1.Therapeutic effects of combined vitamin K2 with arsenic trioxide on proliferation of HL-60 cells
Yan, QU ; Lan, CHEN ; Li-hong, REN ; Hui, QU
Chinese Journal of Endemiology 2013;(3):258-262
Objective To investigate the therapeutic effects of arsenic trioxide(ATO) plus vitamin K2(VK2) on proliferation of HL-60 cells from acute promyelocytic leukemia cell line and explore the possible mechanism.Methods ①HL-60 cells were exposed to ATO(0.0,0.5,1.0,2.0,4.0 μmol/L),VK2(0.0,2.5,5.0,10.0,20.0μmol/L),or both of different concentrations (0.5 μmol/L ATO + 2.5 μmol/L VK2,1.0 μmol/L ATO + 5.0μmol/L VK2,2.0 μmol/L ATO + 10.0 μmol/L VK2,4.0 μmol/L ATO + 20.0 μmol/L VK2) for 24,48 or 72 h,respectively.The method of CCK-8 was used to assess the proliferation of HL-60 cells and the half inhibitory concentration(IC50) of ATO or VK2 was calculated,respectively.②Combination index (CI) was used to evaluate the combinative effect of the two treatments:CI < 1,=1 or > 1 indicated synergistic,additive,or antagonistic effect,respectively.③After HL-60 cells were treated with 1.0 μmol/L ATO or 5.0 μmol/L VK2 individually or simultaneously for 48 h,Annnexin V/PI staining was performed to identify the apoptosis rate of each group.Untreated cells were used as control group.Results ①ATO or VK2 alone inhibited the proliferation of HL-60 cells in a concentration and time dependent manner.The IC50 of ATO or VK2 at time of 24,48,72 h were (22.86 ± 2.44),(6.66 ± 0.34),(4.14 ± 0.41) and (18.40 ± 1.12),(13.48 ± 0.73),(8.95 ± 0.40) μmol/L,respectively; ②The combination of ATO and VK2 illustrated a synergistic effect with CI < 1.③No statistical difference was found among control group [(4.38 ± 0.56)%],1.0 μmol/L ATO group [(5.76 ± 1.63)%] and 5.0 μmol/L VK2 group [(6.38 ± 1.42)%] in the apoptosis rate(all P > 0.05).However,the apoptosis rate of combined group did rise to (44.18 ± 8.42)%,with a significant improvement to that of VK2 or ATO group alone (all P < 0.01).Conclusions The combination of VK2 and ATO exhibits an enhanced synergistical inhibitive effect on proliferation of HL-60 cells,and apoptosis may be involved in this synergy in part.
2.Determination of Forsythiaside and Forsythin from Fruits of Forsythia Suspensa and Forsythia Viridissima Lindl Collected in Different Periods
Huanhuan QU ; Baixue LI ; Fei YAN
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(08):-
Objective To study the content of forsythiaside and forsythin from fruits of Forsythia suspensa and Forsythia viridissima Lindl collected in different periods. Methods Samples were dealt by HPLC-PDA with Diamonsil-C18 (4.6 mm?250 mm,5 ?m) column. The mobile phase consisted of methanol-water,gradient elution,the flow rate was 1.0 mL/min. The UV detection was set at 270 nm. Results The content of forsythiaside from fruits of Forsythia suspensa was much higher than that from fruits of Forsythia viridissima Lindl. And there was no forsythin from fruits of Forsythia viridissima Lindl. Conclusion There was much difference in content of forsythiaside and forsythin from the two kinds of fruits above. Forsythia viridissima Lindl should not be used as Forsythia suspensa.
3.Evaluation of the Antioxidative Capacity of Different Parts of Forsythia Suspensa by DPPH Free Radical Elimination Method
Huanhuan QU ; Baixue LI ; Fei YAN ; At EL
Chinese Journal of Information on Traditional Chinese Medicine 2008;0(S1):-
Objective To study the antioxidant activities of different parts of Forsythia suspensa and two components isolated from it. Methods The antioxidant activities were assayed through scavenging effects to DPPH radical. The content of forsythiaside and forsythin of Forsythia suspensa was determined by HPLC- PDA. Results Both different parts of Forsythia suspensa and two components isolated from it had antioxidant activity. Conclution Forsythiaside showed much higher antioxidant activity than forsythin. It is an effective natural free radical scavenger.
