Objective To evaluate the feasibility and value of quantitatively assessing left ventricular systolic function in rats with myocardial infarction (MI) by myocardial contrast echocardiography (MCE) based on mathematical morphology algorithm.Methods SD rats model of MI were mede,then MCE were performed before MI and at 1,3 weeks after MI.In vitro study:based on the principle of binary morphology,segmentation of endocardium was acquired using dilation,erosion,closed operations and connected domain object tag-based.The automatic segmentation of endocardial contour was compared with the manual segmentation boundary.In vivo study:the left ventricular area was calculated using the software,and the area variation fraction (AVF),a new index of left ventricular systolic function,were acquired.The correlation between AVF and left ventricular ejection fraction (LVEF) was analyzed.The value of AVF for diagnosis for left ventricular dysfunction was evaluated by ROC curve.Results ① The cross-correlation coefficient (CCC) and percent error (PE) between automatic contours and manual boundary were more than 0.90 and less than 9％,respectively.② AVF correlated positively with LVEF (r =0.934,P ＜ 0.001).③ ROC analyses showed the area under curves for AVF diagnosing left ventricular dysfunction was 0.834.The best cutoff value was 27.5 ％ for diagnosing left ventricular dysfunction with the sensitivity of 85 ％ and specificity of 60 ％.Conclusions Left ventricular endocardium can be identificated automatically and rapidly by MCE based on mathematical morphology algorithm.AVF can assessed quantitatively left ventricular systolic function in rats with myocardial infarction.

OBJECTIVETo investigate the action targets of ginseng (GS) and Panax notoginseng (PN), Chinese herbs for benefiting qi and activating blood circulation, on angiogenesis signaling pathway (VEGFR-2-Ras-MAPK) in human umbilical vein endothelial cells (HUVEC).

METHODSTo block the signal pathway by turns, at the first, added the IC50 of VEGFR-2 inhibitor, SU5416, and detected its downstream signaling protein Ras, MAPK expression using Western Blot. Secondly, added the IC50 of the Ras inhibitor, FPP, and detected its downstream signaling protein MAPK expression.

RESULTSCompared with the control group, adding SU5416 made the Ras, MAPK expression significantly decreased (P < 0.05, P < 0.01); and adding FPP made the expression of MAPK significantly decreased (P < 0.01). Compared with the group treated by SU5416 alone, the expression of downstream signaling protein Ras, MAPK were significantly higher in the group treated by SU5416 plus GS and PN (P < 0.05, P < 0.01); same state also found in comparison MAPK expression between groups treated with FPP alone and FPP plus GS and PN (P < 0.05).

CONCLUSIONThe angiogenesis mechanism of GS and PN on HUVEC may be realized by increasing the protein expression of three key signals, VEGFR-2, Ras and MAPK, respectively.

Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Mitogen-Activated Protein Kinase Kinases ; metabolism ; Neovascularization, Physiologic ; drug effects ; Panax ; chemistry ; Panax notoginseng ; chemistry ; Signal Transduction ; drug effects ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism ; ras Proteins ; metabolism

The present study aimed to investigate the effect and the mechanisms of Bu-zhong-yi-qi-tang (BZYQ) on Spleen-Qi deficiency rat's model using nuclear magnetic resonance (NMR) metabolomics and multivariate statistical analysis methods. The rat Spleen-Qi deficiency model was established as follows: oral administration of Radix Rhei extract, loaded swimming and starvation for 24 h. The body weight and motor behavior of the rats were measured and recorded once a week. BZYQ could significantly improve body weight and behavioral of Spleen-Qi deficiency model rats compared with the model group (P < 0.05, 0.01). After drug administration, the changes in the levels of endogenous metabolites in the spleen including decreasing lactate, taurine and hypoxanthine, increasing glutamate and scyllo-inositol compared with the model group. The metabolomics approach is an effective tool for the investigation of the pharmacologic mechanism of BZYQ and it is helpful to further research.
Administration, Oral ; Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Medicine, Chinese Traditional ; Metabolomics ; Proton Magnetic Resonance Spectroscopy ; Qi ; Rats ; Spleen

