Objective To investigate the effects of siRNA against survivin combined with the neoadjuvant chemotherapy (paclitaxel and epirubicin) on the apoptosis in breast cancer MCF-7 cell and reverse drug resistance.Methods Molecular cloning technique was applied to construct the expression vector of siRNA against survivin,and effectene transfection reagent was used to transfect MCF-7 cell.Survivin expression was detected by qPCR and Western blot methods.The effects of paclitaxel or epirubicin combined with or without siRNA against survivin,on the drug susceptibility of MCF-7 cell were detected by CCK-8 assay and that of including MCF-7 cell apoptosis were detected by flow cytometry.Results Survivin-siRNA-2 effectively inhibited the expression of survivin mRNA (0.30±0.03) and protein (0.37±0.09) (P ＜ 0.05).Both paclitaxel [IR 24,48 h =(38.5±4.0) ％,(41.7±6.3) ％] and epirubicin [IR 24,48 h =(37.0±8.6) ％,(40.6± 12.1) ％] can suppress the proliferation of MCF-7 cell and induce its apoptosis,and siRNA against survivin combined with chemotherapy drugs enhanced both effects significantly[paclitaxel IR 24,48 h =(76.0±2.9) ％,(85.1±4.0) ％; epirubicin IR 24,48 h =(74.6±5.6) ％,(82.5±4.8) ％] (P ＜ 0.05).The inhibition rate of MCF-7 was concentration and time dependent.Conclusions Blocking survivin gene expression by RNAi,and combining with the appropriate chemotherapy can significantly enhance the sensitivity of MCF-7 cell to drugs and cell apoptosis.This technology has important potential clinical value in the therapy of breast cancer.

Objective To synthesize poly asparagine derivatives and to evaluate its safety at the cellular level, which provide research platform for its potential application as drug carrier. Methods Polysuccinimide was synthesized by ther?mal polymerization of L-polyaspartic acid, and the target product of PSI-Phe-EA was obtained by the ring-opening reaction of polysuccinimide using L-phenylalanine methyl ester hydrochloride and ethanol amine. The structure of PSI-Phe-EA were characterized by 1H NMR. The rate of ring-opening of PSI was calculated by internal standard method of 1H NMR. The change of hydrophilicity was studied by the comparison of solubility. The cytotoxicity and morphology modification by PSI-Phe-EA at designate concentrations was investigated by MTT method and inverted microscopy respectively. The effects on cell cycles were analyzed by flow cytometry after propidium iodide (PI) staining. Results 1H NMR results confirmed the structure of PSI-Phe-EA and the ring-openning rate of PSI was 40%. The hydrophilicity of PSI-Phe-EA was greatly in?creased upon ring opening using ethanol amine. MTT test showed that the cell survival rates of NIH 3T3 and HepG2 cells were higher than 80%under the examined concentration (<100 mg/L). Inverted microscopy showed that 50 mg/L of PSI-Phe-EA treatment had no adverse effects on cell morphology. Cell cycle analysis indicated that PSI-Phe-EA treatment had no in?fluence on cell cycles of NIH 3T3 and HepG2 cell lines. Conclusion PSI-Phe-EA showed high hydrophilicity without sig?nificant effects on the cells survival, cells morphology and cell cycles. It is a kind of safe polymer material.

Objective To observe the effect of intermedin(IMD) on microvascular injury of renal fibrosis in unilateral ureteral obstruction (UUO) rat model.Methods Seventy-two male Wistar rats were randomly divided into two groups:the sham-operation group (n=24) underwent the left ureteral dissection,the other 48 rats were made as unilateral ureteral obstruction models and subdivided into model group(UUO,n=24) and IMD group (n=24).At the 7,14,21,28 day after the operation,6 randomly-selected rats from each of the three groups respectively were blooded by abdominal arotic and their obstructive kidneys were taken out.The renal histopathological changes were observed through HE and Masson staining,the contents of BUN,Scr and cystatin C (CysC) of the obstructive kidneys were determined,the expressions of transforming growth factor-β1 (TGF-β1),α-SMA,bone morphogenetic protein-7 (BMP-7),E-cadherin,thrombospondin 1 (TSP-1) and vascular endothelial growth factor (VEGF) were detected by RT-PCR and immunohistochemistry.Results Compared with the sham-operated group,the pathological changes of kidney in the model group showed that the degree of fibrosis was obvious,tubular interstitial damage aggravated,the levels of BUN,Scr,CysC in the model group increased (P ＜ 0.05),the mRNA expression and protein content of TGF-β1,oα-SMA,TSP-1 increased (P ＜ 0.05),while the levels of BMP-7,E-cadherin and VEGF decreased (P ＜0.05).Compared with the UUO group,renal tubular damage,interstitial fibrosis in the IMD group were lighter,the levels of BUN,Scr,CysC in the IMD group were lower (P ＜ 0.05),the mRNA expression and protein content of TGF-β1,α-SMA,TSP-1 were down-regulated (P ＜ 0.05),while the levels of BMP-7,E-cadherin and VEGF were up-regulated (P ＜ 0.05).Conclusion IMD can ameliorate the renal interstitial fibrosis,and the mechanism may be related to the fact that VEGF mediated by IMD can reduce vascular injury.

