Objective:To explore the effect of low intensity pulsed ultrasound(LIPUS)on inhibiting the abnormal cell phenotype of fibroblast-like synoviocytes in rheumatoid arthritis(RA-FLS)and possible mechanism. Method:Synoviocytes were isolated by using enzyme digestion,and the morphology of cells was observed un-der microscope.At the same time,the expression of Vimentin protein was detected by immunofluorescence method to identify RA-FLS.Cells cultured in vitro were divided into four groups:control group,LIPUS group,tumor necrosis factor(TNF-α)group and TNF-α+LIPUS group or three groups:control group,interleu-kin-6(IL-6)group and IL-6+LIPUS group.The effects of LIPUS on RA-FLS cell viability and proliferation were detected by CCK8 and EDU assay respectively,and the effects of LIPUS on RA-FLS migration were ob-served by scratch test and Transwell migration assay.RT-qPCR was used to detect the gene expression of im-portant cytokines,chemokines and matrix metalloproteinases(MMPs)in RA-FLS.ELISA was used to further detect the effect of LIPUS on the expression of IL-6,a key effector of RA-FLS,and the effects of LIPUS on mitogen-activated protein kinase(MAPK)signaling pathway in RA-FLS were detected by Western Blot. Result:Purified RA-FLS were obtained.Firstly,LIPUS could suppress the cell activity(P＜0.00l)and prolifer-ation(P=0.007)induced by TNF-α in RA-FLS cultured in vitro.However,the migration and the transcription levels of MMPs related to migration(MMP2 and MMP9)were not significantly different between groups(P＞0.05).LIPUS could inhibit the high expression of IL-6 and interleukin-8(IL-8)at the mRNA level in the in-flammatory environment induced by TNF-α(P＜0.001),but there was no significant difference in the suppres-sion of interleukin-1β(IL-1β),MMP1 and MMP13(P＞0.05).In addition,compared with untreated group,LI-PUS could inhibit the secretion of IL-6 in RA-FLS induced by TNF-α(P＜0.001),and also inhibited the pro-liferation of RA-FLS induced by IL-6(P-0.003).Finally,LIPUS could inhibit the phosphorylation of p38 MAPK and c-Jun N-terminal kinase(JNK)in MAPK signaling pathway(P=0.033),but the effect on the phosphorylation of extracellular signal-regulated kinas 1/2(ERK1/2)was not significantly(P＞0.05). Conclusion:LIPUS could reduce the abnormal proliferation of RA-FLS in inflammatory state without affecting its migration,which might be related to the inhibition of p38/JNK-IL-6 signaling pathway.