Aim To observe the intervention effects of sodium aescinate on acute lung injury model of rats induced by oleate. Methods Fifty four male SD rats were randomly divided into five groups: normal control group, sodium aescinate control group (without oleate) , oleate model control group,medrol interventional group and sodium aescinate interventional group. Acute lung injury models of rats were made by injecting oleate (OA, 0. 1 ml · kg~(-1) ) through caudal veins, and then rats were observed and killed to detect correlated in-dice. The observation indice were the histomorphology of lung, the wet and dry weights of lung ( W/D), score of injury of lung under light microscope (IQA ) , partial pressure of oxygen in artery ( PaO_2) , the levels of SOD and MDA in blood plasma and lung tissue. Results ① Histomorphology of lung: Lung surface hyperemia relieved obviously and pink secretion from trachea of rats in sodium aescinate interventional group and medrol interventioal group decreased significantly compared with oleate model control group. Under light microscope , compared with oleate model control group, effusion of inflammatory cells in alveolar space of rats in sodium aescinate interventional group and medrol interventional group decreased. ② The wet and dry weights of lung ( W/D ) ; W/D of rats in oleate control model group increased obviously compared with those in normal control group, W/D of rats in sodium aescinate interventional group and medrol interventional group decreased obviously compared with those in oleate model control group. ③ Score of injury of lungs under light microscope (IQA) ; IQA of rats in oleate model control group advanced obviously compared with that in normal control group. IQA of rats in sodium aescinate interventional group and medrol interventional group lowered significantly compared with that in oleate model control group.④ Partial pressure of oxygen in artery (PaO_2) : PaO_2 of rats in oleate model control group lowered significantly compared with that in normal control group. PaO_2 of rats in sodium aescinate interventional group and medrol interventional group improved significantly compared with that in oleate model control group. ⑤ The levels of SOD and MDA in blood plasma and lung tissue:The levels of SOD in plasma and lung tissue of rats in oleate model control group lowered significantly compared with those in normal control group. SOD in plasma and lung tissue of rats in sodium aescinate in-terventional group and medrol interventional group increased significantly compared with that in oleate model control group. The levels of MDA in plasma and lung tissue of rats in oleate model control group lowered significantly compared with those in normal control group. MDA in plasma and lung tissue of rats in sodium aescinate interventional group and medrol interventional group increased significantly compared with that in oleate model control group. Conclusion Sodium aescinate can improve W/D, IQA and PaO_2 by adjusting oxidization of the acute lung injury model of rats, which may provide a possible path for treating acute lung injury in clinical practice.

OBJECTIVETo explore the inhibition effect of N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) on myofibroblast differentiation of MRC-5 human fetal lung fibroblasts induced by angiotensin (Ang) II.

METHODSThe study was divided into 2 step: (1) MRC-5 human fetal lung fibroblasts was induced for 48 h at different dose of Ang II and at different time point by 100 nmol/L Ang II. Then the expression of collagen type I and α-smooth muscle actin (α-SMA) were mesaured by western blot. (2) MRC-5 human fetal lung fibroblasts were divided into 4 group: (1) control, (2) Ang II, (3) Ang II+Ac-SDKP, (4) Ang II+8-Me-cAMP (a specific activator of Epac). The α-SMA expression was observed by immnocytochemical stain. The protein expression of collagen type I, α-SMA, serum response factor (SRF), myocardin-related transcription factor (MRTF)-A, exchange protein directly activated by cAMP (Epac) 1, 2 were measured by Westen blot.

RESULTSMyofibroblast differentiation could be induced by Ang II from MRC-5 cells with a dose- and time-dependent manner. The up-regulation of SRF and MRTF-A were observed in MRC-5 cells induced by Ang II and accompanied with collagen I and α-SMA increased. Pre-treatment with 8-Me-cAMP or Ac-SDKP could attenuated all this changes induced by Ang II, and promoted the expression of Epac1.

CONCLUSIONAc-SDKP can inhibit the myofibroblast differentiation of MRC-5 cells induced by Ang II via Epac1 activating.

