AIM: To investigate the synergistic effect of resveratrol and 5-fluorouracil on osteosarcoma CD133+ cell subset.METHODS: Human osteosarcoma cell line MG-63 CD133+ cell subset and the corresponding CD133-cell subset were treated with resveratrol and 5-fluorouracil.After treatment, the viability of MG-63 cells was measured by MTT assay.The apoptosis of MG-63 cells was analyzed by flow cytometry.Activation of caspase-9 and caspase-3, the expression of Apaf-1, and the release of cytochrome C were evaluated by Western blot.The interaction between Apaf-1 and pro-caspase-9 was detected by co-immunoprecipitation.RESULTS: The cell death and apoptosis of MG-63 CD133+ cell subset induced by 5-fluorouracil were significantly weaker than those in the corresponding MG-63 CD133-cell subset.However, co-treatment with resveratrol significantly enhanced the effect of 5-fluorouracil on inhibiting the viability of MG-63 CD133+ cell subset.Mechanically, treatment with resveratrol upregulated the expression of Apaf-1.Transfection with Apaf-1 siRNA abolished the synergistic effect of resveratrol and 5-fluorouracil in MG-63 CD133+ cell subset.In addition, the results of co-immunoprecipitation indicated that the combination of resveratrol and 5-fluorouracil significantly induced the formation of Apaf-1/pro-caspase-9 complex, leading to the activation of caspase-9 in MG-63 CD133+ cell subset.CONCLUSION: Resveratrol enhances 5-fluorouracil-induced apoptosis of osteosarcoma CD133+ cell subset by promoting the formation of Apaf-1/caspase-9 complex.

Ebola virus(EBOV)and Marburg virus(MARV),belonging to the Filoviridae family,emerged four decades ago and caused severe viral hemorrhagic fever in human and other primates.As high as 50-90% mortality,filoviruses can cause significant threats to public health.However,so far no specific and efficient vaccine has been available,nor have other treatment methods proved to be effective.It is of great importance to detect these pathogens specific,rapidly and sensitively in order to control future filovirus outbreaks.Here,recent progresses in the development of detection and diagnosis methods for EBOV and MARV are summarized.

Objective To study the incidence rate and risk factors of catheter-related infection (CRI) in preemie with peripherally inserted central catheter (PICC),in order to facilitate the implementation of control strategies and control infection.Methods Two hundred and fifty-eight PICC preemie of birth weight less than 2.5 kg,according to whether the incidence of CRI were divided into CRI group and non CRI group,two groups of gestational age,birth weight,PICC time,parenteral nutrition time and PICC parenteral nutrition time were statistically analyzed and compared different indwelling time of CRI incidence.Results Diagnosis of CRI in 24 cases,the occurrence rate of 9.3％ (24/258).There were significant differences in gestational age,birth weight,PICC time,parenteral nutrition time,PICC parenteral nutrition time between CRI group aad non CRI group (29.5 weeks vs.33.0 weeks,1.4 kg vs.1.8 kg,31.5 days vs.14.6 days,40.1 days vs.16.7 days,28.8 days vs.13.4 days,P ＜ 0.01 or ＜ 0.05).There was significant difference in incidence rate of CRI among different indwelling time (P ＜ 0.01).The risk factors of CRI including gestational age (P =0.007),birth weight (P=0.000),PICC time (P=0.001),parenteral nutrition time (P=0.035),PICC parenteral nutrition time (P =0.001).Multivariate Logistic regression analysis showed the greatest impact on the 3 factors were birth weight,parenteral nutrition time,gestational age.Conclusion Gestational age,birth weight,PICC time,parenteral nutrition time are the risk factors of CRI in preemie with PICC.

Objective To explore the oxidative damage and genotoxicity of organic chemical pollutants from running water and the source water in Baotou reach of the Yellow River to mice. Methods The Kunming mice were treated with the organic extract solutions of the Yellow River water and running water by gavage at different concentration,once a day and the activity of the superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and the content of malondialdehyde (MDA) in the liver and kidney and the rate of micronucleus were determined. Results SOD and GSH-Px activity in the liver and kidney in the groups treated with the water of three sections of Yellow River in Baotou City decreased as the exposure dose increased. Except the low concentration group of running water,the MDA content in the liver and kidney and the micronucleus rate increased significantly compared with the control (P

