Objective To investigate the effect of chondroitinase ABC (ChABC) on the expression of growth associated protein 43 (GAP-43) and gl ial fibrillary acidic protein (GFAP) after spinal cord injury (SCI) in rats. Methods A total of 150 adult female SD rats, weighing 250-300 g, were randomly divided into ChABC treatment group (group A), sal ine treatment group (group B), and sham operation group (group C) with 50 rats in each group. In groups A and B, the rats were made the SCI models and were treated by subarachnoid injection of ChABC and sal ine; in group C, the rats were not treated as a control. At 1, 3, 7, 14, and 21 days after operation, the Basso, Beattie, and Bresnahan (BBB) score system was used toevaluate the motion function, and immunofluorescent histochemical staining was used to observe the expressions of GAP-43 and GFAP. Results At different time points, the BBB scores of groups A and B were significantly lower than those of group C (P lt; 0.05); there was no significant difference in BBB score between groups A and B after 1, 3, and 7 days of operation (P gt; 0.05), but the BBB score of group A was significantly higher than that of group B after 14 and 21 days of operation (P lt; 0.01). At different time points, the GAP-43 and GFAP positive neurons of groups A and B were significantly higher than those of group C (P lt; 0.05). After 14 and 21 days of operation, the GAP-43 positive neurons of group A were more than those of group B (P lt; 0.01). After 7, 14, and 21 days of operation, the GFAP positive neurons of group A were significantly less than those of group B (P lt; 0.01). Conclusion ChABC can degrade gl ial scar, improve the microenvironment of the injured region and enhance the expression of GAP-43, which promotes axonal growth and extension.
Objective To evaluate the characteristics, classification, treatment methods, and cl inical outcomes of the spoke heel injuries in children. Methods From June 2001 to June 2008, 289 children with bicycle or motorcycle spoke heel injuries were treated, including 179 males and 110 females aged 2-12 years old (average 3.9 years old). There were 179 cases of skin contusion and laceration (type I), 83 cases of skin and soft tissue defect with Achilles tendon exposure (type II), and 27 cases of wide skin and soft tissue defect with the Achilles tendon defect and rupture (type III). The defect size of the skin or the soft tissues ranged from 3 cm × 2 cm to 11 cm × 7 cm in type II and type III injury. The time between injury and hospital admission was 1-53 days (average 14.5 days). Child patients with type I injury were managed with dressing or suturing after debridement. For the child patients with type II injury, the wound was repaired with the regional fascia flap in 53 cases, the reverse sural neurocutaneous vascular flap in 19 cases, the reverse saphenous neurocutaneous vascular flap in 9 cases, and the lateral supramalleolar flap in 2 cases. For the child patients with type III injury, 6 cases underwent primary repair of the Achilles tendon followed by the transposition of the reverse sural neurocutaneous vascular flap, 3 cases received primary repair of the wound with the reverse sural neurocutaneous vascular flap and secondary reconstruction of the Achilles tendon with the upturned fascia strip or the ipsilateral il iotibial tract transplant, and 18 cases underwent primary repair of the wound and the Achilles tendon with the sl iding bi-pedicled gastrocnemius musculocutaneous flap. The flap size ranged from 4 cm × 2 cm to 30 cm × 12 cm. All the donor sites were closed bypartial suture and spl it-thickness skins graft. The lower l imbs were immobil ized with plaster spl ints after operation. Results All the flaps survived except for 1 case of type II suffering from distal flap venous crisis 3 days after operation and 6 cases of type III suffering from distal flap necrosis 3-5 days after operation. All those flaps survived after symptomatic treatment. All the skin grafts at the donor site survived uneventfully. All the wounds healed by first intention. All child patients were followed up for 15-820 days (average 42 days). Child patients with type I and type II injury had a full recovery of ankle functions. While 25 cases of type III injury had ankle dorsal extension degree loss (10-30°) and unilateral plantar flexion strength decrease 3 months after operationwithout influence on walking, and 2 cases recovered well. Conclusion Spoke heel injury in children has special mec hanisms of injury, and the choice of proper treatment method should be based on the types of injury.
