550 between 3–14 days after SCI,whereas 2418 DEGs were identified in GSE450061–56 days after SCI.Moreover,1263,195,and 75 overlapping DEGs were identified between these two expression profiles,3,7/8,and 14 days after SCI,respectively.Additionally,16 overlapping DEGs were obtained in GSE450061–14 days after SCI,including Pank1,Hn1,Tmem150c,Rgd1309676,Lpl,Mdh1,Nnt,Loc100912219,Large1,Baiap2,Slc24a2,Fundc2,Mrps14,Slc16a7,Obfc1,and Alpk3.Importantly,3882 overlapping DEGs were identified in GSE932491–6 months after SCI,including 3316 protein-coding genes and 567 long non-coding RNA genes.A comparative analysis between GSE93249 and GSE45006 resulted in the enrichment of 1135 overlapping DEGs.The significant functions of these 1135 genes were correlated with the response to the immune effector process,the innate immune response,and cytokine production.Moreover,the biological processes and KEGG pathways of the overlapping DEGs were significantly enriched in immune system-related pathways,osteoclast differentiation,the nuclear factor-κB signaling pathway,and the chemokine signaling pathway.Finally,an analysis of the overlapping DEGs associated with both acute and chronic SCI,assessed using the expression profiles GSE93249 and GSE45006,identified four

关键词： bioinformatics differential expression factor gene immune response injury pathways protein spinal cord

Short-term observations of the regenerative potential of injured proximal sensory nerves crossed with distal motor nerves

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Neural Regeneration Research 2017年第7期12卷 1172-1176页

作者： Xiu-xiu Zhang yu-hui kou Xiao-feng Yin Bao-guo Jiang Pei-xun Zhang Department of Orthopedics and Trauma Peking University People's Hospital Beijing China

Motor nerves and sensory nerves conduct signals in different directions and function in different ways.In the surgical treatment of peripheral nerve injuries,the best prognosis is obtained by keeping the motor and sensory nerves separated and repairing the nerves using the suture method.However,the clinical consequences of connections between sensory and motor nerves currently remain unknown.In this study,we analyzed the anatomical structure of the rat femoral nerve,and observed the motor and sensory branches of the femoral nerve in the quadriceps femoris.After ligation of the nerves,the proximal end of the sensory nerve was connected with the distal end of the motor nerve,followed by observation of the changes in the newly-formed regenerated nerve fibers.Acetylcholinesterase staining was used to distinguish between the myelinated and unmyelinated motor and sensory nerves.Denervated muscle and newly formed nerves were compared in terms of morphology,electrophysiology and histochemistry.At 8 weeks after connection,no motor nerve fibers were observed on either side of the nerve conduit and the number of nerve fibers increased at the proximal end.The proportion of newly-formed motor and sensory fibers was different on both sides of the conduit.The area occupied by autonomic nerves in the proximal regenerative nerve was limited,but no distinct myelin sheath was visible in the distal nerve.These results confirm that sensory and motor nerves cannot be effectively connected.Moreover,the change of target organ at the distal end affects the type of nerves at the proximal end.

关键词： nerve regeneration nerve remodeling peripheral nerve acetylcholinesterase staining muscle denervation neural anastomosis nerveconduit neural regeneration

Wnt5a Affects Schwann Cell Proliferation and Regeneration via Wnt/c-Jun and PTEN Signaling Pathway

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Chinese Medical Journal 2018年第21期131卷 2623-2625页

作者： Fei yu Jian Weng yu-Song yuan yu-hui kou Na Han Bao-Guo Jiang Pei-Xun Zhang Department of Orthopedics and Trauma Peking University People's Hospital Beijing 100044 China

To the Editor： Peripheral nerve injury has been a common clinical disease, and delayed treatment may ultimately lead to long-term physical dysfunction. The repair of peripheral nerve injury has become a major challenge in clinical practice. Different from the central nervous system, the peripheral nervous system has the capacity to regenerate after injury. However, the peripheral nerve regeneration after injury is a slow process, and more than 3 months is usually needed for the axons regenerating to the distal target organs or tissues, when the distal nerve stump and skeletal muscle usually become atrophy and the regenerated axons lost the ability to remodel in next few months.

关键词： GROWTH

Advance of Peripheral Nerve Injury Repair and Reconstruction

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Chinese Medical Journal 2017年第24期130卷 2996-2998页

作者： Bao-Guo Jiang Na Han Feng Rao Yi-Lin Wang yu-hui kou Pei-Xun Zhang Department Trauma and Orthopedics Peking University People's Hospital Beijing 100044 China Center Laboratory Peking University People's Hospital Beijing 100044 China

Peripheral nerve injury is a common clinical problem because the dysfunction of distal limb seriously affects patients＇ quality of life,and it results in a huge social and economic burden.[1] Repair of peripheral nerve injury is a very complex pathological process.Due to slow nerve regeneration,Wallerian degeneration of nerve stump,tissue adhesion,atrophy of muscles and motor end plates and other constraints,functional rehabilitation of the damaged nerve is always restricted.

