Effectiveness of contralaterally controlled functional electrical stimulation vs. neuromuscular electrical stimulation for recovery of lower extremity function in patients with subacute stroke: A randomized controlled trial.

Objective: This study aimed to compare the efficacy of contralaterally controlled functional electrical stimulation (CCFES) vs. neuromuscular electrical stimulation (NMES) for motor recovery of the lower extremity in patients with subacute stroke. Materials and methods: Seventy patients within 6 months post-stroke were randomly assigned to the CCFES group (n = 35) and the NMES group (n = 35). Both groups underwent routine rehabilitation plus 20-min electrical stimulation (CCFES or NMES) on ankle dorsiflexion muscles per day, 5 days a week, for 3 weeks. Ankle AROM (dorsiflexion), Fugl-Meyer assessment-lower extremity (FMA-LE), Barthel Index (BI), Functional Ambulation Category scale (FAC), 10-meter walking test, and surface electromyography (sEMG) were assessed at the baseline and at the end of the intervention. Result: Ten patients did not complete the study (five in CCFES and five in NMES), so only 60 patients were analyzed in the end. After the 3-week intervention, FMA-LE, BI, Ankle AROM (dorsiflexion), and FAC increased in both groups (p < 0.05). Patients in the CCFES group showed significantly greater improvements only in the measurement of Fugl-Meyer assessment-lower extremity compared with the NMES group after treatment (p < 0.05). The improvement in sEMG response of tibialis anterior by CCFES was greater than NMES (p < 0.05). Conclusion: Contralateral controlled functional electrical stimulation can effectively improve the motor function of the lower limbs better than conventional neuromuscular electrical stimulation in subacute patients after stroke, but the effect on improving the ability to walk, such as walking speed, was not good. Clinical trial registration: http://www.chictr.org.cn/, identifier: ChiCTR2100045423.

Constraint-induced movement therapy enhances AMPA receptor-dependent synaptic plasticity in the ipsilateral hemisphere following ischemic stroke.

Constraint-induced movement therapy (CIMT) can promote the recovery of motor function in injured upper limbs following stroke, which may be associated with upregulation of α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor (AMPAR) at synapses in the ipsilateral sensorimotor cortex in our previous study. However, AMPAR distribution is tightly regulated, and only AMPARs on the postsynaptic membrane can mediate synaptic transmission. We speculated that synaptic remodeling induced by movement-associated synaptic activity can promote functional recovery from stroke. To test this hypothesis, we compared AMPAR expression on the postsynaptic membrane surface in a rat model of ischemic stroke induced by middle cerebral artery occlusion (MCAO) with versus without CIMT, which consisted of daily running wheel training for 2 weeks starting on day 7 after MCAO. The results showed that CIMT increased the number of glutamate receptor (GluR)2-containing functional synapses in the ipsilateral sensorimotor cortex, and reduced non-GluR2 AMPARs in the ipsilateral sensorimotor cortex and hippocampal CA3 region. In addition, CIMT enhanced AMPAR expression on the surface of post-synaptic membrane in the ipsilateral sensorimotor cortex and hippocampus. Thus, CIMT promotes the recovery of motor function of injured upper limbs following stroke by enhancing AMPAR-mediated synaptic transmission in the ischemic hemisphere. These findings provide supporting evidence for the clinical value of CIMT for restoring limb movement in stroke patients. All experimental procedures and protocols were approved by the Department of Laboratory Animal Science of Fudan University, China (approval No. 201802173S) on March 3, 2018.

Effectiveness of Contralaterally Controlled Functional Electrical Stimulation versus Neuromuscular Electrical Stimulation on Upper Limb Motor Functional Recovery in Subacute Stroke Patients: A Randomized Controlled Trial.