Objective To construct and screen neurite outgrowth inhibitory 66-samll interfering RNA (nogo66-siRNA) eukaryotic expression vectors of effective interference, so as to lay a foundation for further reconstruction of related viral vector. Methods? The nogo66-siRNA fragments were designed and cloned into pGenesil-1.1, 4 plasmids of pGenesil-nogo66-siRNA-1, pGenesil-nogo66-siRNA-2, pGenesil-nogo66-siRNA-hk, and pGenesil-nogo66-siRNA-kb were obtained, sequenced and identified, then were transfected into C6 cell l ine. The transfection efficiency was measured by fluorescence microscope. RT-PCR and Western blot were used to detect the expression of nogo gene and select the plasmid of effective interference. Results DNA sequencing results showed interference sequences were correct. The bands of 800 bp and 4.3 kb were detected when pGenesil-nogo66-siRNAs were digested by Kpn I /Xho I. The expression of green fluorescent protein could be detected under fluorescence microscope, and the transfection efficiency was about 73%. RT-PCR and Western blot results showed that compared to non-transfected cells, the transfection of pGenesil-nogo66-siRNA-1 made the expression of nogo gene decl ine 22% and the expression of nogo protein decl ine 73%; the transfection of pGenesil-nogo66-siRNA-2 made the expression of nogo gene decl ine 28% and the expression of nogo protein decl ine 78%; the differences were significant (P lt; 0.05); and the transfection of pGenesil-nogo66-siRNA-hk and pGenesil-nogo66- siRNA-kb did not make the expressions of nogo gene and nogo protein decrease significantly (P gt; 0.05). Conclusion Nogo66-siRNA eukaryotic expression vector is successfully constructed, it lays an experimental foundation for repair of spinal cord injury.
Objective To assess the relationship between the change in fluid overload at 48 h after initiation of continuous renal replacement therapy (CRRT) and 28-day mortality in critically ill patients with acute kidney injury (AKI). Methods A retrospective cohort study was performed using data from the MIMIC-IV database from 2008 to 2019. Patients who received CRRT for AKI for more than 24 h within 14 d of admission to the intensive care unit were included. The exposure variable was the proportion of change of fluid overload (ΔFO%, defined as the difference between body weight normalized fluid input and output) at 48 h after CRRT initiation, and the endpoint was 28-day mortality. Generalized additive linear regression models and logistic regression models were used to determine the relationship between the exposure and endpoint. Results A total of 911 patients were included in the study, with a median (lower quartile, upper quartile) ΔFO% of ?3.27% (?6.03%, 0.01%) and a 28-day mortality of 40.1%. Generalized additive linear regression model showed that the ΔFO% at 48 h after CRRT initiation was associated with a J-shaped curve with 28-day mortality. After adjusting for other variables, as compared with the second quartile of ΔFO% group, the first quartile group [odds ratio (OR)=1.23, 95% confidence interval (CI) (0.81, 1.87), P=0.338] was not associated with higher risk of 28-day mortality, while the third quartile group [OR=1.54, 95%CI (1.01, 2.35), P=0.046] and the fourth quartile group [OR=2.05, 95%CI (1.32, 3.18), P=0.001] were significantly associated with higher risk of 28-day mortality. There was no significant relationship between ΔFO% groups and 28-day mortality in the first 24-hour after CRRT initiation (P>0.05), but there was a linear relationship between ΔFO% and 28-day mortality in the second 24-hour after CRRT initiation, the larger the ΔFO%, the higher the mortality rate [OR=1.10, 95%CI (1.04 1.16), P<0.001 for per 1% increase]. ConclusionIn critically ill patients with AKI, the ΔFO% greater than ?3.27% within 48 h after CRRT initiation is independently associated with an increased risk of 28-day mortality, and the goals of CRRT fluid management may be dynamical.
