0.05);Compared to Gram-negative bacilli group of sputum culture,Gram-positive cocci group had significant diffe-rence in the incidence of gastrointestinal dysfunction and microcirculatory disorders(Pa

Background The in vitro culture of retinal vascular endothelial cells is the foundation of experimental study of retinal vascular disease. Shortage of human donor eyeballs is a main limiting for the laboratory work. The culture method of rat-derived vascular endothelial cells has been established. However, this method is not enough effective because of severer cellullar injury. Objective Present study was to establish a simple and high effective method for the culture of vascular endothelial cells in vitro. Methods The retinas from 5 SPF SD rats was digested by 0. 1% collagenase and cultured with explant culture method. 20% fetal bovine serum, vascular endothelial growth factor ( VEGF) , insulin-transferrin-selenium( ITS) were composed into the endothelial cell culture medium, and enough blowing was performed to get the cells and fragments from retinal tissue. The cellular suspension was prepared and cultured consequently on human fibronectin-coated culture flasks. Cultured vascular endothelial cells were identified by anti-von Willebrand staining factor. Results The cells emerged from the tissue mass,and cells and some tissue fragments attached to the wall after 24 hours of seeding. The cells grew to show the fusiform in 4 days and merged together in 5 to 6 days,and a cell monolayer was seen in the 14th day after culture. The endothelial cells showed the positive response for von Willebrand factor. After passage, the merging-growth statue of the cells was regained in 2 hours after culture. Conclusion Use of retinal pieces and collagenase-digestion can get the vascular endothelial cells with better activity in vitro. The culture method based on highly selective endothelial cell culture medium associated to FN adhesion-promoting is helpful for gaining the purified of endothelial cells.

OBJECTIVETo investigate the protective effects of oxymatrine on chronic heart failure induced by isoproterenol (ISO) and to observe its effects on ADMA metabolism pathway in ISO-induced chronic heart failure in rats.

METHODMale Sprague-Dawley rats were given oxymatrine (100,50 mg kg-1) orally for 14 days. Heart failure was induced in rats by subcutaneous injection of isoproterenol (5 mg kg-1 d-1 ) at the 8th day for 1 week. Serum parameters, haemodynamic parameters, Heart weight, and histopathological variables were analysed. Expression of protein levels were measured by Western blot.

RESULTOxymatrine (100,50 mg kg-1) significantly attenuated serum content of cTn I, improved left ventricle systolic and diastolic function and left ventricular remodeling, reduced the ISO-induced myocardial pathological changes compared with ISO group. In addition, oxymatrine (100,50 mg kg-1) significantly reduced serum level of ADMA (P <0. 01), normalize the reduced dimethylarginine dimethylaminohydrolase 2 (DDAH2) expression (P <0. 01) , but had no effect on the isoproterenol-induced upregulated protein arginine methyltransferases 1 expression.

CONCLUSIONOxymatrine could ameliorate the experimental ventricular remodeling in ISO-induced chronic heart failure in rats and the mechanism involved in reducing serum content of ADMA and increased DDAH2 expression.

Alkaloids ; pharmacology ; therapeutic use ; Amidohydrolases ; metabolism ; Animals ; Arginine ; analogs & derivatives ; blood ; metabolism ; Chronic Disease ; Gene Expression Regulation, Enzymologic ; drug effects ; Heart Failure ; drug therapy ; metabolism ; pathology ; physiopathology ; Hemodynamics ; drug effects ; Isoproterenol ; adverse effects ; Male ; Organ Size ; drug effects ; Quinolizines ; pharmacology ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Troponin I ; metabolism

AIMTo observe the relationship between amyloid beta-protein (Abeta) and oxidative stress and the protective role of pituitary adenylate cyclase activating polypeptide (PACAP, PACAP-27) against damage induced by oxidative stress (H2O2) in neurem-2a cells.