Purpose: To compare the effectiveness of contralaterally controlled functional electrical stimulation (CCFES) versus neuromuscular electrical stimulation (NMES) on motor recovery of the upper limb in subacute stroke patients. Materials and Methods: Fifty patients within six months poststroke were randomly assigned to the CCFES group (n = 25) and the NMES group (n = 25). Both groups underwent routine rehabilitation plus 20-minute stimulation on wrist extensors per day, five days a week, for 3 weeks. Fugl-Meyer Assessment of upper extremity (FMA-UE), action research arm test (ARAT), Barthel Index (BI), and surface electromyography (sEMG) were assessed at baseline and end of intervention. Results: After a 3-week intervention, FMA-UE and BI increased in both groups (p < 0.05). ARAT increased significantly only in the CCFES group (p < 0.05). The changes of FMA-UE, ARAT, and BI in the CCFES group were not greater than those in the NMES group. The improvement in sEMG response of extensor carpi radialis by CCFES was greater than that by NMES (p = 0.026). The cocontraction ratio (CCR) of flexor carpi radialis did not decrease in both groups. Conclusions: CCFES improved upper limb motor function, but did not show better treatment effect than NMES. CCFES significantly enhanced the sEMG response of paretic extensor carpi radialis compared with NMES, but did not decrease the cocontraction of antagonist.

[Effect of Modified Constraint-induced Movement Therapy on Neurotransmitter Levels of Motor Cortex in Cerebral Ischemia-Reperfusion Injured Rats].

OBJECTIVE: To study the effect and mechanism of modified constraint-induced movement therapy (mCIMT) on motor function recovery in cerebral ischemia-reperfusion rats. METHODS: The rats were randomly divided into the control group and the mCIMT group, with 12 rats in each group. The left middle cerebral artery occlusion (MCAO) model was established by the Longa suture method. In the mCIMT group, the rats started continuous training for 14 d on the 7 th day after modeling. The unaffected limb was tied to the chest with elastic bandages, and the affected limb was trained in the compulsory runner equipment. In the control group, rats moved freely in the cage. The body mass of rats was recorded within 20 d after modeling, and behavior was assessed by the foot-fault test. Some of the rats were euthanized 18 d after modeling, and high performance liquid chromatography (HPLC) was used to detect monoamine neurotransmitters (5-hydroxytryptamine (5-HT), 5-hydroxyindoleacetic acid (5-HIVV), homovanillic acid (HVA) ), and amino acid neurotransmitters (glutamic acid (Glu), asparaginic acid (ASP), glutamine (Gln), glycine (Gly), taurine (Tau), gamma aminobutyric acid (GABA) ) in the motor cortex and striatum, respectively. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of total P70 ribosomal protein S6 kinase (p70s6k) and p70s6k phosphorylated protein (p-p70s6k) in motor cortex and striatum, respectively. RESULTS: Compared with the control group, the body mass of rats in the mCIMT group was comparable (P >0.05) within 21 d after modeling, foot-fault rate of the mCIMT group was significantly lower at 17 d after modeling (P<0.05). At 18 d after modeling, compared with the control group, the level of 5-HIVV in the motor cortex increased significantly (P<0.05), and the relative content of amino acid neurotransmitters (the ratio of Glu) in the motor cortex including Gln, Gly, Tau and GABA to Glu increased significantly (P<0.05 or P<0.01) except for decreased ASP/Glu (P<0.05). Moreover, compared with the control group, the expression of p-p70s6k in the motor cortex of the mCIMT was significantly decreased (P<0.05). There were no significant differences in monoamine neurotransmitters and amino acid neurotransmitters in the striatum between two groups (P>0.05). CONCLUSION: mCIMT improved the motor function of MCAO rats, and the mechanism might be related to the increase of amino acid neurotransmitters and 5-HIVV and decrease of p-p70s6k expression in the motor cortex.

Paired associated magnetic stimulation promotes neural repair in the rat middle cerebral artery occlusion model of stroke.

