ABSTRACT
SUMMARY OBJECTIVE: One of the most important factors that adversely affects the outcome of peripheral nerve surgery is the formation of epineural and extraneural scar tissue after surgery. Many surgical methods and pharmacological and chemical agents have been used to prevent the formation of epineural scar tissue, but satisfactory results have not been achieved in clinical applications. The purpose of this study was to investigate the combined effect of fat graft and platelet-rich fibrin on the formation of epineural scar tissue and on nerve healing in the mature rat model. METHODS: A total of 24 female Sprague-Dawley rats were used. A circumferential segment of epineurium was excised from both bilateral sciatic nerves. The epineurectomized right nerve segment was wrapped with a combination of fat graft and platelet-rich fibrin (experimental group), while the left nerve segment did not receive any surgical procedure other than the epineurectomy (sham group). Notably, 12 randomly selected rats were sacrificed in the fourth week for histopathological examination of early results. The other 12 rats were sacrificed in the eighth week for late results. RESULTS: The formation of fibrosis, inflammation, and myelin degeneration were less common in the experimental group, while nerve regeneration was found to be higher at both 4 and 8 weeks. CONCLUSION: The intraoperative application of a combination of fat graft and platelet-rich fibrin appears to be effective on nerve healing after surgery at both the early and late periods.
ABSTRACT
@#[Abstract] Objective: To investigate the effect of miR-93/EphA4 (Eph receptor A4) axis on the proliferation and migration of nonsmall cell lung cancer (NSCLC) H460 and H1299 cells via regulating extracellular regulated protein kinases (ERK) pathway. Methods: The expression levels of miR-93 in H460 and H1299 cells was detected by qPCR. miR-93 mimics and EphA4 overexpression plasmids were transfected into H460 cells and miR-93 inhibitor was transfected into H1299 cells respectively, after which MTT assay and Transwell assay were used to detect the effects of miR-93 on proliferation and migration of transfected cells. The targeted regulatory relationship betweenmiR-93andEphA4wasverifiedbyDual-luciferasereportergeneassay.Theexpression levels of PCNA(proliferating cell nuclear antigen), EphA4, ERK and p-ERK were detected by Westernblotting.The effects of simultaneous overexpression of miR-93 and EphA4 on proliferation and migration of H460 cells were detected by MTT assay and Transwell assay. Results: The expression of miR-93 in H1299 cells was higher than that in H460 cells (P<0.01). Overexpression of miR-93 promoted proliferation and migration of H460 cells (all P<0.01), and knockdown of miR-93 inhibited proliferation and migration of H1299 cells (all P<0.01). The Dualluciferase reporter gene assay confirmed that miR-93 could target EphA4. Overexpression of miR-93 down-regulated the mRNA and protein expression levels of EphA4(allP<0.05), and promoted proliferation and migration of H460 cells through targeted regulation of EphA4 and activation of ERK pathway (all P<0.01). Conclusion: miR-93 promotes the proliferation and migration of NSCLC cells, and its mechanism may be related to the targeted regulation of EphA4 and activation of the ERK pathway.
ABSTRACT
Objective To investigate the effect of Yanghe decoction serum on the proliferation of breast cancer stem cells HMLER90hi and its mechanism. Methods Twenty female Sprague-Dawley rats were randomly divided into low,medium and high dose groups containing drug serum and control group, in order to prepare the Yanghe decoction serum and blank serum. After 24 hours of drug intervention,the effects of each group on the proliferation of HMLER90hi cells at 24 h,48 h,and 72 h were detected by MTT assay. The expression of EphA4 and p50 mRNA in each group were detected by real-time fluorescence quantitative PCR. Results Compared with the blank control group,the cell proliferation activity of each dose group of Yanghe decoction significantly decreased at 24 h (0.818 ± 0.061, 0.706 ± 0.073, 0.587 ± 0.052 vs. 0.928 ± 0.075), 48h (0.760 ± 0.047, 0.638 ± 0.056, 0.510 ± 0.059 vs. 0.973 ± 0.095), and 72 h (0.672 ± 0.102, 0.508 ± 0.092, 0.448 ± 0.048vs.1.023 ± 0.099) (P<0.05 orP<0.01), respectively. After 24 h of drug intervention, compared with the control group, the expression of EphA4 mRNA (0.54 ± 0.07, 0.54 ± 0.07, 0.33 ± 0.04 vs.0.68 ± 0.09) significantly decreased, and p50 mRNA (0.69 ± 0.10, 0.54 ± 0.08, 0.41 ± 0.06vs. 0.85 ± 0.13) significantly decreased in each dose group of Yanghe decoction (P<0.05 orP<0.01).ConclusionsTheYanghe decoction can inhibit the proliferation of breast cancer stem cell HMLER90hi,and its mechanism may be related to its inhibition of the conduction of the juxtacrine pathway of monocyte macrophage.