METHODSWith cultured neuro-2a cells the cell survival and apoptosis were measured by MTT assay, Hoechest33258 staining, DNA ladder and the percentage of small DNA fragment.

RESULTSConcentration-dependent toxicity was induced with H2O2 treatment for 24 h. The neurotoxicity of H2O2 was increased by about 10 times with cotreatment neurons with amyloid beta-protein fragment 25-35 (Abeta(25-35)). While decrease the percentage of small DNA fragmentation the cell survival was increased with co-treatment with PACAP-27(which were added to the culture everyday). The effect of PACAP was not reversed with antagonist of PACAP receptor, PACAP(6-27).

CONCLUSIONAbeta and H2O2 can promote each other's neurotoxicity. Cultured neurons were protected by PACAP27 from the neurotoxicity of H2O2 but not through the activation of PACAP-27 receptor.

Amyloid beta-Peptides ; toxicity ; Apoptosis ; Cell Survival ; Cells, Cultured ; Humans ; Hydrogen Peroxide ; pharmacology ; Neurons ; cytology ; drug effects ; Oxidative Stress ; Pituitary Adenylate Cyclase-Activating Polypeptide ; pharmacology

OBJECTIVEThis study aims to investigate relationship between daily concentration of PM(10), SO(2), NO(2) and daily mortality due to circulatory system diseases in Chaoyang district, Beijing.

METHODSThe time-series data of daily mortality from circulatory system diseases of registered residents in Chaoyang were obtained from Chaoyang District Center for Disease Control and Prevention. The daily concentration of sulfur dioxide (SO(2)), nitrogen dioxide (NO(2)), and particulate matter (PM(10)) were collected from Beijing Municipal Environmental Monitoring Center. And the routine monitoring meteorological data were collected from Beijing Meteorological Bureau, including daily mean temperature and daily mean relative humidity. The time-series analysis was then conducted to determine the relationship of mortality from circulatory system diseases with daily concentrations of SO(2), NO(2) and PM(10) by using Poisson regression with generalized additive model (GAM).

RESULTSDuring January 2004 to September 2008, the cumulative death number from circulatory system diseases of registered residents in Chaoyang district of Beijing was 19 241, the daily average concentration of SO(2), NO(2), PM(10) was 48.7, 63.9, 146.1 µg/m(3), respectively. The single pollutant model showed an increase of daily concentration of PM(10), SO(2) and NO(2) by 10 µg/m(3) will augment the mortality from circulatory system diseases by 0.20% (95%CI: 0.01% - 0.39%), 0.36% (95%CI: -0.13% - 0.85%) and 0.30% (95%CI: -0.34% - 0.94%), respectively. In the multiple air pollutants models, combinatorial effects of PM(10) and SO(2) still positively correlated with increased mortality from circulatory system diseases (P < 0.05), whereas changes of the concentration of NO(2) had no significant effect on mortality from circulatory system diseases (P > 0.05).

CONCLUSIONOur findings in this study elucidated that changes of the concentration of PM(10) and SO(2) had a positive correlation with daily mortality from circulatory system diseases among the local residents in Chaoyang District, whereas the daily concentration of NO(2) was irrelevant with that.

Aged ; Air Pollutants ; analysis ; Air Pollution ; analysis ; Cardiovascular Diseases ; epidemiology ; mortality ; China ; epidemiology ; Environmental Monitoring ; Female ; Humans ; Male ; Models, Theoretical ; Nitrogen Dioxide ; analysis ; Particulate Matter ; analysis ; Sulfur Dioxide ; analysis ; Survival Rate ; Time Factors

OBJECTIVETo investigate the adjuvant effect of intraoperative and postoperative low-dose ketamine administration to remifentanil consumption in patient-controlled analgesia (PCA) for lower limb fracture.