Paired associative stimulation has been used in stroke patients as an innovative recovery treatment. However, the mechanisms underlying the therapeutic effectiveness of paired associative stimulation on neurological function remain unclear. In this study, rats were randomly divided into middle cerebral occlusion model (MCAO) and paired associated magnetic stimulation (PAMS) groups. The MCAO rat model was produced by middle cerebral artery embolization. The PAMS group received PAMS on days 3 to 20 post MCAO. The MCAO group received sham stimulation, three times every week. Within 18 days after ischemia, rats were subjected to behavioral experiments-the foot-fault test, the balance beam walking test, and the ladder walking test. Balance ability was improved on days 15 and 17, and the foot-fault rate was less in their affected limb on day 15 in the PAMS group compared with the MCAO group. Western blot assay showed that the expression levels of brain derived neurotrophic factor, glutamate receptor 2/3, postsynaptic density protein 95 and synapsin-1 were significantly increased in the PAMS group compared with the MCAO group in the ipsilateral sensorimotor cortex on day 21. Resting-state functional magnetic resonance imaging revealed that regional brain activities in the sensorimotor cortex were increased in the ipsilateral hemisphere, but decreased in the contralateral hemisphere on day 20. By finite element simulation, the electric field distribution showed a higher intensity, of approximately 0.4 A/m2, in the ischemic cortex compared with the contralateral cortex in the template. Together, our findings show that PAMS upregulates neuroplasticity-related proteins, increases regional brain activity, and promotes functional recovery in the affected sensorimotor cortex in the rat MCAO model. The experiments were approved by the Institutional Animal Care and Use Committee of Fudan University, China (approval No. 201802173S) on March 3, 2018.

Constraint-induced movement therapy improves functional recovery after ischemic stroke and its impacts on synaptic plasticity in sensorimotor cortex and hippocampus.

Constraint-induced movement therapy (CIMT) has proven to be an effective way to restore functional deficits following stroke in human and animal studies, but its underlying neural plasticity mechanism remains unknown. Accumulating evidence indicates that rehabilitation after stroke is closely associated with synaptic plasticity. We therefore investigated the impact of CIMT on synaptic plasticity in ipsilateral and contralateral brain of rats following stroke. Rats were subjected to 90 minutes of transient middle cerebral artery occlusion (MCAO). CIMT was performed from 7 days after stroke and lasted for two weeks. Modified Neurology Severity Score (mNSS) and the ladder rung walking task tests were conducted at 7,14 and 21 days after stroke. Golgi-Cox staining was used to observe the plasticity changes of dendrites and dendritic spines. The expression of glutamate receptors (GluR1, GluR2 and NR1) were examined by western blot. Our data suggest that the dendrites and dendritic spines are damaged to varying degrees in bilateral sensorimotor cortex and hippocampus after acute stroke. CIMT treatment enhances the plasticity of dendrites and dendritic spines in the ipsilateral and contralateral sensorimotor cortex, increases the expression of synaptic GluR2 in ipsilateral sensorimotor cortex, which may be mechanisms for CIMT to improve functional recovery after ischemic stroke.

Modified constraint-induced movement therapy alters synaptic plasticity of rat contralateral hippocampus following middle cerebral artery occlusion.

