Objective To investigate influence of Ang-(1-7) on the inducible nitric oxide synthase (iNOS) activity and gene expression following focal cerebral ischemia/reperfusion in rats. Methods Cerebral ischemia/reperfusion injury was induced by intraluminal thread occlusion of middle cerebral artery in the adult male Sprague-Dawley (SD) rats. Ang-(1-7) or artificial cerebrospinal fluid (aCSF) was continuous administrated by implanted Alzet osmotic minipumps into lateral cerebral ventricle after reperfusion. Experimental animals were divided into sham-operated group ( sham operation + aCSF), aCSF treatment group(MCAO+aCSF)and ang-(1-7)treatment groups(MCAO+Ang-(1-7))at low(1 pmol·0.5 μl-1·h-1),medium (100 pmol·0.5 μl-1·h-1)or hith(10 nmol·0.5 μl-1·h-1)dose levels.The activity of iNOS in ischemic tissues were measured by iNOS detection kits. Reverse transcription( RT)-PCR was used to determine messenger RNA (mRNA) of the iNOS in ischemic tissues. Results The cerebral ischemic lesion resulted in a significant increase of iNOS expression compared with sham operation group. The high-dose Ang-(1-7) markedly enhanced (iNOS) activity ( 160. 83 vs 116. 75 U/mg, F = 19. 22,P＜0.01; 151.87 vs 113.07 U/mg, F=63.52,P＜0. 01) and gene expression(0.43 vs 0.38, F=21.83,P ＜ 0. 01; 0. 40 vs 0. 35, F = 19.49, P ＜ 0. 01 ) compared with aCSF treatment group at 24 hours and 48hours after reperfusion, whereas medium and low-dose Ang-( 1-7 ) didn't stimulate iNOS activation.Conclusions The obtained results suggest that high-dose Ang-(1-7) upregulate iNOS expression following ischemic stroke.Moreover,overdose Ang-(1-7)(10 nmol·0.5 μl-1·h-1)may have Ang Ⅱ-like effects in iNOS expression increase.

Objective: To explore the relationship between ambulatory arterial stiffness indexes (AASI), AASI-blood pressure variability (AASI-BPVR) and left ventricular mass index (LVMI) left atrium diameter (LAD) in patients with hypertension.
Methods: A total of 286 hypertensive patients were enrolled in this study. The AASI, AASI-BPVR were calculated from 24-hour ambulatory blood pressure monitoring. Left ventricular internal dimension in diastole (LVIDd), interventricular septal thickness in diastole (IVSd), posterior wall thickness in diastole (PWd), LAD were detected by echocardiography and LVMI, relative wall thickness (RWT) were calculated. The fasting blood glucose, blood lipids were examined. According to 24 h AASI, the patients were divided into 2 groups, Group A, the patients with AASI > 0.51, n=133 and Group B, the patients with AASI ≤ 0.51,n=153. Pearson and multi regression analysis were conducted to analyze the relevant correlations.
Results: Group A had increased LVMI than that in Group B,P<0.05, the left ventricular masses were similar between 2 groups,P=0.384. Pearson correlation analysis indicated that LVMI and LAD were not related to arterial stiffness index, the coefifcient between 24 h-AASI and LAD was atr=0.111,P=0.057.
Conclusion: AASI and AASI-BPVR were not the independent factors for left ventricular hypertrophy and left atrial enlargement, therefore, they were not the predictors for cardiac damage in patients with hypertension at present time.

Objective To improve the recognition of rapid eye movement(REM) sleep behavior disorder(RBD) as an early marker for ?-synucleinopathies.Methods By studying a typical case of RBD followed with multiple system atrophy-P,the clinical features,pathogenesis and its correlation with ?-synucleinopathies of RBD were elucidated.Results This case manifested a serial of paroxysmal increased activities of the limbs and behavioral disturbances during his REM sleep,and parkinsonism features appeared 9 years later.His cranial MRI showed the abnormal long T1 and T2 signals at bilateral centrum ovale,corona radiate and basal ganglia area of the cerebral hemisphere.Conclusions RBD is clinically characterized with paroxysmal behavioral disorder in the REM sleep,the changes of the brain stem,striatum and cortical perfusion are attributed to the RBD pathogenesis.Closely linked to a-synucleinopathies,RBD may be clinical harbinger of those disorders.

