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Objective:To confirm the reuse of mismatched regenerated motor axons of brachial plexus and explore the effect of target organs on their regeneration in a rat model.Methods:This study was carried out between January 2021 and December 2021 at the research laboratory of the Department of Microsurgery, Orthopaedic Trauma and Hand Surgery, the First Affiliated Hospital of Sun Yat-sen University. Animals were randomly assigned into 2 groups, as a regeneration group (RGen) with 5 subgroups and a reuse group (RUs) with 3 subgroups. There were 6 rats per subgroup with 42 rats in total. It was observed that in the groups of RGen1-4, after the transection and suture of the musculocutaneous nerve, the motor axons of the proximal end could accurately grow into the distal corresponding endoneural tube. It was also observed that in the mismatched regenerated group, motor axons were the axons that grew into the endoneurial tube of the lateral forearm cutaneous nerve (LFCN), and other non-target organ contacts were made to the regenerated nerves after mismatch. It was specifically further divided into RGen1, the group without an organ for nerve to make contact with; RGen2, the group with skin as the target organ with nerves contact by neurorrhaphy; RGen3, the group with skin as the target organ with originally reserved natural nerve contact; RGen4, the group with muscle as the target organ with nerves contact by neurorrhaphy and RGen5, a control group. After 8 weeks, the positive area (PA), mean density (MD) and integral optical density (IOD) were measured, with AChE and ChAT fluorescence staining of the medial branch of LFCN, to evaluate the regenerated nerves after mismatch. Of the RUs group, firstly, the innervating branches of the flexor carpi radialis (FCR) were dissected and exposed, then further assigned according to initially innervated FCR (RUs1), contacted with regenerated nerves after mismatch (RUs2) and denervated (RUs3), respectively. After 8 weeks, compound muscle action potential (CMAP) and wet weight ratio of FCR were taken. Masson staining of FCR was also performed to evaluate muscle reinnervation by the regenerated nerves after mismatch. Data analysis with One-Way ANOVA and Bonferroni 0.05 indicated a statistically significant difference.Results:In the RGen groups, after AChE staining, the PA, MD and IOD of RGen3 and RGen4 were higher than that of RGen1 and RGen5, and PA of RGen4 were higher than that of RGen2, with a statistically significant difference ( P<0.05). After ChAT staining, the values of PA and IOD of RGen3 and RGen4 were higher than that of RGen1 and RGen5, and PA of RGen4 were higher than that of RGen2, with a statistically significant difference ( P<0.05). In the RUs, electrophysiological assessment showed that no CMAP was observed in RUs3, there was no significant difference in Latency of RUs1 and RUs2. The difference was statistically significant ( P<0.05). Wet weight rate of muscle of RUs1 (98.91%±3.86%) was higher than that of RUs3 (86.67%±4.68%) with a statistically significant difference ( P<0.01), but no significant difference when compared with RU2 (92.74%±3.88%). Masson staining showed that the CVF value of RUs2 (8.61%±1.16%) was significantly higher than that of RUs1 (3.17%±0.76%), and statistic significantly lower than that of RUs3 (16.44%±2.26%)( P<0.01). Conclusion:Target organ contact can promote the regenerated nerves after mismatched regeneration, and the muscle target organs exhibit greater facilitation than the cutaneous target organs. Besides, regenerated nerves after mismatch can establish effective innervation with muscle target organs, comfirming their effective reuse.
