AIM:To investigate the effect of NF-κB binding element deletion on transcriptional regulation of (NOX1).METHODS:pGL3-Basic vector inserted with the NOX1 proximal promoter, and the same vector inserted with the NOX1 proximal promoter in the absence of the positive NF-κB binding element, were constructed.After cloning, diges-tion and purification, NOX1 proximal promoter (≈1 415 bp) was inserted into the multicloning site of the pGL3-Basic vec-tor and then sequenced ( pGL3-NOX1-1415) .NF-κB binding elements in the NOX1 promoters were predicted by Alibaba 2.1 software.The positive element was deleted by overlapping PCR.The deletion mutant was inserted into the pGL3 vector in the same way (pGL3-NOX1-1327).The plasmids were transfected into A549 cells, and then the cells were stimulated with TNF-α.The luciferase activity was monitored on MD SpectraMax M5 enzyme-labeled instrument.RESULTS:The se-quences of pGL3-NOX1-1415 and pGL3-NOX1-1327 were identified to be correct.Compared with control group, the lucif-erase activity was significantly higher in the cells transfected with pGL3-NOX1-1327 (P<0.05), but it was significantly lower than that in the cells transfected with pGL3-NOX1-1415 (P<0.05).CONCLUSION: NF-κB plays an essential role in the transcriptional regulation of NOX1 in TNF-α-induced A549 cells.Activated NF-κB binds to specific elements in NOX1 promoter regions to control the transcription.

Objective To systematically review the clinical efficacy of botulinum toxin A for the treatment of upper limb spasticity after stroke. Methods PubMed,EMbase,Cochrane Library,CBM, China National Knowledge Internet (CNKI),and Wanfang Data were retrieved by a computer. The randomized controlled trials and Case-control study of comparing botulinum toxin A combined with rehabilitation training and routine rehabilitation treatment for upper limb spasticity after stroke were collected. The retrieval time was from the foundation of the database to September 1,2016. At least two reviewers conducted literature screening,data extraction,and quality evaluation according to the inclusion and exclusion criteria. The Rev-Man 5. 3 software provided by the Cochrane Collaboration was used to conduct the Meta-analysis. Results A total of 356 articles were retrieved. Finally,11 articles met the inclusion criteria. The modified Jadad score showed that the scores of 9 articles were more than 4,which belonged to high-quality literature. Eleven articles included 614 patients,in which 316 were treated with botulinum toxin A. There were 298 patients in the control group. The results of Meta-analysis indicated that the Fugl-Meyer score (SMD =0. 94,95% CI 0. 75 to 1. 12),modified Ashworth score (SMD = - 1. 59,95% CI - 1. 78 to - 1. 40),and modified Barthel index (SMD = 0. 86,95% CI 0. 65 to 1. 08)in the botulinum toxin treatment group were superior to the control group. There were significant differences (all P < 0. 05). The incidence of adverse events (RR = 1. 06,95% CI 0. 78 to 1. 45)was similar in both groups. There was no significant difference (P > 0. 05). Conclusion The efficacy of treatment of upper limb spasticity after stroke with botulinum toxin A is exact,and the security is high. The appropriate rehabilitation training can effectively improve the limb motor function,muscle tension,and quality of life of patients.

Objective To screen the variation in NeuroD1 gene and to study its function in vitro and its clinical phenotypes and genetic characteristics in Chinese early-onset type 2 diabetic probands. Methods PCR-direct sequencing of NeuroD1 gene was performed in 85 early-onset type 2 diabetic probands, 95 late-onset type 2 diabetics with strong diabetic history and 87 non-diabetic control subjects. Distributions of the identified variation were calculated and compared among the three groups. Expression vectors with mouse NeuroD1 (mND1)cDNA wild type or mutant type and reporter vectors with human insulin promotor-linked luciferase were constructed. Then the above vectors were co-transfected into rat INS-1 cells. Relative luciferase activities were measured to compare transcriptional activities of insulin gene between WT and MT. Results S159P (T→C), a new mutation was identified in a proband, which was co-segregated with diabetes in 4 carriers from the paternal side. The functional study showed that the S159P mutant exhibited a 25% reduction in transcriptional activity of insulin gene as compared with the wild type. A45T (G→A), a common variation was identified. The AA + GA genotypic frequencies were markedly increased in early-onset type 2 diabetic probands as compared with late-onset type 2 diabetic probands and non-diabetic control subjects (P=0.006 and P=0.014, respectively). Conclusion The novel S159P mutation in the NeuroDl gene seems to contribute to the development of diabetes in the Chinese early-onset type 2 diabetic family. The A45T variation may increase susceptibility to or be in disequilibrium with early-onset type 2 diabetes mellitus in Chinese population. In addition, the A45T variation may affect the onset pattern of type 2 diabetes mellitns, such as early-onset but not late-onset type 2 diabetes mellitus.

