Aim To investigate the effects of EDT on anoxia and ischemic injury in cultured PC12 cells. Methods Cultured PC12 cells were treated with 1 mmol?L -1 Na 2S 2O 4 and 20 mmol?L -1 NaCN in combination with glucose deprivation. The protective effects of EDT on these two models were evaluated by lactate dehydrogenate (LDH) efflux assay and colormetric MTT assay.ResultsEDT, within the range of 10 -8～ 10 -6 mol?L -1, significantly antagonized LDH efflux induced by two models and increased the optical density at 570 nm tested by colorimetric MTT assay in concentration-dependent manner. 10 -6 mol?L -1 EDT might time-dependently inhibit two injuries and reach maximal level at 48 h. Conclusion EDT can protect PC12 cells from anoxia and ischemic injury.

AIM: To study the effects of propofol on membrane fluidity and intracellular free Ca2+ concentration ([Ca2+]i) in PC12 cells and discuss its relevant mechanism. METHODS: PC12 cell lines were divided into seven groups: control, solvent and propofols (1, 3, 10, 30, 100 mg·L-1). Fluorescence depolarization method was used to measure dynamically microviscosity in PC12 cells and [Ca2+]i was detected using calcium fluorescentprobe Fluo-3/AM and a laser scanning confocal microscope. RESULTS: 1 Acute administration of various doses of propofol induced a significant decrease of microviscosity in PC12 cells dose-dependenty. 2 Solvent, propofol at dose of 10 mg·L-1 had no effect on [Ca2+]i in PC12 cells, however, after 30 and 100 mg·L-1 administration, [Ca2+]i increased markedly at 20-30 seconds (increase percentage were 119% and 140% respectively) and then recovered to their pre-administration levels within 50 seconds. CONCLUSION: The propofol can significantly increase membrane fluidity in PC12 cells in a dose-dependent manner and elevate [Ca2+]i in PC12 cells at doses of 30 and 100 mg·L-1. These changes are consistent with each other and related closely with anesthetic effect of propofol.

Objective To assess left ventricular function early and late improvement of patients with acute myocardial infarction (AMI) treated by percutaneous coronary intervention (PCI) by speckle tracking imaging (STI).The clinical values of the assessment of STI for the prognosis and heart function improvement of AMI patients treated by PCI were discussed.Methods 73 AMI patients who had AMI for the first time and had been treated by primary PCI from September 2010 to July 2011 and were examined in the follow-ups from December 2010 to February 2012 in our hospital were enrolled.Dynamic images were acquired before PCI,at 3 months and 6 months after PCI and analyzed by STI.Dynamic images were analyzed for longitudinal peak systolic strain (LPSS),radial peak systolic strain (RPSS) and circumferential peak systolic strain (CPSS) values by STI.According to the comparison of left ventricular ejection fraction (LVEF) before PCI and 6 months after PCI,patients were divided into left ventricular function improved group (ΔLVEF6＞5％) and not-improved group.According to the comparison of LVEF before PCI and 3 months after PCI,improved group were divided into left ventricular function early-improved group (ΔLVEF3 ＞5％) and late-improved group.Results The values of all STI parameters before PCI,3 months and 6 months after PCI in improved group were higher than those in not-improved group (P ＜0.001,all).LPSS before PCI and at the follow-ups and RPSS at 3 months after PCI in early-improved group were higher than those in late-improved group (LPSS at 3 months after PCI:P＜0.001;Other parameters:P ＜0.05).There were significant correlations between all STI parameters and both ΔLVEF3 and ΔLVEF6.LPSS before PCI was more closely related to ΔLVEF3 (r =-0.781,P ＜0.001).CPSS at 6 months after PCI was more closely related to ΔLVEF6 (r =-0.834,P ＜ 0.001).Conclusions Early and late function improvement of left ventricle in AMI patients who is treated by PCI are accurately assessed by STI.The precise analyses of longitudinal and circumferential movements in STI are important for clinical diagnosis.