Objective To investigate the correlation between QRS amplitudes and left ventricular wall thickness in autopsy specimens of elderly men.Methods The data of autopsy cases in our hospital since 1990 were retrospectively analyzed.The cases with QRS duration≥0.12 s and the pacing electrocardiogram were excluded.QRS amplitudes of standard 12-lead electrocardiography in 3 months before death were measured and the correlation between QRS amplitudes and left ventricular wall thickness was analyzed in the elderly men.Results Correlations were found between the amplitudes of the R waves in leads V5 ,V6, Ⅰ ,aVL[(1.1±0.7) mV, (0.95±0.6) mV, (0.44±0.3)mV and(0.35±0.3)mV] and left ventricular wall thickness[(13.6±5.4)mm;r=0.22,0.14,0.22,0.23,all P<0.05], and between the combination of QRS amplitudes SV1 +RV5 or RV6(1.9±1.2) mV] and left ventrieular wall thickness [(13.8± 5.4) mm; r = 0.23, P < 0.05].The correlationbetween the combination of QRS amplitudes (SV1 + RV5 or RV6 ) and left ventricular wall thickness was the strongest in 60-79 years old cases (r=0.48, P<0.01) ,and was decreased in 80-89 years old cases (r= 0.23, P<0.05).There was no correlation between the combination of QRS amplitudes (SV1+RV5or RV6) and left ventricular wall thickness in 90-101 years old cases (r= 0.03, P> 0.05).Conclusions Electrocardiogram is a reliable method for diagnosis of left ventricular hypertrophy in elderly men aged < 90 years.

Objective To explore the relationship between intermedin (IMD) and renal interstitial capillary loss in IgA nephropathy (IgAN) patients.Methods Renal biopsy specimens collected from primary IgAN patients in our hospital (n=80) were compared with normal renal tissues.Expressions of IMD,CD31 and VE-cadherin were examined by immunohistochemical method,and plasma concentrations of IMD and TGF-β1 in 37 cases from the 80 cases were compared.The relationship between IMD and renal interstitial capillary loss in IgAN patients was analyzed.Results IMD and VE-cadherin in renal tubule interstitium expressions increased compared to the control group at the early stage of IgAN (P ＜ 0.05).CD31 expression remained unchanged at the early stage of pathological lesions of IgAN (P ＞ 0.05),but decreased at the early stage of clinical stage of IgAN compared to the control (P ＜ 0.05).Expressions of IMD,CD31 and VE-cadherin were reducing as the disease progressed,and the correlations of CD31 and VE-cadherin (r=0.517,P ＜ 0.01),IMD and CD31(r=0.655,P ＜ 0.01) or IMD and VE-cadherin (r=0.576,P ＜ 0.01) were positive.Plasma concentrations of IMD and TGF-β1 were higher than those of the control group at the early stage of IgAN (P ＜ 0.05),and the changes of IMD and TGF-β1were correlated positively (r=0.582,P ＜ 0.01).Conclusion Compared with the control group,expression of IMD in kidney tubules increases at the early stage of IgAN,and change of IMD correlates closely with the renal interstitial capillary loss.Plasma concentrations of IMD and TGF-β1 increase compared with the control group at the early stage of IgAN,and the changes of IMD and TGF-β1 are related closely.

