目的:探讨脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)对有氧运动促进心肌梗死后心肌血管生成、改善心功能的协同效应及其机制.方法:细胞实验中,利用平行板流动小室建立12 dyn/cm2的层流剪切力(laminar shear stress,LSS)以模拟运动对血管壁的生理效应.Western blot分别检测非循环流体、循环流体、循环流体加人重组BDNF蛋白(50 ng/mL)处理的人脐静脉内皮细胞HUVEC中BDNF蛋白表达水平、BDNF高亲和力受体TrkB及其下游Akt通路磷酸化水平,Transwell细胞迁移实验和小管形成实验检测HUVEC的体外血管生成能力.动物实验中,60只12周龄雄性Sprague-Dawley(SD)大鼠随机分为假手术组(Sham)、心肌梗死组(MIC)、心肌梗死+BDNF组(MICB)、心肌梗死+运动组(MIE)、心肌梗死+运动+BDNF组(MIEB);造模术前及术后1周分别进行心彩超检查,随后开始为期4周的有氧运动训练,MIEB组大鼠在运动前10 min尾静脉注射人重组BDNF蛋白(0.4 μg/kg),MICB组仅注射等量BDNF;训练结束行心彩超检查后腹腔麻醉取材,免疫组化检测心肌梗死周围区血管密度.结果:循环流体产生的LSS可诱发HUVEC中BDNF蛋白表达增加,TrkB及其下游Akt通路磷酸化程度持续增强,循环流体中BDNF水平随干预时间的延长而升高.非循环流体产生的LSS可诱发HUVEC上BDNF蛋白表达增加,但TrkB及其下游Akt通路均处于未激活状态.增加循环流体中BDNF浓度可提高TrkB及其下游Akt通路活化程度,进一步提高HUVEC体外血管生成能力.心肌梗死大鼠有氧运动结合尾静脉注射BDNF较单纯注射BDNF和单纯运动组大鼠心肌血管密度显著增高、心功能改善.结论:运动通过LSS以BDNF浓度依赖性模式诱发HUVEC上TrkB持续磷酸化并激活其下游Akt信号通路,提高细胞迁移及小管形成能力;外源性补充BDNF可协同有氧训练强化运动的促心肌血管生成效应,进一步改善心功能.

功能解剖学是我国康复治疗学本科专业的重要基础学科课程之一，是研究正常人体解剖结构与机体功能关系的一门 学科，具有重要的学科地位和指导作用。随着现代社会需求的不断增加以及我国康复治疗学专业的飞速发展，如何 对康复治疗学专业的功能解剖学课程进行教学标准、教学模式、教学质量考核的整体改革，从而有效提升教学效 果，激发学生的主观能动性，值得思考和探讨。

目的 研究脑源性神经营养因子(BDNF)对内皮细胞Sestrin2表达的影响及机制,并探讨其在血管新生中的作用.方法 用100 μg/L的BDNF分别处理人脐静脉内皮细胞(HUVEC)1 h、2h、4h、6h、8h,实时荧光定量PCR检测Sestrin2 mRNA水平,免疫荧光和Western blot检测Sestrin2蛋白表达.将HUVEC分为6组:对照组(Control组)、BDNF组(加BDNF 100 μg/L)、BDNF+TrkB-Fc(1 mg/L)组、BDNF+KT-5823 (500 nmol/L)组、BDNF+L-NAME(10-4 mol/L)组、BDNF+DMSO组,共干预4h,Western blot检测Sestrin2蛋白表达.将HUVEC分为4组:对照组(Control组)、BDNF组(加BDNF 100μg/L)、BDNF+ Sestrin2 siRNA组和BDNF+Control siRNA组,共干预6h,细胞迁移实验和小管成形实验分别检测HUVEC迁移能力和血管生成能力.结果 与0h和1h组比较,100 μg/L BDNF分别干预HUVEC2、4及6h时段,Sestrin2 mRNA水平显著增高(P＜0.001),在2、4及8h时段Sestrin2蛋白表达显著增高(P＜0.05);阻断NO/PKG通路可抑制BDNF诱导的Sestrin2表达上调(P＜O.001);抑制Sestrin2表达后,HUVEC迁移及小管形成能力较BDNF干预组显著降低(P＜0.01).结论 BDNF通过NO/PKG通路促进内皮细胞表达Sestrin2,从而提高内皮细胞血管生成能力.

