Objective To investigate the effects of mirror therapy supplemented with motor imaging training on the upper extremity functions of hemiplegic stroke patients and on their ability in the activities of daily living (ADL).Methods Forty-four stroke patients were divided into a control group and an experimental group with 22 patients in each.All were given routine rehabilitation treatment.In addition,the experimental group was given 30 minutes of mirror therapy daily supplemented with 15 minutes of motor imaging training.The control group was given routine occupational therapy.All the patients were assessed with Brunnstrom staging,the functional test for the hemiplegic upper extremity-Hong Kong (FTHUE-HK) and the Barthel index (BI) scale before therapy and after 6 weeks.Results After 6 weeks of treatment,both groups showed significant improvements in upper limb function,hand function,and ADL ability.There were significant intra-group differences and significant differences between the experimental and control groups in terms of Brunnstrom staging,FTHUE-HK scores and BI scores.Conclusions Mirror therapy supplemented with motor imaging training can benefit patients suffering hemiplegia in improving their upper limb functioning and their ADL ability.

Objective To observe any effect of transcutaneous electrical nerve stimulation (TENS) on pul monary function,airflow obstruction,dyspnea,exercise capacity and levels of tumor necrosis factor-α (TNF-α) in the sputum of patients with chronic obstructive pulmonary disease (COPD).Methods Thirty patients with stage Ⅰ or Ⅱ COPD were randomly divided into a treatment group and a control group with 15 cases in each.TENS applied at the Feishu acupuncture point (BL13) was used in the treatment group,while sham stimulation of the same point without current output was used in the control group.The treatment was administered 40 minutes once daily,5 days a week for a total of 4 weeks.The outcome measures were measured before and after 4 weeks of treatment.Results FVC,FEV1,FEV1％,and peak expiratory flow rate (PEFR) all improved significantly in the treatment group after 4 weeks of treatment.After treatment,the average TNF-α level in induced sputum and exercise capacity as indicated by the average BODE index had both improved significantly in the treatment group.Compared with the control group,pulmonary function,TNF-α and the BODE index in the treatment group were all significantly better.There was no significant difference in pulmonary function,BODE index or TNF-α before and after treatment in the control group.Conclusion TENS at the Feishu acupoint can reduce airway inflammation,improve pulmonary function and exercise capacity,and decrease TNF-α levels in patients with stage Ⅰ or Ⅱ COPD.This treatment should be very useful for enhancing their ability in the activities of daily living.

Objective To compare the effects of repetitive transcranial magnetic stimulation (rTMS) at various low frequencies on upper limb function after cerebral infarction.Methods Fifty patients were randomly assigned to a control group (10 cases),a sham rTMS group (10 cases) or an rTMS group which had three sub-groups treated at 0.25 Hz,0.5 Hz and 0.75 Hz with 10 cases in each.All of the patients were treated with conventional medical treatment and rehabilitation training.The sham and true rTMS groups received rTMS applied over the M1 area of the unaffected hemisphere,5 days per week for 4 weeks.Motor evoked potential (MEP) cortical latency,and central motor conduction time (CMCT) were measured and the Fugl-Meyer assessment (FMA),motricity index (MI) and a Hong Kong functional test for the hemiplegic upper extremity (FTHUE-HK) were evaluated beforehand and at Post 1 after 2 weeks of treatment and Post 2 after 4 weeks of treatment.Results The average CMCT and FMA scores of the control and sham rTMS groups both had improved significantly at Post 2.There was no significant difference in any of the indices between those 2 groups at any time point.At Post 1,the average MEP cortical latencies of the 0.25 Hz and 0.5 Hz subgroups had improved to be significantly better than those of the control and sham rTMS groups.The average CMCTs of the 0.25 Hz and 0.5 Hz rTMS subgroups were significantly shorter after treatment,and significantly better than those of the control and sham rTMS groups.At Post 2,the average MEP cortical latency of all groups except the control group showed significant improvement compared with pre-treatment.The 2 indices of the 0.25 Hz and 0.5 Hz subgroups were again significantly shorter than those of the control and sham rTMS groups,and the average CMCTs were significantly better than that of 0.75 Hz subgroup.At Post 1 the average FMA and MI scores of the rTMS subgroups had all improved significantly.In the 0.25 Hz and 0.5 Hz subgroups the average MI scores were significantly higher than those of the control and sham rTMS groups.The FTHUE-HK scores of those 2 subgroups had also improved significantly.At Post 2,the average FMA and MI scores of all groups and the FTHUE-HK scores of rTMS group had improved significantly.In the 0.25 Hz and 0.5 Hz subgroups,all of the indices were significantly better than in the control and sham rTMS groups.The average FTHUE-HK score of the 0.25 Hz subgroup was significantly superior to that of the 0.75 Hz subgroup.In the 0.75 Hz subgroup the average MI score was significantly higher than in the control and sham rTMS groups.Conclusions rTMS at either 0.25 Hz or 0.5 Hz applied to the unaffected hemisphere provides effective treatment for enhancing the excitability of the motor cortex and the motor function of a paretic upper limb after stroke.Compared with others,the total number of stimulus pulse in 0.25 Hz subgroup was the least,and priority consideration should be given to the frequency of 0.25 Hz when using rTMS in clinical treatment of cerebral infarction.

