Aim To investigate the protective effects of nicorandil, glutamine and metoprolol used in single and combination on the antiapoptosis ability of cardio-cytes and the expression of HSP70 after myocardial is-chemia/reperfusion injury in rats. Methods Male Wistar rats were divided randomly into seven groups:( 1 ) Sham group:in which the coronary artery was not roped;(2) I/R group:in which only 30 min ischemia and 120 min reperfusion of LAD was executed; ( 3 ) Nicorandil group; ( 4 ) Glutamine group; ( 5 ) Meto-prolol group; ( 6 ) Nic + Glu + Met ( NGM ) group;(7) low dose of Nic+Glu+Met ( NGML) group. In the pharmacological precondition groups, correspond-ing drugs were administered 30 min before I/R proto-col, and the initial drug dose in each group was 1 mg ·kg-1 . Myocardial apoptotic rates were measured with TUNEL ( terminal dexynucleotidyl transferase-mediated dUTP nick end labeling) , and the expression of apop-tosis related proteins-Bcl-2/Bax and protective pro-tein-HSP70 was detected by Western blot. ResultsIn I/R group, the apoptotic rate in ischemic region was very high ( 36.9% ± 10.3%) , and Bcl-2/Bax was very low ( 0.14 ±0.08 ) . Compared with I/R group, the apoptotic rate of pharmacological precondition groups was decreased significantly ( P <0.01 ) , and the ratio of Bcl-2 to Bax was raised ( P<0.01 ) . Com-pared with Sham group, the expression of HSP70 in-creased significantly ( P<0.01 ) in I/R group. Com-pared with IR group, the expression of HSP70 of phar-macological precondition groups was elevated ( P <0.01 ) , and the expression in NGM group was higher than in the single drug group ( P<0.01 ) . Conclusion Compared with single therapy after I/R in rat, the combination therapy of nicorandil, glutamine, meto-prolol can decrease the apoptotic rate, and the expres-sion of apoptosis related proteins is developed. In the mean time, the expression of protective protein HSP70 is elevated.

Objective To investigate the effects of pulsed electromagnetic fields(PEMFs)on proliferation and differentiation of endothelial progenitor cells(EPCs).Methods EPCs were isolated from rat bone marrow by density gradient centrifugation.EPCs were exposed to PEMFs from the 5th day to the end of culture.MTT was used to measure the proliferation of EPCs.The expression ofⅧ-related antigen and NOS_3 was evaluated by flow cytometry. Results Compared with the control,the proliferating ability of EPCs exposed to PEMFs was stronger;the number ofⅧ-related antigen and NOS_3 positive cells increased significantly in EPCs exposed in PEMFs.Conclusion PEMFs promotes the proliferation and differentiation of rat bone marrow EPCs.

AIM: To investigate the effect of Z,E-butylidedephthalide (Bdph) on laser-induced experimental choroidal neovascularization (CNV) in rat model and choroid blood flow in rabbits'eyes.METHODS: Male Brown Norway rats were treated with Nd:YAG laser to break Bruch's membrane. Thirty mg/kg and 15 mg/kg Bdph were given daily through intraperitoneal injection for 4 weeks after laser treatment. Fluorescein angiography (FA) and choroidal flat mount were used to measure the development of CNV. Female New Zealand white rabbits' eyes were instilled with 10g/L Z,E-butylidenephthalide solution,and ocular blood flow was measured with colored microsphere technique. RESULTS: The intensity of fluorescein leakage, indicating the ocular lesion, decreased significantly in group Bdph 30mg/kg and 15mg/kg, as compared to the control at P＜0.01.The area of neovascularization checked by FA in both groups of Bdph, at 30mg/kg and 15mg/kg decreased significantly compared to the control group at P＜0.05. On the choroid flat mount, the areas of CNV were also smaller in both Bdph groups than in control group. One percent Z,E-butylidenephthalide solution instilled into rabbits' eyes could improve the choroid blood flow at 30 and 60 minutes after drug instillation (P＜0.05).CONCLUSION: Z,E-butylidedephthalide could inhibit the development of CNV in the rat eyes and increase the choroid blood flow in the rabbit eyes. These results suggest that Z,E-butylidedephthalide may be a good agent for the treatment of age-related macular degeneration(ARMD).

Objective To analysis the etiology and molecular classification of brucella strains isolated in Guangdong province in 2010.Methods The strains of 19 brucella were verified and identified by some methods including traditional biology phenotype confirmation,PCR amplification and pulsed field gel electrophoresis (PFGE).Results On phenotype level,4 strains were brucella melitensis biovar 1,2 strains were brucella suis biovar 3,and the rest were brucella melitensis biovar 3,which were specific B genes positive strains,and the PFGE typing similar values ranging from 67.9％ to 100％.In addition to the four strains from Zhuhai for the outbreak,the homology was 100％,and the rest were sporadic cases.Conclusions Brucella cases,in Guangdong province,are highly sporadic and dispersed outbreaks.Compared with a few years ago,it shows species diversification,and brucella melitensis biovar 3 is still the dominant serotype.PFGE can be used to distinguish the three species of brucella,but it can't effectively distinguish the allotypes.