Objective:To explore the effects of different duration of exercise preconditioning on changes in cerebral blood flow and microglia activation related proteins in rats with vascular dementia. Method:Sixty SPF SD male rats were used to prepare vascular dementia rat models by permanent ligation of bilateral common carotid arteries.They were randomly divided into the model group,sham-operated group,ex-ercise preconditioning 4-week model group,exercise preconditioning 4-week sham-operated group,exercise pre-conditioning 2-week model group and exercise preconditioning 2-week sham-operated group,with 10 rats in each group.The exercise preconditioning 4-week rats received 30 minutes of moderate intensity non-weight-bear-ing swimming training 5 times a week for 4 weeks before modeling,while the exercise preconditioning 2-week rats received the same training for 2 weeks.Morris water maze was used to detect the spatial learning and memory ability of rats,laser speckle imaging technique was used to observe the changes of cerebral blood flow and the opening of collateral circulation of rats at different time point before and after the model-ing,and Western Blotting was used to detect the expression of TLR4 and Iba 1 protein in hippocampus. Result:Compared with the sham-operated group and the exercise preconditioning 2-week sham-operated group,the average escape latency time of rats in the exercise preconditioning 4-week sham-operated group,the model group,the exercise preconditioning 4-week model group and the exercise preconditioning 2-week model group was significantly prolonged(P＜0.05).Compared with the exercise preconditioning 4-week sham-operated group,the average escape latency time of rats in the model group and the exercise preconditioning 4-week model group was significantly prolonged(P＜0.05).Compared with the model group and exercise preconditioning 4-week model group,the average escape latency time of rats in exercise preconditioning 2-week model group was significantly decreased(P＜0.05).The simple effect of repetitive measurement deviation analysis suggested that the average cerebral blood flow before modeling,2h after modeling,3d after modeling and 7d after model-ing was statistically significant between the groups(P＜0.05).The simple effect of time factor on average cere-bral blood flow of the model group,the exercise preconditioning 4-week model group and the exercise precon-ditioning 2-week model group was statistically significant(P＜0.01).The opening of collateral circulation of rats in each group was observed.Compared with the model group,less reduction in microvessel diameter was ob-served in the exercise preconditioning 2-week model group(P＜0.05).Compared with the sham-operated group,the exercise preconditioning 4-week sham-operated group and the exercise preconditioning 2-week sham-operat-ed group,Ibal and TLR4 protein expressions in the model group were significantly increased(P＜0.01).Com-pared with the model group,Ibal and TLR4 protein expressions in the exercise preconditioning 2-week model group were decreased(P＜0.05). Conclusion:Moderate intensity exercise preconditioning for 2 weeks can improve the learning and memory abili-ty of vascular dementia rats,but exercise preconditioning for 4 weeks has no obvious effect on the improve-ment of learning and memory ability.The mechanism may be related to the improvement of cerebral blood flow status and the inhibition of microglia activation.

Objective:To establish a reliable,stable and clinically realistic model of blunt contusion injury to the rat skele-tal muscle. Method:A model of the rat gastrocnemius muscle blunt contusion injury was established by using a weight of 170g,a height of 70cm and a spring with a diameter of 0.5cm at the bottom to buffer the polymeric strik-ing part and guide the fall to hit for 5 consecutive times.The gait changes of rats were evaluated by CatWalk analysis system at 0h,12h,24h,3 days,7 days and 14 days after modeling;local histopathological changes were observed by HE staining and local ultrastructural changes were observed by transmission electron micros-copy.Enzyme-linked immunosorbent assay(ELISA)was used to detect the changes of tumor necrosis factor-α(TNF-α)in local tissues. Result:Compared with the blank group,the mean speed and maximum speed change rate of the model group were significantly decreased,and the difference was statistically significant(P＜0.001);compared with the normal foot group,the maximum contact area,footprint area and standing phase duration of the limb of the injured limb group were significantly decreased,and the difference was statistically significant(P＜0.05).A large infiltration of inflammatory cells and massive lysis necrosis of muscle fibers were observed on HE stain-ing after 3 days.By 14 days,although scar tissue was still present,tissue repair with HE staining was almost indistinguishable from that of the blank group.Vacuolation was observed in part of the sarcoplasmic reticulum at 0h after the blow,and lysis of muscle fibers and part of the sarcoplasmic reticulum at 3 days with in-creased inflammatory response which basically returned to normal by 14 days.The TNF-α level began to rise gradually and reached a peak from 0h to 12h after injury,and remained higher level from 12h to 72h,and the difference was statistically significant compared with the blank group(P＜0.05),and basically returned to the normal level by 14 days. Conclusion:The model can cause moderate damage of skeletal muscle in rats,with a 14-day natural recovery time at least.It can provide as one of the methods for for the establishment of the rat skeletal muscle blunt contusion model.