Aim Tongluoxingnao effervescent tablets ( TLXNET) ,based on the ancient formula of QiongGui Tang, can improve cognitive dysfunction in different AD models. This research is aimed to study the effects of TLXNET on the Aβmetabolism and explore the anti-AD mechanism in SH-SY5 Y-APP and SH-SY5 Y-C99 cells. Methods Cells were incubated for 48h in dif-ferent concentrations of medicated serum ( containing 0% ~40% of TLXNET in the serum ) . Firstly, the non-toxic concentration was measured by MTT assay, the activity of cells was detected by LDH methods, and ELISA was used to measure the levels of Aβ1-40 and Aβ1-42 . Then, the gene and protein expressions of APP were detected via RT-PCR and Western blot of which the expressions of shearing fragments such as sAPPα and sAPPβ were investigated by Western blot, the same as the expressions of IDE and NEP. Addi-tionally, the level of mRNA, protein and activity of BACE1 were measured by qRT-PCR, Western blot and fluorescence detection kits respectively. Eventually, MTT assay was performed to detect the cell viability after the SH - SY 5 Y cells were treated with various concentrations of medicated serum and Aβ25-35 for 48h. Results There was no significant toxicity of TLXNET medicated serum in SH-SY5 Y-APP and SH-SY5 Y-C99 cells ( P >0 . 05 ) . TLXNET could signifi-cantly inhibit Aβ secretion in SH-SY5 Y-APP cells ( P<0 . 05 ) , but had no effect on Aβ secretion in SH-SY5Y-C99 cells, the same as the level of APP mRNA and protein in SH-SY5 Y-APP cells and the expressions of IDE and NEP (P>0. 05). Additionally, TLXNET could still notably inhibit the expression of sAPPβ pro-tein in a dose-dependent way, with statistical signifi-cance ( P <0. 05 ) . Meanwhile, the level of mRNA, protein and activity of BACE1 were also significantly decreased by TLXNET (P <0. 01). Moreover, the medicated serum of TLXNET had a protective effect on SH-SY5 Y apoptosis induced by Aβ25-35 ( P <0 . 01 ) . Conclusion TLXNET could obviously inhibit β-secretase enzyme, and has an antagonistic effect a-gainst Aβneurotoxicity, which suggests that the inhibi-tion of β-secretase enzyme and antagonism against Aβneurotoxicity are the main anti-AD mechanism of TLX-NET .

Background and purpose:Liquid biopsy is a kind of blood, urine and other non-solid biologi-cal tissue sampling analysis, mainly for malignant tumor diagnosis, monitoring and predicting its prognosis. In this research, we optimized the extraction of miRNA in urine, established a standardized means of liquid biopsy, screened and verified the miRNA markers in patients with bladder cancer.Methods:From Jan. 2014 to Sept. 2015, we used miRNA microarray in six patients with bladder cancer and six healthy controls. Samples of 78 cases of bladder cancer and 23 healthy controls were tested by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) to verify the relationship between miRNA markers in liquid biopsy and clinical pathological parameters. The diagnostic value of miRNA markers was also analyzed and compared.Results:We screened 10 miRNAs differential expression in urine. Combined with previous literature, we selected 20 miRNAs to verify their expression levels in bladder cancers and healthy controls. miR-509-5p/miR-124 ratio in the urine was found higher in patients with bladder cancer than in healthy controls (P<0.0001). With the rise of miR-509-5p/miR-124 ratio in urine, tumor stage and grade were also increased (P=0.003). When the cutoff was set at 0.41, the diagnostic sensitivity and specificity of miR-509-5p/miR-124 ratio were 73.1% and 82.6%, respectively. The AUC of miR-509-5p/miR-124 ratio to detect bladder cancer was 0.864, higher than that of urinary exfoliated cells (P=0.0002).Conclusion:We optimized the extraction of miRNAs in urine,established a standardized liquid biopsy of miRNA markers. The miR-509-5p/miR-124 ratio could be an ideal diagnos-tic marker for bladder cancer.