关键词： Functional Remodeling Peripheral Nerve Injury Repair

Comparison of commonly used retrograde tracers in rat spinal motor neurons

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Neural Regeneration Research 2015年第10期10卷 1700-1705页

作者： You-lai yu Hai-yan Li Pei-xun Zhang Xiao-feng Yin Na Han yu-hui kou Bao-guo Jiang Department of Trauma and Orthopedics Peking University People’s Hospital Department of Gastroenterology Weifang People’s Hospital

The purpose of this study was to investigate the effect of four fluorescent dyes, True Blue（TB）, Fluoro-Gold（FG）, Fluoro-Ruby（FR）, and 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate（Di I）, in retrograde tracing of rat spinal motor neurons. We transected the muscle branch of the rat femoral nerve and applied each tracer to the proximal stump in single labeling experiments, or combinations of tracers（FG-Di I and TB-Di I） in double labeling experiments. In the single labeling experiments, significantly fewer labeled motor neurons were observed after FR labeling than after TB, FG, or Di I, 3 days after tracer application. By 1 week, there were no significant differences in the number of labeled neurons between the four groups. In the double-labeling experiment, the number of double-labeled neurons in the FG-Di I group was not significantly different from that in the TB-Di I group 1 week after tracer application. Our findings indicate that TB, FG, and Di I have similar labeling efficacies in the retrograde labeling of spinal motor neurons in the rat femoral nerve when used alone. Furthermore, combinations of Di I and TB or FG are similarly effective. Therefore, of the dyes studied, TB, FG and Di I, and combinations of Di I with TB or FG, are the most suitable for retrograde labeling studies of motor neurons in the rat femoral nerve.

关键词： nerve regeneration tracing efficacy fluorescent tracers retrograde tracing femoral nerve motor neurons

Intraoperative single administration of neutrophil peptide 1 accelerates the early functional recovery of peripheral nerves after crush injury

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Neural Regeneration Research 2020年第11期15卷 2108-2115页

作者： yu-Song yuan Su-Ping Niu Fei yu Ya-Jun Zhang Na Han Hao Lu Xiao-Feng Yin Hai-Lin Xu yu-hui kou Department of Trauma and Orthopedics Peking University People’s HospitalPeking UniversityBeijingChina Key Laboratory of Trauma and Neural Regeneration(Peking University) Ministry of EducationBeijingChina Office of Academic Research Peking University People’s HospitalBeijingChina National and Local Joint Engineering Research Center of Orthopaedic Biomaterials Department of Bone&Joint SurgeryPeking University Shenzhen HospitalShenzhenGuangdong ProvinceChina National Center for Trauma Medicine BeijingChina Diabetic Foot Treatment Center Peking University People’s HospitalPeking UniversityBeijingChina

Neutrophil peptide 1 belongs to a family of peptides involved in innate immunity. Continuous intramuscular injection of neutrophil peptide 1 can promote the regeneration of peripheral nerves, but clinical application in this manner is not convenient. To this end, the effects of a single intraoperative administration of neutrophil peptide 1 on peripheral nerve regeneration were experimentally observed. A rat model of sciatic nerve crush injury was established using the clamp method. After model establishment, a normal saline group and a neutrophil peptide 1 group were injected with a single dose of normal saline or 10 μg/mL neutrophil peptide 1, respectively. A sham group, without sciatic nerve crush was also prepared as a control. Sciatic nerve function tests, neuroelectrophysiological tests, and hematoxylin-eosin staining showed that the nerve conduction velocity, sciatic functional index, and tibialis anterior muscle fiber cross-sectional area were better in the neutrophil peptide 1 group than in the normal saline group at 4 weeks after surgery. At 4 and 8 weeks after surgery, there were no differences in the wet weight of the tibialis anterior muscle between the neutrophil peptide 1 and saline groups. Histological staining of the sciatic nerve showed no significant differences in the number of myelinated nerve fibers or the axon cross-sectional area between the neutrophil peptide 1 and normal saline groups. The above data confirmed that a single dose of neutrophil peptide 1 during surgery can promote the recovery of neurological function 4 weeks after sciatic nerve injury. All the experiments were approved by the Medical Ethics Committee of Peking University People's Hospital, China(approval No. 2015-50) on December 9, 2015.

关键词： crush injury defensin 1 gait analysis intraoperative administration nerve conduction velocity nervous system neutrophil peptide 1 peripheral nerve injury peripheral nerve regeneration sciatic nerve tibialis anterior muscle trauma