Objective To observe the effects of five flavonoids include rutin(RU),dihydromyricetin(DMY),hesperetin(HP),daidzein(DA)and hydroxysaffor yellow A(HYSA)on myocardiocyte apoptosis induced by H2O2 and to explore their relationships with Keshan disease and the possible mechanism.Methods Primary cultured cardiocytes of neonatal rats were randomly divided into control group,model group,and flavonoids preincubation group.The cardiocyte apoptosis was examined by fluorescent staining,the rates of apoptosis were detected by flow cytometry,the expression of Bcl-2 family proteins associated with apoptosis were observed:by Western blot.Results Compared with model group[(24.33±6.51)%],RU[(13.95±3.80)%],DA[(11.82±3.50)%],HYSA[(12.33±3.78)%]could decreased the rate of apoptosis(P＜0.05).The five flavonoids could up-regulate Bcl-2 expression,down-regulate Bax expression,and increase the Bcl-2/Bax ratio[RU(0.989±0.094),DMY(0.931±0.280),HP(0.980±0.095),DA(1.049±0.092),HYSA(1.031±0.039),vs model(0.490±0.046),the difference had statistical significances(P＜0.05)],but the Bcl-xl did not significantly changed(P＞0.05).Conclusions RU,DMY,HP,DA and HYSA have antiapoptotic effects on cardiomyocyte via regulating Bcl-2 and Bax,which gives us a hint in prevention and treatment of Keshan disease.

OBJECTIVERecent studies have found that the variation of G894T on the region of T786C and 7th exon promoted by endothelial nitric oxide synthase (eNOS) gene is associated with cardiovascular disease. This research explored possible correlations between eNOS gene polymorphisms and orthostatic intolerance (OI) in children through linkage disequilibrium analysis between eNOS genes T786C and G894T and OI.

METHODSPCR, Macrorestriction Map and other molecular biotechnology were used to determine the genotypes of eNOS/T786C and G894T in 60 OI probands and their parents. Correlation analysis and transmission disequilibrium test (TDT) between T786C, G894T and OI were performed.

RESULTSThere was linkage disequilibrium of eNOS/T786C and G894T gene polymorphisms in the occurrence of childhood OI (P<0.05).

CONCLUSIONSeNOS genes T786C and G894T may be associated with the pathogenesis of OI.

Adult ; Child ; Female ; Humans ; Linkage Disequilibrium ; Male ; Nitric Oxide Synthase Type III ; genetics ; Orthostatic Intolerance ; genetics ; Polymorphism, Genetic

In order to clarify the chemical constituents of Si-Wu Decoction rapidly and holistically, we analyzed the ethanol extract of Si-Wu Decoction by UPLC/Q-TOF-MSE and UNIFI which based on traditional Chinese medicine database, the probable structures of 113 compounds were identified. The results show that this method can rapidly and effectively characterize the chemical compounds of Si-Wu Decoction and provide a new solution for identification of components from complex TCM extract.
Chromatography, High Pressure Liquid ; Databases, Pharmaceutical ; Drugs, Chinese Herbal ; chemistry ; Medicine, Chinese Traditional

BACKGROUNDThe results of clinical trials of rapamycin-eluting stents reduce restenosis have been quite promising. The main purpose of this study was to characterize the in vivo pharmacokinetics of high dose rapamycin (Rapa)-eluting stents in a miniswine coronary model.

METHODSTen miniswines underwent placement of 18 high dose Rapa-eluting stents in the left anterior descending and right coronary arteries. At the planned times of the 1.5th, 12th, 24th hour, 3th, 7th and 28th day, the animals (n = 1, 1, 2, 2, 2, and 2, respectively) were euthanized after completion of coronary angiography. Blood samples were obtained at 0, 10, 20, 30 minutes; 1, 2, 6, 24 hours; and 3, 7, 28 days to determine systemic Rapa levels. Rapa levels in whole blood, arterial wall, heart, renal and liver tissues were determined by high-performance liquid chromatography/mass spectroscopy.

