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 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.

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

The rate of corneal graft rejection is still high for high-risk keratoplasty although immune suppression drug is routinely used.The role of traditional Chinese medicine in corneal transplantation is concerned gradually.Heijingpaichitang on the prevention and treatment of rats with high-risk corneal allograft rejection needs further study.Objective This study was to investigate the inhibitory effect of heijingpaichitang on high-risk corneal transplantation immune rejection in rats.Methods Sixteen female SD rats were used as the donors and 32female Wistar rats were served as recipients.The high-risk corneal trasplantation models were established by corneal suture in 32 Wistar rats,and then homogeneity variant SD-Wistar corneal transplantation was performed.The recipients were randomized into model control group,cyclosporinc A (CsA)group,heijingpaichitang group and CsA +heijingpaichitang group.CsA,heijingpaichitang and CsA + heijingpaichitang was orally administered 4 days after operation once per day for 15 days,and normal saline solution was used at the same way in the model control group.Ocular anterior segment reaction was examined under the slit lamp and corneal opacification,edema and neovasculation were scored based on Larkin' s criteria.Rejection index of the corneal graft was recorded and the graft survival time was calculated.The pathological examination of the corneal graft was carried out in all rats,and the inflammatory cells in the corneas and CD4+ cells in the periphery blood were assayed using flow cytometry.The use of the animals complied with ARVO Statement.Results Corneal graft rejection occurred in 10 days after operation in the model control group,12-13 days in the CsA group and heijingpaichitang group and 22 days in the CsA +heijingpaichitang group.Compared with model control group,the scores of the corneal opacification,corneal edema and neovascularization were significantly lower in the CsA group,heijingpaichitang group and CsA+heijingpaichitang group (P＜0.05),and all the scores were declined in the CsA+ heijingpaichitang group compared with CsA group and heijingpaichitang group(P＜0.01),but no significant differences were seen in the scores between the CsA group and heijingpaichitang group(P＞0.05).The mean survival time of grafts was (10.38 ±1.69)days in the model control group,(22.50 ± 3.07) days in the CsA + heijingpaichitang group,with the significant difference (t =-9.790,P =0.000).The pathological examination of graft showed that the lymphocytes and new blood vessels were less in the CsA+heijingpaichitang group compared with CsA group and heijingpaichitang group 15 days after operation.Flow cytometry verified that the number of lymphocytes in graft,CD4+cells and CD4+/CD8+ in periphery blood were significantly lower in the heijingpaichitang group,CsA group and CsA+heijingpaichitang group compared with model control group (P＜0.05).Conclusions Heijingpaichitang can inhibit immune rejection to certain extent in high-risk corneal transplantation rat and has a similar effect to 0.1％ CsA.Heijingpaichitang and 0.1％ CsA have a synergistic effect.

Objective To increase the transfection of EGFP-N3 plasmids into 293T cells using ultrasound-targeted microbubbles delivery(UTMD) mediated a peptide nucleic acid (PNA) binding nuclear localization signal (NLS).Methods Antibody-targeted microbubbles were used in the experiments which can specifically recognize the SV40T antigen receptor.The SV40T antigen receptors were expressed on the membranes of 293T cells.The PNA containing the NLS were inserted in the EGFP-N3 plasmid DNA,which increased nuclear localization.Ultrasound-targeted microbubble delivery (UTMD) and the PNA binding NLS were utilized to improve the cytoplasmic import of plasmids and the nuclear intake of the plasmid from the cytoplasm,respectively.The study was divided into five groups:Contrast (group A),Common microbubble + DNA (group B),Antibody-targeted microbubbles + DNA (group C),Common microbubbles + NLS-PNA-DNA (group D),Antibody-targeted microbubbles + NLS-PNA-DNA (group E).Fluorescence microscope was used to observe the fluorescent light in each group;flow cytometry to test the transfection;RT-PCR and Western blot to detect genes' mRNA and protein expression level.Results Ultrasound and antibody-targeted microbubble delivery (UTMD) significantly enhanced the cytoplasmic intake of exogenous genes and maintained high cell viability(＞80％).Fluorescent microscope showed that the quantities of green fluorescence in cells were increased successfully.The transfection results of flow cytometry were 0,(9.30 ± 0.46)％,(26.46 ± 2.01)％,(29.54 ± 0.62)％,(45.72 ± 1.86)％,respectively,and the differences were statistically significant(P ＜0.05).The relative mRNA and protein expression in group E were greater than those in group C and D respectively (P ＜0.05).Conclusions UTMD combined with antibody-targeted microbubbles and a PNA binding NLS plasmid can significantly improve transfection efficiency of exogenous genes by enhancing both cytoplasmic and nuclear DNA import.

Objective To evaluate the left ventricular synchrony after myocardial infarction (MI) by ultrasound targeted microbubbles destruction (UTMD)-mediated angiogenin 1 (Ang1) gene transfection in canine.Methods Twenty-one dogs were divided into three groups (n =7 in each group):①control group (healthy dogs);②MI group (MI dogs without treatment);③UTMD group (MI dogs with UTMD treatment).One month later,the size and systolic function of heart were measured by echocardiography.The synchronization parameters derived from two dimensional-speckle tracking imaging(2D-STI) included the standard deviation and maximum difference of time to peak strain for all left ventricular segments (Tls-SD,Trs-SD,Tcs-SD,Tls-Dif,Trs-Dif and Tcs-Dif).CD31 and α-SMA were applied for quantifying capillary and arteriolar density.The Ang1,SERCA2a and PLB protein were detected by Western blotting.Results ① One month later,the conventional ultrasonic parameters were compared among three groups,the LVEDD,LVESD and E/e'increased and LVEF,e'and E/A reduced in MI group than those in control group,all of them partially recovered in UTMD group than those in MI group,but were still lower than those in control group (P ＜0.05);②The left ventricular synchrony parameters of Tls-SD,Tls-Dif and Trs-SD showed significant differences among the three groups(P ＜0.05),the degree of dyssynchrony increased in MI group than control group,they were lower in UTMD group than those in MI group.The Tcs-SD,Tcs-Dif and Trs-Dif showed no significant difference among three groups (P ＞ 0.05);③ The immunohistochemistry showed the higher blood vessel density in UTMD group than that in MI group(P ＜ 0.05);④The relative quantity of Ang1 was significantly higher in UTMD group.The relative quantity of SERCA2a protein was lower in MI group than that in control group,increased in UTMD group,the trend of PLB was contrary to it.The differences were statistically significant (all P ＜0.05).Conclusions The UTMD-mediated Ang1 gene transfection can promote angiogenesis after MI,reverse left ventricular remodeling and improve left ventricular synchrony.The myocardial synchrony may be related to the expression of calcium ions key protein SERCA2a and PLB.

Objective To develop simple theories and methods for health quality evaluation in moderately prosperous society, and to establish a health quality index (HQI) model to assess the improvement in health quality. Methods Four health quality - related indexes were selected following screening. HQI was then calculated. In 5 provinces, empirical analysis of HQI was conducted. Results In F test, there was significant difference in HQI between different regions of China in 2005. Conclusions The study provides a scientific, comprehensive, sensitive, and intuitive evaluation index, HQI, which may be helpful in assessing the health level.