OBJECTIVE:To study the vasodilatation effects of the alcohol extract from Uighur medicine Ziziphora clinopodioi-des(EEZ)on isolated rat thoracic aorta vessel rings. METHODS:Isolated rat thoracic aorta vessel rings were prepared. There was a group with intact vessel ring endothelium and a group with vessel ring endothelium removed in the test. After preshrinking the ves-sel rings with phenylephrine(PE),EEZ of 100,300,500,700,900 and 1 100 mg/L was added gradually,concentration-vasodila-tation curve was drawn and maximal vasodilatation rate (Emax) and median effective concentration (EC50) were calculated. For the group with vessel ring endothelium removed,after the vessel ring was pretreated with EEZ at EC50 in calcium-free solution or calci-um-free high-potassium solution,CaCl2 of 0.4,0.8,1.2,1.6,2.0 and 2.4 mmol/L was added gradually,and calcium concentra-tion-tension curve was drawn;following the pretreatment of the vessel ring with EEZ at EC50,1 μmol/L PE was added and shrink tension was recorded and vasoconstriction percentages was calculated. RESULTS:EEZ had vasodilatation effect on the vessel ring preshrunk with PE in a concentration and smooth muscle-dependent manner. The group with intact vessel ring endothelium and the group with vessel ring endothelium removed respectively had Emax of (58.18 ± 16.23)% and (73.54 ± 17.21)%,and EEZ EC50 of 773.27 mg/L. For calcium-free high-potassium solution,EEZ could cause the calcium ion-vasoconstriction curve to move to the right obviously;for calcium-free solution,EEZ could inhibit the vasoconstriction caused by PE. CONCLUSIONS:EEZ has vasodi-latation effects by a mechanism which may be related to inhibiting calcium influx and intracytoplasmic calcium release through the inhibition of voltage-dependent calcium channels(VDCCs),and thus interfering intracytoplasmic calcium ion balance.

OBJECTIVE:To establish a method for the simultaneous determination of baicalin,baicalein and wogonin in Hua-tan pingchuan tablet. METHODS:HPLC was performed on the column was Diamonsil C18 with mobile phase of methanol-0.1%phosphoric acid(gradient elution)at a flow rate of 1.0 ml/min,detection wavelength was 277 nm,column temperature was 30 ℃, and the injection volume was 10 μl. RESULTS:The linear range was 0.069 04-0.690 4 μg for baicalin(r=0.999 4),0.054 56-0.5456 μg for baicalein(r=0.999 6)and 0.030 36-0.303 6 μg for wogonin(r=0.999 1);RSDs of precision,stability and reproduc-ibility tests were lower than 2%;recoveries were 97.64%-99.06%(RSD=0.57%,n=6),98.31%-100.64%(RSD=0.85%,n=6) and 97.53%-99.48%(RSD=0.76%,n=6). CONCLUSIONS:The method is simple,stable and reproducible,and can be used for the contents determination of baicalin,baicalein and wogonin in Huatan pingchuan tablet.

Objective To investigate the relationship between the polymorphism of adiponectin -11 ,377C> G gene and the risk of coronary heart disease(CHD). Methods A total of 126 CHD patients and 130 healthy controls were enrolled and the frequency of each genotypes and allele gene of adiponectin -11 ,377C > G were detected by polymerase chain reaction fragment length polymorphism (PCR-RFLP). Results (1) The adiponectin gene -11,377C > G sites existed gene polymorphism and the three genotypes were GG, CG and CC. (2) There was statistical difference between CHD group and control group; The G allele frequency of CHD group was significantly higher than that in control group (P < 0.05); The frequency of the C allele gene in CHD group was significantly decreased (P < 0.05). (3) There was no statistical difference of frequency distribution of each genotype and allele gene of adiponectin -11,377C > G between acute coronary syndrome (ACS) group and stable angina group . ( 4 ) The risk of CHD were increased in CHD patients with G allele gene of adiponectin-11,377C > G (P < 0.05). Conclusions The polymorphism of adiponectin -11,377C > G is associated with the increased risk of CHD. The increased G allele gene frequency may represent the increased risk of CHD.

Objective To study the effects of high concentration oxygen exposure on the Sox17 expression of vascular endothelial cells of neonatal mice lungs,and to explore the pathogenesis of blocked lung vascular development.Methods Thirty two C57B1/6J newborn mice within six hours after birth were randomly divided to hyperoxia group (n=16) and room air group (n=16).Mice of hyperoxia group were exposed to 85％ oxygen.Eight mice of either group were sacrificed at 7 and 14 days after birth respectively to observe the lung morphology and calculate radial alveolar counts (RAC),which is the number of alveoli on the straight line from the center of respiratory bronchioles to the nearest fibrous septa or the pleura.Sox 17 expression in the pulmonary vessels was detected by immunohistochemical staining.Sox17 mRNA was measured by reverse transcription polymerase chain reaction.Sox17 protein level was measured by Western blot.Two independent samples t-test was used for statistical analysis.Results Compared with day 7,the lung structures matured with more uniformed alveoli and the septas became thinner on day t4 in room air group.However,the lungs developed slowly with simplified and non-uniformed alveoli on day 14 in hyperoxia group.The Sox17 protein was positive on endothelial cells of pulmonary arteries,veins and alveolar capillarys,as well as the alveolar epithelial cells.The RAC on day 7 and day 14 in hyperoxia group were both lower than that in room air group (3.7±0.7 vs 5.0±0.8,5.3±0.6 vs 8.3±0.9,respectively,t=3.057 and 8.148,both P ＜ 0.01).Sox17 mRNA on day 7 and day 14 in hyperoxia group were both lower than that in room air group (0.62±0.10 vs 0.88±0.11,0.44±0.06vs 0.90±0.15,t=3.607 and 6.926,both P ＜ 0.01).Sox17 protein level on day 7 and day 14 in hyperoxia group were both lowered than that in room air group (0.32±0.04 vs 0.76±0.04,0.36±0.07 vs 0.96±0.06,t=3.102 and 8.421,both P ＜ 0.01).Conclusions Exposure of high concentration of oxygen may cause impairment of lung vascular development by inhibiting Sox17 expression in lungs of neonatal mice.