METHODSA total of 200 patients with lower limb fracture receiving the surgery were randomly divided into 4 groups. In Groups A, B and C, patients received 0.5 mg/kg ketamine infusion under general anesthesia, and ketamine in a dose of 0.1 mg/ kg, 0.05 mg/kg, 0.01 mg/kg per hour continuously for 24 hours after surgery, respectively. The control group (Group D) received an equivalent volume of normal saline only. With 20 microgram/ml remifentanil in normal saline, postoperative PCA was administered with a background infusion at 2 ml/h following 2 ml as a loading dose and 1ml demand dose with a 3-minute lockout period. Remifentanil consumption, 11-point visual analog scale (VAS) scores, global satisfaction score (GSS), and side effects were also recorded by the acute pain service.

RESULTSCumulative PCA remifentanil consumption in Groups A and B were (1378+/-77) microgram and (1531+/-402) microgram, significantly lower than (1807+/-510) microgram and (1838+/-523) microgram in Groups C and D (P<0.01). VAS scores in Groups A and B were significantly lower than those in Groups C and D (P<0.01). In the first 12 hours after operation, GSS was improved (P<0.01). No respiratory depression was observed. No significant difference in side effects was observed among groups.

CONCLUSIONLow-dose ketamine can relieve postoperative pain and moderately decrease remifentanil consumption for PCA, with no obvious side effects of ketamine.

Adult ; Aged ; Analgesia, Patient-Controlled ; Double-Blind Method ; Female ; Fractures, Bone ; surgery ; Humans ; Ketamine ; administration & dosage ; Lower Extremity ; injuries ; Male ; Middle Aged ; Pain, Postoperative ; drug therapy ; Piperidines ; administration & dosage

OBJECTIVETo investigate the multiple iron metabolism-related genes expression, its regulation by iron and the expression correlation among the genes in rat tissues.

METHODSTwo groups (n=30) of Sprague-Dawley female weanling rats were fed with a control diet and an iron deficient diet respectively for 4 weeks. All rats were then sacrificed, and blood and tissue samples were collected. The routine blood examination was performed with a veterinary automatic blood cell analyzer. Elemental iron levels in liver, spleen and serum were determined by atomic absorption spectrophotometry. The mRNA expression of genes was detected by real-time fluorescence quantitative PCR.

RESULTSAfter 4 weeks, the hemoglobin (Hb) level and red blood cell (RBC) count were significantly lower in the iron deficient group compared with those in the control group. The iron levels in liver, spleen and serum in the iron deficient group were significantly lower than those in the control group. In reference to small intestine, the relative expression of each iron-related gene varied in the different tissues. Under the iron deficiency, the expression of these genes changed in a tissue-specific manner. The expression of most of the genes significantly correlated in intestine, spleen and lung, but few correlated in liver, heart and kidney.

CONCLUSIONFindings from our study provides new understandings about the relative expression, regulation by iron and correlation among the mRNA expressions of transferrin receptors 1 and 2, divalent metal transporter 1, ferritin, iron regulation proteins 1 and 2, hereditary hemochromatosis protein, hepcidin, ferroportin 1 and hephaestin in intestine, liver, spleen, kidney, heart, and lung of rat.

