Objective To establish luciferase-labeled mesenchymal stem cells in vitro.Methods From May,2013 to January,2014,recombinant lentiviral vectors containing luciferase gene was transfected in to mesenchymal stem cell line C3H10T1/2.Puromycin was used to filter the cells.Western-blot and immunofluorescenceimaging were used to detect the transfection efficiency.Results Western blotting analysis showed that the size of this protein in cells expressing about 60KD.Showed that infected C3H10T1/2 cells expressing luciferase protein.I mmunofluorescence,compared with the uninfected control group,virus-infected cells group of about 100％ of the cells were positive for red fluorescence.Show that had built a good stable expression of luciferase C3H10T1/2 cells.Conclusion A monoclonal cell line stably and highly expressing luciferase is obtained with luciferase-labeled mesen chymal stem cells.

Animals ; Extremities ; blood supply ; Ischemia ; metabolism ; Male ; Myocardial Reperfusion Injury ; metabolism ; Nitric Oxide ; blood ; Oxidative Stress ; Rats ; Rats, Wistar

Objective To investigate the effects of heart displacement on hemodynamics during off-pump coronary artery bypass grafting (OP-CABG) while the sites for anastomosis were being exposed. Methods Forty-seven patients of both sexes (36 male, 11 female) aged 50-82 years undergoing OP-CABG were enrolled in the study. Preoperative cardiac function was assessed : class Ⅱ in 22 patients; Ⅲ in 23 and Ⅳ in 2 according to NYHA classification.The mean ejection fraction was 0.55?0.14 before surgery.They received on average 3.2 grafts. Premedication consisted of intramuscular morphine 10 mg, midazolam 3-5 mg and scopolamine 0.3 mg.Before induction of anesthesia ECG and SpO2 were monitored and radial artery was cannulated for continuous direct BP monitoring. Anesthesia was induced with midazolam 0.1 nig?kg-1 , fentanyl 4?g?kg-1 and pancuronium 0.1 mg?g-1 iv.The patients were mechanically ventilated after tracheal intubation and PETCO2 was maintained at about 40 mm Hg. Anesthesia was maintained with isoflurane and 50%-60% N2O in O2 and intermittent intravenous boluses of fentanyl and pancuronium. Swan-Ganz catheter which can continuously monitor mixed venous blood O2 saturation (SvO2) was placed in pulmonary artery via right internal jugular vein. SvO2, cardiac output (CO), BP, pulmonary arterial pressure (PAP) and HR were continuously monitored. Right atrial pressure (RAP) and PAWP were measured intermittently. Cardiac index (CI),stroke index (SI),systemic vascular resistance index (SVRI),PVRI, left and right ventricular work index (LVWI,RVWI) and left and right ventricular stroke work index (LVSWI,RVSWI) were calculated. The hemodynamic parameters were recorded after induction of anesthesia before surgery (T1,baseline),before heart displacement (T2), while anastomosis to anterior descending branch was being made (T3), while anastonosis to right coronary artery or posterior descending branch (T4) and to left circumflex artery or diagonal branch (T5) was being made, after normal heart position was resumed (T6) and at the end of operation (T7). Results While anastomosis to the anterior descending branch was being made (T3) SI and LVSWI significantly decreased as compared with the baseline (P

Traditional mineral Chinese medicine is a characteristic part of Chinese medicine, in the development of traditional Chinese medicine has its unique role. With the development of science and technology and the increase of the medical level, traditional mineral medicine research is gradually thorough and wide-ranging. In recent years, traditional mineral Chinese medicine research mainly includes the physical properties of mineral medicine, structure, chemical composition, pharmacology and treatment mechanism research. The above several aspects of research in the mineral medicine has important practical and theoretical significance. The above several aspects of research status and the problems existing in the research were briefly summarized and reviewed in this paper, and its development were discussed, to provide reference for further studies in the future.
Animals ; Humans ; Medicine, Chinese Traditional ; methods ; Minerals ; chemistry ; pharmacology ; toxicity ; Physical Phenomena