Objective:To explore the mechanism of olfactory three needle therapy on Parkinson's disease dementia(PDD)by observing its effects on expression of apolipoprotein E(ApoE)、glial fibrillary acidic protein(GFAP)and related core pathology substrates in the hippocampus of PDD model mice. Method:Male C57BL/6 mice were randomly divided into four groups:control group(Control),sham opera-tion group(Sham),model group(Model)and acupuncture electrotherapy group(AE),with 10 mice in each group.The PD model was established by injecting 6-OHDA into the medial forebrain tract(MFB)and PDD mice were selected.After successful modeling and selection,the AE group received"olfactory three needle"electro acupuncture treatment.After 14 days of intervention,Morris water maze test and shuttle box test were used to evaluate the learning and memory ability of mice in each group.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of hippocampal CA1 region.Western blotting was used to detect the expression of α-syn,Aβ and ApoE proteins in hippocampal CA1 region.The co-location rate of ApoE and GFAP in hippocampal CA1 region was observed by double immunofluorescence markers. Result:Compared with Model group,the AE group exhibited a shortened escape latency in water maze(P＜0.01),increased platform crossing(P＜0.05),increased active escape times of shuttle box(P＜0.05),and reduced the average total time of electric stimulation(P＜0.01).In the Model group,the neurons in the hippocampal CA1 area were sparsely arranged and showed signs of degeneration and necrosis;and cell nuclei were small,hyperchromatic and had unclear structures,indicating the appearance of nuclear pyrosis.In contrast,the AE group showed significant improvements in neuronal pathology,with most cells regularly arranged,round and large nuclei,lightly stained and clearly shaped.Compared with the Model group,the expression levels of α-syn,Aβ,ApoE protein and the co-localization rate of ApoE and GFAP in hippocampal CA1 region in AE group were decreased(P＜0.01,P＜0.01,P＜0.01,P＜0.05). Conclusion:The"Olfactory three needle"acupuncture can improve the cognitive ability and restore the morpho-logical structure and function of neurons in PDD mice.The mechanism may be related to the inhibition of ApoE expression in astrocytes and the reduction of α-syn and Aβ deposition in hippocampal CA1 region.

Objective:To explore the effectiveness and feasibility of strengthening inspiratory muscle training program matching the preoperative waiting time for patients with valvular heart disease in China with limited resources in preventing postoperative pulmonary complications(PPCs). Method:Patients undergoing elective heart valve surgery were randomly divided into control group and inter-vention group,with 65 patients in each group.All the enrolled patients received a routine physical therapy pro-gram before surgery,and the intervention group received an additional 3 days of enhanced inspiratory muscle training program before surgery.The incidence of postoperative complications was recorded for all patients with-in 14 days after surgery,respiratory function and motor capacity(6-minute walking test)were tested for all patients on the day of admission and discharge,and the length of postoperative hospital stay was counted. Result:Compared with the control group,the incidence of PPCs was lower in the intervention group(OR 2.18,95％CI 1.07-4.47,P-0.03),and the walking distance was longer(P=0.01).There was no statistical difference in lung function and length of hospital stay(P＞0.05),and no adverse events occurred during train-ing.There was no significant difference in general demographic characteristics between the two groups before treatment.The incidence of PPCs in the intervention group was lower than that in the control group(P＜0.05),and the 6-minute walking distance at discharge was higher than that in the control group(P＜0.05).There were no statistically significant differences in lung function and postoperative hospital stay between the intervention group and the control group(P＞0.05),and no adverse events occurred during training. Conclusion:Enhanced inspiratory muscle training 3 days before surgery is safe and feasible in patients with elective heart valves and can reduce postoperative pulmonary complications and improve the level of motor function at discharge.