4.Carbapenemase genes and homology of Acinetobacter baumannii in two hospitals of Qingdao
Qian LI ; Qingshu LI ; Zhi LI ; Yan QU ; Dan HU
Chinese Journal of Infection Control 2015;(7):437-442
Objective To investigate antimicrobial resistance,distribution,and carriage of carbapenemase genes of Acinetobacterbaumannii(AB)from two hospitals in Qingdao.Methods 145 AB isolates collected from two hospi-tals (78 from hospital A,67 from hospital B)were performed antimicrobial susceptibility testing,carbapenemase genes were amplified by polymerase chain reaction (PCR);homology analysis were conducted with enterobacterial repetitive intergenic consensus (ERIC)-PCR.Results AB from hospital A were generally resistant to 16 commonly used antimicrobial agents,with the lowest resistant rate of 3.85% to cefoperazone/sulbactam,followed by resist-ance rate of 16.67% to minocycline,resistant rates to the other antimicrobial agents were all>73% . AB from hos-pital B were generally resistant to 23 commonly used antimicrobial agents,but the resistance rates to minocycline and tigecycline were both 0,resistance rates to amikacin and levofloxacin were 23.88% and 38.81% respectively, resistant rates to the other antimicrobial agents were all >64% . All strains carried OXA-5 1 gene,the carriage rates of OXA-23 gene in carbapenem-resistant group were 86.76% (59/68)and 56.67% (34/60)in hospital A and B re-spectively,the difference was significant(χ2= 14.53,P<0.001);OXA-58 gene was detected in 3 isolates in hospi-tal A but not detected from hospital B. 145 AB strains were classified into 8 types,the major prevalence types were type A (n= 71)and E(n= 37);the major prevalence types in hospital A were type A (46.15% )and E(41.03% ), hospital B were type A (52.24% )and C (17.91% ).Conclusion Antimicrobial resistance of clinically isolated AB is serious and prevailed in two hospitals. OXA-23 and OXA-51 genes play an important role in AB resistance to car-bapenems.
6.Effects of lentivirus-mediated heat shock protein 70 gene on calcium homeostasis in PC12 cells undergone ischemia and hypoxia
Yanning HU ; Qingshu LI ; Zhi LI ; Dan HU ; Yan QU
Chinese Critical Care Medicine 2015;(4):295-299
ObjectiveTo investigate the effects of lentivirus-mediated heat shock protein 70 (HSP70) gene on calcium homeostasis in PC12 cells undergone ischemia and hypoxia, and the mechanism involved.Methods PC12 cells at logarithmic phase were collected, and were divided into recombination lentivirus infection group (infected by lentivirus containing HSP70 and fluorescent gene), lentivirus control group (infected by lentivirus containing fluorescin without HSP70 gene) and non-infection group. HSP70 gene and protein expressions in PC12 cells were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot. After being challenged with ischemia and hypoxia for 4 hours, the viability of cells was detected by methyl thiazolyl tetrazolium (MTT), the levels of lactic acid dehydrogenase (LDH) in cell supernatant were determined by LDH measurement test kit. The concentration of intracelluer calcium ([Ca2+]i) was assayed by flow cytometer. The activities of Na+-K+-ATPase, Ca2+-Mg2+-ATPase and total ATPase were measured by ATPase test kits.Results The expressions of exogenous HSP70 gene and protein were found by RT-PCR and Western Blot in the recombination lentivirus infection group. After being challenged with ischemia and hypoxia, the viability of cells in the recombination lentivirus infection group was increased significantly as compared with the lentivirus control group and non-infection group (A value: 0.575±0.020 vs. 0.395±0.014, 0.363±0.045,t1= 17.996,t2= 10.600, bothP< 0.001), the levels of LDH in culture medium and the concentration of [Ca2+]i were decreased significantly [LDH (U/L): 743.46±23.68 vs. 935.43±34.77, 962.89±26.68,t1= 11.179, t2= 15.044, bothP< 0.001; [Ca2+]i relative fluorescence: (31.60±2.43)% vs. (41.48±3.33)%, (40.40±3.05)%, t1= 5.853,t2= 5.502, bothP< 0.001], and the activities of Na+-K+-ATPase, Ca2+-Mg2+-ATPase and total ATPase were increased significantly [Na+-K+-ATPase (μmol·mg-1·h-1): 8.608±0.307 vs. 6.728±0.173, 6.450±0.091,t1=9.237,P1= 0.001,t2= 11.675,P2< 0.001; Ca2+-Mg2+-ATPase (μmol·mg-1·h-1): 10.523±0.036 vs. 7.910±0.209, 8.064±0.195,t1= 9.718,P1= 0.001,t2= 11.535,P2<0.001; total ATPase (μmol·mg-1·h-1): 17.041±0.324 vs. 14.150±0.182, 13.983±0.085,t1= 16.113,t2= 17.602, bothP<0.001]. There was no statistical difference in above indexes between lentivirus control group and non-infection group.Conclusion HSP70 can maintain the PC12 cells calcium homeostasis, which may be one of the important mechanisms of anti-apoptosis.