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@#Objective To analyze the clinical efficacy of transthoracic occlusion via a right subaxillary incision and conventional surgery in the treatment of ventricular septal defect (VSD). Methods The clinical data of patients with congenital VSD undergoing right subaxillary incision surgery in our hospital from January 2017 to January 2020 were retrospectively analyzed. According to the surgical methods, the patients were divided into two groups: a conventional surgery group (conventional group) and a transthoracic occlusion group (occlusion group). There were 221 patients in the conventional group, including 97 males and 124 females, with an average age of 2.6±2.2 years and an average weight of 13.4±6.2 kg; there were 185 patients in the occlusion group, including 90 males and 95 females, with an average age of 3.2±2.6 years and an average weight of 14.7±6.6 kg. The clinical effectiveness was compared. Results The success rate of surgery was 100% in both groups. The intraoperative blood loss was less in the occlusion group (P<0.05). The incision length, operation time, postoperative mechanical ventilation time, retention time in the intensive care unit, the time to resume normal diet and normal activities after operation were all shorter than those in the conventional group (P all <0.05). The total cost during hospitalization of the conventional group was less than that of the occlusion group (P<0.001). There was no statistical difference in the incidence rate of perioperative complications between the two groups (P>0.05). During the follow-up (15.8±8.8 months), the incidence of complications in the conventional group was higher than that in the occlusion group with a statistical difference (P<0.001). Conclusion Compared with conventional surgery, transthoracic occlusion for VSD via right subaxillary incision has the advantages of smaller incision, shorter operation time, less blood loss, shorter postoperative recovery time and less long-term complications. However, the total hospitalization cost is relatively high, mainly because of the high consumables cost, and the long-term effects still need further comparative observation.
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In all the carpal fractures, scaphoid fracture is the most common in clinic (from 60% to 70% in proportion) and likely leads to nonunion. If nonunion is not treated in time, it probably causes instability of the scaphoid lunate joint, further leading to scapholunate advanced collapse. Its comprehensive manifestations include degenerative arthropathy of radial styloid process, radial scaphoid joint, capitulolunate joint and even total wrist joint, eventually leading to wrist joint dysfunction. Therefore, more and more attention has been paid to treatments of scaphoid fracture nonunion. Bone grafting is the most common practical treatment, but new surgical procedures have emerged in recent years. This article thus reviews the research advances in bone grafting for scaphoid fracture nonunion, commenting on the advantages and disadvantages of various techniques.
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Objective:To observe the effects of Etifoxine on proliferation and migration of RSC96 (Schwann cells of rat) and its potential molecular mechanisms.Methods:From March, 2020 to October, 2020, cultured RSC96 were treated with 20 μmol/L Etifoxine and saline respectively for 48 h. Cell proliferation was tested by EdU assay using Cell-Light EdU DNA Cell Proliferation Kit and the capability of migration was determined by wound healing assay and a transwell system. To investigate the effects of Etifoxine on CELSR2 protein expression, after treated with different concentrations of Etifoxine at 0-20 μmol/L for 48 hours, cells were subject to Western blot analysis to verify the expression of CELSR2 protein. To explore whether CELSR2 would be a potential target of Etifoxine, siRNA targeting CELSR2 and control siRNA groups were transfected into 20 μmol/L Etifoxine-treated RSC96 using Lipo3000. Again, the cell proliferation and migration of were investigated after 48 hours with the same procedures. The two-tailed Mann-Whitney U test was employed in statistical assessment.Results:EdU results showed a significant higher percentage of Edu-positive (proliferating) cells in the 20 μmol/L Etifoxine-treated group than the control group[(36.30±3.09)% vs (19.40±2.50)%, P<0.05]. Transwell migration assay demonstrated that the number of 20 μmol/L Etifoxine-treated RSC96 which migrated through the transwell membrane was higher than saline group, with significant statistical difference [(132.30±6.77) vs(65.33±7.37), P<0.05]. The percentage of reduction of wound area measured at 24 hours and 36 hours after the scratch also showed the similar results [(30.67±2.16)% vs (23.00±2.61)%; (86.00±2.19)% vs (49.67±2.81)%, respectively, P<0.05]. Besides, with increase of the concentration of etifoxine, the expression of CELSR2 showed an trend of increase in RSC96 ( P<0.05), but no significant statistical difference was found between 10 μmol/L and 20 μmol/L groups ( P>0.05). Interestingly, the rate of cell proliferation, the number of migrating cells and the percentage of wound area reduction of RSC96 in which were treated by Etifoxine and transfected with CELSR2 siRNA were significantly decreased compared with the control siRNA treatment ( P<0.05). Conclusion:Etifoxine could promote proliferation and migration of RSC96. Upregulation of CELSR2 protein expression in RSC96 is associated with the Etifoxine-induced enhancement of cell proliferation and migration.