Objective. In this study, a hybrid method combining hardware and software architecture is proposed to remove stimulation artefacts (SAs) and extract the volitional surface electromyography (sEMG) in real time during functional electrical stimulations (FES) with time-variant parameters. Approach. First, an sEMG detection front-end (DFE) combining fast recovery, detector and stimulator isolation and blanking is developed and is capable of preventing DFE saturation with a blanking time of 7.6 ms. The fragment between the present stimulus and previous stimulus is set as an SA fragment. Second, an SA database is established to provide six high-similarity templates with the current SA fragment. The SA fragment will be de-artefacted by a 6th-order Gram–Schmidt (GS) algorithm, a template-subtracting method, using the provided templates, and this database-based GS algorithm is called DBGS. The provided templates are previously collected SA fragments with the same or a similar evoking FES intensity to that of the current SA fragment, and the lengths of the templates are longer than that of the current SA fragment. After denoising, the sEMG will be extracted, and the current SA fragment will be added to the SA database. The prototype system based on DBGS was tested on eight able-bodied volunteers and three individuals with stroke to verify its capacity for stimulation removal and sEMG extraction. Results. The average stimulus artefact attenuation factor, SA index and correlation coefficient between clean sEMG and extracted sEMG for 6th-order DBGS were 12.77 ± 0.85 dB, 1.82 ± 0.37 dB and 0.84 ± 0.33 dB, respectively, which were significantly higher than those for empirical mode decomposition combined with notch filters, pulse-triggered GS algorithm, 1st-order and 3rd-order DBGS. The sEMG-torque correlation coefficients were 0.78 ± 0.05 and 0.48 ± 0.11 for able-bodied volunteers and individuals with stroke, respectively. Significance. The proposed hybrid method can extract sEMG during dynamic FES in real time.

NLRP3 inflammasome-mediated vascular EC pyroptosis is a key event in the pathogenesis of atherosclerosis. Dysregulation of glucose metabolism is involved in EC dysfunction. Although BDNF plays a protective role in vascular endothelium physiological activity, the mechanisms underlying this activity are not yet clear. In this study, we investigated the role of BDNF in NLRP3 inflammasome-mediated EC pyroptosis and its associated reprogramming of glucose metabolism. HUVECs were treated with human rBDNF under ox-LDL stimulation. rBDNF alleviated ox-LDL-induced NLRP3 inflammasome formation and HUVEC pyroptosis, as evaluated by NLRP3, caspase1-p10, interleukin-18, and interleukin-1β protein levels, co-localization of NLRP3 and apoptosis-associated speck-like protein, and lactate dehydrogenase release. These effects were prevented by tropomyosin receptor kinase B inhibition and KLF2 silencing. The hyper-activation of glycolysis induced by ox-LDL-induced was mitigated by rBDNF via KLF2 as assessed by glucose uptake, lactate production, and extracellular acidification rate. In addition, the BDNF/KLF2 pathway preserved the mitochondrial membrane potential, intracellular reactive oxygen species generation, electron transport chain processing, oxygen consumption rate, and adenosine triphosphate production. Furthermore, KLF2 interacted with HK1 and HK1 overexpression evoked NLRP3 inflammasome formation. At the clinical level, plasma BDNF and lactate levels were measured in 274 patients who underwent computed tomography and coronary angiography for CAD diagnosis. Patients with CAD had lower BDNF and increased lactate levels than those without CAD. In 94 patients with CAD, circulating BDNF levels were inversely associated with lactate levels. In the receiver operating characteristic analysis of CAD, the areas under the curves for 1/BDNF, lactate, and 1/BDNF+lactate were 0.707, 0.702, and 0.753 respectively. These results indicate that BDNF and lactate are linked in atherosclerotic patients, and BDNF inhibits ox-LDL induced NLRP3 inflammasome formation and pyroptosis in HUVECs via KLF2/HK1-mediated glucose metabolism modulation and mitochondrial homeostasis preservation.

