Objectives To explore the clinical effect of coupling meglumine cyclic adenylate (MCA)and the human granulocyte colony-ostimulating factor (G-CSF)on rat with diastolic heart failure(DHF).Methods Totally 60 rats of DHF model were evenly divided into 4 groups according to random number:Control group(n=15,control),Model group(n=15,DHF model),MCA group(n =15,treated with MCA)and MCA+GCSF group(n=15,treated with MCA plus G-CSF).MCA group were administered by intragastric injection of MCA 30 mg/kg/d for 15 d,MCA+G-CSF group were administered by intragastric injection of MCA 30 mg/kg/d and plus G-CSF 100 μg/kg/d for 15 d,while Control group and Model group were given same volume of saline solution.BIOPAC SYSTEM was used to analyze the model establishment.The mRNA levels of GATA-4 and Cx43 were measured by RT-PCR.The protein expressions of GATA-4,Cx43,cTNI and c-kit were measured with western blotting.ELISA and flow cytometry were used to detect cAMP and differentiation rate of bone marrow mesenchymal stem cells (BMSCs),respectively.Results Compared with MCA group,the denaturation degree of myocardial tissues in DHF rat was significantly improved than in MCA+G-CSF group.Moreover,the level of GATA-4 (1.62 ± 0.09),Cx43 (1.02 ± 0.07),cTNI (1.42 ± 0.12),c-kit (0.65±0.02),cAMP(283.67± 18.09)nmol/L and BMSCs cell differentiation rate(38.62 ± 1.52)％ in MCA + GCSF group were significantly promoted (all P＜ 0.05)than in MCA group,GATA-4 (0.82±0.07),Cx43 (0.52±0.05),cTNI(0.86 ± 0.13),c-kit (0.48 ± 0.03),cAMP(198.83 ± 16.03) nmol/L and BMSCs cell differentiation rate (19.82 ± 0.89)％.Conclusions The combination of MCA with G-CSF is significantly improved DHF than single MAC treatment,which may regulate BMSCs differentiation though cAMP/PKA signaling pathways.

Objective To construct the eukaryotic expression vector for human hypoxia inducible factor-1? gene (pSNAV-HIF-1?), and to investigate the apoptosis and intracellular calcium concentration of the transfected A?25-35 induced hippocampal neurons of primary culture. Methods Human hypoxia inducible factor-1? gene from pBSKhHIF1?T7 was inserted into pSNAV2.0 by the method of gene recombination, then the constructed vector was transfected into hippocampal neurons of primary culture and followed by A?25-35 for treatment. Empty pSNAV2.0 vector was treated the same way as pSNAV-HIF-1? as a control. The expression level of HIF-1? protein was assayed by Western blotting. Apoptosis was detected by flow cytometry. Intracellular calcium concentration was determined by laser scanning confocal microscopy with Fluo-3/AM as the fluorescent dye. Results It was shown that pSNAV-HIF-1? was successfully constructed by restriction enzyme digestion, PCR and DNA sequencing. The expression level of HIF-1? protein was significantly increased in transfected hippocampal neurons of primary culture (P

Objective To observe pre-syncopal limited tolerance and cardiovascular responses to head-up tilt combined with lower body negative pressure (HUT/LBNP) following exposure to head-down tilt (HDT, -1 Gz). Method Exposures to HUT/LBNP (-60 mmHg) in control session (without preceding 30 s -1 Gz treatment) and in simulated push-pull effect (PPE) session (with preceding 30 s -1 Gz treatment) were performed in 8 healthy adults. The changes of hemodynamic parameters were monitored by electrical impedance instrument during the experiments. Result The mean endurance time in presyncopal symptom limited HUT/LBNP in control session and in simulated PPE session were 8.4±2.1 min and 4.5±2.4 min, respectively, the two means were significantly different (P< 0.01). In simulated PPE session, as compared with baseline, heart rate (HR) during HDT was significantly lowered (P<0.01), while stroke volume (SV) and cardiac output (CO) were increased significantly (P<0.01). During HUT/LBNP, the increased percentage (relative to baseline) of HR in PPE session was lower than these in control session (P<0.05); the decreased percentages of SV and CO during HUT/LBNP in PPE session were both higher than those in control session (P<0.05). During HUT/LBNP, arterial pulse pressure (PP) of control session was significantly decreased than the value of baseline value (P<0.05); Total peripheral resistance (TPR) of PPE session was significantly increased than baseline value (P<0.05). Conclusion Tolerance time before the appearance of presyncopal symptoms during HUT/LBNP decreases and cardiovascular responses to HUT/LBNP are impaired, preceding exposure to HDT.