Objective:To investigate the effect of a visual walking training based on wearable cueing devices on ground reaction force of Parkinson's disease patients with freezing of gait. Method:Twenty-three PD with FOG(PD+FOG)were selected and twenty healthy controls(HC)were tested in gait laboratory.The experimental group was trained with wearable walking guide devices for 20 minutes,twice a day for one week.The Vicon 3D gait analysis system was used to collect the spatiotemporal parame-ters and the ground reaction force parameters of the non-dominant side and the dominant side including front-back peak,medial-lateral peak and the first vertical peak)of the experimental group and the control group be-fore and after training. Result:①After the intervention,the bilateral step length and velocity of patients in the experimental group in-creased signi ficantly(P＜0.05),while cadence,double stance phase decreased significantly(P＜0.05).After train-ing,there was no significant difference in the dominant step length,cadence and double stance phase between experimental group and control group(P＞0.05).②After the training,bilateral parameters(landing angles of hip joints,the maximum angles of ankle dorsiflexion and plantar flexion,and the ranges of motion of lower limb joints)in the experimental group increased significantly,while the landing angles of knee joints de-creased,and the stage of the first peak of ankle joints was delayed,with statistical significance(P＜0.05).Af-ter the training,knee joint landing angle,maximum ankle dorsiflexion angle,the first peak stage of ankle joint,lower limb joint range of motion had no significant difference between the experimental group and the control group in both sides(P＞0.05).③After the intervention,the front-back peak value and the first vertical peak of the bilateral ground reaction force in the experimental group were significantly increased(P＜0.05),while the medial-lateral peak value of the ground reaction force was significantly decreased(P＜0.05).Compared with the control group,there was no significant difference in the backward peak,medial peak of non-domi-nant side and parameters of the dominant side(P＞0.05). Conclusion:The visual walking training based on wearable walking guide devices improve the stability and consistency of gait of PD+FOG.

Objective:To construct the regulatory network of competitive endogenous RNA(ceRNA)and explore the mo-lecular mechanism of ischemic stroke(IS)by using bioinformatic analysis to screen the differentially-expressed genes. Method:The expression profiles of miRNA,mRNA and lncRNA in IS were downloaded from the NCBI GEO database.Differentially-expressed miRNAs,lncRNAs,and mRNAs were identified by the limma package in R software.The prediction of the relationship of lncRNA-miRNA and miRNA-mRNA were performed by starBase,miRDB and miRwalk databases.The results of prediction and differential analysis were taken to inter-sect and screened out differentially-expressed target mRNA(DETmRNAs),Kyoto Encyclopedia of Genes and Genomes(KEGG)and gene ontology(GO)enrichment analysis was performed by using the DAVID database.The protein-protein interaction(PPI)network was constructed by using the STRING database and the core tar-get genes in the network were identified by Cytoscape software. Result:A total of 20 differentially-expressed miRNAs,1512 lncRNAs,and 278 mRNAs were identified,and a ceRNA network was successfully constructed with the interactions of 5 lncRNAs-6 miRNAs-102 mRNAs in IS.The 285 DETmRNAs functions are mostly involved in the biological process such as chromatin organization,negative regulation of phosphoprotein phosphatase activity,or cell cycle.KEGG mainly enriched the signaling pathway including leukocyte trans-endothelial migration and platelet activation.The top 10 core genes were CREB1,MAPK1,GSK3B,SP1,PIK3R1,NR3C1,NCOR1,NFATC1,SETD2,and NSD1. Conclusion:The construction of the ceRNA network can help to further understand the molecular mechanism of IS and screen potential biomarkers,providing clues to further define rehabilitation targets and develop reha-bilitation strategies.

Objective:To evaluate the reliability of the Edinburgh visual gait score(ECGS)and assess its validity by com-parihg the results to three-dimensional gait analysis.Additionally,the study examines the correlation between EVGS and gross motor function classification system(GMFCS).The findings are expected to support the clini-cal application of EVGS. Method:Twenty children with cerebral palsy(CP)were randomly selected from 727 children who underwent three-dimensional gait analysis at Yangzhi Rehabilitation Hospital between 2019 and 2021.The walking videos of CP were evaluated using EVGS by four evaluators with different experience backgrounds,and repeated by the same evaluators after one month.Intragroup correlation coefficient(ICC)was used to test inter-and intra-evaluator reliability.Spearman was used to test EVGS,three-dimensional gait analysis data and GMFCS. Result:The ICC of the first assessment among evaluators was 0.947(P＜0.001),the ICC of the second as-sessment was 0.952(P＜0.001),indicating high inter-and intra-evaluator reliability.The Spearman correlation analysis results showed a moderate relationship between ECGS and three-dimensional gait data,with poor valid-ity in coronal items.A significant correlation was found between EVGS and GMFCS.Further,the intra-item correlation analysis of EVGS showed that the foot,pelvis and trunk significantly influenced the overall EVGS outcomes. Conclusion:The inter-and intra-evaluator reliability among the four evaluators were highly reliable.The validi-ty of EVGS was lower when compared with three-dimensional gait analysis.However,EVGS were correlated with the motor dysfunction levels in CP.Overall,EVGS can be used as a substitute tool for gait analysis,which is the most reliable visual gait score at present.