In this study, a wearable prototype system was developed for multiple-gesture rehabilitation using electrical stimulation controlled by a volitional surface electromyography (sEMG) scan of a healthy forearm. The purpose of the prototype system is to reconstruct multiple gestures of a paralysed limb and to simplify the positioning of sEMG detection sites on a healthy forearm. A self-designed eight-channel sEMG detection armband was used to detect the sEMG signal distributions of the muscle groups in healthy forearms. Linear discriminant analysis (LDA) was used to classify the sEMG signal distributions corresponding to different gestures, and then the classification results were mapped to corresponding stimulation channels. The sEMG signal with the maximum root mean square (RMS) was used as the source of stimulus coding for each gesture. Our proposed mean absolute value (MAV)/number of slope sign changes (NSS) dual-coding (MNDC) algorithm was used to encode the sEMG signal into an electrical stimulus with a dynamic pulse width and frequency. The constant-current stimulation armband electrically stimulated multiple muscles in the affected forearm by means of a circuit designed with a time-division multiplexed stimulation channel. An experiment involving 6 able-bodied volunteers showed that when the detection armband was located near the middle of the forearm, the gesture classification accuracy was greater than 90%, and each active sEMG signal was high. Gesture bridge experiments, including grasping, wrist flexion, wrist extension and finger extension, were carried out among six hemiplegic subjects and between one able-bodied volunteer acting as a controller and each of six stroke patients as the controllee. Both sets of results show that the proposed system can reconstruct these four gestures in the controlled subject with a delay of at most 360 ms and with a correlation coefficient of >0.72.

Endothelial cell dysfunction and diabetic vascular complications are intrinsically linked. Although BDNF plays a protective role in cerebral microvascular complications caused by diabetes, the mechanisms of this activity are not fully clear. In this study, we investigated the role of BDNF in the hyperglycemic injury of BMECs and its associated intracellular signal transduction pathways. BMECs were treated with 33 mM glucose to imitate the endothelium under hyperglycemic conditions. The high-glucose treatment caused cell dysfunction, as evaluated by oxidative stress and cell apoptosis, which could be alleviated by BDNF. In addition, BDNF preserved mitochondrial function as assessed by mPTP opening, mitochondrial membrane potential, calcium content, and mitochondrial biogenesis markers. Western blot analysis of LC3-II, p62, and TOMM20 and the detection of mRFP-GFP-LC3 adenovirus for autophagy flux revealed that BDNF enhanced autophagy flux. Furthermore, BDNF activated mitophagy, which was confirmed by the observed colocalization of LC3-II with BNIP3 and from transmission electron microscopy observations. The HIF-1α/BNIP3 signaling pathway was associated with BDNF/TrkB-induced mitophagy. In addition, BDNF-induced mitophagy played a protective role against BMEC damage under hyperglycemia. Thus, the results of this study suggest that BDNF/TrkB/HIF-1α/BNIP3-mediated mitophagy protects BMECs from hyperglycemia.