Objective To investigate the prevalence and main genotypes of carbapenemases in carbepenem‐resistant Enterobacteriaceae (CRE) .Methods A total of 114 strains of CRE were isolated in Shanghai Ruijin Hospital from May 2011 to June 2013 .The diameter of inhibition zone of imipemen or meropenem for these strains was not larger than 22 mm .PCR method was used to screen for the main carbapenemase genes (blaKPC ,blaIMP ,blaVIM ,blaOXA‐48 and blaNDM ) with previously described primers followed by nucleotide sequencing analysis . Conjugation experiments were performed to examine the transferability of plasmids .Pulsed‐field gel electrophoresis (PFGE) was used to show the relatedness of KPC‐2‐producing Enterobacteriaceae .Results Most of the 114 isolates were K lebsiella pneumoniae and Escherichia coli .Of the 114 isolates ,98 was positive for carbapenemases ,specifically ,78 blaKPC‐2‐positive ,15 blaIMP‐4‐positive ,2 blaIMP‐8‐positive ,1 positive for both blaKPC‐2 and blaIMP‐4 and 4 blaNDM‐1‐positive .None of the strains was positive for blaOXA‐48 or blaVIM .About 21 .4% (21/98) of the isolates were conjugated successfully .The 49 blaKPC‐2‐positive K .pneumoniae isolates were grouped into 12 types according to PFGE patterns .Majority (34/49) of these isolates belonged to the same type A .Conclusions BlaKPC‐2 was the primary epidemic genotype of Enterobacteriaceae in Ruijin Hospital ,followed by blaIMP‐4 .NDM‐1 carbapenemase was produced in 4 strains of CRE . Meanwhile , clonal spread of KPC‐2‐producing K . pneumoniae was observed in some departments of our hospital , such as surgical ICU , respiratory medicine and thoracic surgery . Appropriate measures should be taken timely and effectively to prevent the in‐hospital spread of resistant genes .

Objective To investigate the relaxant effects of ethanol extract fromRhodiola crenulata (Hook.f.et Thoms.) H.Ohba in isolated rat thoracic aorta and its possible mechanisms.MethodsThe thoracic aortas from male Sprague-Dawley rats were isolated and cut into 3- to 4-mm-wide transverse rings, and the tension in endothelium-intact or endothelium-denuded aortic rings was recorded. The relaxant effects of ethanol extract fromRhodiola crenulata in various concentrations (100, 300, 500, 700, 900, and 1 100 mg/L) in aortic rings precontracted with phenylephrine (1 μmol/L) were observed. The effects of pretreatment with soluble guanylyl cyclase inhibitor ODQ (10 μmol/L) on the relaxant effects of ethanol extract fromRhodiola crenulata were observed. The effects of ethanol extract fromRhodiola crenulataon calcium-induced contractions in calcium-free high-potassium medium and phenylephrine-induced contractions in calcium-free medium were also observed.ResultsEthanol extract fromRhodiola crenulatashowed a concentration-dependent relaxation in aortic rings precontracted with phenylephrine. The maximal relaxations of ethanol extract fromRhodiola crenulataln endothelium-intact or endothelium-denuded aortic rings were 57.4% ± 21.81% and 60.51% ± 0.34%, respectively, with a half-maximal effective concentration of 1 062.88 mg/L. The relaxant effects of ethanol extract fromRhodiola crenulatawere not statistically inhibited by pretreatment with ODQ (P>0.05). Contractions of aortic rings resulted from phenylephrine-induced Ca2+ release from intracellular stores in calcium-free medium or high potassium-induced influx of Ca2+ in calcium-free high-potassium medium were significantly inhibited by ethanol extract fromRhodiola crenulata(allP<0.05).ConclusionEthanol extract fromRhodiola crenulatahas a concentration-dependent vasorelaxation, its mechanisms may be involved in blocking extracellular Ca2+ influx and intracellular Ca2+ release.