Objective To evaluate the effects of cardiac resynchronization therapy with pacemaker (CRT- P) and cardiac resynchronization therapy with defibrillator (CRT -D) on the outcomes of patients with heart failure.Methods MEDLINE,Cochrane Controlled Trials Register,EMBASE and Chinese Wan Fang,CNKI database were searched to collect data from randomized controlled trials and cohort trials of CRT - P versus CRT - D for the treatment of heart failure from January 1,1990 through September 30,2011.Meta - analysis of data including all causes leading to mortality and mortality of sudden cardiac arrest and heart failure was carried out by using the RevMan 5.0 package.Results A total of 3 404 patients were collected from seven studies.Pooled analysis demonstrated CRT - D significantly reduced mortalities of all causes in comparison with CRT - P [ odds ratio (OR) =0.61,95％ confidence interval (CI) =0.47 ～0.79,P =0.0001 ].Sub - group analysis showed that an increased benefit was seen after extended follow -up period ( after 1 year,OR =0.56,95 ％ CI =0.41 ～ 0.77,P =0.0004),but not after relatively short follow - up period ( within one year,OR =0.76,95 ％ CI =0.54 ～ 1.06,P =0.11 ).Mortalities of sudden cardiac arrest ( OR =0.20,95％ CI =0.07 ～ 0.59,P =0.003) and heart failure ( OR =0.72,95％ CI =0.54 ～ 0.96,P =0.02) of patients treated with CRT - D were lower than those of patients treated with CRT- P.Conclusions This study suggests that CRT - D is superior over CRT - P in respect of better outcome of patients with heart failure.

Objective To assess the change of left ventricular wall motion at ventricular pacing(VP)by using global circumferential strain (GCS) and segmental circumferential strain of speckle tracking imaging(STI). Methods Seven adult pigs were sedated,followed by median sternotomy and opening of the pericardium. Temporary pacing leads were placed on the right ventricular (RV) epicardium and LV epicardium for RV or LV pacing. A LV short axis view at the mid-level LV was acquired at sinus and VP rhythm. Analysis of circumferential strain by STI was performed offline. The onset of QRS wave was used to characterize the beginning of the cardiac cycle, and the ratio of beginning systolic time, end systolic time and total systolic time to cardiac cycle dyssynchrony was analyzed at sinus and VP rhythm. Results Compared with sinus rhythm,in VP rhythm all of the ratio of the beginning systolic time,end systolic time and total systolic time to cardiac cycle were increased (all P ＜0.05), and the construction of six segments were dyssynchrony,even segmental paradoxical movement. At VP ≥130 beats/min,both LV global and LV segments sometimes showed irregular motion while the EKG remained normal, which included: ( 1 ) multiple construction and diastole in one heart cycle; (2) A large variability of construction amplitude in several consecutive cardiac cycles. Conclusions VP can lead to several kinds of disturbance of left ventricular wall motion, including delay and lengthening of LV systole, dyssynchrony, and irregularity of systole and diastole.

Objective In order to know the effect of rehabilitation of pelvic organs by pelvic floor muscle training and electrical stimulation. Methods Divided 330 pregnant women into the integrated group(120 cases),the experimental group(100 cases) and the control group(100 cases) randomly. Rou-tine nursing cares was used in the control group, pelvic floor muscle training and electrical stimulation were used in the integrated group in addition, pelvic floor muscle training was used in the experimental group. Evaluated the rehabilitation condition of pelvic organs in the 42nd day after delivery among the three groups. Results All the indexes which can indicated the rehabilitation condition of pelvic or-gans in the integrated group and the experimental group were significant better than those of in the con-trol group, while there was no significant differences about the above indexes between the integrated group and the experimental group. Conclusions Pelvic floor muscle training combined with electrical stimulation can effective promote the rehabilitation of function of pelvic organs, and then prevent the in-cidence of related diseases.

Objective: To study the effect of flavonoids from seed residues of Hippophae rhamnoides L.( FSH) and Hippophae rhamnoides L. oil(HRO)on serum lipid in female rats with obesity. Methods Female aged rats with obesity were given FSH and HRO by oral administration for 6 w. Serum triglyceride(TG) and cholesterol(TC) were determined every 2 w. The serum free fatty acid (FFA), high-density lipoprotein cholesterol (HDL-C),body weight and Lee’s index were measured after 6 w. Results: Serum TG and FFA level decreased significantly in rats given FSH. At the same time, FSH could inhibit the increase in body weight and Lee’s index. HRO could regulate the lipid metabolism and decrease serum TG level significantly in female aged rats with obesity. Conclusion: Both FSH and HRO can improve lipid metabolism in female aged rats with obesity.