To observe the change of morphology and neuropeptide in the spinal neurons in order to clarify the functional state after injury of peripheral nerves is especially in the late stage. Sciatic nerves were cut with their proximal segments in the preparation of a model of peripheral nerve injury. Combination of horseradish peroxidase retrograde tracing immunohistochemistry and computer image analysis the changes in the morphometry of the perikarya of ventral horn neurons of the spinal cord, the quantitative changes of substance P (SP). Calcitonin gene-related peptide (CGRP) in dorsal horn and CGRP and choline acetyransferase (CHAT) in ventral horn of the spinal cord were examed. The results showd: (1) At the 3rd week after injury, swollen perikarya of the ventral horn neurons were observed, subseauently the swelling of perikarya was decreased tile the 6th week the neurons recovered to their normal size. At the 12th week the neurons were generally stable in their size, shortening of the dendrites was seen in 27% of the neurons. (2) The dendrites of the neurons progressively contracted till at the 12th week 53% of them were degenerated. The results of the 24th week were similar to the that at the 12th week. (3) CGRP in the ventral horn of the spinal cord was elevated to the highest point after 1 week of injury, that lasting for 4 weeks and 8 weeks later, the lever of CGRP returned to normal. From 20th to 24th week, there was no obvious changes of CHAT in the ventral horn of the spinal cord during observation. (4) SP went to the lowest point in the dorsal horn during 2-6 weeks, then recovered slowly, and beiny normal again after 16 weeks, however, CGRP was changed slightly. The results indicated that although a series of degenerating changes occurred in the neurons of the spinal cord during the late peripheral nerve injury, but the functional activity of the central meurons still was maintained at a certain level.
ObjectiveTo observe the effect of transplantation of neural stem cells (NSCs) induced by all-trans-retinoic acid (ATRA) combined with glial cell line derived neurotrophic factor (GDNF) and chondroitinase ABC (ChABC) on the neurological functional recovery of injured spinal cord in Sprague Dawley (SD) rats.
MethodsSixty adult SD female rats, weighing 200-250 g, were randomly divided into 5 groups (n=12): sham operation group (group A), SCI model group (group B), NSCs+GDNF treatment group (group C), NSCs+ChABC treatment group (group D), and NSCs+GDNF+ChABC treatment group (group E). T10 segmental transversal injury model of the spinal cord was established except group A. NSCs induced by ATRA and marked with BrdU were injected into the site of injury at 8 days after operation in groups C-E. Groups C-E were treated with GDNF, ChABC, and GDNF+ChABC respectively at 8-14 days after operation;and group A and B were treated with the same amount of saline solution. Basso Beattie Bresnahan (BBB) score and somatosensory evoked potentials (SEP) test were used to study the functional improvement at 1 day before remodeling, 7 days after remodeling, and at 1, 2, 5, and 8 weeks after transplantation. Immunofluorescence staining and HE staining were performed to observe the cells survival and differentiation in the spinal cord.
ResultsFive mouse died but another rats were added. At each time point after modeling, BBB score of groups B, C, D, and E was significantly lower than that of group A, and SEP latent period was significantly longer than that of group A (P<0.05), but no difference was found among groups B, C, D, and E at 7 days after remodeling and 1 week after transplantation (P>0.05). BBB score of groups C, D, and E was significantly higher than that of group B, and SEP latent period was significantly shorter than that of group B at 2, 5, and 8 weeks after transplantation (P<0.05);group E had higher BBB score and shorter SEP latent period than groups C and D at 5 and 8 weeks, showing significant difference (P<0.05). HE staining showed that there was a clear boundary between gray and white matter of spinal cord and regular arrangement of cells in group A;there were incomplete vascular morphology, irregular arrangement of cells, scar, and cysts in group B;there were obvious cell hyperplasia and smaller cysts in groups C, D, and E. BrdU positive cells were not observed in groups A and B, but could be found in groups C, D and E. Group E had more positive cells than groups C and D, and difference was significant (P<0.05). The number of glial fibrillary acidic protein positive cells of groups C, D, and E was significantly less than that of groups A and B, and it was significantly less in group E than groups C and D (P<0.05). The number of microtubule-associated protein 2 positive cells of groups C, D, and E was significantly more than that of groups A and B, and it was significantly more in group E than groups C and D (P<0.05).
ConclusionThe NSCs transplantation combined with GDNF and ChABC could significantly promote the functional recovery of spinal cord injury, suggesting that GDNF and ChABC have a synergistic effect in the treatment of spinal cord injury.