Objective: To establish an analytical method for serum prolactin (PRL) based on the photoinduced chemiluminescence technology, and evaluate its performance. Methods: A pair of PRL monoclonal antibodies were labeled with luminescent nanospheres and biotin respectively, and the double antibody sandwich detection system was formed with the serum prolactin and streptavidin-labeled photosensitive microspheres (universal photosensitive solution) under homogeneous conditions. The performance index and correlation of the detection system were evaluated. Results: The precision of intra-assay and within-day (coefficient of variation) of the developed assay were 4.60% and 5.25%, respectively. The functional sensitivity was 2.48 μIU/mL, and its reportable results were ranged from 2.48 to 4 240 μIU/mL. The recovery rates of different PRL calibrators (42.2, 424, 4 240 μIU/mL) added to human sera were ranged from 96.25% to 102.93%. There was no interference from bilirubin＜20 mg/dL, hemoglobin＜200 mg/dL, triglyceride＜3 000 mg/dL and biotin＜20 ng/mL. Also, the light-initiated chemiluminescent assay for PRL (PRL-LICA) correlated well with Beckman Unicel Dxi 800 Access 2. Conclusions LICA showed effective performance for detecting PRL in human serum, and it could meet the basic requirements of clinical diagnosis.

The study is to investigate the effect of angiotensin II (Ang II) and its receptor blockers on migration and endothelin-1 (ET-1) expression of rat vascular adventitial fibroblast subpopulations. Vascular adventitial fibroblasts were individually expanded by using cloning rings, and the effects of Ang II on the migration of adventitial fibroblast subpopulations were evaluated by Transwell. Fluorescence quantitative-PCR detected the expression of preproET-1 mRNA induced by Ang II, and its receptor antagonists losartan and PD-123319. The concentration of ET-1 was determined by ELISA. It showed that spindle shaped and epithelioid shaped cells were isolated by using cloning rings, named as spindle cells and round cells. RT-PCR showed that fibroblast subpopulations did not have leukocytes, endothelial cells and smooth muscle cells, namely pure cell lines. Compared with respective control cells, two subpopulations had transferring ability. Ang II significantly improved round cells migration in a concentration-dependent manner, and had no obvious influence on spindle cells migration. Ang II (1 x 10(-8) - 1 x 10(-6) mol x L(-1)) significantly increased the expression of preproET-1 mRNA in round cells (P < 0.01), and had no significant effect on the expression of preproET-1 mRNA in spindle cells. Losartan blocked the expression of preproET-1 mRNA induced by Ang II in round cells, and had no significant effect on the expression of preproET-1 mRNA in spindle cells. The effects of Ang II and ET-1 receptor inhibitors on the release of ET-1 were similar to the expression of preproET-1 mRNA. The results indicate that there are two cell subpopulations: round cells and spindle cells in rat vascular adventitial fibroblasts. Ang II significantly improved cells migration, and increased the expression of ET-1 in round cell subpopulation. It suggested that there may be different migratory mechanisms in two cell subpopulations, and the two subpopulations may play a different role in vascular remodeling and reparative process.

Objective To explore factors influencing the prognosis of hepatocellular carcinoma (HCC) patients after hepatectomy. Methods From May 1994 to January 1998, 189 patients who underwent hepatectomy for HCC were enrolled for reviewing their clinical characteristics, treatment and prognosis. Twenty-two parameters contributing to long-term survival rate (SR) and disease-free SR were analysed. Results The 3,5-year cumulative SR of the whole group was 63%,45%, respectively. The 3,5-year SR and disease-free SR in the curative resection (CR) group ( n =162) were 67%,47% and 45%,26% respectively. Results showed that the way by which a tumor was found, tumor size, portal tumor thrombi, satellite nodule, TNM stage, cirrhosis type, recurrent and treatment, blood transfusion, differentiation grade,and CR were risk factors by individual variable analysis( P =0.0000～0.0034); A multivariable analysis showed that CR, tumor size, tumor finding mode and reoperation were significant factors associating with prognosis( P =0.0000～0.0024). Blood transfusion and type of cirrhosis were closely correlated with tumor-free survival ( P =0.0001). Conclusions Curative resection, tumor size, reoperation for recurrence were important factors for recurrence by multivariate analysis. The severity of concomittant liver cirrhosis and perioperative blood transfusion were closely correlated with postoperative tumor free survival.

Ventricular myocytes from hearts of the neonatal SD rats were treated with 10-7,10-6,and 10-5 mol/L simvastatin for 72 hours under high glucose condition. Compared with control group,the viability of cadiomyocyte was significantly lower in high glucose group (P＜0.01 ).The activity of lactate dehydrogenase,the relative expressions of NADPH oxidase subunits p22phox,p47phox mRNA,and reactive oxygen species level in the high glucose group were higher than those of control group ( all P＜0.05).lndexes mentioned above were significantly improved by simvastatin treatment in a dose-dependent manner.These results suggest that simvastatin ameliorates high glucose-induced injury of cardiomyocytes via increasing the expression of NADPH oxidase mRNA.

