Objective:To investigate the species and number of glial cell clusters, main signaling pathway and progression of glial cell clusters after peripheral nerve injury (PNI) in rats using single-cell RNA sequencing (scRNA-seq).Methods:Twenty-seven SD rats were randomly divided into sham-operated group, group of 3 d after PNI and group of 7 d after PNI ( n=9). Thrice squeezing the right sciatic nerves in the group of 3 d after PNI and group of 7 d after PNI and no damage in the sham-operated group were performed. Species and number of glial cell clusters in the right sciatic nerve samples were detected by scRNA-seq. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to analyze the signal pathway enriched by differentially expressed genes (DEGs) of glial cells in the right sciatic nerve samples. Pseudo time analysis was used to simulate the progression of glial cell clusters in the right sciatic nerve samples. Results:(1) ScRNA-seq revealed a total of 1 609 glial cells (mainly cluster 6 [ n=1 388]) in the sciatic nerve samples of sham-operated group; 6 176 glial cells were observed in the sciatic nerve samples of group of 3 d after PNI, mainly cluster 2 ( n=3 124) and cluster 3 ( n=959); 8 975 glial cells were observed in the sciatic nerve samples of group of 7 d after PNI, mainly cluster 1 ( n=3 071), cluster 4 ( n=1 696), and cluster 5 ( n=1 389). (2) GO and KEGG analysis showed that compared with those in the sham-operated group, biological processes such as protein translation, cadherin binding and ribosome composition were up-regulated in glial cells in the group of 3 d after PNI and group of 7 d after PNI. Compared with those in the group of 3 d after PNI, glial cells enriched in biological processes such as axonal regeneration, myelination and focal adhesion, and in upregulated mitogen activated protein kinase (MAPK) signaling pathway and phosphoinositide 3-kinase (PI3K)-protein kinase B (PKB) signaling pathway in the group of 7 d after PNI. (3) Pseudo time analysis showed that glial cells in the sciatic nerve samples were mainly cluster 6 (marker genes: Atp1a2 and Sparcl1) in the sham-operated group, progressed into cluster 2 (marker genes: Mapt and Slc7a11) and cluster 3 (marker genes: Esco2 and Neil3) mainly in the group of 3 d after PNI, and progressed into cluster 1 (marker genes: Bcas1 and Prx), cluster 4 (marker genes: Ccn2 and Gap43), and cluster 5 (marker genes: Cd24 and Atxn1) mainly in the group of 7 d after PNI. Conclusion:In rats after PNI, glial cells can up-regulate MAPK signaling pathway and PI3K-PKB signaling pathway; with prolonged injury time, glial cells can progress into clusters with marker genes Bcas1, Prx, Cd24 and Atxn1 mainly.

Objective:To explore the effect of sciatic nerve derived exosomes(SN-EXO) on axon regeneration and functional recovery after peripheral nerve injury(PNI).Methods:From March 2021 to October 2022, the Department of Orthopedics of the First Affiliated Hospital of Zhengzhou University studied the effect of SN-EXO on the proliferation of Schwann cells(SCs) through EdU cell proliferation experiment. Twenty-one healthy male SD rats were randomly divided into 3 groups of sham operation, peripheral nerve injury(PNI) and SN-EXO treatment, with 7 rats in each group. The right sciatic nerves of rat models in sham group were exposed without injury. In the rat in PNI group and SN-EXO treatment group, PBS and SN-EXO were injected under the epineurium of right sciatic nerves following sciatic nerve crush. Sciatic nerve function index(SFI) was performed at 28 days after operation, and then sacrificed. Right sciatic nerves were removed for further exploration of nerve regeneration. The histopathological changes and axon arrangement of sciatic nerves were evaluated by HE staining. Regeneration efficiency of neurofilaments and SCs were obserred by NF200 and S100β double staining of sciatic nerve. The data obtained were statistically analyzed, and P<0.05 was statistically significant. Results:It was found that SN-EXO can significantly enhance the proliferation ability of SCs, with statistically significant difference( P<0.05). SFI in SN-EXO treatment group and PNI group were(-27.65±4.36) and(-57.33±7.49), respectively, and the difference was statistically significant( P<0.05). Axons in SN-EXO treatment group were arranged more closely and orderly than those in the PNI group at 28 days after operation, and there were less injury induced axon disintegration and vacuolation. Immunofluorescence assay indicated that NF200 and S100β fluorescence intensity in SN-EXO treatment group was significantly higher than that in the PNI group, and the difference was statistically significant( P<0.05). Conclusion:SN-EXO could enhance the proliferation of SCs to promote axon regeneration following peripheral nerve injury.