Objective To explore the effect and mechanism of renal denervation combined with Losartan on cardiac hypertrophy in spontaneous hypertensive rats. Methods Spontaneous hypertensive rats with cardiac hypertrophy underwent renal denervation combined with following 4 weeks drug treatments. Transthoracic two-dimensional guided M-mode echocardiography was performed. The expression of hypertrophy-related gene was detected. Left ventricularmass index was calculated and histological sectionsstained with hematoxylin and eosin (HE). Results The characteristics of cardiac hypertrophy was observed in left ventricular tissues with HE staining in12 weeks old spontaneous hypertensive rats.The thickness was increased in those ventricular (IVSs, IVSd, LVPWs and LVPWd). After4 weeks after treatment, histological improvements were observed in both RDN and R+L groups compared with sham group. The improvement was more obvious in R+L group. Similar results were observed in histological sections, ventricular thicknesses (IVSs, IVSd, LVPWs and LVPWd), related-hypertrophy genes (ANP and β-MHC) expression, left ventricular mass idex (LVMI) and the protein expression of inflammatory cytokines (TNF-α and IL-6). Conculsion Renal denervation therapy can improve hypertensive-induced cardiac hypertrophy in spontaneous hypertensive rats. The effect was more significant when combined with Losartan. The mechanism might be involved in inhibiting inflammatory cytokines.

Objective To explore the effects of miR-34a and the alteration of AMPK/mTOR signal pathway on angiotensin (Ang)Ⅱ-stimulated cardiomyocytes hypertrophy. Methods Neonatal rat cardiomyocytes were cultrued in vitro and were divided into 3 groups:negative control for lentivirus carrying miR-34a group (NC), AngⅡplus lentivirus carrying negative control group (AngⅡ+NC) and AngⅡplus lentivirus carrying miR-34a group (AngⅡ+miR-34a). The relative cell area was detected by confocal microscopy. The expression of miR-34a and hypertrophy-related genes (ANP and β-MHC) were analyzed by real time PCR. The AMPK/mTOR signal pathway was measured by Western blot assay. Results Compared to NC group, the relative cell area was increased in AngⅡ+NC group (P<0.05). But compared with AngⅡ+NC group, the relative cell area was decreased in AngII+miR-34a group (P<0.05). Moreover, compared with NC group, the expression of miR-34a was decreased, and the expression of hyperthophy-related genes(ANP and β-MHC) was up-regulated in AngⅡ+NC group. However, the expression of miR-34a was decreased, and the expression of hyperthophy-related genes (ANP and β-MHC) was down-regulated (P<0.05). Finally, compared to NC group, the ratio of phosphop-AMPK/AMPK was significantly induced in AngII + NC group, but the ratio of phosphop-mTOR/mTOR was significantly decreased (P<0.05). However, compared to AngⅡ+NC group, the ratio of phosphop-AMPK/AMPK was significantly decreased in AngII + miR-34a group, but the ratio of phosphop-mTOR/mTOR was significantly increased (P<0.05). Conclusion miR-34a is able to inhibit myocardial hypertrophy of neonatal rat cardiomyocytes, and its mechanism is partly carried out by the alteration of the signal pathway of AMPK/mTOR.