RESULTSPeak whole blood concentration (Cmax), time to peak concentration (tmax), elimination half-life (t1/2beta), area under the curve (AUC), and apparent systemic clearance (Cl/F) were (10.91 +/- 1.28) ng/ml, (2.0 +/- 0.2) hours, (7.25 +/- 0.63) hours, (1.15 +/- 0.11) ng x h x ml(-1), and (180 +/- 12) ml x h(-1) x kg(-1), respectively. More than 95% Rapa detected is localized in the coronary artery surrounding the stent and heart.

CONCLUSIONStent-based delivery of Rapa via a copolymer stent is feasible and safe. This strategy holds promise for the prevention of stent restenosis.

Animals ; Chromatography, High Pressure Liquid ; Coronary Restenosis ; prevention & control ; Male ; Mass Spectrometry ; Sirolimus ; administration & dosage ; pharmacokinetics ; Stents ; Swine ; Swine, Miniature ; Tissue Distribution

AIMTo determine the mechanisms of glucocorticoids in inhibiting advanced prostate cancer growth.

METHODSThe cell proliferation and cell cycle of prostate cancer DU145 cells following dexamethasone treatment were determined by proliferation assay and fluorescence-activated cell sorter. Western blot analysis was carried out to evaluate the effects of dexamethasone on phosphorylation of extracellular signal-regulated kinase (ERK)1/2 and expression of cyclin D1 in DU145 cells with or without glucocorticoid receptor (GR) antagonist RU486. Reverse transcription-polymerase chain reaction verified the expression of GR mRNA in DU145 cells.

RESULTSDexamethasone significantly inhibited DU145 cell proliferation at the G(0)/G(1) phase. Western blot analysis showed a dramatic reduction of ERK1/2 activity and cyclin D1 expression in dexamethasone-treated cells. The decreased phosphorylation of ERK1/2 in dexamethasone-treated cells was attenuated by GR blockade. Additionally, the effects of dexamethasone in inhibiting cyclin D1 expression were altered by GR blockade.

CONCLUSIONDexamethasone suppresses DU145 cell proliferation and cell cycle, and the underlying mechanisms are through the inhibition of phosphorylation of ERK1/2 and cyclin D1 expression. The inhibition of ERK1/2 phosphorylation and cyclin D1 expression is attenuated by GR blockade, suggesting that GR regulates ERK1/2 and cyclin D1 pathways. These observations suggest that dexamethasone has a potential clinical application in prostate cancer therapy.

Antineoplastic Agents, Hormonal ; pharmacology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin D1 ; drug effects ; metabolism ; Dexamethasone ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Male ; Mitogen-Activated Protein Kinase 1 ; drug effects ; metabolism ; Mitogen-Activated Protein Kinase 3 ; drug effects ; metabolism ; Prostatic Neoplasms ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Receptors, Glucocorticoid ; metabolism ; Signal Transduction ; drug effects

Adaptor Proteins, Signal Transducing ; genetics ; B-Cell CLL-Lymphoma 10 Protein ; Chromosomes, Human, Pair 11 ; genetics ; Chromosomes, Human, Pair 18 ; genetics ; Gene Rearrangement, B-Lymphocyte ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Lymphoma, B-Cell, Marginal Zone ; genetics ; Paraffin Embedding ; Translocation, Genetic