Objective　It is important to distinguish between motor and sensory fascicles of the peripheral nerves for nerve alignment in surgery.No biomarkers currently are available for identification of motor or sensory fascicles.The objective of this study is to search the specific proteins between sensory and motor fascicles of peripheral nerves and provide biomarkers for the identification of functional fascicles of peripheral nerves.Methods　The normal state of motor branch and saphenous nerve of femoral nerve in Wistar rats,and at 8 hours and 8 days after Sunderland V injury were respectively sampled.Five mm long samples were taken from the distal side of the broken end,and a total of 18 groups of proteins were isolated from 6 samples.After purification and quantification,differential gel electrophoresis (DIGE) was used to label the proteins,gel image was scanned,and image analysis software (DeCyder) was used to compare and identify the differentially expressed proteins in each group.Protein spots with more than 1.5 times of difference in expression were selected to prepare glue-cutting,enzyme-cutting and spot target.PMF chromatogram was analyzed and identified by MALDI-TOF-PRO mass spectrometer,and the results of proteomics were analyzed and compared by RT-PCR.Chi-square tests and t-tests were performed for comparison between motor or sensory nerve groups.Results　The data identified 6 proteins that were differentially expressed between motor and sensory fascicles (>1.5-fold,P<0.05),including Annexin V,neurofilament light polypepticle,TEC kinase,serine protease inhibitor A3N,Peroxiredoxin-2,and TPM1.The proteomic results were consistent with the mRNA expression levels of these genes as determined by quantitative reverse transcription polymerase chain reaction.Conclusion　There were significant differences in proteomic expression between the peripheral sensory and motor fascicles,and Annexin V can be used as a high-difference marker protein to distinguish the peripheral sensory from motor fascicles.

Peripheral nerve injuries, the worldwide difficult and complicated diseases, cause serious dysfunction but without ideal strategies to assist the successful treatment and recovery. The primary strategy to repair the peripheral nerve injuries is to bridge the lesions by promoting axon regeneration. The importance of mechanical factors in the nervous system has been appreciated only recently. It has been demonstrated that the application of external mechanical force is able to direct axonal outgrowth and to stimulate axonal elongation. For this purpose, to direct axonal growth cones in vivo with accurate force appears to be a challenge that must be resolved. Bionanotechnology is promising a new tool that offers the possibility to influence the process of axonal regeneration. The magnetic force generated by the interaction between Magnetic nanoparticles (MNPs) and magnetic fields provides remote, non-invasive, accurate, and controllable lead and control of neuronal axon regeneration.

Objective To discuss the clinical application of reverse dorsal metatarsal artery perforator flap for reconstruction of forefoot soft tissue defects.Methods Since January,2008 to March,2016,43 patients with forefoot soft tissue defect due to various causes were reconstructed with the flap based on dorsal metatarsal artery perforator.Dorsal pedis was used as a donor site with dorsal metatarsal artery perforator as the donor artery.The flap size varied from 2.0 cm × 4.0 cm to 6.0 cm × 8.0 cm.Among them,27,8,4 and 4 cases were reconstructed with flap based on first,second,third and fourth dorsal metatarsal artery perforator flap respectively.Results All the patients were followed-up which ranged from 3 months to 24 months.Six flaps suffered from post operative venous congestion.Out of 6,4 survived with early emergency management while the remaining 2 suffered epidermal necrosis which survived with regular dressing change.Eventually,all the flaps survived.They had good texture and they were elastic,good-looking and very wear-resisting.Walking function was normal.Conclusion Reverse dorsal metatarsal artery perforator flap is an ideal choice in reconstruction of small to medium sized soft tissue defects of forefoot.

Objective To investigate the surgical indications and repair methods for the treatment of electrical bums in the limbs.Methods Thirty-eight cases since May,1997 to June,2014 who underwent flap treatment in 7-14 days after electrical bums was summarized.Among these patients,4 cases were treated by local flaps,1 case was treated by thenar flap,2 cases were treated by dorsal island skin flap of the index finger,1 case was treated by cross-leg flap,1 case was treated by medial plantar skin flap;2 cases were treated by pedicle latissimus dorsi flaps,4 cases were treated by antebrachial flap with a pedicle of the posterior interosseous artery,4 cases were treated by metacarpal artery reversed island flaps,10 cases were treated by groin flaps;1 case was treated by free radial artery flap,1 case was treated by free latissimus dorsi flap,3 cases were treated by free anterolateral thigh flaps;1 case was treated by reverse island forearm posterior interosseous flap,2 cases were treated by posterior perforator reversed island flaps of the lower leg,1 case was treated by anterolateral perforator reversed island flaps of the lower leg.The area of flap was from 42.0 cm × 13.0 cm to 6.0 cm × 3.0 cm.The survival rate of the flap,the appearance and function of limb were observed.Results All these patients were followed up from 4 months to 3 years 6 months,the mean time was 6 months and 20 days.Thirty-five flaps survived completely,accounting for 92.1％,and 3 cases suffered partial necrosis at the distal end,accounting for 7.9％.Secondary healing was achieved by dress changing.The appearance of the limbs was satisfactory.Among the 28 cases whose bums were on the hands,the total number of excellent and good was 15,medium was 8,and 5 poor cases.The excellent and good rate was 57.1％.Conclusion Early repairing of the electrical burns with flaps is the effective method for protecting the deep structures of the limb,preventing and control the secondary infections and preserve the function of the limbs.