Objective To analysis the relationship between gestational age and perinatal outcomes in patients complicated with early onset severe preeclampsia.Methods Retrospective study was conducted on clinical documents of 221 patients with early onset severe preeclampsia( ＜ 34 weeks) who delivered after 28 gestational weeks in Peking University First Hospital from July 1999 to June 2009.Patients were divided into three groups based on gestational weeks at delivery: group Ⅰ (n = 81 ) delivered at 28 -31 weeks+6,group Ⅱ (n = 78) at 32 -33 weeks+6 and group Ⅲ (n = 62) after 34 weeks.The clinical characteristics and perinatal outcomes were compared among those three groups.Results ( 1 ) Outcome of neonates:Among 221 neonates, 13 neonates lost follow-up, including 9 in group Ⅰ , 3 in group Ⅱ, 1 in group Ⅲ.The incidence of neonatal respiratory distress syndrome ( RDS ) of 26% ( 19/72 ) in group Ⅰ were significantly higher than 7% (5/75) in group Ⅱ and 10% (6/61) in group Ⅲ (P ＜ 0.05 ).The neonatal mortality rate of (43% ,31/72) in group Ⅰ were significantly higher than 3% (2/61) in group Ⅲ and 28%(21/75) in group Ⅱ (P ＜0.05 ).The incidence of maternal complications showed no statistical difference among three groups.(2) Neonatal death analysis: all neonatal death were due to parents' give up, including 26%(8/31) in group Ⅰ, 67% (14/21)in group Ⅱ and 1/2 in group Ⅲ, which reached statistical difference(P＜0.05).Conclusions The incidence of neonatal RDS in mother with early onset severe preeclampsia was decreased if delivered after 32 weeks, and the perinatal mortality was remarkably decreased if delivered after 34 weeks.Therefore, the perinatal survival rate in women with early onset severe preeclampsia can be improved by minimizing the impact of social factors.

Objective To establish the sperm specific Sleeping Beauty ( SB ) transposase-expression transgenic mouse for the study of the genetic modification mediated by transposon system in mouse .Methods Prm1 promoter was cloned from mouse genomic DNA to drive the expression of SB transposase .The transgenic mice were generated by microinjection .The gene type of transgenic line was identified by PCR .The expressing level in testis was determined by western blot and immunohistochemistry (IHC) staining.Results Five lines of transposase transgenic mice were obtained by microinjection and three can be germline .One mouse line with higher expression level of transposase in the testis was obtained.Conclusion One transgenic mouse model with Sleeping Beauty transposase - expression was successfully established .This model will greatly contribute to the research of genetic modification mediated by transposon in mouse.

Objective To observe the clinical efficacy of moxibustion plus tuina in treating 0-grade diabetic foot.Method Seventy patients with 0-grade diabetic foot were randomized into a treatment group and a control group, 35 cases in each group. The control group was intervened by conventional medication, while the treatment group was by moxibustion plus tuina in addition to the interventions given to the control group. The Ankle Brachial Index (ABI) was observed before intervention and after 2 treatment courses, and the clinical efficacies were compared between the two groups.Result The ABI scores were changed significantly after intervention in both groups (P<0.05). After intervention, there was a significant difference in comparing the ABI score between the two groups (P<0.05). The total effective rate and recovery rate were respectively 94.3% and 71.4% in the treatment group, versus 74.3% and 20.0% in the control group, and the differences were statistically significant (P<0.05).Conclusion Moxibustion plus tuina is an effective approach in treating 0-grade diabetic foot.