Cardiovascular disease (CVD) is a leading cause of mortality and morbidity among patients with diabetes. Endothelial dysfunction is an early physiological event in CVD. Metformin, a common oral antihyperglycemic agent, has been demonstrated to directly affect endothelial cell function. Brain-derived neurotrophic factor (BDNF), originally discovered in the brain as a neurotrophin, has also been reported to play a protective role in the cardiovascular system. In our study, we demonstrated that high glucose (HG) reduced cell proliferation and induced cell apoptosis via changes in BDNF expression and that metformin reversed the effects of HG injury by upregulating BDNF expression. Furthermore, we found that cyclic AMP response element binding (CREB) phosphorylation was reduced in HG-treated human umbilical vein endothelial cells (HUVECs), and this effect was reversed by the metformin treatment. However, the metformin effect on BDNF levels in HG-incubated HUVECs was blocked by a CREB inhibitor, which indicated that BDNF expression is regulated by metformin through CREB activation. In addition, we found that adenosine monophosphate-activated protein kinase (AMPK) activation is involved in CREB/BDNF regulation in HG-incubated HUVECs treated with metformin and that an AMPK inhibitor impaired the protective effects of metformin on HG-treated HUVECs. In conclusion, this study demonstrated that metformin affects cell proliferation and apoptosis via the AMPK/CREB/BDNF pathway in HG-incubated HUVECs.

Background Brain-derived neurotrophic factor (BDNF) is a neurotrophin involved in angiogenesis and maintenance of endothelial integrity. Whether circulating BDNF levels are associated with von Willebrand factor (vWF) levels, which are indicators of endothelial dysfunction is not known. This study investigated the association between plasma BNDF and vWF levels and whether these biomarkers could predict cardiovascular events at a 12-month follow-up in patients with stable coronary artery disease (CAD). Methods We recruited 234 patients with suspected angina pectoris. Subjects were divided into CAD ( n = 143) and control ( n = 91) groups based on coronary angiography. Plasma BDNF and vWF levels were measured using ELISA. Patients were followed-up for one year, and information on adverse cardiac events was collected. Results CAD patients exhibited significantly lower plasma BDNF and higher vWF levels than those of control patients. High vWF levels were associated with low BDNF levels even after adjustment for age, gender, low-density lipoprotein (LDL) levels, and the presence of diabetes mellitus. A receiver operating characteristic curve was used to determine whether low BDNF and high vWF levels could predict adverse cardiovascular events. The area under the curve for vWF and the inverse of BDNF were 0.774 and 0.804, respectively. Conclusions These findings suggest that endothelial dysfunction is an important determinant of the impaired circulating BDNF levels, and they further reflected cardiovascular prognosis in stable CAD patients.

Exercise induces a number of benefits, including angiogenesis in post-myocardial infarction (MI); however, the underlying mechanisms have not been fully clarified. Neurotrophic brain-derived neurotrophic factor (BDNF) serves a protective role in certain adult cardiac diseases through its specific receptor, BDNF/NT-3 growth factors receptor (TrkB). The present study explored the mechanisms by which exercise improves cardiac function, with a focus on the involvement of the BDNF/TrkB axis. MI rats were assigned to Sham, sedentary, exercise, exercise with K252a (a TrkB inhibitor), and exercise with NG-nitro-L-arginine methyl ester (L-NAME) groups. The exercise group was subjected to 8 weeks of treadmill running. The results demonstrated that the rats in the exercise group exhibited increased myocardial angiogenesis and improved cardiac function, which was attenuated by K252a. Exercise induced activation of the BDNF/TrkB axis in the ischaemic myocardium and increased serum BDNF levels were abated by exposure to L-NAME. Improvements in angiogenesis and left ventricular function exhibited a positive association, with changes in serum BDNF. In the in vitro experiments, human umbilical vein endothelial cells were exposed to shear stress (SS) of 12 dyn/cm(2) to mimic the effects of exercise training on vascular tissue. An increased tube-forming capacity, and a nitric oxide (NO)-dependent prolonged activation of the BDNF/TrkB-full-length axis over 12 h, but not the TrkB-truncated axis, was observed. The SS-related angiogenic response was attenuated by TrkB inhibition. Overall, these results demonstrate that exercise confers certain aspects of its cardioprotective effects through the activation of the BDNF/TrkB axis in an NO-dependent manner, a process in which fluid-induced SS may serve a crucial role.