Objective To assess the left ventricular function of patients with acute myocardial infarction (AMI) treated by percutaneous coronary intervention (PCI) by speckle tracking imaging (STI).Methods 75 AMI patients who had AMI for the first time and have been treated by primary PCI were enrolled.Dynamic images were acquired before PCI,at 6 months after PCI and analyzed by STI.Dynamic images were analyzed for longitudinal peak systolic strain (LPSS),radial peak systolic strain (RPSS) and circumferential peak systolic strain (CPSS) values by STI.According to the comparison of left ventricular ejection fraction (LVEF) before PCI and 6 months after PCI,patients were divided into left ventricular function improved group (△LVEF≥5％) and not-improved group.Results Compared to non-improved group,LPSS (P ＜0.001),RPSS (P ＜0.05,P ＜0.001) and CPSS (P ＜0.001) of improved group were all higher before and 6 months after PCI.LPSS (r =-0.578,P ＜0.001) and CPSS (r =-0.817,P ＜0.001) before PCI were both closely related to △LVEF.In single parameter mode of ROC curve analysis,the area under the ROC curve (AUC) (0.867),sensitivity (94.7％) and specificity (74.4％) of CPSS are relatively higher than other STI parameters.In multiple parameters united mode of ROC curve analysis,AUC (0.897),sensitivity (94.7％) and specificity (74.4％) of LPSS,RPSS and CPSS united were the highest among all the combinations of all STI parameters.Conclusions Left ventricular function improvement of patients with AMI 6 months after PCI is accurately assessed and predicted by STI.CPSS is a strong predictor for left ventricular function improvement 6 months after PCI of AMI patients among all the STI parameters and is an effective indicator for the assessment of left ventricular function improvement of AMI patients.

Objective To create a three-dimensional speckle tracking imaging (3DSTI) multiparameter analysis model to improve the diagnostic efficiency of obstructive coronary artery stenosis.Methods One hundred and four patients with chest pain were divided into two groups:coronary heart disease(CHD) group (61 patients) and control group (43 patients) according to the result of selective coronary angiography (SCA).The two groups' clinical data and echocardiographic parameters were aquired,including mitral flow E and A velocities,E peak deceleration time,isovolumic relaxation time (IVRT),mitral annulus velocity e' and a' peak in diastolic and s' peak in systolic by tissue Doppler imaging (TDI),and left ventricular ejection fraction (LVEF).The global longitudinal peak strain (2D-GLPS) by two dimensional speckle tracking,global longitudinal peak strain (3D-GLPS),circumferential peak strain(3D-GCPS),radial peak strain (3D-GRPS) and area peak strain (3D-GAPS) were acquired.Results For conventional parameters,there were no significant difference between the two groups.Compared with control group,TDI e' peak,3D-GRPS decreased,significantly E/e',2D-GLPS,3D-GLPS,3D-GAPS,3D-GCPS increased significantly (P ＜0.01).For single parameter,area under the ROC curve (AUC) were successively 3D-GAPS(0.766) ＞ 3D-GLPS(0.746) ＞ 2D-GLPS(0.746) ＞3D-GRPS(0.727) ＞ s' (0.703) ＞E/e' (0.688)＞3D-GCPS(0.686).AUC for single and multi technology were successively p-Union(0.856)＞p-3DSTI(0.772) ＞ p-TDI (0.757) ＞ p-2DSTI (0.746).Conclusions 3DSTI together with multi-parameter analysis model can significantly improve the diagnostic efficiency of obstructive coronary artery stenosis.Area strain is an independent predictor of obstructive coronary artery stenosis.

OBJECTIVETo observe the effect of Zhibai Dihuang Decoction (ZDD) on mRNA and protein expressions of transient receptor potential family vanilloid subtype 1 (TRPV1) and transient receptor potential family vanilloid subtype 5 (TRPV5) in Ureaplasma urealyticum (UU)-infected rat semens and spermatogenic cells, and to explore the pathomechanism of UU-infected infertility and the intervention of ZDD.

METHODSTotally 45 were randomly selected from 60 4-5 months old SD rats. UU testicular infected animal models were set up after bladder inoculation of UU suspension. The remaining 15 rats were simultaneously injected with normal saline as a normal control group. After a successful modeling, UU infected model rats were randomly divided into the model group, the azithromycin group, and the ZDD group, 15 in each group. Rats in the ZDD group were administered with ZDD at the daily dose of 1 g/kg by gastrogavage. Rats in the azithromycin group were administered with azithromycin suspension at the daily dose of 0. 105 mg/kg by gastrogavage. Equal volume of normal saline was administered to rats in the normal control group and the model group. All medication was performed once daily for 21 successive days. Testes and epididymis were extracted after rats were killed and UU positive rates were compared among all groups. Sperm cells were separated using a mechanical separation technique. Sperm motility parameters were detected using color sperm motion detection system. mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells were determined by real-time quantitative PCR and Western blot.