Modified constraint-induced movement therapy is an effective treatment for neurological and motor impairments in patients with stroke by increasing the use of their affected limb and limiting the contralateral limb. However, the molecular mechanism underlying its efficacy remains unclear. In this study, a middle cerebral artery occlusion (MCAO) rat model was produced by the suture method. Rats received modified constraint-induced movement therapy 1 hour a day for 14 consecutive days, starting from the 7th day after middle cerebral artery occlusion. Day 1 of treatment lasted for 10 minutes at 2 r/min, day 2 for 20 minutes at 2 r/min, and from day 3 onward for 20 minutes at 4 r/min. CatWalk gait analysis, adhesive removal test, and Y-maze test were used to investigate motor function, sensory function as well as cognitive function in rodent animals from the 1st day before MCAO to the 21st day after MCAO. On the 21st day after MCAO, the neurotransmitter receptor-related genes from both contralateral and ipsilateral hippocampi were tested by micro-array and then verified by western blot assay. The glutamate related receptor was shown by transmission electron microscopy and the glutamate content was determined by high-performance liquid chromatography. The results of behavior tests showed that modified constraint-induced movement therapy promoted motor and sensory functional recovery in the middle cerebral artery-occluded rats, but had no effect on cognitive function. The modified constraint-induced movement therapy upregulated the expression of glutamate ionotropic receptor AMPA type subunit 3 (Gria3) in the hippocampus and downregulated the expression of the beta3-adrenergic receptor gene Adrb3 and arginine vasopressin receptor 1A, Avpr1a in the middle cerebral artery-occluded rats. In the ipsilateral hippocampus, only Adra2a was downregulated, and there was no significant change in Gria3. Transmission electron microscopy revealed a denser distribution the more distribution of postsynaptic glutamate receptor 2/3, which is an α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor, within 240 nm of the postsynaptic density in the contralateral cornu ammonis 3 region. The size and distribution of the synaptic vesicles within 100 nm of the presynaptic active zone were unchanged. Western blot analysis showed that modified constraint-induced movement therapy also increased the expression of glutamate receptor 2/3 and brain-derived neurotrophic factor in the hippocampus of rats with middle cerebral artery occlusion, but had no effect on Synapsin I levels. Besides, we also found modified constraint-induced movement therapy effectively reduced glutamate content in the contralateral hippocampus. This study demonstrated that modified constraint-induced movement therapy is an effective rehabilitation therapy in middle cerebral artery-occluded rats, and suggests that these positive effects occur via the upregulation of the postsynaptic membrane α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor expression. This study was approved by the Institutional Animal Care and Use Committee of Fudan University, China (approval No. 201802173S) on March 3, 2018.

Fluoxetine adjunct to therapeutic exercise promotes motor recovery in rats with cerebral ischemia: Roles of nucleus accumbens.

The study aimed to explore the molecular mechanism of fluoxetine as an adjunct to therapeutic exercise to improve motor recovery using a rat cerebral ischemic model with middle cerebral artery occlusion (MCAO). We hypothesized that the nucleus accumbens (NAc) may be one of the responding areas to fluoxetine where relevant elevations in 5-hydroxytryptamine (5-HT) and ΔFosB were associated with motor behavioral recovery. Male Sprague-Dawley rats were randomly divided into five groups: rats without intervention; rats that underwent MCAO without exercise or fluoxetine; rats that underwent MCAO treated only with fluoxetine; rats that underwent MCAO treated only with exercise; and rats that underwent MCAO treated with both exercise and fluoxetine. Motor function and motivation were assessed by the fault footsteps test and the forced swimming test. 5-HT level in the bilateral NAc and the expression of 5-HT2C receptor (5-HT2CR) and ΔFosB in the ipsilesional (left) NAc were measured. Correlation was explored by Pearson correlation analysis. Our results indicated that either treatment helped improve the grasp dexterity of the affected limb, motor motivation, and resilience to adverse environment in MCAO rats. The dual treatment with fluoxetine and exercise may hasten the recovery process. The dual treatment helped restore the balance of 5-HT level between the bilateral NAc by significantly increasing its level in the ipsilesional side. Either treatment could resume the expression of 5-HT2CR in the ipsilesional side of the NAc close to the normal level, which was correlated with motor recovery. The dual treatment significantly increased the expression of ΔFosB in the ipsilesional side of the NAc, which was correlated with the balance of 5-HT in the bilateral NAc, but not directly with motor recovery. In conclusion, the NAc may play an important role in driving physical motivation, which was possibly related to motor recovery after stroke. Fluoxetine may hasten the effectiveness of therapeutic exercise, possibly via regulating 5-HT and its receptors in the NAc.

Constraint induced movement therapy promotes contralesional-oriented structural and bihemispheric functional neuroplasticity after stroke.