Actins ; Angiotensin II ; Cell Differentiation ; drug effects ; Collagen ; Collagen Type I ; Cyclic AMP ; analogs & derivatives ; Fetus ; cytology ; Fibroblasts ; cytology ; Guanine Nucleotide Exchange Factors ; Humans ; Lung ; cytology ; Myofibroblasts ; drug effects ; Oligopeptides ; pharmacology ; Serum Response Factor ; Trans-Activators

OBJECTIVETo study the safety, biodistribution, and dosimetry of myocardial perfusion imaging agent 99Tc(m)N-NOET in 10 healthy volunteers.

METHODS744-792 MBq of 99Tc(m)N-NOET was injected to each volunteer. Safety parameters and adverse event was measured in 24 hours of injection. Biodistribution was studied by whole-body imaging 1, 30 minutes, 1, 2, 4, 8, and 24 hours after the injection of 99Tc(m)N-NOET. The estimation of dosimetry was based on the standard medical internal radiation dose method using MIRDOSE 3.0 analysis program. Myocardial single photon emission computed tomography (SPECT) imaging was performed at 1 and 4 hours after injection.

RESULTSNo undesirable effects were reported by the subject during 24 hours after injection of 99Tc(m)N-NOET. No clinically significant changes were found in vital signs (heart rate, blood pressure, and electrocardiogram). No biochemical aspects and serology changes were measured. The myocardial SPECT imaging was clear. Cardiac uptake of 99Tc(m)N-NOET was as high as 2.68% at 2 hours after injection. The heart to lung ratio was more than 1 from 30 minutes after injection, reaching a maximum of 1.91 +/- 0.53 at 2 hours after injection. Radiation dosimetry calculations indicated an effective absorbed dose of 1.28 x 10(-5) Sv/MBq. The dosimetry in each main organ is lower then 50 mGy given 740 MBq of 99Tc(m)N-NOET in once imaging.

CONCLUSIONS99Tc(m)N-NOET exhibits high cardiac uptake and low estimated effective absorbed dose. It's a safe myocardial perfusion imaging agent.

Heart ; diagnostic imaging ; Humans ; Myocardium ; metabolism ; Organotechnetium Compounds ; adverse effects ; pharmacokinetics ; Radiation Dosage ; Radiopharmaceuticals ; adverse effects ; pharmacokinetics ; Thiocarbamates ; adverse effects ; pharmacokinetics ; Tissue Distribution ; Tomography, Emission-Computed, Single-Photon

OBJECTIVETo investigate the differentially expressed genes in rat in the process of regression of vascular calcification by using the suppression subtractive hybridization (SSH).

METHODS24 SD male rats which aged 6 weeks and specific pathogen free grade were selected and randomly divided into 3 groups (n = 8): control group, calcification group and regression group respectively. Vascular calcification model (vitamin D3 plus nicotine, VDN) were made from rats in calcification group and regression group, and rats in control group were intragastric administered with normal saline and lavaged with peanut oil. Rats were bred for 8 weeks in calcification group and control group, while rats in regression group were fed for 16 weeks. All rats were killed to measure concentration of calcium in the arterial tissue and examine the pathological lesion changes. Subtractive hybridization among vascular cDNA sequences from calcification group and regression group were established. The cDNA fragments which expressed higher or lower in regression group than those in calcification group were isolated. Differentially expressed genes with cDNA fragment were inserted into PMD18-T plasmid vector and transformed competent DH-5alpha, cDNA libraries of differentially expressed gene between calcification group and regression group were then constructed. Recombinant vectors were analyzed by colony PCR, positive genes were randomly selected for sequencing and analyzed by BLAST. 4 genes were randomly selected for RT-PCR certification combined with semi-quantitative analysis of DNA bands.

RESULTSVDN model of rats were successfully constructed. Concentration of tissue calcium in calcification group (15.34 mg/g +/- 2.51 mg/g) was significantly increased compared to that in control group (5.20 mg/g +/- 0.75 mg/g, P < 0.001), while in comparison with calcification group (15.34 mg/g +/- 2.51 mg/g), calcium in regression group was relatively lower (12.73 mg/g +/- 1.89 mg/g, P < 0.05). 28 up-regulated genes and 22 down-regulated genes were gained through sequencing and BLAST analysis among positive clones. RT-PCR validation indicated that 4 genes such as prdx3 and Ank2 had increasedly expressed in regression group than those in calcification group, the average fold change was 1.7.

