BACKGROUND: Studies confirm that ischemia/reperfusion (I/R) injury can induce myocardial apoptosis. The loss of mitochondrial membrane potential (MMP) after reperfusion is the inevitable pathway of apoptosis. Protection of MMP may reduce apoptosis.OBJECTIVE: To observe the effect of naloxone on MMP of hypoxic myocardial cells and apoptosis in neonatal rats, and investigate the protective effect of naloxone on hypoxic myocardial cells.DESIGN: Observation and controlled trial.SETTING: Department of Cardiology, the General Hospital of Chinese PLA.MATERIALS: Detection of apoptosis of myocardial cells was carried out in the Laboratory of Pathophysiology, General Hospital of Chinese PLA in December 2004. Ten neonatal rats were used, and detection of MMP of myocardial cells was carried out in the same laboratory in March 2006 and 20 rats were used. All the involved rats were provided by the Animal Center of the General Hospital of Chinese PLA on the day of birth, were involved in this trial. Reagents: Naloxone hydrochloride injection (0.4 g/L, Beijing Sihuan Pharmaceutical Factory, Batch No. 0206272); the lowest and essential medium (Dulbecco, DEME,GIBCO company); phosphate buffer solution and fetal bovine serum (PBS and FBS, SIGMA Company).METHODS: The neonate rats were cut open along the median line of chest bone after local sterilization with iodine tincture on the day of birth. 1/3 ventricular myocardium before cardiac apex was harvested. On the 4th day of the culture,culture flasks of cells in good growth status ( ＞ 106 cells/bottle) were selected and divided into 3 groups: control group (normal culture, n =3 bottles), hypoxia group (hypoxia/reoxygenation, n =15 bottles) and naloxone group (hypoxia/reoxygenation, and treated by naloxone, n =15 bottles). Three time points were set in hypoxia group and naloxone group according to different time of hypoxia and reoxygenation: hypoxia 2 hours/reoxygenation 0 hour; hypoxia 2 hours/reoxygenation 2 hours; hypoxia 2 hours/reoxygenation 4 hours, 5 bottles at each time point. In the hypoxia group, DEME medium, which was pre-filled with 0.95 volume fraction of N2 and 0.05 volume fraction of CO2, containing 0.01 volume fraction of FBS, was used and 0.95 volume fraction of N2 and 0.05 volume fraction of CO2 was also filled to replace the air in the culture flasks. The culture flasks were enveloped for incubation at 37 ℃. The cells in the hypoxia group were incubated at normal condition (0.95 volume fraction of air and 0.05 volume fraction of CO2) at set time. In the naxolone group, hypoxia/reoxygenation treatment was the same as above, and naloxone hydrochloride was added at the sametime and the final concentration of naloxone in culture flasks was 5 μmol/L (The volume for naloxone ≤ 0.5% of the total medium). In the control group, hypoxia/reoxygenation and naloxone treatment were not given, but the same volume of normal saline was added, and this time served as time point of hypoxia 0 hour/reoxygenation 0 hour. After intervention,myocardial MMP changes and apoptosis were detected with fluorescent staining-flow cytometer at different time after hypoxia/reoxygenation.MAIN OUTCOME MEASURES: ① MMP changes of hypoxia group and naloxone group at different time points; ② Comparison of survival, apoptosis and necrosis of cells at hypoxia 2 hours/reoxygenation 4 hours in each group.RESULTS: ① After hypoxia, MMPs of the cells in hypoxia group and naloxone group were decreased. MMP of the cells in the naloxone group was higher than that in the hypoxia group at each time point (P ＜ 0.01). The great amplitude of decrease of MMP occurred in hypoxia period, but not in reoxygenation period. ② At hypoxia 2 hours/reoxygenation 4 hours, the apoptotic and necrotic rates in the hypoxia group were significantly higher than those in the naloxone group [(9.88±0.98)% vs. (2.41±0.52)%; (5.10±0.29)% vs. (3.56±0.56)%, both P ＜ 0.01]. The apoptotic rate was significantly higher than the necrotic rate in the hypoxia group (P ＜ 0.05).CONCLUSION: Early application of naloxone can significantly alleviate and postpone the decrease of myocardial MMP after I/R, and reduce apoptosis and necrosis.