Objective:To explore the protective effect of vagus nerve stimulation (VNS) on the prognosis of rats suffering from cardiac arrest/cardiopulmonary resuscitation (CA/CPR) under different treatment timings.Methods:The method of percutaneous epicardial electrical stimulation was used to establish CA model of rat. Fifty-three male SD rats were randomly (random number) divided into the sham group ( n=5), CPR group ( n=12), PRE group ( n=12), POST5 group ( n=12) and POST30 group ( n=12). The sham group did not experience CA/CPR. VNS treatment was started at 30 min before CA (PRE group, n=12), 5 min after recovery of spontaneous circulation (ROSC) (POST5 group, n=12), and 30 min after ROSC (POST30 group, n=12) in different VNS-treated group, respectively. The electrical stimulation was applied to the vagus nerve for 30 min with a unified parameter. The neurological deficit scores at 24, 48, and 72 h after ROSC were recorded, and the survival rate in each group was observed. TUNEL staining was used to detect the apoptosis of cortical area and the expression of α7 nicotinic acetylcholine receptor (α7nAChR) in brain tissue was measured by immunofluorescence at 72 h after ROSC. Variables were compared with one-way analysis of variance, and survival for Kaplan-Meier curves were tested with the log-rank test. A P value less than 0.05 was considered statistically significant. Results:Compared with the CPR group (survival rate 33.33%), both pre-treatment (survival rate 75%) and post-treatment of VNS (POST5 group survival rate 75% and POST30 group survival rate 83.33%) significantly improved the 72 h survival rate after CPR ( P<0.05), mitigated neurological deficits after ROSC, reduced the positive rate of apoptosis neurons, and up-regulated the expression of α7nAChR in cerebral cortex. There was no significant difference among the VNS-treated groups (all P>0.05). Conclusions:Both pre-treatment and post-treatment of VNS can play a protective role in rats after CA/CPR, which may be related to the activation of α7nAChR-mediated anti-inflammatory and anti-apoptosis effects.

Objective: To investigate the incidence of coutrast media iodixanol-induced delayed adverse reaction with the risk factors in general clinical practice.
Methods: A total of 20,185 patients with contrast iodixanol were recruited from 95 medical centers in China. The risk factors for adverse drug reaction as hypertension, asthma, previous contrast reaction were assessed;the administrative processes as route, injection manner, lfow rate of injection, prior heating of iodixanol were monitored and the demographic information was documented. The immediate adverse reaction within 1 hour of media administration and the delayed adverse reaction from 1 hour to 7 days after administration were recorded. The risk factors for iodixanol-induced delayed adverse reaction were studied by singlevariate and multivariate logistic regression analysis.
Results: The overall iodixanol-induced adverse reaction rate was 1.52%, of which the immediate reaction was 0.58%and delayed reaction was 0.97%. The major delayed reaction was mild and it mostly happened in skin (0.68%) including rash, pruritus and urticaria. Multivariate logistic regression analysis revealed that male gender (OR=0.71, P=0.036), age (OR=0.82, P=0.001), route of administration (OR=0.21, P<0.001), prior heating of iodixanol (OR=1.44, P=0.036), lfow rate of injection (OR=1.28, P=0.001) and previous contrast reaction (OR=16.04, P<0.001) were the independent risk factors for delayed adverse reactions.

[ ABSTRACT] AIM:To investigate the effects of pathological products, urinary proteins and advanced glycosyla-tion end products ( AGE) produced in the progression of chronic kidney disease ( CKD) , on the structure and function of lysosomes in renal tubular epithelial cells ( TECs ) , and try to find a novel approach for preventing or delaying CKD. METHODS:The renal specimens of the untreated patients with minimal change nephrotic syndrome (MCNS), diabetic nephropathy (DN) or normal kidney were collected.The expression of lysosomal-associated membrane protein 1 (LAMP1) and cathepsin B ( CB) was studied in TECs by indirect immunofluorescent staining.Human renal tubular epithelial cell line HK-2 was incubated with 8 g/L urinary proteins or 100 mg/L AGE.The expression of LAMP1 and CB was investigated by indirect immunofluorescence and the activity of CB and cathepsin L ( CL) was measured by biochemical and enzymatic as-says.The degradation of DQ-ovalbumin was also determined.RESULTS: The lysosomal membrane permeabilization oc-curred in the TECs of MCNS and DN patients.After treatment with urinary proteins or AGE-BSA, the lysosomal membrane permeabilization of the HK-2 cells was increased.The activity of CB and CL and degradation of DQ-ovalbumin were de-creased as compared with normal control group.CONCLUSION:The digestive function of lysosome was decreased and ly-sosomal membrane permeabilization occurred in the TECs exposed to urinary proteins and AGE, which might be a key factor to induce the tubulointerstitial fibrosis.