CONCLUSIONRat vascular calcification tissue had characteristic of active regression. Genes in relation to pyrophosphoric acid synthesis, glutamate signal peptides, anti-oxidant and ant-apoptosis were up-regulated, at the same time many genes related to ossification and oxidation activity were down-regulated in the process of calcification regression. Increased expression of calcification suppressor genes accompanying decreased expression of calcification promoting genes might be the intrinsic mechanisms which initiated the active regression of calcified tissues.

Animals ; Aorta ; metabolism ; pathology ; Gene Expression Profiling ; Gene Expression Regulation ; Male ; Rats ; Rats, Sprague-Dawley ; Vascular Calcification ; genetics ; physiopathology

OBJECTIVETo evaluate whether human mesenchymal stem cells (hMSCs) administration alter the clinical course of hyperoxia-induced lung injury.

METHODShMSCs were obtained from bone marrow aspirates from healthy donors after informed consent was signed, hMSCs were separated, cultured, amplified, identified and labeled with BrdU. For BrdU labeling, a sterile stock solution was added to the culture medium 48 h before the end of culture, at a final concentration of 10 micromol/L. Thirty-two 3-day old SD rats from four litters were randomly divided into four groups, as hyperoxia exposed + hMSC group (A), air-exposed + hMSC group (B), hyperoxia exposed group (C), and air-exposed group (D). The rats from the group A and the group C were placed in a sealed Plexiglas chamber with a minimal in- and outflow, providing six to seven exchanges per hour of the chamber volume and maintaining O2 levels above 95%, while the rats in the group B and the group D were only exposed to room air. Seven days later, all of them were taken out of the chamber, rats in the group A and B were injected intraperitoneally with hMSCs (1 x 10(5) in 50 microl of PBS) immediately, while the rats in the group C and D were only treated with 50 microl of PBS 3 days later. All the animals were sacrificed by an injection of sodium pentobarbital (120 mg/kg), perfused with cold 0.9% NaCl, and the left lungs were removed, the upper lobes of which were ground as tissue homogenates and used for ELISA, while the inferior lobes were stored at -70 degrees C until use for RT-PCR. The right lungs were fixed in situ for 2 h by the intratracheal instillation with 10% neutral formalin and then postfixed for 24 h. Sagittal sections (4-microm) of paraffin-embedded middle lobe and upper lobe of the right lung were used for immunohistochemistry and histology, respectively.

RESULTS(1) There was a significant difference in the value of RAC (raditive alveoli coant) among the 4 groups (11.145 +/- 1.331, 13.941 +/- 0.985, 9.595 +/- 0.672, 14.819 +/- 1.080, F = 43.234, P = 0.000). RAC in group A and C were significantly reduced compared with subjects in group D (P < 0.05, P < 0.05); and there was also a significant difference between group A and group C (P < 0.05), but not between group B and D subjects (P > 0.05). (2) There were significant differences in the levels of both TNFalpha and TGFbeta(1) in the homogenate of lungs among the 4 groups (142.933 +/- 24.017, 79.033 +/- 11.573, 224.088 +/- 41.915, 76.500 +/- 10.373, F = 59.970, P = 0.000; 1726.484 +/- 91.086, 1530.359 +/- 173.441, 2047.717 +/- 152.057, 1515.777 +/- 131.049, F = 24.977, P = 0.000). The levels of TNFalpha and TGFbeta1 were significantly elevated in both group A and group C when compared with subjects in group D (P < 0.05 for both). Concentrations of TNFalpha and TGFbeta1 were both significantly decreased in group A versus group C (P < 0.05 for both). There was no significant difference between group B and D subjects in the fields of TNFalpha and TGFbeta(1) (P > 0.05 for both). (3) BrdU-labelled cells were observed at alveolar wall and bronchioles in both group A and group B, and there was a significant difference in BrdU-labeled cells between two groups (0.230 +/- 0.026, 0.190 +/- 0.015; t = 3.769, P = 0.002), but none was found in group C and group D. Electrophoresis of the PCR products showed a 224 bp band, specific for Alu mRNA, in 7 of 8 rats of group A and 5 of 8 rats of group B, respectively, but no such band was found in group C and group D.