Biocontrol bacteria 308R(pKSH) with hrpN of Erwinia amyloyora. It can secrete the Harpin protein which can induce plant resistance. After successive growth in antibiotic-free LB medium for 50 generations, only 23.1% of the cell still retained the plasmid pKSH, in comparison with 4.75% of the cosmid pCPP430. Sprayed onto the leaves of tomato, the recombinant strain maintained a population density of over 10~ 5 cfu/cm~ 2 when kept under high humidity in thirteen days, but if without humidity, the strain keep 10~ 4 cfu/cm~ 2 over in five days. On the tomato leaves, the stability of the recombinant strain 308R(pKSH) was higher than the strain 308R(pCPP430). The experiment indicated that the recombinant strain 308R(pKSH) had higher stability compared with the strain 308R(pCPP430), but that was not enough. In this paper reasons for unstability and strategies for improving were discussed for the recombinant strain.

AIM: To observe the effect of ischemic preconditioning (IPC) on the subsequent permanent ischemic rat retina. METHODS: A model of two-vessel occlusion (2VO) was used to give an ischemic insult to rat retina. Bilateral carotid arteries were occluded directly in the simple ischemic groups. A procedure of double 2 min ischemia- 3 min reperfusion was applied to IPC groups before their carotid arteries were occluded. Experimental control groups received the same operation except that their exposed vessels were not occluded. Other 6 normal rats served as immunohistochemical controls. Eyeballs were taken out after being subjected to 1, 3 and 7 days of ischemia. The morphometry of retina was measured by stereologic methods. The apoptosis during the retinal ischemia was detected by TUNEL. Immunohistochemistry staining was used for the localization of bcl-2 in retina. RESULTS: In IPC groups, the thickness of retina was thinner than that in simple ischemic groups. The apoptosis was less when compared with the simple ischemic groups at the same ischemic time. The apoptotic cells show only in INL and the apoptosis of RGCs was not appeared until 7 days postischemia. There was no significant difference of the numerical density of RGCs at the different time after the operation. The bcl-2 immunoreactivity was weaker than that of the simple ischemic groups too. CONCLUSION: IPC reduced the injury caused by ischemia in retina, showing a protective effect.

Objective To systematically the efficacy of transcatheter arterial chemoembolization (TACE) conbined with percutaneous microwave coagulation therapy (PMCT) in treatment of advanced liver cancer.Methods A search was performed by retrieving the domestic and foreign literature database,including WanFang Data,VIP,CNKI,PubMed,Cochrane Library,CBM,EMBASE,Medline,between January 2005 and May 2015.These documents were about the analysis of the efficacy of TACE combined with PMCT in treatment of advanced liver cancer,including complete response(CR),partial response(PR),total effective rate,the levels of AFP declining,1,2,and 3 year survival rate and all the trials must be randomized controlled trials.Meta-analyses were conducted using the Cochrane Collaboration's RevMan 5.2 software.Results Fourteen documents were retrieved,including 989 patients,conbined treatment group 470 patients,simple treatment group 519 patients.The results of Mete-analysis shows that the total effective rate of TACE combined with PMCT is higher than TACE alone in treatment of advanced liver cancer.AFP declining ＞ 50％ of TACE combined with PMCT more obvious than TACE alone.1,2,and 3 year survival rate of TACE combincd with PMCT higher than TACE alone.These differences were statistically significant.Conclusion TACE combined with PMCT might be more effective than TACE alone in treatment of advanced liver cancer.