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@#Objective To reveal the risk factors for delayed recovery and complications in infants with weight≤ 5.0 kg after surgical ventricular septal defect (VSD) closure. Methods We retrospectively reviewed a consecutive series of 86 patients with weight≤5.0 kg who were admitted to our institution for surgical VSD closure between January 2016 and July 2019, including 31 males and 55 females with an age of 17-266 (80.3±40.4) d and a weight of 2.5-5.0 (4.4±0.6) kg. The VSDs were divided into perimembranous (n=65, 75.6%), subaortic (n=17, 19.8%) and subaortic combined muscular types (n=4, 4.7%). Mechanical ventilation (MV) time≥24 h or ICU stay≥72 h were defined as delayed recovery. Death, sudden circulatory arrest, complete heart block requiring a permanent or temporary pacemaker implantation, neurological complications, reoperation (for residue shunt or valvular regurgitation), reintubation and diaphragmatic paralysis were considered as significant major adverse events. Results There was no death, reoperation due to residual VSD or neurological complication. Totally 51 (59.3%) patients had MV timec≥24 h and 51 (59.3%) patients stayed in the ICU≥ 72 h. Two (2.3%) patients required temporary pacemaker and six (7.0%) patients required reintubation. During the follow-up of 3-36 (15.8±8.8) months, 1 patient died of pneumonia after discharge, 5 patients suffered mild tricuspid valve regurgitation and 1 patient suffered decreased left ventricular systolic function in the follow-up. No aortic valve injuries occurred. Conclusion For patients whose weight≤5.0 kg, short-term results of surgical VSD closure are excellent. Low weight and age may prolong MV time; low birth weight and pulmonary hypertension may prolong ICU stay, but are not independent risk factors.
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Objective To investigate the anatomy characteristics of saphenous artery(SA) and its perforator vessels in mini pig,and to provide a new perforator flap animal model.Methods Between May,2018 and August,2018,6 7-months-old mini pigs weight 25 kilograms were sacrificed by blooding under general anesthesia.The bilateral external iliac arteries were injected with a mixture of latex-lead oxide mixture and underwent CT-scanning to provide 3-dimensional reconstruction.The origin,diameter,courses and distribution of the SA and perforators of both hind limbs were observed.Results One of the hind limbs was abandoned due to the leakage of perfusion fluid.The other 11 hind limbs were available.The femoral artery passed through the medial femoral muscle and sartorius muscle in the middle of thigh,and run between the medial femoral muscle and gracilis muscle above the knee.Below the knee,it run on the surface of the medial leg muscles.A number of small branches were distributed in the tarsal joint and calcaneus at the medial malleolus,and there were abundant communicating branches with the posterior tibial artery.The average length of SA was (14.86±0.76) mm.The outer diameters of SA at initiative,medial tibial condyle and medial malleolus were(1.73±0.15) mm,(1.50±0.12) mm and (1.30±0.13) mm,respectively.There were a total of 79 perforators which were identified,with a mean of (7.09±1.16) perforators per hind limbs.And the average outer diameter was 0.10-0.78 (0.40±0.13) mm.Conclusion The SA constantly exist with good outer diameter.The number of perforator is abundant.SA can be used as a reliable animal model for studying and training of perforator flap.
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We aimed to express and purify three rabies virus glycoproteins with different tags and sizes. After analyzing their binding function, we wish to obtain a rabies virus glycoprotein with higher affinity and ability to specifically bind memory B cells. Experiments were carried out to express full length, as well as the ectodomain RVG by gene engineering method. Combined with the antibody of CD19 and CD27, the candidate protein labeling with fluorescence was used to analyze its binding function. Flow cytometry was used to detect the anti-rabies virus specific memory B cells in PBMCs, and confirm the binding ability between the candidate proteins and anti-rabies virus-specific memory B cells. We successfully constructed three expression vectors pGEX-5X-1-RVG, pET28a-RVG and pET30a-G. Three glycoproteins GST-RVG, His-RVG and His-G were obtained by optimized expression and purification conditions. The antigen specificity of purified GST-RVG, His-RVG and His-G were identified by Western blotting and ELISA. The affinity of these three purified glycoproteins to anti-rabies virus antibody were detected by competitive ELISA. Anti-rabies virus specific memory B cells in positive PBMCs gained from people who had ever been injected with the vaccine can be detected by flow cytometry. Thus, we got a recombinant rabies virus glycoprotein that had high-affinity and could sort antigen specific memory B cells.