Objective By sequenceing the Cj1136,Cj1138 and Cj1139 gene of Campylobacter jejuni(C. Jejuni) strains associated with Guillain-Barre Syndrome(GBS),features of Cj1136,Cj1138 and Cj1139 gene were studied.Results were compared with the C.jejuni strain NCTC11168, to find the mutations in sequence of C.jejuni which inducing GBS and their polygenetic relationship was analyzed.Methotis Three GBS-associated C.jejuni strains were isolated from stools of GBS patients from Hebei province who had been diagnosed as clinical AMAN pattern and electrophysiological tests were performed.After distilling and sequencing Cj1136,Cj1138 and Cj1139 genes,results were spliced and assembled into a complete sequence by the terminals overlapped with each other.Sequences of Cj1136,Cj1138 and Cj1139 genes were compared with NCTC11168,to find the mutations and gene feature.Results The Cj1136,Cj1138 and Cj1139 gene of the three GBS-associated C.jejuni strains were composed by 1173 base pairs,1170 base pairs,912 base pairs respectively. The alignment with the related sequence of NCTC11168 showed that there were two same mutations in the Cj1138 gene of the three C.jejuni stains.Data from phylogenetic analysis demonstrated that the three C.jejuni strains were genetically closed to NCTC11168,with the biggest phylogenetic distance between the three of them as 2.1%.Conclusion When compared with NCTC11168 the Cj1138 gene of the three GBS-associated C.jejuni strains had the same mutations which might be related to the development of GBS.Relation between the variation and GBS-pathogenesis remained to be confirmed.The mutations found in the three C.jejuni strains established the foundation for exploring the biological characteristics of GBS-associated C.jejuni strains and demonstrated that the GBS-associated C.jejuni strains of Hebei province having its regional features.

Acute respiratory distress syndrome (ARDS) is severe respiratory failure in clinical practice, with a mortality rate as high as 40%. Injury of pulmonary endothelial cells and alveolar epithelial cells occurs during ARDS, and pulmonary endothelial injury results in endothelial barrier disruption, which usually occurs before epithelial injury. Especially, when harmful factors enter the blood, such as sepsis and hemorrhagic shock, the pulmonary endothelial cells are affected firstly. The injured endothelial cells may loss cell-to-cell connections and even die. After the endothelial barrier is disrupted, fluid and proteins cross the endothelial barrier, causing interstitial edema. The alveolar epithelium is more resistant to injury, and when the tight barrier of the epithelium is broken, fluids, proteins, neutrophils, and red blood cells in the interstitium enter the alveolar space. From this process, it is easy to find that the endothelium is the first barrier to prevent edema, therefore, the protection of endothelium is the key to the prevention and treatment of ARDS. In addition, the injured endothelial cells express selectin and cell adhesion molecules, promoting the recruitment of immune cells, which exacerbate the inflammatory response and pulmonary endothelial cell injury. Mesenchymal stem cells (MSCs) can be derived from umbilical cord, bone marrow, adipose and so on. Because of low immunogenicity, MSCs can be used for allogeneic transplantation and have great application potential in tissue repairing. Through paracrine effect, MSCs can promote cell survival and balance inflammatory response. MSCs infused intravenously can locate in lungs rapidly and interact with endothelial cells directly, thus MSCs have advantages in protecting pulmonary microvascular endothelial cells. Animal experiments and clinical trials have found that MSC transplantation can significantly improve the symptoms of ARDS and reduce inflammatory reactions and endothelial permeability. Mechanically, MSCs acts mainly through paracrine and immunomodulatory effects. Paracrine cytokines from MSCs can not only promote pulmonary endothelial proliferation, but also reduce inflammatory response and promote cell survival to maintain endothelial integrity. In addition to paracrine cytokines, extracellular vesicles of MSCs are rich in RNAs, proteins and bioactive substances, which can protect pulmonary endothelial cells by intercellular communication and substance transport. Furthermore, MSCs may protect pulmonary endothelial cells indirectly by regulating immune cells, such as reducing the formation of extracellular trapping network of neutrophils, regulating macrophage polarization and regulating Th17/Treg cell balance. Although the beneficial effects of MSCs are verified, much work still needs to be done. MSCs from different tissues have their own characteristics and the scope of application. Different lung diseases possess different endothelial injury mechanisms. Thus, determining the indications of MSCs derived from different tissues is the direction of pulmonary disease clinical trials. From the perspective of transplantation route, intravenous injection of MSCs may have better clinical application in pulmonary endothelial injury caused by endogenous harmful factors in blood. Previous reviews mostly focused on the protective effects of MSCs on alveolar epithelium. In this article, we focused on endothelial cells and reviewed the direct protective effects and mechanisms of MSCs on endothelium through paracrine cytokines and extracellular vesicles, and summarize the mechanisms by which MSCs may indirectly protect pulmonary endothelial cells by regulating immune cells.