Objective To study the relationship of insulin receptor substrate-1 (Irs1) and metabolic disease, we generated Irs1 gene knockout rat by CRISPR/Cas9 system.Methods Two sgRNA targeting sites were designed for Irs1 targeting.The Cas9 and sgRNAs were transcribed by T7 RNA polymerase in vitro.Cas9 mRNA and sgRNA mixtures were pooled and microinjected into one-cell fertilized eggs of SD rats to generate rats with targeted mutation .Results Five rats with the mutations were detected with the efficiency of 83%.Conclusion The Irs1 gene knockout rats generated in this study can be transmitted by germline .

Objective To study the effects of NRG2 on cardiac structure and function , we established the cardiac-specific human NRG2 transgenic mice and investigate the effect of NRG2 on cardiac structure and function under pressure overload situation .Methods The transgenic vector was constructed by insertion of the human NRG2 gene under the α-MHC promoter.The transgenic mice were generated by microinjection and were all maintained on a C57BL/6J genetic background .The genotype of transgenic mice was identified by PCR and the expression level of target gene was determined by western blot .Transverse aortic constriction ( TAC) was applied to prepare the pressure overload induced cardiomyopathy mice model .The cardiac structure and function of the transgenic mice were compared and analysized by echocardiographic and pathological observation .Results Transgenic mice with high level of NRG2 in heart tissues were established.The left ventricular wall thickness (LVPWD) was increased, and to 15.6% at 3 months old compared with that of the non transgenic ( NTG) mice.The hypertrophy of left ventricular wall caused by pressure overload was removed due to the expression of NRG2 .Meanwhile, cardiac disarray and fibrosis were increased obviously compared with that of the NTG mice.Conclusion The transgenic expression of NRG2 in heart tissues could shorten the pathological process of hypertrophy, but accelerated the process of heart failure (HF).

Hepatic myelopathy (HM)is a rare complication of chronic liver diseases.One case of HM treated by was reported.A 40-year-old male recipient of liver transplantation (LT) complained of lower extremity paralysis.Muscle strength of lower extremity recovered partially at Week 3 post-transplantation and completely at Month 3 post-discharge.

Objective:To investigate the predictive value of serum hypersensitive C-reactive protein (hs-CRP) for stroke-associated pneumonia (SAP) in patients with acute ischemic stroke (AIS) who received intravenous thrombolysis.Methods:From May 2015 to April 2017, the clinical data of the patients with AIS treated with intravenous thrombolysis in Nanjing First Hospital were collected retrospectively. Multivariate logistic regression analysis was used to determine the independent risk factors for SAP in patients with AIS after intravenous thrombolysis. Receiver operating characteristic (ROC) curve and nomogram-based methods were used to analyze the predictive value of hs-CRP for SAP. Results:A total of 243 patients with AIS who received intravenous thrombolysis were included, and 63 (34.6%) of them had SAP. There were significant differences in age ( P=0.006), leukocyte count ( P=0.044), fasting blood glucose level ( P=0.003), serum hs-CRP level ( P=0.001), hs-CRP classification ( P=0.001) and dysphagia rate ( P=0.035) between the SAP group and non-SAP group. Multivariate logistic regression analysis showed that after adjusting for the confounding factors, taking the first quartile of serum hs-CRP level as a reference, the third quantile (odds ratio [ OR] 18.790, 95% confidence interval [ CI] 4.771-74.007; P=0.001) and the fourth quantile ( OR 54.054, 95% CI 12.248-324.088; P=0.001) of hs-CRP were the independent predictors of SAP. The area under the ROC curve of the baseline serum hs-CRP level for predicting SAP was 0.805 (95% CI 0.742-0.868; P<0.001). When the optimal cut-off value of hs-CRP was 5.54 mg/L, the sensitivity and specificity of predicting SAP were 76.11% and 76.19%, respectively. The analysis of nomogram also showed that hs-CRP was an independent predictor of SAP (consistency index 0.862, 95% CI 0.738-0.986; P<0.001). Conclusions:The increased serum hs-CRP was an independent predictor of SAP in patients with AIS receiving intravenous thrombolysis, and had a higher predictive value.