Objective To explore the effects of miRNA-204 targeted LC3B expression on Ang Ⅱ induced cardiomyocytes hypertrophy.Methods The primary neonatal rat cardiomyocytes served as the research objects and divided into the control group,AngⅡ group,combination-treated group 1 (cardiomyocytes were given Ang Ⅱ stimulation,meanwhile infected by negative control lentivirus vector),combination-treated group 2 (cardiomyocytes were given Ang Ⅱ stimulation,meanwhile infected by lentivirus carrying miRNA-204 overexpression vector) according to different treatments.About 48 h to 72 h after intervention treatment,the cardiomyocyte hypertrophy change was detected by confocal microscopy,the expression of miRNA-204 was analyzed by real time PCR,the protein expression of LC3B was measured by Western blot and targeted gene of miRNA-204 was demonstrated by dual-luciferase reporter assay system.Results Compared with the control group,the cardiomyocyte relative surface area in the Ang Ⅱ group was significantly enlarged,the protein expression of LC3B was significantly increased,the expression of miRNA-204 was upregulated,the differences were statistically significant (P＜0.05).Whereas comparing the combination-treated group 1 with combination-treated group 2,the protein expression of LC3B in the latter was down-regulated and the cell area was reduced (P＜0.05).The further luciferase activity report gene experiment results suggested that miRNA-204 was able to bind to LC3B 3'-UTR and decreased the luciferase activities (P＜0.05),but not to bind its mutated fragment for inactivating luciferase activity(P＞0.05).Conclusion miRNA-204 is able to inhibit Ang Ⅱ induced cardiomyocytes hypertrophy,its action is realized by targeting the expression of LC3B.

Objective:To analyze the expression of circular RNA (circRNA) in peripheral blood mononuclear cells (PBMCs) of patients with gout and to explore the possible mechanism of circRNA in the pathogenesis of gout.Methods:Peripheral blood samples of 24 patients with acute gout (AG), 24 patients with intermittent gout (IG) and 24 healthy control subjects (HC) were collected. Three cases of AG, IG, and HC were randomly selected, and the differentially expressed circRNA in PBMCs was screened by human circNA microarrays. The 6 circRNAs with large differences between the two comparison groups were selected, and the relative expression levels of 6 circRNAs in all the collected 72 PBMCs of the study subjects were detected by real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). The significantly differentially ex-pressed circRNA (fold change>1.5, P<0.05) was analyzed by GO analysis, Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis, and its interaction with microRNA (miRNA) was predicted. The median (interquartile range) was used to describe the data, and the Mann-Whitney U test was used for comparison between groups. Results:(1) The microarray analysis results showed that compared with the HC group, the AG group and the IG group had 116 and 41 significantly differently expressed circRNAs, respectively; com-pared with the IG group, the AG group had 105 significantly differently expressed circRNAs. (2) Among the 6 circRNAs verified by PT-qPCR, the expression trends of 5 were consistent with the microarray results. The expression of hsa_circRNA_105034 in the AG group [5.17(4.60)] was statistically significantly different com-pared to the IG [1.68(2.39)] and HC [0.90(0.73)] groups (AG vs IG: Z=-4.413, P<0.01; AG vs HC Z=-5.052, P<0.01). (3) Bioinformatics analysis: ① GO analysis found that differential circRNA swere mainly involved in DNA transcriptional regulation, positive cell regulation and protein modification, etc. ② KEGG pathway analysis revealed that differential circRNA might be involved in the immune response mediated by the mitogen-activated protein kinase signaling pathway. ③ CircRNA might affect its inflammatory response by targeting molecules such as miRNA-146a, miRNA-302b and miRNA-23a. Conclusion:There are differentially expressed circRNAs in PBMCs of patients with gout, which may be closely related to the occurrence and development of gout.

Objective To construct vector expressing soluble mPDL1-hIgGFc and study its effect on the proliferation and apeptosis of cells in vitro. Methods The extrncellular domain of mPDL1 gene was amplified from pmPDL1 vector by PCR and inserted into phIgGFc vector. The recombinant pmPDL1-hIgGFc was transfected into CHO cells by LipofectAMINETM2000, and the transfected cells were named as CHOp. The expression of mPDL1-hIgGFc in the culture supernatants of CHOp was assayed by ELISA and Western blot. The effects of CHOp culture supernatants on mixed lymphocyte culture(MLC) was analysed by Flowm-etry. Results The extracellular domain of mPDL1 gene were obtained from PCR. DNA sequencing and the identification of digestion by HindⅢ and KpnⅠ indicated the recombinant plasmid pmPDL1-hIgGFc was suc-cessfully constructed. ELISA and Western blot analysis proved that the CHOp could express mPDL1-hIgG-Fc. CHOp culture supernatants could inhibit lymphocyte proliferation and induce the apoptosis of the activa-ted T cells in MLC in vitro in a dose-dependent manner. Conclusion The mPDL1-hIgGFc protein could in-hibit lymphocyte proliferation and induce the apoptosis of the activated T cells.