Objective:To evaluate the predictive value of gait kinematic combined with total MRI burden for the risk of falls in patients with gait disorder in cerebral small vessel disease(CSVD)using logistic regression analysis and ROC curve. Method:Forty-three patients diagnosed with CSVD and presenting primarily with gait disorder at Rehabilitation Department of Gansu Province Hospital from March 1,2019 to March 30,2020 were selected.The statistical clinical data were collected,and according to the TUG test time,all patients were divided into a group with high risk of fall(high risk of falling,HRF,TUG≥15s)and a group of low risk of falls(low risk of falling,LRF,TUG＜15s).Logistic regression analysis and ROC curve were used to assess the predictive value of gait kinematic characteristic combined with total MRI burden for the risk of falls in CSVD patients. Result:A total of 43 patients were included with average age(71.07±8.17)years.Among them,there were 26 female(60.4％)and 30 hypertension patients(69.8％),After adjusting for age and TUG,the logistic regres-sion analysis showed that the step length(OR 0.821,95％CI 0.702-0.959,P=0.013)was an independent protec-tive factor against the risk of falls in CSVD patients,while the total MRI burden(OR 4.217,95％CI 1.444-12.317,P=0.009)was an independent risk factor for the falls in CSVD.The ROC curve analysis showed that the combination of step length and total MRI burden had a high predictive value for the risk of fall in CSVD patients with gait disorder(AUC=0.904),with a sensitivity of 82.6％and a specificity of 90％. Conclusion:Step length combined with total MRI burden has a high predictive value for the risk of falls in CSVD patients with gait disorder.

Objective:To analyze the consistency and repeatability of ultrasonography in diaphragm function evaluation. Method:Sixty healthy subjects participated in the present study,using a PA12A portable color doppler untra-sound diagnostic system.The diaphragm data were recorded during calm breathing and deep breathing,includ-ing diaphragm muscle mobility during calm breathing and deep breathing,diaphragmatic thickness during calm exhalation and calm inspiration,and diaphragmatic thickness during deep exhalation and deep inspiration.The in-tra-test reliability,inter-tester reliability and retest reliability of the above indexes were analyzed respectively. Result:For intra-test reliability,the intra-class correlation coefficient(ICC)values for diaphragm muscle mo-bility during calm breathing,diaphragm muscle mobility during deep breathing,diaphragmatic thickness during calm exhalation,diaphragmatic thickness during calm inspiration,diaphragmatic thickness during maximal exha-lation and diaphragmatic thickness during maximal inspiration were 0.99,0.99,0.99,0.99,0.99,0.99 and 0.99,respectively.The ICC values for the six indicators were 0.84,0.80,0.82,0.82,0.83,and 0.77 for in-ter-tester reliability.For retest reliability,the ICC values for the six indicators were 0.98,0.86,0.98,0.97,0.94,and 0.86 for relative reliability.The standard error of measurement(SEM)for the absolute reliability of the six indicators were 0.09,0.39,0.01,0.02,0.02,0.03,SEM％were 5.23％,8.40％,5.38％,5.7％,8.17％,8.77％,and the minimal detectable change(MDC)were 0.25,1.08,0.03,0.04,0.04,0.09,and MDC％were 14.49％,23.28％,14.9％,15.8％,22.64％,24.31％,respectively.Bland-Altman graphical analy-sis showed no systematic errors. Conclusion:Ultrasound has good reliability in diaphragm function evaluation and can provide objective evi-dence in clinical application.