目的 观察电刺激结合咽部肌群训练对脑卒中并发阻塞性睡眠呼吸暂停综合征(OSAS)患者临床症状、睡眠呼吸参数及认知功能的影响.方法 2014年12月1日至2017年11月30日,脑卒中并发OSAS住院患者50例,随机分为对照组和研究组各25例.两组均接受常规药物治疗及康复训练,研究组在此基础上接受VocaStim?-Master电刺激,包括咽部神经肌肉电刺激和舌下神经分支刺激,以及咽部肌群训练,共4周.记录干预前后临床症状变化,采用多导睡眠呼吸监测(PSG)及简明精神状况检查(MMSE)进行评定.结果 治疗后,研究组临床症状较治疗前及对照组改善(P<0.05),睡眠呼吸紊乱指数(AHI)下降(t>2.270,P<0.05),平均血氧饱和度、最低血氧饱和度明显增高(t>3.095,P<0.01),氧减指数显著下降(t>3.044,P<0.01),MMSE评分明显增高(t>2.859,P<0.01).对照组治疗前后AHI、平均血氧饱和度、最低血氧饱和度、氧减指数和MMSE评分均较治疗前无显著性差异(P>0.05).结论 VocaStim?-Master电刺激结合咽部肌群训练能有效缓解脑卒中并发OSAS患者的临床症状,改善呼吸功能和认知功能.

In this presentation the motor function rebuilding of paralyzed limbs of the paraplegic patients caused by spinal cord injury and the hemiplegic patients after stroke and SCI is concerned. The biomedical methods and the traditional physical methods for the rehabilitation of two kinds of paralyses are reviewed. The core part is to discuss the neural and muscular signal regeneration and the limb function rebuilding based on the principles of communication and functional electrical stimulation - a novel concept developed by the speakers. For the communication, a microwave transmission system is incorporated. The construction of the whole bio-electronic system, the animal experiments, and the elementary experiments on healthy and paralyzed patients will be demonstrated.

In this paper, we report our novel idea on the function rebuilding for hemiplegic limbs and the primary experiments. The main concept is to connect the control-lost nerves or neuromuscular junctions by using a multi-channel micro- electronic neural bridge (MENB), regenerate the nervous signal, and rebuild the motor functions of the related limb. Since the injured nervous system in stroke-related hemiplegia is located in the brain and difficult to be identified and operate on, we use another nervous system functioning as a new signal source to supply similar neural signals. In these cases, that means, two independent nervous systems are connected by a MENB. As preclinical experiments, we have made a series of tests on bodies of animals and healthy human. The principle, the system construction and the experimental results will be given.

Purpose: To evaluate the effect of aerobic cycling training with lower limb weights on cardiovascular fitness (peak VO2) and walking ability in chronic stroke survivors, and to investigate the relationship between changes in these parameters. Methods: 133 Chinese patients with chronic hemiparetic stroke (mean age 58 years) were randomized to either 8-week (5×/week) aerobic cycling training with lower limb weights group (n = 68) or a low-intensity overground walking group (n = 65). Peak VO2, 6-minute walk distance (6MWD), knee muscle strength, balance and spasticity were measured before and after intervention. Results: Cycling training increased peak VO2 (24% vs. 3%, p < 0.001), 6MWD (2.7% vs. 0.5%, p < 0.001), paretic (11% vs. 1.6%, p < 0.001) and nonparetic knee strength (16% vs. 1.0%, p < 0.001). In the cycling group, percent changes in peak VO2 were positively associated with those in paretic (r = 0.491, p < 0.001) and nonparetic knee strength (r = 0.432, p < 0.001). Increased 6MWD correlated significantly with improved balance, spasticity and paretic knee strength by the stepwise regression analysis (r2 = 0.342, p = 0.004), but not fitness gains. Conclusions: The enhanced cardiovascular fitness after aerobic cycling training in Chinese patients with chronic stroke is not associated with the increased walking ability. Unparallel improvements in these parameters related different determinants may have implications for intervention strategy.Implications for RehabiliationPatients with chronic stroke have residual physical impairments secondary to low activity levels and are often severely deconditioned.Exercise therapy is important to improve recovery in mobility and optimize cardiovascular health after stroke.The randomized control study shows that the intensive aerobic cycling training with lower limb weights improves both cardiovascular fitness and walking ability in Chinese patients with chronic stroke. However, these improvements after intervention were not parallel, indicating the determinants of improvements in these two parameters are different and may have implications for therapy prescription.