ABSTRACT
<p><b>Background</b>MicroRNAs (miRNAs) have been reported to play vital roles in liver regeneration. Previous studies mainly focused on the functions of intracellular miRNAs, while the functions of circulating exosomal miRNAs in liver regeneration remain largely unknown. The aim of this study was to identify the key exosomal miRNA that played vital roles in liver regeneration.</p><p><b>Methods</b>The Sprague-Dawley male rats were assigned to 70% partially hepatectomized group (n = 6) and sham surgery group (n = 6). The peripheral blood of both groups was collected 24 h after surgery. The exosomal miRNAs were extracted, and microarray was used to find out the key miRNA implicated in liver regeneration. Adenovirus was used to overexpress the key miRNA in rats, and proliferating cell nuclear antigen (PCNA) staining was applied to study the effect of key miRNA overexpression on liver regeneration. Western blotting was used to validate the predicted target of the key miRNA.</p><p><b>Results</b>Exosomal miR-10a was upregulated more than nine times in hepatectomized rats. The level of miR-10a was increased in the early phase of liver regeneration, reached the top at 72 h postsurgery, and decreased to perioperative level 168 h after surgery. Moreover, enforced expression of miR-10a by adenovirus facilitated the process of liver regeneration as evidenced by immunohistochemical staining of PCNA. Erythropoietin-producing hepatocellular receptor A4 (EphA4) has been predicted to be a target of miR-10a. The protein level of EphA4 was decreased in the early phase of liver regeneration, reached the bottom at 72 h postsurgery, and rose to perioperative level 168 h after surgery, which was negatively correlated with miR-10a, confirming that EphA4 served as a downstream target of miR-10a. Moreover, inhibition of EphA4 by rhynchophylline could promote the proliferation of hepatocytes by regulating the cell cycle.</p><p><b>Conclusion</b>Exosomal miR-10a might accelerate liver regeneration through downregulation of EphA4.</p>
ABSTRACT
Objective To investigate the influences of sodium salicylate on the auditory brainstem response (ABR)and expression of EphA4mRNA in rat inferior colliculus and its effects on salicylate ototoxicity.Methods A total of 30 healthy SPF rats were randomly divided into five groups:the control group (without any treatment),S7 (i.m.injection of sodium salicylate,175mg/kg,twice daily for 7 days),S14(the same method as S7,twice daily for 14 days),S14+R14(the same method as S14,twice daily for 14 days and recovered for 14 days),and S14+R28(the same method as S14,twice daily for 14 days and recovered for 28 days).After the ABR assessment,rats were sacri-ficed after deep anesthesia and the inferior colliculus tissues were dissected.Real-time PCR was used to detect the expression of EphA4mRNA.Results Compared with the control group,ABR thresholds in the S7 group and S14 group were increased significantly (P0.05).The inferior colliculus EphA4mRNA expression level of S7 group was signifi-cantly decreased than the control group (P0.05).Conclusion Long term injection of sodium salicylate can cause changes in the inferior colliculus of EphA4mRNA which are related closely with synaptic plasticity.It may lead the alteration of the inferior colliculus synaptic plasticity,which is associated with the changes of the hearing failure and the tinni-tus behavior.This indicates that EphA4 which is considered as a related protein in the inferior colliculus may play an important role in the pathology of tinnitus.
ABSTRACT
Objective To investigate the effects of locomotor training on improving locomotor function after spinal cord injury(SCI)and the mechanism of spinal cord plasticity.Methods A model of complete thoracic cord transection was established using 84 adult female rats divided into sham,SCI and treadmill training(BWSTT) groups.The hind limb locomotor function of all the rats was evaluated.The fluorescence intensities due to (EphA4),vesicular glutamate transporter 2(VGluT2)and EphA4/VGluT2 double-positive neurons in the ventromedial area of the anterior horn of the lumbar COrd were detected using immunofluorescence double labeling and laser confocal microscopy.Results The rats in the BWSTT group showed better functional recovery in their hind limbs than those in the SCI group.BWSTT was correlated with markedly increased EphA4.VGIuT2 and EphA4/VGluT2 intensities in the ventromedial area.Conclusion BWSTT improves hind limb locomotor function in rats with thoracic cord transections by elevating the expression of EphA4/VGluT2,promoting neuronal plasticity in the lumbar anterior horn.