RESULTSThe UU positive rate was obviously higher in the model group than in the normal control group [(86.7% (13/15 cases) vs. 0] P < 0.05). It was lower in the ZDD group [33.3% (5/15 cases)] and the azithromycin group [26.7% (4/15 cases)] than in the model group (P < 0.05). Compared with the normal control group, class A and B sperms were reduced, the linear velocity and the average velocity were significantly lowered, mRNA and protein expressions of TRPV1 and TRPV5 in spermated genic cells significantly decreased in the model group (P < 0.05). Compared with the model group, class A and B sperms were increased, linear and curve velocities and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group and the azithromycin group (P < 0.05, P < 0.01). Compared with azithromycin group, class A and B sperms were increased, the linear velocity and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group (P < 0.05, P < 0.01).

CONCLUSIONZDD could fight against UU infection and elevate semen quality, which might be associated with up-regulating mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells.

Animals ; Calcium Channels ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Infertility ; Male ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Semen Analysis ; Sperm Motility ; Spermatozoa ; TRPV Cation Channels ; metabolism ; Testis ; Ureaplasma Infections ; Ureaplasma urealyticum

Objective To evaluate the function and synchronization in patients with paroxysmal atrial fibrillation(PAF) by two dimensional speckle tracking imaging(2D-STI) measuring the strain of left atrial.Methods Thirty healthy controls and thirty-five patients with PAF were studied.The PAFs were divided into group PAF1 during the diapause and group PAF2 during the stage of attack.From the strain-time curve of 2D-STI workstation,the peak velocity(Vs,Ve and Va),the peak amplitude of longitudinal strain (Ss,Se and Sa) and strain rate (SRs,SRe and SRa) were acquired in the course of systole,the early and the late diastole respectively.The synchronization of left atrial (Ts-SD,Te-SD and Ta-SD) could be gotten from the time to the peak strain.The correlation were analysed between the synchronization indices and the strain rate parameters.Results ①No significant differences were found at the age,heart rate,E/A,IVS,LVEDD and LVEF.Compared with the control group,the LAD increased and DT decreased in PAF1 and PAF2 groups (P ＜0.05).②The peak velocity were slower,the peak strain were shorter and the peak strain rate were lessened in the PAFs than the controls(P ＜0.05),except the SRa(P ＞0.05).③The Ts-SD and Te-SD were much more different during the three groups(P ＜0.05),the PAF2 showed fewer synchronization; the Ta-SD was higher in the PAF1 than the controls(P ＜0.05).④The Ta-SD had a positive correlation with SRa(r =0.61,P ＜0.01).Conclusions The reservoir and conduit function of left atrial are impaired in PAFs during the diapause,the booster pump could recover partially,however,there are dyssynchronization.The booster pump function disappear during the stage of attack and there are worse synchronization.2D-STI may paly important roles in the assessment of function and synchronization of left atrial.

AIM: To establish a method of in vitro donor heart perfusion in murine cardiac transplantation during preservation and apply it in adenovirus mediated gene transfection for donor heart. METHODS: Donor heart was transfected with recombinant adenovirus and stored for 2 hours after harvest, then it was transplanted heterotopically in abdomen. The grafts were appraisal by palpitation. Marked gene products were determined by X-Gal staining, aod T cell infiltration was determined by immunohistochemistry. The activation markers of recipients' lymphocytes were examed by cytometry. RESULTS: The grafts survival rate is 100% after perfusion and cold storage. The LacZ staining became strong 1 week after transplantation. The grafts remained an intact structure and no apparent T cell infiltration. The activation status of recipients' lymphocytes were not enhanced by transfected cardiac graft. CONCLUSION: In vitro perfusion during graft cold preservation is feasible for adenovirus mediated gene transfection. [