Objective To observe the concentration effects of alcohol in perineurial block on the structure and function of sciatic nerve and it's innervated muscles in rats so as to provide the basis for clinical application of chemical neurolysis.Methods One hundred and fifty female Sprague-Dawley rats were used and randomly assigned into a blank group(BG,n =6),a control group(CG,n =36),a 50％ alcohol group(50G,n =36),a 75％ alcohol group(75G,n =36)and a 99.9％ alcohol group(99.9G,n =36).The CG received physiological saline injection,the 50G,75G,99.9G received corresponding concentrations alcohol perineurial block,respectively.Changes of motor function was assessed,electrophysiological and histomorphological observations of sciatic nerve and its innervated muscles were conducted before and at 24 h,72 h,1 week,4 week,12 week after block.Results(① Hypokinesia and decrease of motor conduction velocity(MCV)were observed at 24 h after block,and peaked at 72 h after block; at 1 week after block,the motor function and MCV improved,the improvement persisted to the 12th week (P ＜ 0.05); but at the 12th week,MCV was still slower than that before block(P ＜ 0.01).②There were significant differences with regard to motot function and MCV of sciatic nerve among 99.9G and the other groups at e ery time point after block(P ＜0.05); ③ Reversible dengeneration of sciatic nerve and hind limb muscle,nonreversible necrosis of muscle occurred more seriously with the increase of alcohol concentration,and the cicatrization in 99.9G was more obvious than that in 50G and 75G at the 12th week; ④Structural lesion of sciatic nerve occurred at 24 h after block and peaked at the 72th h,myelinated axonal sprouts appeared at 1 week after block and persisted to the 12th weck.Conclusions ① The effects of 99.9 ％ alcohol perineural block on the structural lesion,motor function and MCV of sciatic nerve and its related muscle were more obvious than those of 50％ and 75％ alcohol; ② The destructive effect of alcohol block would maintain more than 12 week and neural restoration would maintain more than 12 weeks too.

Aim To explore the role of advanced glycation end products(AGE) in pathogenesis and development of diabetic complications.Methods Indirect ELISA and Western blotting were used to analyse the immunological property of McAb and the epitope which would be combined with McAb.AGE in serum of human and aortas,renal,heart from diabetic rats were also detected by McAb.Results McAb reacted with AGE specially and was combined with non-carboxymethyl lysine(non-CML).AGE in renal and heart of diabetic rats had been detected,but were not obviously detected in normal rats.Conclusion McAb(non-CML)reacted with AGE specially,and might be of value for AGE measurement.

Objective:To prepare and identify the monoclonal antibodies against AGEs.Methods:BALB/c mice were immunized with purified AGE-HSA protein, McAb against AGE-HSA was produced by hybridoma technique. Indirect ELISA and Western blot were used to identify the immunoglobin subtype and specificity.Results:Three novel murine monoclonal antibodies only against AGE-HSA were harvested. The subclass of these monoclonal antibodies were determined as IgM.Conclusion:The McAb against AGE-HSA protein may be a useful tool for studying on the biological properties of AGEs and for its detection clinical laboratory.

Aim To explore the role of advanced glycation end products (AGEs)in pathogenesis and development of diabetic complications, and analyse the immunological property of monoclonal antibodies (McAb)against AGEs. Methods Indirect ELISA and western blotting were used to analyse the immunological property of McAb and the epitope which would combine with McAb. AGEs in serum of human and aortas from diabetic rats were also detected by McAb. Results McAb reacted with AGEs specially and combined with non-carboxymethyl lysine (non-CML). AGEs in serum of human and aortas from diabetic rats had been detected, however, there had been no AGEs detected in aortas from normal rats. Conclusion McAb(non-CML)reacted with AGEs specially,and might be of value for AGEs measurement.