OBJECTIVETo compare the effects of perindopril and enalapril on the development of atherosclerotic lesions in ApoE knockout mice.

METHODSApoE knockout mice were treated with perindoprilor (1.5 mg.kg(-1).d(-1), n = 20), enalapril (7.5 mg.kg(-1).d(-1), n = 20) or saline (0.2 ml saline/d, n = 20) per gavage for 20 weeks. Blood pressure and lipids were measured at the study end. Aortic root atherosclerotic plaque was then quantified and the content of collagen and the size of lipid core in the plaque assessed. Cryostat sections were used to quantify the expressions of monocyte/macrophage-2 (MOMA-2), intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and matrix metalloproteinases-9 (MMP-9) in the plaque by immunofluorescence method.

RESULTSBlood pressure and lipid profiles were similar among different groups. Compared with control group, the plaque areas of perindopril group and enalapril group displayed significantly decrease (25.33% and 22.86%, respectively, both P < 0.01). However, no significant different were observed in the plaque size between the different ACE inhibitors groups. Perindopril group and enalapril group also significantly decreased the size of lipid core (52.98% and 38.98%, respectively, both P < 0.01) and the expression of MOMA-2 (88.38% and 52.16%, respectively, both P < 0.01), ICAM-1 (80.87% and 49.59%, respectively, both P < 0.01), VCAM-1 (77.56% and 56.44%, respectively, both P < 0.01) and MMP-9 (86.93% and 55.56%, respectively, both P < 0.01), and increased the plaque collagen content (298.36% and 168.14%, respectively, both P < 0.01) and the effects of perindopril was superior to those of enalapril (all P < 0.05).

CONCLUSIONSACE inhibitors significantly suppressed tissue inflammation and attenuated the development of atherosclerosis in ApoE knockout mice independent of their effects on the lipid and blood pressure. Perindopril is superior to enalapril in stabilizing the plaques and has similar effect on reducing the plaque size as that of enalapril.

Angiotensin-Converting Enzyme Inhibitors ; therapeutic use ; Animals ; Apolipoproteins E ; genetics ; Atherosclerosis ; drug therapy ; pathology ; Collagen ; metabolism ; Enalapril ; therapeutic use ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Perindopril ; therapeutic use

AIMTo investigate the possible role of manganese in the regulation of mitochondrial aconitase (mACON) activity human prostate carcinoma cell line PC-3 cells.

METHODSThe mACON enzymatic activities of human prostate carcinoma cell line PC-3 cells were determined using a reduced nicotinamide adenine dinucleotide-coupled assay. Immunoblot and transient gene expression assays were used to study gene expression of the mACON. The putative response element for gene expression was identified using reporter assays with site-directed mutagenesis and electrophoretic mobility-shift assays.

RESULTSIn vitro study revealed that manganese chloride (MnCl2) treatment for 16 h inhibited the enzymatic activity of mACON, which induced the inhibition of citrate utility and cell proliferation of PC-3 cells. Although results from transient gene expression assays showed that MnCl2 treatment upregulated gene translation by approximately 5-fold through the iron response element pathway, immunoblot and reporter assays showed that MnCl2 treatments inhibited protein and gene expression of mACON. This effect was reversed by co-treatment with ferric ammonium citrate. Additional reporter assays with site-directed mutagenesis and electrophoretic mobility-shift assays suggested that a putative metal response element in the promoter of the mACON gene was involved in the regulation of MnCl2 on the gene expression of mACON.

CONCLUSIONThese findings suggest that manganese acts as an antagonist of iron, disrupting the enzymatic activity and gene expression of mACON and citrate metabolism in the prostate.

Aconitate Hydratase ; antagonists & inhibitors ; genetics ; Actins ; genetics ; Adenosine Triphosphate ; metabolism ; Cell Line, Tumor ; Chlorides ; pharmacology ; Citrates ; metabolism ; DNA Primers ; Gene Expression Regulation, Enzymologic ; drug effects ; Gene Expression Regulation, Neoplastic ; drug effects ; Genes, Reporter ; Humans ; Iron ; metabolism ; Male ; Manganese Compounds ; pharmacology ; Mitochondria ; enzymology ; Prostatic Neoplasms ; enzymology

OBJECTIVETo observe the pathological changes and morphological alterations of ECV304 cells after the infection by herpes simplex virus type 2 (HSV-2) in vitro.

METHODSPassaged ECV304 cells were infected with HSV-2, TCID50 and morphological changes were observed by optical microscopy and tissue staining.

RESULTSOne day after HSV-2 infection, swelling, rounding, and increase of thickened cytoplasmic granules occurred in the ECV304 cells, and on day 2, cell fusion was observed with weakened nuclear staining.

CONCLUSIONECV304 cells mostly undergo necrosis after HSV-2 infection without obvious evidence of cell apoptosis.

