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.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.
3.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.
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.Observation on Treatment of Advanced Esophageal Carcinoma with Three-Dimensional Conformal Radiotherapy Combined with Chemotherapy and Traditional Chinese Medicine
Manling TIAN ; Xiuling WANG ; Lei QU ; Yan LI ; Guangzhong FU
International Journal of Traditional Chinese Medicine 2009;31(1):43,48-
Objective To observe the clinical effect of treating advanced esophageal carcinoma with three-dimensional conformal radiotherapy combined with chemotherapy and traditional Chinese medicine.Methods 36 cases with advanced esophageal carcinoma were randomly divided into a eontrol group and a treatment group.The control group was treated with conventional radiotherapy combined with chemotherapy,and the treatment group was treated with three-dimensional conforrnal radiotherapy combined with chemotherapy and traditional Chinese medicine.The short-term effect,toxicity and long-term effect were observed,and cause of death was analyzed.Results The complete remission rate in the treatment group was significantly higher than that of the control group(P<0.05).The toxicity of the treatment group was markedly decreased.One and three years'survival rate between the two groups had a significant difierent(P<0.05),but five years'survival rate between the two groups had no significant difference(P>0.05).Local uncontrolled rate and recurrence rate in the treatment group were significantly lower than those in the control group(p<0.05).Conclusion The treatment of three-dimensional conformal radiotherapy combined with chemotherapy and traditional Chinese medicine on advanced esophageal carcinoma was better than conventional radiotherapy combined with chemotherapy.
9.Immunophenotyping detected by flow cytometry and its clinical significance in acute myeloid leukemia
Jinjing ZHANG ; Pingping WANG ; Yi QU ; Yan LI
Journal of Leukemia & Lymphoma 2013;22(10):597-599
Objective To investigate the immunophenotypic characteristics of acute myeloid leukemia (AML)and its relationship with chemotherapy response.Methods The immunophenotyping was performrd by flow cytometry in 96 newly diagnosed AML patients.Results In 96 patients with AML,the myeloid antigens were mainly expressed including CD13(94/96,97.92%),CD117(86/96,89.58%),CD33(79/96,82.29%)respectively.The positive expression of stem/progenitor cell differentiation antigens were CD38 (92/96,95.83%),HLA-DR(72/96,75.00%)and CD34(60/96,62.50%).While CD34 and HLA-DR were fewly expressed in M3.The CD14 was only expressed in M4 and M5.The lymphocytic antigens had positive expression in the patients with AML,in which CD4 was 37.5%(36/96),CD7 was 22.92%(22/96)and CD2 was 10.42 %(10/96).In 46 patients for treatment response analysis,total CR rate was 67.39%(31 /46).There was no significant difference of CR rates between the antigens positive group and negative group including CD33,CD117,CD11b,CD14,CD64,CD4,CD34 and HLA-DR.However,the CR rate was significantly lower in CD7 positive cases than CD7 negative ones,with statistically significance difference(P=0.01).Conclusions CD13,CD33and CD117 are the most common antigens in AML.The patients with the positive expression of CD7 have poor response to chemotherapy in AML.
10.Correlation between the biofilm-forming ability, biofilm-related genes and antimicrobial resistance of Acinetobacter baumannii
Xinxin HAN ; Qingshu LI ; Liting SHEN ; Dan HU ; Yan QU
Chinese Critical Care Medicine 2014;26(9):639-643
Objective To study the biofilm-forming ability and the distribution of biofilm-related genes in Acinetobacter baumannii clinical isolates as well as antimicrobial resistance,to analyze their relationships with the bacterial resistance phenotype.Methods A prospective study was conducted.Biofilm models of 70 strains Acinetobacter baumannii collected in Chengwu County People's Hospital from October 2012 to October 2013 were constructed using 96-well polystyrene plate.In order to analyze the biofilm-forming ability,a qualitative and quantitative analysis was conduct by crystal violet staining assay.And the antimicrobial resistance of different biofilm-forming ability strains was compared including imipenem,amikacin,meropenem,cefepime,sulbactam cefoperazone,trimethoprim,levofloxacin,gentamicin,ciprofloxacin,cefotaxime,ceftizoxime,aztreonam,piperacillin,ceftriaxone,cefuroxime.In addition,the expressions of biofilm-related gene Bap,bfs and intI1 were tested with polymerase chain reaction (PCR) assay.Results Among 70 strains Acinetobacter baumannii,40 strains were multi-drug resistant (57.14%) and 6 strains were pan-drug resistant (8.57%); 68 strains had biofilm-forming ability (97.14%),14 of which were weakly positive,20 were positive and 34 were strongly positive.The antimicrobial resistant rate of Acinetobacter baumannii to imipenem,amikacin,meropenem and cefepime was decreased,it was 30.00%,32.86%,38.57% and 41.43%,respectively.However,the antimicrobial resistant rates to other commonly used antibiotics were all higher than 50%.The drug resistance of Acinetobacter baumannii to levofloxacin (85.71%,45.00%,38.24%,x2=9.225,P=0.010),cefepime (71.43%,45.00%,29.41%,x2=7.222,P=0.027),gentamicin (78.57%,55.00%,38.24%,x2 =6.601,P=0.037) was significantly decreased when biofilm-forming ability reinforced (weakly positive,positive,hadro-positive).Bap gene positive rate of weakly positive,positive and strong positive biofilm-forming strains Acinetobacter baumannii was 50.00%,65.00% and 79.41% (x2=4.244,P=0.120),respectively.Bfs gene positive rate was 35.71%,65.00% and 88.24%,respectively (x2=13.602,P=0.001) and intI1 gene positive rate was 42.86%,75.00% and 91.18%,respectively (x2 =12.902,P=0.002).Moreover,the antimicrobial resistances of biofilm-related gene positive strains were higher than the negative,of which the drug resistance of intI1 positive group to amikacin was significantly higher than the negative group (40.38% vs.11.11%,x 2=5.194,P=0.023).Conclusions The Acinetobacter baumannii collected from the hospital had strong multi-drug resistance as well as strong biofilm-forming ability.The drug resistance of Acinetobacter baumannii decreased when biofilm-forming ability reinforced.In addition,genes,such as Bap,bfs,and intI1,contributed to biofilm formation.