OBJECTIVETo evaluate the clinical function and significance of establishing a regional active neonatal transport network (ANTN) in Beijing.

METHODThe authors retrospectively studied intensive care and the role of ANTN system in management of critically ill neonates and compared the outcome of newborn infants transported to our NICU before and after we established standardized NICU and ANTN system (phase 1: July 2004 to June 2006 vs phase 2: July 2006 to May 2008).

RESULTThe number of neonatal transport significantly increased from 587 during phase 1 to 2797 during phase 2. Success rate of transport and the total cure rate in phase 2 were 97.85% and 91.99% respectively, which were significantly higher than those in phase 1 (94.36% and 88.69%, respectively, P < 0.01). The neonatal mortality significantly decreased in phase 2 compared with that in phase 1 (2.29% vs 4.31%, P < 0.01). The capacity of our NICU was enlarged following the development of ANTN. There are 200 beds for level 3 infants in phase 2, but there were only 20 beds in phase 1. Significantly less patients in the phase 2 had hypothermia, acidosis and the blood glucose instability than those in phase 1 (P < 0.01, 0.05, 0.01 and 0.05, respectively). The proportion of preterm infants transported to our NICU were higher in phase 2 compared with that in phase 1, especially infants whose gestational age was below 32 weeks. The proportions of asphyxia and respiratory distress syndrome were lower in phase 2 than that in phase 1, but the total cure rates of these two diseases had no significant changes between the two phases. The most important finding was that the improvement of outcome of premature infants and those with asphyxia and aspiration syndrome was noted following the development of ANTN.

CONCLUSIONEstablishing regional ANTN for a tertiary hospital is very important to elevate the total level in management of critically ill newborn infants. It plays a very important role in reducing mortality and improving total outcomes of newborn infants. There are still some problems remained to solve after four years practice in order to optimize the ANTN to meet needs of the development of neonatology.

Humans ; Infant Mortality ; Infant, Newborn ; Infant, Premature ; Intensive Care Units, Neonatal ; standards ; Transportation of Patients ; standards

OBJECTIVEThrough systematic monitoring of the number and strain types of O1 and O139 Vibrio cholerae in the Pearl River estuary waters to analyze it's relevance with the temperature of environment, and the relevance between strains in water and isolates during outbreaks and epidemics as well as to estimate the methods used for environmental water detection and the potential role in cholera surveillance program.

METHODSTwenty-four stations along the Pearl River were selected and the water samples were collected monthly from March 2006 to February 2007. V. cholerae O1 and O139 strains were isolated from the samples. Real-time PCR established in our laboratory was used to detect V. cholerae O1 and O139. Air temperature and water temperature were collected during sampling. Pulsed field gel electrophoresis (PFGE) was applied in molecular typing of the isolates.

RESULTS862 water samples were collected during the study period. A total number of 77 O1 and O139 V. cholerae were isolated in 67 water samples and the positive rates were 7.77% for isolation and 26.33% for real-time PCR. Seasonal trend of positive rates by month were approximately coincident with the change of water temperature. The positive rates in the stations in urban area were higher than those in other areas. Toxigenic O139 strains were found in one station located in downstream of a marine market. Most of the O1 and O139 isolates were non-toxigenic. No trend of seasonal variation of the strains was noticed. Within these 75 isolates, 49 PFGE patterns were identified and the patterns differed widely with the similarity of 57.4% - 100%.

CONCLUSIONV. cholerae existed as the natural habitat in estuary water of the Pearl River and showed obvious genetic diversity. Data from monitoring waters might show the separation of strains with certain seasonal association. But the crowd did not show the relationship between the infections. Results from water surveillance program might provide indicators on the appearance of cholera pathogen which might be used in assessing the environmental risk of cholera epidemics as well as the alert of cholera.

Electrophoresis, Gel, Pulsed-Field ; Environmental Monitoring ; Phylogeny ; Polymerase Chain Reaction ; Seasons ; Temperature ; Vibrio cholerae O1 ; classification ; genetics ; isolation & purification ; Vibrio cholerae O139 ; classification ; genetics ; isolation & purification