7.Dose-effect relationship of cassia seed decoction with its inhibition on increased body mass in nutritional obese rats
Jianzhuang YANG ; Junbao YAN ; Tianzhen ZHENG ; Songyi QU ; Wei LI
Chinese Journal of Tissue Engineering Research 2005;9(31):226-228
BACKGROUND: Cassia seed acts on decreasing blood pressure and blood lipid, protecting liver and inhibiting bacteria. It is worth to carry on a further discussion on its effect of weight loss.OBJECTIVE: To observe the influence of cassia seed decoction drunk naturally on body mass of nutritional obese rats in physiological state.DESIGN: Completely randomized grouping was designed, in which, control experiment, analysis of variance and q test were applied in comparison among groups.SETTING: Cardiovascular Institute, Second Affiliated Hospital, Henan University of Science and Technology.MATERIALS: The experiment was performed in Cardiovascular Instutute,Second Affiliated Hospital, Henan University of Science and Technology from March 2004 to September 2004, in which, 27 male SD rats were employed and randomized into 3 groups, named normal control group, model group and cassia seed group, 9 rats in each one.METHODS: [1] In normal control, the rats were bred with basic forage(the contents of protein, fat, carbohydrate were 18.2%, 4.5% and 55.2%successively, with 14.54 kJ caloric each gram) and drank water naturally.In model group, the rats were bred with high nutritive forage (the contents of protein, fat, carbohydrate were 23.7%, 21.6% and 39.0% successively,with 19.56 kJ caloric each gram) and drank water naturally. In cassia seed group, the rats were bred with high nutritive forage and drank cassia seed decoction of various concentration naturally. The concentration of cassia seed decoction started at 10 g/L (equally contained 10 mg raw cassia seed each milliliter) and was increased by 100% concentration each day (10 g/L)till to 60 g/L on the 6th day. Since the 7th day, the concentration of 60 g/L was maintained till to the 7th weekend. [2] It was to record appetite and drinking quantity at definite time every day and calculate absorbed caloric(intake mass × caloric contained each gram). It was to measure body mass at definite time each week. On the 7th weekend, the body length of rat was measured and Lee's index was calculated [ 3√body mass (g)×103/body length (cm)]MAIN OUTCOME MEASURES: Influences of cassia seed on body mass, Lee's index, appetite, caloric and drinking quantity in nutritional obese rats.RESULTS: Twenty-seven rats all entered result analysis. [1] Body mass:that in model group from the 3rd to 7th week in experiment group was higher remarkably than normal control group (P < 0.05-0.01). That in cassia seed group from the 2nd to 7th week was lower remarkably than that in the model group (P < 0.05-0.01). [2] Lee's index: that in model group and cassia seed group on the 7th week of experiment was higher remarkably than that in the normal control group [(358.60±8.55), (341.84±7.29), (322.00±6.89) g/cm, P < 0.05-0.01] and that in cassia seed group was lower remarkably than that in the model group (P < 0.05). [3] Appetite: that in model group and cassia seed group was lower remarkably than that in the normal control group (P < 0.05-0.01) and that in cassia seed group was near to the control group (P > 0.05). [4] Absorbed caloric: that in model group and cassia seed group was higher remarkably than that in the normal control group (P < 0.05-0.01) and that in cassia seed group was near to the control group (P > 0.05). [5] Drinking quantity: that in cassia seed group was basically near to that in the model group and the control group (P > 0.05) and that in model group was near to the control group. It was indicated that cassia seed decoction at mass concentration of 60 g/L did not affect appetite.CONCLUSION: Cassia seed decoction at mass concentration of 60 g/L inhibits remarkably the increased body mass of nutritional obese rats and is free from influence on appetite.