The mechanism behind constraint-induced movement therapy (CIMT) in promoting motor recovery after stroke remains unclear. We explored the bilateral structural and functional reorganization of the brain induced by CIMT after left middle cerebral artery occlusion (MCAO) in rats. CIMT started on the 8th day (D8) after MCAO surgery and lasted for 3 weeks. Skilled walking was assessed by Foot-Fault tests. The efferent neuron network innervating the paralyzed forelimb was labeled by pseudorabies virus (PRV) to explore neuron recruitment. Synapsin Ⅰ was used as an indicator of the number of synapses. Additionally, C-fos expression 1 h after walking was detected to explore the activation of the brain. As a result, CIMT significantly improved skilled walking and elicited more neuron recruitment into the innervating network of a paralyzed forelimb in the contralesional rather than the ipsilesional motor cortex and red nucleus. CIMT also increased the synapse number in the contralesional cortex but there was no corresponding effect in the intact ipsilesional cortex. Furthermore, MCAO decreased ipsilesional motor cortex activation, but CIMT partially compensated for this by increasing the number of activated neurons (c-fos+) in both the left and right motor cortex. In conclusion, the contralesional motor cortex and red nucleus might play more important roles than corresponding ipsilesional regions in structural reorganization during CIMT-induced motor recovery after stroke. However, CIMT promotes bilateral motor cortex activity without a side preference.

Constrained-induced movement therapy promotes motor function recovery by enhancing the remodeling of ipsilesional corticospinal tract in rats after stroke.

Constraint-induced movement therapy (CIMT), which forces the use of the impaired limb by restraining the unaffected limb, has been used extensively for the recovery of limb motor function after stroke. However, the underlying mechanism of CIMT remains unclear. Diffusion tensor imaging (DTI) is a well-known neuroimaging technique that reflects the microstructure of white matter tracts and potential changes associated with different treatments. The aim of this study is to use DTI imaging to determine how corticospinal tract (CST) fibers remodel in ischemic rats with CIMT. In the present study, rats were randomly divided into three groups: a middle cerebral artery occlusion group (MCAO), a therapeutic group (MCAO + CIMT), and a sham-operated group (sham). A plaster cast was used to restrict the unaffected limb of the rats in the MCAO + CIMT group for 14 days. The Catwalk system was used to assess the limb motor function of rats. Fractional anisotropy (FA) and the average diffusion coefficient (ADC) of the CST were quantified through DTI. The expression of the c-Jun-N-terminal kinase signaling pathway (JNK) was examined after 14 days of CIMT. We found that CIMT could accelerate and enhance motor function recovery, and the MCAO + CIMT group showed significantly increased FA values in the ipsilesional posterior limb of internal capsule (PLIC) compared with the MCAO group. In addition, we found no significant difference in the ratio of phosphorylated-JNK/total-JNK among the three groups, whereas the expression of P-JNK decreased significantly in the chronic phase of stroke. In conclusion, CIMT-induced functional recovery following ischemic stroke through facilitation of the remodeling of ipsilesional CST, and restoration after ischemic stroke may be associated with the declining value of the ratio of P-JNK/JNK.

Early wheel-running promotes functional recovery by improving mitochondria metabolism in olfactory ensheathing cells after ischemic stroke in rats.

Olfactory ensheathing cells (OECs) has been widely studied in stroke. The present study was aimed at examining the role of wheel-running treatment (WR) on rat olfactory ensheathing cells (rOECs) functions. Thirty adult male Sprague Dawley rats were randomly divided into two groups: the middle cerebral artery occlusion (MCAO) group and WR + MCAO group. Motor behavior was assessed through the footfault test, and the results showed that WR training markedly improved the neurobehavioral outcome. The glucose metabolic status of the brain was assessed with the micro-PET. This training significantly enhanced the glucose uptake of olfactory bulb in the early stage of WR treatment. The function of rOECs mitochondrial was significantly enhanced after 10 days of treatment. Body weight of rats in both of the two groups decreased and then increased slowly following the days. But the growth trend of the WR + MCAO group was no significantly higher than that of the WR group. This training significantly enhanced the glucose uptake, improved the proliferation of rOECs and increased the expression level of cytochrome C (Cyt-c). The mechanism may be associated with the facilitation of mitochondrial function of rOECs cells. Including facilitation of mitochondrial fusion, fission, and accompanying increased quantities of mitochondria. Obtained results indicate that early WR treatment may exert enhanced function on rOECs in vivo and increased mitochondrial amounts, and improved the expression level of Cyt-c after ischemic stroke.