Cells, Cultured ; Endothelium, Vascular ; pathology ; virology ; Herpesviridae Infections ; pathology ; Herpesvirus 2, Human ; Humans ; Necrosis ; Umbilical Veins ; pathology

OBJECTIVETo investigate the mechanisms for the cytopathic effect (CPE) of human cytomegalovirus (HCMV) in ECV304 endothelial-like cells.

METHODSPCR and indirect immunofluorescence were used to detect HCMV infection by examining immediate-early (IE) gene and protein expression of the virus in ECV304 cells. Phase-contrast and electron microscopies were performed to observe the morphological changes of the infected and uninfected cells, and DNA ladder analysis and flow cytometry were carried out to study HCMV-induced cell apoptosis.

RESULTSIn HCMV-infected ECV304 cells, cytopathic effects were first observed at approximately 72 h post-infection. The cells with CPE changes exhibited detachment from the monolayer, cell rounding and shrinkage. The expression of the IE gene was detected. Chromatin condensation and nuclear fragmentation along with dramatic changes of the mitochondria were observed by electron microscopy at 96 h post-infection. Cellular DNA fragmentation was observed in the infected cells, which had cells apoptotic rates of 4.1% and 45.7% at 96 h and 144 h post-infection, respectively.

CONCLUSIONHCMV can induce apoptosis of ECV304 endothelial-like cells.

Antigens, Viral ; genetics ; metabolism ; Apoptosis ; physiology ; Cell Line ; Cytomegalovirus ; genetics ; growth & development ; metabolism ; DNA, Viral ; analysis ; Endothelial Cells ; cytology ; ultrastructure ; virology ; Fluorescent Antibody Technique, Indirect ; Humans ; Immediate-Early Proteins ; genetics ; metabolism ; Microscopy, Electron ; Microscopy, Phase-Contrast ; Polymerase Chain Reaction ; Umbilical Veins ; cytology ; virology

OBJECTIVETo investigate whether adiponectin plays a role in the protection of myocardium in the rat myocardial ischemia preconditioning (IPC) model.

METHODInfarct size was measured by Masson's Trichrome staining, the expression of protein and mRNA of adiponectin at 0, 6, 12 and 24 h after IPC was examined by immunohistochemistry and quantitative real time RT-PCR, plasma levels of adiponectin at above mentioned four time points after IPC were detected by ELISA in IPC and MI rats.

RESULTInfarct size was smaller in IPC than in MI rats (20% ± 2% vs. 31% ± 3%, P < 0.05). The expression of adiponectin mRNA at 6 h and 12 h after IPC was 2.2 and 2.1 times higher than in Sham rats at respective time points (P < 0.05). Immunohistochemistry staining evidenced increased adiponectin expression in the ischemic area and weak expression of adiponectin in non-ischemic area (P < 0.05). Compared to the sham group, the plasma level of adiponectin increased significantly at 0, 6 and 12 h after IPC (0 h: 7.40 ± 0.47 vs. 10.90 ± 1.74; 6 h: 8.18 ± 1.41 vs. 10.98 ± 1.74; 12 h: 6.97 ± 1.02 vs. 9.31 ± 0.96, P < 0.05).

CONCLUSIONIPC reduced infarction size, upregulated the myocardial expression of adiponectin at mRNA and protein levels, and increased plasma adiponectin concentration, suggesting that the adiponectin may play a critical role in the protective effect of IPC.

Adiponectin ; metabolism ; Animals ; Ischemic Preconditioning, Myocardial ; Male ; Myocardial Infarction ; metabolism ; prevention & control ; Myocardial Ischemia ; metabolism ; prevention & control ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo evaluate curative effect of plate and xenogenic bony plate were applied in refracture in plate-screw fixation of femoral shaft.

METHODSThirteen cases of refracture in plate-screw fixation of femoral shaft included 8 males and 5 females, average age was 31.2 years ranging from 14 to 57. Fracture type was comminuted fracture in 7 cases, oblique fracture in 4 cases, transverse fracture in 2 cases. Fixation type used eight holes femoral LC-DCP in 5 cases, eight holes epipodite LC-DCP in 2 cases, six holes femoral LC-DCP in 2 cases, 8 holes La-Plate in 4 cases. All the patients were treated by femoral LC-DCP and xenogenic bony plate.

RESULTSAll of the patients were followed up from 16 to 40 months with average of 32 months. All cases had undergone only one operation and achieved bony union. Average time of bony union was 9 months. The lower limbs resumed walk and beared a heavy burden. According to criterion of Merchan, the results were excellent in 7 cases,good in 4,fair in 1 and poor in 1, the excellent and good rate of knee function was 84.6% (11/13) in one year after operation.

CONCLUSIONTreatment of refracture in plate-screw fixation of femoral shaft with armor plate and xenogenic bony plate is a reliable treatment.

Adolescent ; Adult ; Bone Plates ; Bone Screws ; Female ; Femoral Fractures ; pathology ; physiopathology ; prevention & control ; surgery ; Follow-Up Studies ; Fracture Fixation, Internal ; instrumentation ; Humans ; Male ; Middle Aged ; Recurrence ; Time Factors ; Young Adult