Objective To demonstrate the clinical outcome of bilateral gluteus maximus musculocutaneous flap in conjunction with continuous postoperative negative pressure wound therapy in reconstruction of sacral soft tissue defects.Methods From January, 2008 to April, 2013, 18 patients (8 males and 10 females, aged from 34 to 78 years old) with full-thickness sacral soft tissue defects were treated.The size of the defects after initial debridement ranged from 3.0 cm × 2.0 cm to 18.0 cm × 14.0 cm, with the exposure of sacrum or ligament.Bilateral gluteus maximus musculocutaneous flap were applied in all the patients.Two drainage tubes were placed on each side of the flaps during the surgery and suck for 10 to 12 consecutive days after the operation.Results The size of the harvested flaps ranged from 12.0 cm × 8.0 cm to 18.0 cm × 12.0 cm, and all the donor sites of the flaps were closed with primary suture.Thirty-five flaps in 17 patients survived without any complication.Partial necrosis of one flap was found in 1 patient and managed successfully with conservative dressing change.Fourteen patients were followed-up ranged from 8 months to 2.5 years (mean follow-up was 18 months).Color and texture of the flaps were satisfactory and no recurrence of sacral defect was noted.Conclusion Bilateral gluteus maximus musculocutaneous flap in conjunction with continuous postoperative negative pressure wound therapy may serve as a useful option for fullthickness sacral soft tissue defects.

Objective To analysis the clinical diagnosis and treatment of the scaphoid and lunate bone fracture with (or) wrist dislocation.Methods A retrospective analysis of the clinical data included 77 cases,which got admitted according to the inclusion and exclusion criteria and followed up completely from 2005 to 2013.In this group,there were 64 males and 13 females whose age arranged from 15 to 73 with a mean age of 31.9.There were 41 cases of simple fracture,32 cases of bone fracture with dislocation and 4 cases of simple dislocation.Treatment of cases was divided into the conservative treatment group (9 cases) and the surgery group (68 cases).The surgery group was further divided into four subgroups according to surgical method (i.e.compression screw fixation,Kirschner wire fixation,proximal row of carpal resection,and lunate excision head shaped bone transposition).Clinical outcome was evaluated based on modified Mayo wrist score and X-ray.Results All of 77 patients were followed up completely.Follow-up time arranged from 6 months to 8 years,with an average follow-up time of 3 years and 5 months.9 cases were treated conservatively.In the surgical treatment group,there were 36 cases of compression screw fixation,34 cases of Kirschner wire fixation,1 case of proximal row of carpal resection and 1 case of the lunate bone resection heads transposition.The average healing time of compression screw group was 6.1 weeks and the Kirschner group was 7 weeks.According to the modified Mayo wrist score,excellent:51 cases (66.2％),good:11 cases (14.3％),fair:9 cases,and poor:6 cases.The rate of excellent and good was 80.5％.The cure rate of compression screw treatment was 97％,and 85％ of Kirschner.The difference of cure rate was statistically significant.After surgery,there were four cases that got scaphoid malunion.The degree of deformity was not serious with only one case happened in the dominant hand,and the wrist pain got better after symptomatic treatment.1 case of scaphoid fracture with dislocation had the problem of wrist instability.18 weeks later,with ligament tightening surgery,postoperative Mayo score went back to excellent.Conclusion Lateral X-ray film of wrist is a preferred method for the examination of scaphoid and lunate fractures with (or) dislocation.Since it's easy to misdiagnose,films of special position are needed to be added.Surgery is preferred for treatment,and the selecting of surgical methods are based on the situation of fracture and dislocation.The advantages of screw fixation are obvious.

10.3969/j.issn.2095-4344.2013.25.013

Objective To introduce our experiences of reconstruct the first web space using flaps.Methods One hundred and twelve patients sustained contracture of the first, summarized the indications of different flaps in coverage of the first webspace and their prognosis of these flaps were retrospectively reviewed.Results The local flaps, such as double Z-plasty or Z-plasty, were suitable to cover minor contracture of the first webspace; the dorsal flap arising from the index finger was suitable for the coverage of the medium contracture, and the posterior interosseous artery flap or the reverse antebrachial island flap based on the dorsal branch of the anterior interosseous artery suitable for the severe cases. Conclusion The usage of flaps is important to resume the cosmetic shape and function of the first webspace after release of the contracture of the first webspace.