Post-stroke Constraint-induced Movement Therapy Increases Functional Recovery, Angiogenesis, and Neurogenesis with Enhanced Expression of HIF-1α and VEGF.

BACKGROUND: Constraint-Induced Movement Therapy (CIMT) is one efficient approach to improve functional recovery after ischemic stroke. The underlying molecular mechanism remains unclear. In the current study, we investigated the effects of CIMT on angiogenesis and neurogenesis. To start linking our findings to molecular mediators, we further examined the expression of Hypoxia-Inducible Factor-1α (HIF-1α), Factor Inhibiting HIF-1 (FIH-1) and Vascular Endothelial Growth Factor (VEGF). METHODS: Rats were randomly assigned into three groups: a Middle Cerebral Artery Occlusion group (MCAO), a therapeutic group (CIMT+MCAO), and a sham middle cerebral artery occlusion group (Sham). Seven days after surgery, a plaster cast was placed around the unimpaired upper limb of the rats in the CIMT+MCAO group for 14 days. CIMT was performed on a horizontal ladder. Neurobehavioral consequences were evaluated using the Open-Field Test (OFT) and the Foot-Fault Test (FFT). The number of new neurons, the length of vessels as well as the expression of HIF-1α, FIH-1, and VEGF were examined before and after 14 days of CIMT. RESULTS: The CIMT+MCAO group showed a significant increase in the total length of microvessels and increased number of Bromodeoxyuridine+ (BrdU+)/NeuN+ double-labeled cells. These changes were correlated with an increase in HIF-1α and VEGF expressions and a decrease in FIH-1expression. FFT showed that the CIMT+MCAO group exhibited marked improvement in neurobehavioral outcome when compared to the MCAO group. Adverse effects on total activities or anxiety were not observed using open field analysis. CONCLUSION: CIMT-induced neuroprotection and functional recovery following cerebral ischemia were possibly mediated by an increase in endogenous HIF-1α and VEGF expression with subsequent neurogenesis and angiogenesis.

Two phases of response to long-term moderate heat: Variation in thermotolerance between Arabidopsis thaliana and its relative Arabis paniculata.

Long-term moderate heat is often experienced by plants and will become even more common in the future due to global warming. However, the responses of plants to this stress have not been characterised. In the present study, growth between Arabidopsis thaliana and its relative Arabis paniculata upon long-term exposure to moderate heat was compared. It was found that the latter was more tolerant than the former, and the patterns of physiological and biochemical responses of both plants presented two phases. The early phase involved no significant visible morphological and physiological changes. It occurred during the first third of the heat treatment and was extended when the stress was attenuated. During the later phase, the plants died or were damaged. Heat shock proteins were dramatically induced at the early phase and gradually decreased at the later phase in A. thaliana. By contrast, the levels were induced and maintained in A. paniculata. Profiling of membrane lipids found that the two plants exhibited opposite patterns of lipid remodelling at the early phase: A. paniculata synthesised phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol and showed a rapid decrease in the degree of lipid unsaturation, while A. thaliana degraded its lipids at the early phase and showed an accelerated degradation at the later phase. These biochemical adjustments during the early phase could favor the thermotolerance of A. paniculata. These results suggest that this species could thus be a model for the study of resistance to long-term moderate heat, through a strategy by which plants can adapt to long-term moderate heat.