ObjectiveTo summarize the mechanism of neutrophil extracellular traps (NETs) in hepatic ischemia-reperfusion injury (HIRI) and the research progress in targeting NETs to reduce HIRI, providing valuable reference for reducing HIRI. MethodThe related literatures at home and abroad about the role of NETs in the pathogenesis of HIRI and target NETs to alleviate HIRI were retrieved and reviewed. ResultsHIRI usually appeared in the process of liver surgery and was a common clinical problem, which occured in situations such as liver surgery, organ transplantation, liver ischemia and so on. This kind of injury would lead to tissue necrosis, inflammatory response and oxidative stress, which was a major cause of hepatic dysfunction and multiple organ failure after hepatic surgery, greatly increases the complications and mortality after hepatic surgery. NETs played a crucial role in the aseptic inflammatory response induced by hepatic ischemia/reperfusion. During hepatic ischemia-reperfusion, neutrophils promoted inflammatory cascade reactions and cytokine storms by forming NETs, exacerbating damage caused by hepatic ischemia-reperfusion. At present, some experimental and clinical studies had shown that inhibiting the formation of NETs or eliminating the formed NETs could alleviate the hepatic ischemia-reperfusion injury and improve the prognosis. ConclusionsTargeting NETs may become a new method for treating hepatic ischemia-reperfusion injury. In the future, it is foreseeable that more experiments and clinical trials will be conducted on targeted NETs for the treatment of hepatic ischemia-reperfusion injury. And gradually establish more comprehensive and effective treatment strategies, thereby providing new ways to improve the prognosis of hepatic surgery patients in clinical practice.
Objective
To investigate the expressions of heat shock protein 27 (HSP27), Bcl-2, and Bax proteins of the nerve cells after spinal cord ischemia/reperfusion injury (SCII) in rats and their relationship.
Methods
Seventy adult male Sprague Dawley rats (weighing, 200-220 g) were randomly divided into the sham operated group (sham group, n=35) and the SCII group (n=35). Only the left renal artery was exposed with no occlusion of the abdominal aorta in the rats of sham group. The left renal artery was exposed with occlusion of the abdominal aorta for 20 minutes in the rats of SCII group. At 4, 8, and 12 hours and at 1, 2, 3, and 5 days, reperfusion treatment was performed in 5 rats respectively, and then the spinal cord tissue was harvested to detect the expressions of HSP27, Bcl-2, and Bax protein of the nerve cells by using immunohistochemistry staining.
Results
The HSP27 began to express at 4 hours, reached the peak at 3 days, and decreased at 5 days in SCII group; significant differences were found between at 3 and 5 days and at the other time points (P lt; 0.05). The Bcl-2 expression increased at 4 hours, reached the peak at 1 day and maintained a high level at 2 days, and then gradually decreased; significant differences were found between at 1 and 2 days and at the other time points (P lt; 0.05). The Bax expression reached the peak at 12 hours and 3 days, and decreased at 5 days; significant differences were found between at 12 hours and 3 days and at the other time points (P lt; 0.05). A little expression of each protein was observed in sham group at different time points; the expressions of HSP27, Bcl-2, and Bax proteins in SCII group were significantly higher than those in sham group at different time points (P lt; 0.05).
Conclusion
There may be the time window of self repair after SCII. High expression of HSP27 has an obvious protective effect on the SCII in rat, by promoting the expression of the anti-apoptotic protein Bcl-2 and reducing the expression of the pro-apoptotic protein Bax so as to inhibit spinal cord cell apoptosis.
ObjectiveTo investigate the effect of continuous occupational therapy (OT) on the life satisfaction of patients with spinal cord injury (SCI). MethodsFifty-two SCI patients treated in Department of Rehabilitation at People’s Hospital of Mianzhu City between 2008 and 2010 were randomly assigned into two groups with 26 patients in each. Patients in the trial group received OT and rehabilitation nursing both in hospital and after being discharged from hospital, whereas patients in the control group only received treatment in hospital. Life satisfaction was assessed when patients were discharged from hospital and 21 months later. ResultsThe patients were treated for an average of 12 weeks in hospital before being discharged. Twenty-six questionnaires were given out to the patients when they were discharged from hospital, and another 26 were given 21 months later. All the questionnaires were retrieved, with a retrieval rate of 100%. The life satisfaction scores between the trial and control groups were not different from each other when the patients were discharged from hospital (P>0.05). The trial group was more satisfied with their life 21 months after being discharged from hospital (P<0.05). The life satisfaction scores of the control group were not changed (P>0.05). The trial group had higher life satisfaction than the control group 21 months after being discharged (P<0.05). ConclusionContinuous OT instruction on patients can increase their life satisfaction, and the rehabilitation effect of patients is better than rehabilitation intervention at a certain stage.