Objective To analyze longitudinal layer‐specific strain in endocardial ,midcardial and epicardial layers of left ventricle (LV) by two‐dimensional speckle tracking imaging(2D‐STI) in patients with coronary heart disease (CHD) and to evaluate the regional or global systolic function of LV . Methods Seventy‐eight patients with suspected CHD were divided into CHD group (46 subjects) and control group (32 subjects) according to the results of selective coronary arteriography . According to the stenosis degree or number of coronary arteries ,the CHD group was divided into mild stenosis group ( corresponding blood‐supply vessel stenosis 50% -75% ) ,severe stenosis group ( corresponding blood‐supply vessel stenosis ≥75% ) or single‐vessel stenosis group ,multi‐vessel stenosis group . Two‐dimensional images with high frame rate were recorded in apical four‐chamber ,long‐axis ,two‐chamber of LV in all subjects . The strain parameters by EchoPAC analysis software included territorial longitudinal strain ( TLS ) of endocardial , midcardial and epicardial layers ( TLSendo ,TLSmid ,TLSepi) and global longitudinal strain ( GLS) of endocardial ,midcardial and epicardial layers ( GLSendo ,GLSmid ,GLSepi) . Results Regardless of with or without corresponding blood‐supply coronary artery stenosis and range of coronary artery stenosis , TLSendo ,TLSmid ,TLSepi and GLSendo ,GLSmid ,GLSepi all showed a gradient decrease( P < 0 .05) . Compared with the control group ,TLSendo ,TLSepi of mild stenosis group and severe stenosis group decreased significantly( P <0 .05) ,while there was no significant difference about TLSmid between the two groups( P > 0 .05) . Compared with the mild stenosis group ,TLSendo ,TLSepi of severe stenosis group decreased significantly( P < 0 .05) ,while there was no significant difference about TLSmid( P > 0 .05) . Moreover ,compared with the control group ,GLSendo ,GLSmid ,GLSepi of single‐vessel stenosis group and multi‐vessel stenosis group decreased significantly( P < 0 .05) . Compared with the single‐vessel stenosis group ,GLSendo ,GLSmid ,GLSepi of multi‐vessel stenosis group decreased significantly( P < 0 .05) . For predicting corresponding blood‐supply vessel stenosis ≥75% ,the sensitivity and specificity were 55 .6% , 72 .6% and 57 .8% ,76 .4% ,when the cut‐off of TLSendo and TLSepi were 20 .5% and 15 .5% respectively . For predicting multi‐vessel stenosis ,the sensitivity and specificity were 54 .5% ,71 .4% ;50 .0% ,87 .5% and 63 .6% ,87 .5% ;when the cut‐off of GLSendo , GLSmid , GLSepi were 21 .5% , 17 .5% and 16 .5%respectively . Conclusions Systolic dysfunction of LV happened through all layers in CHD patients . The longitudinal strain of layer‐specific by 2D‐STI can accurately evaluate the regional and global systolic function of LV in patients with CHD ,which can also be used to predict the degree or range of coronary artery stenosis .

OBJCETIVETo investigate the method of separation of culture of bone microvascular endothelial cells (BMECs) of human femoral head in vitro.

METHODSFrom October 2013 to January 2014,15 femoral heads without pathologic change from patients resected during hip replacement were selected involving 2 males and 13 females with a mean age of 71.2 years old ranging from 38 to 92. Cancellous bone in femoral head was bited into broken bone grain and transfered into medium in aseptic contidion. Cells were isolated by the methods of enzymic digestion and density gradient centrifugation,purified by differiential attachment. The characteristics of cells was observed by inverted microscope. vWF and CD31 immunofluorescence analysis was applied for identification of cells.

RESULTSThe number of cells was positively correlated with patients' age after 24 hours in primary culture. The older patients had the less cells numbered. After 4 to 5 days' culture, primary cells appeared short spindle,polygon shaped and cobblestone-like morphology. After 7 to 10 days' culture, primary cells proliferated densely, became fusion, arranged in swirl, and contact inhibition appeared significantly. Immunofluorescence staining revealed the cells were 100% positive for vWF and CD31, and it showed that the cultured cells were BMECs.

CONCLUSIONIt was a simple, steady, effective method with good reproducibility, by which highly purified human BMECs can be obtained.

Adult ; Aged ; Aged, 80 and over ; Cell Culture Techniques ; Cell Proliferation ; Cell Separation ; methods ; Cells, Cultured ; Endothelial Cells ; cytology ; Female ; Femur Head ; blood supply ; Humans ; Male ; Microvessels ; cytology ; Middle Aged