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 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.

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 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.

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

AIMTo study the effects of 9-(4-ethoxycarbonylphenoxy)-6,7-dimethoxy-1,2,3,4-tetrahydro acridine (EDT) on free radical induced injury in primary cultured rat cortical neuron and cerebral ischemia in mice.

METHODSIn primary rat cortical neuron, free radical injury model was established by 10 mumol.L-1 H2O2. The content of malondiadehyde (MDA) and activity of superoxide dismutase (SOD) in cells were investigated. Chronic cerebral ischemia model was produced by occlusion of one carotid artery and pneumogastric nerve in mice. The step down test was adopted to investigate the effect of EDT on the memory impairment. The cerebra morphology and MDA, NO content and SOD activity in mice cerebra were detected.

RESULTSIn primary rat cortical culture, 0.01-3 mumol.L-1 EDT concentration-dependently inhibited the formation of MDA and reduction of SOD activity induced by 10 mumol.L-1 H2O2. In chronic cerebral ischemia, EDT 2.5, 5 and 10 mg.kg-1 ig for 5 d greatly improved the memory impairment, reduced NO efflux and MDA content, while increased SOD activity in mice cerebra.

CONCLUSIONEDT was found to protect neurons from H2O2-induced neurotoxicity and inhibit chronic cerebral ischemia mediated injury and memory impairment in mice.

Acridines ; pharmacology ; therapeutic use ; Animals ; Animals, Newborn ; Brain Ischemia ; complications ; etiology ; prevention & control ; Cells, Cultured ; Cerebral Cortex ; cytology ; Female ; Hydrogen Peroxide ; antagonists & inhibitors ; Male ; Malondialdehyde ; metabolism ; Mice ; Neurons ; drug effects ; metabolism ; Neuroprotective Agents ; pharmacology ; therapeutic use ; Nitric Oxide ; metabolism ; Rats ; Rats, Wistar ; Superoxide Dismutase ; antagonists & inhibitors ; metabolism

OBJECTIVETo investigate mRNA expression of integrin beta3 in gastric carcinoma, its correlation with microvascular density (MVD), growth-pattern, invasion, metastasis and prognosis.

METHODSIn situ hybridization and immunohistochemistry techniques were used to study the expression of integrin beta3 mRNA and CD34 protein in 105 gastric carcinoma specimens.

RESULTSIn situ hybridization revealed a 30% (6/20) positive rate of integrin beta3 mRNA in non-tumor gastric mucosa, which was significantly lower than that of the gastric cancer group (61.0%, 64/105, chi(2) = 8.85, P = 0.037); In infiltrating type cases (70.2%, 40/57), stage III - IV (72.1%, 44/61), lymphatic metastasis (68.6%, 48/70) and distant metastasis (85.7%, 36/42), the positive expression rates were significantly higher than those of the expanding type (chi(2) = 14.97, P = 0.002), stage I - II (chi(2) = 15.21, P = 0.015), non-lymphatic metastasis (chi(2) = 17.89, P = 0.025) and non-distant metastasis (chi(2) = 20.22, P = 0.005; chi(2) = 21.35, P = 0.035); its mean MVD was also significantly higher than that of the expanding type (t = 10.105, P = 0.001), stage I approximately II (t = 5.961, P = 0.001), non-lymphatic metastasis (t = 3.819, P = 0.01)and non-distant metastasis (t = 10.578, P = 0.001; t = 7.882, P = 0.001), respectively; The mean MVD (41.02 +/- 8.55)/0.72 mm(2) of the integrin beta3 mRNA positive expression group was significantly higher than (25.26 +/- 11.25)/0.72 mm(2) of the negative expression group. There was a positive relation between MVD and integrin beta3 mRNA expression in the tumor (r(s) = 0.316, P = 0.001). The mean survival time in cases with positive integrin beta3 mRNA expression or MVD value >or= 39.5 was significantly shorter than that in cases with negative expression or MVD value < 39.5.

CONCLUSIONSIntegrin beta3 expression correlates with enhanced tumor angiogenesis and may play a role in the invasion and nodal metastasis of gastric carcinoma, therefore it may serve as a prognostic marker of gastric cancer.

Adult ; Aged ; Disease Progression ; Female ; Humans ; Integrin beta3 ; genetics ; Male ; Middle Aged ; Neovascularization, Pathologic ; etiology ; Prognosis ; RNA, Messenger ; analysis ; Stomach Neoplasms ; blood supply ; metabolism ; pathology ; Survival Rate

Adenocarcinoma ; blood supply ; metabolism ; mortality ; secondary ; Adult ; Aged ; Female ; Humans ; Lymphatic Metastasis ; Male ; Microcirculation ; Middle Aged ; Neovascularization, Pathologic ; metabolism ; pathology ; Prognosis ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Cell Surface ; biosynthesis ; genetics ; Receptors, Urokinase Plasminogen Activator ; Stomach Neoplasms ; blood supply ; metabolism ; mortality ; pathology ; Survival Rate ; Vascular Endothelial Growth Factor A ; metabolism

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