AIM:To investigate the ability of a metal complex ammonium tetrathiomolybdate (ATTM) to re-lease H2 S and its cytoprotective effect on an oxidative injury model .METHODS:Released H2 S was absorbed in a reaction flask from ATTM dissolved in the cell medium .Staining with dichlorodihydrofluorescein diacetate or rhodamine 123 fol-lowed by photofluorography was conducted for the observation of reactive oxygen species ( ROS) and mitochondrial mem-brane potential (ΔΨm) levels, respectively.Cell viability and release of lactate dehydrogenase (LDH) from the cells were measured with commercial kits.RESULTS:Similar to another H2S donor GYY4137, ATTM had an ability to release H2S in the cell medium in a dose-dependent manner .Treatment of human skin HaCaT cells with ATTM at concentrations of 25~400 μmol/L didn’ t significantly alter cell viability .Exposure of the cells to ultraviolet rays or a ROS donor H 2 O2 in-creased the intracellular ROS levels .Treatment with 400 μmol/L H2 O2 significantly reduced the viability of HaCaT cells (P<0.01).However, before the treatment with H2O2, pretreatment with ATTM at 100 and 200 μmol/L markedly pre-vented the H2O2-induced cell injury (P<0.01).In addition, the treatment with H2O2 triggeredΔΨm loss (P<0.01) and LDH release from the cells (P<0.01).Prior to suffering from H2O2 injury, the preconditioning with 200 μmol/L ATTM significantly improved ΔΨm levels ( P<0.05 ) and attenuated LDH release from the cells ( P<0.01 ) .CONCLUSION:ATTM is capable of releasing H 2 S and protecting human skin cells against oxidative injury .

Five compounds (tenuifoliside C, tenuifoliside D, telephiose A, telephiose C and polygalaxanthone III) from polygala tenuifolia wild were incubated together with CYP probe substrate in human liver microsomes to investigate the inhibitory effect towards CYP450 enzyme. Phenacetin (CYP1A2), coumarin (CYP2A6), paclitaxel (CYP2C8), diclofenac (CYP2C9), S-mepheriytoin (CYP2C19), dextromethorphan (CYP2D6), chlorzoxazone (CYP2E1), midazolam (CYP3A) were selected as the isoforfn specific substrate. And the formation of paracetamol, 7-hydroxycoumarin, 6alpha-hydroxy paclitaxel, 4'-hydroxydiclofenac, dextrorphan, 6-hydroxychlorzoxazone, 1'-hydroxymidazolam, 4'-hydroxymephenytoin were detected respectively to measure the effect towards CYP450 by high-pressure liquid chromatography (HPLC). The result shows that five compounds from polygala tenuifolia willd significantly inhibit chlorzoxazone 6-hydroxylation catalyzed by CYP2E1, while showed no effect towards CYP1A2, CYP2A6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A. And IC50 value was 38.73, 54.14, 61.77, 62.22, 50.56 micromol x L(-1), respectively.
Cytochrome P-450 Enzyme System ; metabolism ; Esters ; pharmacology ; Glycosides ; pharmacology ; Humans ; Microsomes, Liver ; drug effects ; enzymology ; Oligosaccharides ; pharmacology ; Polygala ; chemistry ; Xanthones ; pharmacology

To explore a new method for identification of Mongolian patent medicine (MPM) by PCR amplification of specific alleles. Eight kinds of MPM were used to study the identification of "Digeda" raw materials. The total DNA of Lomatogonium rotatum and Corydalis bungeana samples were extracted through modified CTAB method, psbA-trnH sequence was amplified by PCR and sequenced directionally. Specific primer was designed. The DNA of 8 kinds of MPM also was extracted and purified by the commercial DNA purification kits. The rbcL and two pair of specific primers sequences were amplified. The specific amplified products were sequenced in forward directions. All specific sequences were aligned and were analyzed. The results indicated that L rotatum can be identified by specific primers from Digeda-4 Tang, Digeda-8 San, Digeda-4 San, and C. bungeana medicinal materials can be identified by specific primers from Li Dan Ba Wei San, Yi He Ha Ri-12 and A Ga Ri-35. PCR amplification of specific alleles can stably and accurately distinguish raw medicinal materials in MPM.
Alleles ; DNA Primers ; genetics ; DNA, Plant ; genetics ; Medicine, Mongolian Traditional ; Molecular Sequence Data ; Plants, Medicinal ; classification ; genetics ; Polymerase Chain Reaction ; methods