Objective To prepare long-circulating temperature-sensitive liposomes with paclitaxel( LTSLP ),develop methods for determination of paclitaxel,related substances and monostearoyl phosphatidylcholine( MSPC ) in LTSLP,and the haemolysis of LTSLP in vitro. Methods HPLC-UV methods for paclitaxel content and related substances and HPLC-CAD method for MSPC in LTSLP were established and validated. Spectrophotometric method was used to determine hemolysis in vitro. Results There was a good linear relationship between peak area and concentration within the range of 60. 39-181. 17μg·mL-1 . Recovery and precision of the method for determination of paclitaxel content met the requirements. Specificity,sensitivity,and system suitability for related substances were consistent with requirements. There was a good linear relationship between peak area and concentration within the range of 1. 5-50. 0μg·mL-1 for the determination of MSPC with good specificity,sensitivity and recovery. Paclitaxel contents in three batches of self-prepared LTSLP were between 90. 0% and 110. 0%,single related substances were below 0. 5% and total impurities were below 2. 0%. There was almost no hemolysis in vitro. Conclusion The methods for determining paclitaxel content,related substances and haemolysis can be used to assess the quality of LTSLP. Self-produced LTSLP consistently meet the quality standards.

Objective To analyze the antibiotic resistance of the carbapenem no-susceptibility Enterobacteriaceae isolated from pediatric patients and the resistant genes of carbapenemase-producing.Methods In all,46 carbapenem no-susceptibility Enterobacteriaceae strains were isolated from patients at Beijing Children's Hospital between January 2008 and December 2010.Agar dilution method recommended by the Clinical and Laboratory Standards Institute was used to examine the minimum inhibitory concentrations (MICs) of 14 antimicrobial agents.Phenotypic testing for carbapenemase-producing was conducted using Hodge test and double-disk synergy test.PCR was used to detect the expression of the carbapenemase-related genes KPC,GES,IMI/NMC-A,SME,IMP,VIM,GIM,SPM,SIM and OXA.WHONET5.6 was used to perform resistance analysis.Results Among 46 carbapenem no-susceptibility Enterobacteriaceae strains,26 (56.5％) were Klebsiella pneumoniae strains,13(28.3％ ) were Enterobacter cloacae and 7( 15.2％ ) were Escherichia coli.The rates of imipenem and meropenem no-susceptibility Klebsiella pneumoniae were 69.2％ and 80.8％,Enterobacter cloacae were 76.9％ and 100％ and Escherichia coli were 85.7％ and 100％,respectively.40(87.0％ ) strains were positive of Hodge test.41 (89.1％) strains were positive of doubledisk synergy test.38 (82.6％) were positive for the IMP genotype.The carbapenemase-related genes were not found in other 8 strains.Conclusion The prevalence of carbapenem no-susceptibility Enterobacteriaceae strains in Klebsiella pneumoniae isolates is relatively high in children.Resistance to imipenem was lower than that to meropenem from Klebsiella pneumoniae,Enterobacter cloacae and Escherichia coli strains.Many carbapenem no-susceptibility Enterobacteriaceae isolated from pediatric patients carry the blaIMP gene.No the KPC gene was found.

Objective To investigate changes of learning ability and somatostatin (SS) changes after positive acceleration (+Gz) exposures. Mehtod Eighty male SD rats were randomly divided into 3groups: control group(Con), +6 Gz/3 min group ( +6 Gz), and +10 Gz/3 min group ( +10 Gz),8 rats in each group. Changes of learning ability in rats were observed at 0 d, 2 d, 4 d and 6 d after + Gz exposure. SS in hippocampus was measured by RIA at 0 d, 2 d and 4 d after + Gz exposures ( there were 8 rats every time, in each group). Result In Y-maze test,number of correct response decreased significantly (P ＜0.01 ), and total reaction time increased significantly(P ＜0.01 ) in +6Gz and +10 Gz groups as compared with control group; number of correct response and total reaction time in +10 Gz group changed significantly at 0 d(P ＜0.01 or P ＜0.05) as compared with +6Gz group. RIA showed that, content of SS in hippocampus declined at 0 d and 2 d(P ＜0.05 or P ＜0.01) in +6 Gz and + 10 Gz groups as compared with control group. Conclusion + Gz exposure could impair learning ability of rats, and inhibit expression of SS in hippocampus.