Objective: Our aim was to observe the protective effect of propofol in clinical relevant concentration on the peroxidative injured erythrocyte. Methods: Intravenous blood samples taken from 20 healthy adults were prepared for red blood cell (RBC) suspensions and divided equally into 5 groups: groupⅠfor control, group Ⅱ with hydrogen peroxide (H2O2, 100 mmol/L) -induced injury, and group Ⅲ, Ⅳ, Ⅴ with the same injury as the group Ⅱ but being pretreated with 3 different concentrations of propofol (25, 50, 75 μmol/L), respectively. The concentrations of potassium and malondialdehyde (MDA) in RBC suspensions and hemolytic degree after incubation were measured. Results: After 60-minute incubation, the extracellular potassium concentrations (0.16, 0.14, 0.14 mmol/L), MDA concentrations (5.66, 5.57, 6.20 nmol/L), and hemolytic degree (76.89%, 59.84%, 64.22%) decreased significantly in the groups that were pretreated with propofol as compared with the group Ⅱ (0.26 mmol/L, 9.19 nmol/L, and 100%), but no difference has been seen within the groups pretreated with 3 different concentrations of propofol and between the propofol-treated groups and the group Ⅰ(0.10 mmol/L, 4.13 nmol/L, 52.73%). Conclusion: Propofol in clinical relevant concentrations may decrease MDA production, hemolytic degree, and potassium exflux from erythrocyte in response to in vitro oxidative challenge with hydrogen peroxide and enhance erythrocyte antioxidant capacity. The protective effect is not related with concentrations.

ObjectiveTo evaluate the impacts of pereutaneous nephrolithotomy (PCNL) on perioperative renal hemodynamics.MethodsThe hemodynamics of operated renal arteries of 30 patients who underwent unilateral PCNL with single pole access were observed 1 day before and 5-7 days after operation with CDFI.Parameters were analyzed statistically.ResultsAfter operation,resistance index (RI) of renal arteries decreased (P＜0.05).The diastolic flow statistically increased at main renal artery (MRA) of renal hilus,interlobar renal artery and interlobular renal artery (all P＜0.05).After PCNL,in serious hydronephrosis patients,RI decreased (P＜0.05) at segmental renal artery (SRA) and interlobar artery,end-diastolic flow velocity (Vmin) increased at interlobar renal artery (P＜0.05).In moderate hydronephrosis patients,RI decreased at all renal arteries (P＜0.05) after PCNL,Vmin increased at MRA and interlobular renal artery (P＜0.05).In minor hydronephrosis patients,RI decreased at MRA and SRA,Vmin increased at SRA.In patients without hydronephrosis,RI changeed like serious hydronephrosis patients.ConclusionAfter PCNL,ipsilateral renal perfusion improves,renal diastolic flow increases and RI decreases.CDFI can be used to observe the blood perfusion of kidney,and provide quantitative information of renal hemodynamics.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; genetics ; Female ; Genetic Predisposition to Disease ; Humans ; Lung Neoplasms ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Receptor, EphB4 ; genetics

Angioplasty, Balloon ; methods ; Aortic Coarctation ; diagnostic imaging ; pathology ; therapy ; Aortography ; Cardiac Catheterization ; methods ; Child ; Humans ; Male ; Platinum ; Prosthesis Design ; Stents ; Tomography, X-Ray Computed ; Treatment Outcome

Sequences analysis revealed Grass carp reovirus (GCRV) s10 was 909 nucleotides coding a 34 kDa protein denoted as VP7, which was determined to be a viral outer capsid protein (OCP). To obtain expressed OCP in vitro, a full length VP7 gene was produced by RT-PCR amplification, and the amplified fragment was cloned into T7 promoted prokaryotic expression vector pRSET. The recombinant plasmid,which was named as pR/GCRV-VP7,was then transformed into E.coli BL21 host cells. The data indicated that the expressed recombinant was in frame with the N-terminal fusion peptide. The over-expressed fusion protein was produced by inducing with IPTG, and its molecular weight was about 37kDa, which was consistent with its predicted size. In addition, the fusion protein was produced in the form of the inclusion body with their yield remaining steady at more than 60% of total bacterial protein. Moreover,the expressed protein was able to bind immunologically to anti-his-tag monoclonal antibody (mouse) and anti-GCRV serum (rabbit). This work provides a research basis for further structure and function studies of GCRV during entry into cells.

Angioplasty, Balloon ; methods ; Aortic Valve Stenosis ; therapy ; Cardiac Pacing, Artificial ; methods ; Child, Preschool ; Heart Ventricles ; Humans ; Infant ; Male