8.Expression of the hepatic insulin signal transduction proteins in the rats with intrauterine growth retardation during the perinatal period
Yan LIAO ; Haiqi LI ; Bing DENG ; Ping QU
Journal of Peking University(Health Sciences) 2003;0(05):-
Objective:To investigate the effects of maternal malnutrition on the expression of the insulin signal transduction proteins in the liver of perinatal rats.Methods:(1)The dams were semi-starvation from the first day during pregnancy to build the animal model of intrauterine growth retardation(IUGR).On days 15,17,19,21(E15,E17,E19,E21)of gestation,the fetuses were delivered by cesarean section.The fetal rats,placenta,fetal liver and the fetal brain were weighed respectively.And the placenta weight to the body weight ratio(PWR),the liver weight to the body weight ratio(LWR),the brain weight to the body weight ratio(BWR)were calculated;The expressions of the hepatic insulin receptor(IR),insulin receptor substrate-1(IRS-1),insulin receptor subtrate-2(IRS-2),phosphatidyl inositol 3-kinse(PI3K)mRNA in the rats during perinatal period were detected by RT-PCR.Results:(1)The PWR of the IUGR fetus on E15 was lower than that in the control(P0.05).Conclusion:(1)In order to protect the development of brain,the fetus that suffered maternal malnutrition throughout the pregnancy had liver weight and body weight reduced much more than brain weight,which was known as "brain spare effects".(2)The expressions of hepatic IR mRNA and PI3K mRNA of the fetus were in normal ranges;But the expressions of the hepatic IRS-1 mRNA and IRS-2 mRNA of the IUGR fetus were decreased during the perinatal period.These may be related to the growth retardation and insulin resistance of the adult IUGR rats.
9.Study on the mechanism of insulin resistance in rats born with intrauterine growth retardation
Yan LIAO ; Haiqi LI ; Bing DENG ; Ping QU
Journal of Peking University(Health Sciences) 2003;0(06):-
Objective:To investigate the expressions of the insulin receptor(IR),insulin receptor substrate-1(IRS-1),insulin receptor substrate-2(IRS-2),phosphatidyl inositol 3-kinase(PI3K) and insulin-like growth factor-1(IGF-1) of the livers of the male adult rats born with intrauterine growth retardation(IUGR),and to find out the relationship between IUGR and insulin resistance in their adult life.Methods:The foods were available ad libitum throughout the pregnancy to the control group and the rats in the experimental group were fed 50% of the control group to build the IUGR animal model.Liver samples were collected when the male offspring grew up to 12 weeks old.The mRNA expressions of hepatic IR,IRS-1,IRS-2,PI3K,and IGF-1 were detected by RT-PCR.The protein expressions of hepatic IR,IRS-1,and IRS-2 were analyzed by immunohistochemistry.Results:The mRNA expressions of hepatic IR in the adult male rats with IUGR were significantly lower than control group[(0.41?0.06) vs(0.62?0.11),P0.05] in both groups.The mRNA and protein expressions of the hepatic IRS-1[mRNA:(0.77?0.20) vs(1.32?0.42),P0.05].The expressions of hepatic IGF-1 mRNA of the rats with IUGR were significantly lower than those in control group [(0.55?0.12) vs(1.22?0.34),P
10.Preliminary study of intestinal permeability of children with food hypersensitivity
Yan HU ; Haiqi LI ; Wenlin YANG ; Guomin CHEN ; Ping QU
Journal of Third Military Medical University 2003;0(14):-
Objective To evaluate the intestinal barrier function of the patients by two-sugar absorption test with high performance liquid chromatography (HPLC). Methods Nineteen children with confirmed food hypersensitivity (FH) and 19 normal children were included in this study. The average age was (8.1?1.7) months. After 8-hour fasting, subjects drank test solution (5 g lactulose and 2 g mannitol per 100 ml) at the dose of 2 ml/kg. Five-hour urinary excretion ratio of lactulose/mannitol (L/M) were measured using high performance liquid chromatography (HPLC). The condition of HPLC was as follows: Sugar-PakI column with refractive index detector; Mobile phase: pure deionized water; Flow rate: 0.5 ml/min; temperature of column: 85 ℃. Results HPLC chromatogram of urine sample was stable. The retention time of mannitol and lactulose was 13.86 min and 9.27 min respectively. A significant rise in 5 h urinary L/M excretion ratios was found in children with FH (0.18?0.06) as compared to that of controls (0.05?0.03) (P