Animals ; Ferritins ; blood ; Gene Expression ; Hepcidins ; Iron ; Liver ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the characteristics of 99Tcm-ciprofloxacin and explore its feasibility in early detection of secondary infectious foci of severe acute pancreatitis (SAP).Methods Ciprofloxacin was labeled with 99Tcm.The labeling efficiency and radiochemical purity of 99Tcm-ciprofloxacin were calculated and its biodistribution in normal pigs was measured.The recruited baby pigs were divided into three groups:normal control group (6), non-infected group (6) and infected group (16).370-400 MBq of 99Tcm-ciprofloxacin was injected into each pig intravenously.SPECT scanning was performed at 0.5, 1,2, 3, 4 and 6 h after administration.The differences of 99Tcm-ciprofloxacin uptake among groups were calculated and the tracer activity ratio of lesion-to-background was recorded at each time point.The diagnostic value of 99Tcm-ciprofloxacin SPECT imaging for the dectection of secondary infection of SAP was assessed using histopathological results as the gold standard.Variance analysis and least significant difference test were used to analyze the data.Results Both the labehing efficiency and radiochemical purity of 99Tcm-ciprofloxacin were over 90% within 6 h.Organs with rich blood supply, such as kidney, liver and spleen were the target organs for the accumulation of 99Tcm-ciprofloxacin; while no significant uptake was found in gastrointestinal tract or normal pancreas tissue of SAP.Rapid plasma clearance and renal excretion were observed.In the infected group, the lesion was visualized at 1 h after administration.The highest radioactivity ratio of lesion-to-background (3.36 ± 0.33) was at 3 h after administration, which was significantly higher than that of the other time point ( F =99.570, P ＜0.001 ).The sensitivity, specificity, positive and negative predictive values, Youden's index (YI) and Kappa value of 99Tc%ciprofloxacin imaging were 88.2% (15/17), 83.3% (5/6), 93.8% ( 15/16), 71.4% (5/7), 0.715 and 0.667 respectively.Conclusions The biodistribution of99Tcm-ciprofloxacin is suitable for imaging infectious focus of SAP.The optimal imaging time for the detection of secondary infection of SAP is 3 h after administration, with high sensitivity and specificity.

Two female patients with rectal tumor undergoing proctectomy via vagina, namely natural orifice transluminal endoscopic surgery (NOTES), are reported. The operations were performed on June 8 and August 10, 2010, respectively. No Trocar was used in the abdomen except for the transumbilical incision. There were no visible scars in the abdomen. Tubulovillous adenoma and moderately differentiated adenocarcinoma were diagnosed respectively through postoperative pathological examination. Both patients resumed normal work and life at the most recent follow up. Sexual life was satisfactory.
Adult ; Female ; Humans ; Natural Orifice Endoscopic Surgery ; methods ; Rectal Neoplasms ; surgery ; Vagina ; surgery

OBJECTIVETo preliminarily investigate the long-term structural and functional injuries of mitochondria in rat brain caused by sepsis.

METHODSWistar rats were randomly assigned into sepsis and control groups. A rat model of sepsis was prepared by an intraperitoneal injection of 10 mg/kg lipopolysaccharide (LPS) of gram-negative bacteria, and the survival assay was performed. Eight rats in the sepsis group were sacrificed at 12, 24, 48, or 72 hours after LPS injection, while rats in the control group were sacrificed after an intraperitoneal injection of an equal volume of normal saline. Mitochondria were extracted from rat brain tissue. Mitochondrial membrane potential (MMP) and mitochondrial swelling level were determined by flow cytometry, and the activities of electron transport chain complexes (I-V) were measured using enzyme assay kits. Hematoxylin-eosin (HE) staining and electron microscopy were used to observe morphological changes in brain tissue and mitochondria.

RESULTSThe sepsis group had a significantly lower survival rate than the control group (P<0.01). The MMP and activities of electron transport chain complexes (I-V) in the sepsis group, which were significantly lower than those in the control group (P<0.05), were reduced to the lowest levels at 48 hours and partially recovered at 72 hours. The mitochondrial swelling level in the sepsis group, which was significantly higher than that in the control group (P<0.05), increased to the peak level at 48 hours and partially recovered at 72 hours. Hematoxylin and Eosin staining revealed substantial damages in the structure of brain tissue, and electron microscopy showed mitochondrial swelling, and vacuolization in a few mitochondria.

CONCLUSIONSIn the rat model of LPS-induced sepsis, both structural and functional injuries are found in cerebral mitochondria, and achieve the peak levels probably at around 48 hours.

Animals ; Brain ; pathology ; physiopathology ; ultrastructure ; Lipopolysaccharides ; toxicity ; Male ; Membrane Potential, Mitochondrial ; Mitochondria ; physiology ; ultrastructure ; Rats ; Rats, Wistar ; Sepsis ; chemically induced ; mortality ; physiopathology