Objective To investigate the glomerular microvascular injury and repair in patients with IgA nephropathy (IgAN) as well as its relationship with intermedin (IMD).Methods Eighty cases of renal tissue taken from patients first diagnosed as IgAN in Shanxi Provincial People's Hospital Affiliated to Shanxi Medical University and 15 cases of normal renal tissue were detected by the expression of glomerular IMD,CD31,and VE-cadherin through immunohistochemical method.ELISA method was used to detect VEGF and IMD of plasm from 31 normal subjects and 36 cases chosen from the IgAN patients.Their changes and internal relationship were analyzed according to Lee's and chronic kidney disease (CKD) classification.Results (1) Compared with the control group the expressions of CD31,IMD,and VE-cadherin in IgAN patients were statistically significant (P ＜0.01).Compared with the control group the levels of IMD and VEGF in plasma of IgAN patients in early stage of CKD group and late stage of CKD group were statistically significant (P ＜ 0.01).(2)Correlation analysis:the expression of glomerular CD31 and Lee's classification were negatively correlated (r=-0.232,P ＜ 0.05);glomerular IMD was negatively correlated with Lee's classification (r=-0.241,P＜0.05),while positively correlated with glomerular VE-cadherin (r=0.417,P＜ 0.01).VEGF in plasma of IgAN patients was positive correlated with CKD classification,BUN (r=0.458,0.409,P＜0.05),and negatively correlated with serum ALB (r=-0.532,P＜0.01).Conclusion Microvascular injury exists in patients with IgAN.The expression of VE-cadherin and IMD are positively correlated,suggesting that IMD may be involved in the progression of vascular protection and angiogenesis in IgAN.The contents of IMD and VEGF in plasma of IgAN patients increase,indicating that they may play a role in the progression of IgAN.

The comparative analysis of the biological characterization and the genetic background study of EV71 circulating strains is commonly recognized as basic work necessary for development of an effective EV71 vaccine. In this study, we sequenced five EV71 circulating strains, isolated from Fuyang, Hefei, Kunming and Shenzhen city of China and named them FY-23, FY-22, H44, K9 and S1 respectively. The sequence alignment demonstrated their genotypes be C4. The genetic distance of the VP1 gene from these isolates suggested that they were highly co-related with genetic identity similar to other previously reported EV71 strains in China. Additionally, these strains were identified to display some obvious proliferation dynamics and plaque morphology when propagated in Vero cells. However, a distinctive difference in pathogenic ability in neonatal mice was found. Some differences in cross neutralization test & immunogenic analysis were also found. All these results are related to the biological characterization of circulating EV71 strains in China and aid in the development of an EV71 vaccine in the future.

Objective:To compare the efficacy, safety and total clinical benefit between urokinase and reteplase in catheter directed thrombolysis(CDT) of deep venous thrombosis(DVT) patients.Methods:The clinical data of 88 patients receiving IVC filter insertion and CDT in iliofemoral veins was retrospectively analyzed. Fifty-two patients received urokinase(UK), 36 received reteplase(rPA). The efficacy of thrombolysis were evaluated by thrombolytic rate , the change of limb circumferences. All the patients were followed up at the 3rd、6th and 12th month , Villalta score was used to estimate the severity of postthrombotic syndrome(PTS).Results:The thrombolytic rate in the UK group was 75.5% and 84.2% in the rPA( P<0.001); the change of limb circumferences in the UK group was 84.0%, vs. 90.0% in the rPA group( P<0.001); the Villalta score at the 3rd、6th and 12th month in the UK group were 3.03、3.63、4.57 respectively, while in the rPA group were 2.29、3.06、3.70 respectively( P<0.001) . The thrombolytic time and length of hospital stay in the rPA group is shorter than that in the UK′s. Conclusions:The thrombolytic efficacy of rPA is superior to UK. In addition, rPA can reduce the severity of PTS when compared with UK.

Objective To explore the relationship between the level of plasma ADAMTS4 and the vulnerability of carotid atherosclerotic plaque in patients with carotid artery stenosis and the clinical value of evaluating the vulnerability of the carotid atherosclerotic plaque.Methods 60 patients with carotid artery stenosis were divided into stable plaque group and vulnerable plaque group according to the histopathological typing after carotid endarterectomy (CEA).30 patients who had atherosclerosis but no carotid plaque formation served as control.Using ELISA to measure the level of plasma ADAMTS4.Results The level of plasma ADAMTS4 was significantly higher in the vulnerable-plaque group (112.74 ± 17.47) ng/ml compared with the stable plaque group (56.67 ± 22.14)ng/ml and the control group (56.67 ± 22.14)ng/ ml,P ＜ 0.001.Logistic analysis showed that high level of plasma ADAMTS4 was risk factor for vulnerability of carotid atherosclerotic plaque (P ＜ 0.05,OR =8.240,95 ％ CI:1.512-22.915).ROC curve showed that ADAMTS4 =100.935 ng/ml could accurately predict significant vulnerability with accuracy of 94.3％.Conclusion ADAMTS4 is an effective,sensitive and non-invasive biomarker to evaluate the vulnerability of carotid atherosclerotic plaque.