CONCLUSIONhMSCs administered by intraperitoneal injection could be implanted in the lungs of newborn rats, and they could effectively protect the rats against damage to the lungs caused by hyperoxia.

Animals ; Animals, Newborn ; Bone Marrow Cells ; drug effects ; Bromodeoxyuridine ; pharmacology ; Cell Communication ; Cell Differentiation ; drug effects ; Cells, Cultured ; Hematopoietic Stem Cells ; Humans ; Hyperoxia ; metabolism ; Infant, Newborn ; Lung ; pathology ; Lung Injury ; pathology ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; drug effects ; physiology ; Oxygen ; metabolism ; Pulmonary Alveoli ; pathology ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Transforming Growth Factor beta ; analysis ; Tumor Necrosis Factor-alpha ; analysis

Objective To investigate the prevalence of vitiligo in China through a multi-center and larse-population epidemiological survey.Methods A community-based survey was conducted in 6 cities from 6 provinces.Cluster sampling method was used.Subjects were required to fulfill the self-report questionnaires and received physical examination by dermatologists.EpiData and SPSS11.5 were utilized for statistical analysis. Results Totally,19 974 patients participated in this study,and 17 345 valid questionnaires were retrieved with a return rate of 86.84%.Of them,122 were found to have vitiligo.The prevalence and standardized prevalence of vitiligo was 0.70% and 0.56% in all patients,0.95% (75) and 0.69% in male patients and 0.50% (47) and O.45% in female patients.respectively.A significant elevation was observed in the prevalence of vitiligo in males than in females (P<0.01).The prevalence of vitiligo was increased with age and peaked in patients aging from 60 to 69 years and those over 70 years.The age at onset of vitiligo varied from 0 to 19 years in 21.85% of these patients,from 20 to 49 years in 47.05%.The most connnon type was focal vitiligo,which accounted for 36.06%,while the rarest type wag segmental type (2.46%).The pesitivity rate of family history of vitiligo was 9.84% in patients and 1.31% in community population (P<0.01).About 31.97% of the patients complained of negative influence of vitiligo on quality of life.Conclusions The standardized prevalence of vitiligo is 0.56%in 6 provinces from China.Males seem to have a higher prevalence of vifiligo than females.

Objective:To evaluate the relationship between choline acetyltransferase (ChAT) positive neurons in parabrachial nucleus and development of fear memory in mice.Methods:Eighteen healthy male ChAT-ires-cre mice, aged 8-9 weeks, weighing 22-25 g, were divided into 3 groups ( n=6 each) using a random number table method: Cre-dependent AAV-DIO-hM 3Dq-mcherry (Gq) virus/clozapine-N-oxide (CNO) group (group Gq/CNO), Gq/normal saline (NS) group (group Gq/NS) and Cre-dependent AAV-DIO-mcherry (mc) virus/CNO group (group mc/CNO). Gq virus was injected into parabrachial nucleus, and CNO 2 mg/kg was injected intraperitoneally 3 weeks later in group Gq/CNO.Gq virus was injected into parabrachial nucleus, and the equal volume of normal saline was injected intraperitoneally 3 weeks later in group Gq/NS.In group mc/CNO, mc virus was injected into parabrachial nucleus, and CNO 2 mg/kg was injected intraperitoneally 3 weeks later.The fear conditioning test was performed at 30 min after intraperitoneal injection in all the 3 groups.The brains were then removed and sliced.The virus expression and areas of the brain projected by ChAT positive neurons were observed. Results:Compared with group Gq/CNO, the percentage of freezing time was significantly increased during testing phase in Gq/NS and mc/CNO groups ( P<0.05). Gq/mc virus carrying fluorescent protein mcherry was expressed in parabrachial nucleus and was co-expressed with mcherry-ChAT.The fibers of ChAT positive neurons projected to the red nucleus, substantia nigra, central amygdala, anterodorsal thalamic nucleus and bed nucleus of stria terminalis. Conclusion:The ChAT positive neurons in parabrachial nucleus are involved in the regulation of the development of fear memory in mice, which can impair fear memory, and the regulation is carried out probably through central amygdala.