Objective To explore the relationship between obesity phenotypes and adipocytokines in children.Methods Based on the Beijing child and adolescent metabolic syndrome (BCAMS) study,3 508 children (1 788 boys and 1 720 girls) aged 6-18 were recruited.In this study,participants were categorized into four groups:226 cases in general obese group,192 cases in abdominal obese group,1 004 cases in combined obese group and 2 086 cases in non-obese group,according to the sex,age,specific body mass index(BMI),and waist circumference (WC) equal to or greater than the 90th percentile for age and gender of school children in Beijing in 2004.The levels of plasma insulin,serum leptin,resistin and adiponectin were measured by sensitive,specific double-antibody sandwich enzyme-linked immunosorbent assays (ELISA).Analysis of covariance,multivariate linear regression and binary logistic regression analysis were performed.Results There were highest plasma insulin and serum leptin,and lowest adiponectin levels in combined obese group than those in other obese groups and non-obese group and resistin level in abdominal obese group was highest than those in other obese groups or non-obese group.Among subjects with general obesity and conbined obesity,WC was more important factor than BMI for plasma insulin[?(WC)=0.158 P0.05].With covariates adjusted,the odds ratios(OR)and 95% confidence intervals of general obesity,abdominal obesity and combined obesity were 3.46(2.44-4.91),5.41(3.87-7.57) and 10.10(8.26-12.35) for predicting hyperinsulinemia,respectively,5.83(4.02-8.45),7.07(4.97-10.05)and 20.82(16.49-26.28) for hyperleptinaemia,respectively,1.47(1.05-2.07),2.0(1.42-2.80) and 2.66(2.23-3.18) for hypoadiponectinaemia,respectively.Serum resistin was highest in abdominal obesity.Conclusion The levels of adipocytokines in children were correlated with the phenotypes of obesity,especially for abdominal obesity.

Objective To study inflammatory reaction induced by N-protein of severe acute respiratory syndrome(SARS)-coronavirus(CoV)in human alveolar typeⅡepithelial cell(A549). Methods Effects on growth of A549 cell by N-protein of SARS-CoV:activity of A549 cells was determined by thiazylyl blue colorimetry assay at 24,48,72 and 96 h,respectively.Effects on cyto- kine production by A549 cells exposed to N-protein of SARS-CoV:interleukin(IL)-6,IL-10 and transforming growth factor-?1(TGF-?1)concentration in culture supernatant were determined by enzyme-linked immunosorbent assay(ELISA).Effects on mRNA expression of cytokine of A549 cells and matrix metalloproteinases-9(MMP-9)exposed to N-protein of SARS-CoV:total RNA of A549 cells was extracted using Rneasy mini kit;RT-PCR was employed to measure the mRNA expression of IL-6,IL-10,TGF-?1 and MMP-9 semiquantitatively.Results Different concentrations of N-protein could all inhibit the growth of A549 cells(after 48 h)and the inhibition by 20?g/mL pro- tein was the strongest.Compared with the control group(0.737?0.024,0.968?0.007),the A val- ues of experimental groups at 72 h and 96 h(0.672?0.027,0.799?0.092)decreased obviously (P

Objective To search the relationship among blood pressure,blood lipid and fat liver in childrcn with simple obesity.Methods Blood pressure,blood lipid and ultrasound of liver of 40 cases with simple obesity and 20 cases in normal control had been detected.Results there were 11 cases without complication in children with simple obesity,the simple obesity with hypertension complication were 14 cases,and the simple obesity with high blood lipid were 15 cases.The level of total cholestrrol(TC),triglyceride(TG),light density lipoprotein(LDL)and very light density lipoprotein(VLDL)in obesity children were significantly higher than those of control group(P