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.

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.

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.

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.

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.

Chromosome Aberrations ; Cytogenetic Analysis ; standards ; Humans ; Multiple Myeloma ; diagnosis ; genetics

ABSTRACT:Objective To explore the risk factors for maternal adverse pregnancy outcomes.Methods An unmatched case-control study based on hospital was performed.Univariate and multivariate Logistic regression were used to analyze the related factors of maternal adverse pregnancy outcomes,including general condition,history of fertility,abnormal symptoms and diseases during pregnancy.Results Univariate analysis results showed that high education level of gravida might be the protective factors of adverse pregnancy outcomes.The risk factors for adverse pregnancy outcome might include advanced maternal age,intensive workload,frequent pregnancy,history of spontaneous abortion,severe morning sickness,and sickness during pregnancy or progestation.Multivariate Logistic regression analysis showed that high education level of gravida (OR=0.63,95% CI:0.50-0.80)was the protective factor of adverse pregnancy outcomes;severe morning sickness (OR=2 .1 3 ,9 5% CI:1 .6 3-2 .7 9 )and sickness during progestation (OR=2.25,95% CI:1.06-4.77)were the risk factors for maternal adverse pregnancy outcomes.Conclusion The level of maternal education should be improved.We should attach great importance to preventive education and thorough treatment of severe morning sickness. Couples should be encouraged to have physical examination before marriage and pregnancy.Corresponding pregnancy care guidance should be given to pregnant women with different physical conditions so as to effectively reduce the occurrence of adverse pregnancy outcomes.

Objective To study the effect of DNA methylation regulation on the toxic effect of paraquat on the sensitized V79 cells t pretreated with 5-aza-2'-deoxycytidine.Methods V79 cells were treated by 5-aza-2'-deoxycytidine (5-Aza-2'-dc) for 12h,which is a DNA methylation inhibitor,and then treated with paraquat for 12h.The morphological changes of V79 cells were observed by microscopy and the cell viability was determined by MTT assay and trypan blue staining method.Results Microscopic examination showed that the combination of 5-Aza-2'-dc and paraquat had stronger effect in inhibiting the growth of V79 cells(the cells became smaller and poorer adhensive ability) than single 5-Aza-2'-dc or paraquat.MTT assay showed that cell viability in the combination group (54.47 ± 3.04) ％ was significantly lower than the 5-Aza-2'-dc group (95.52 ± 0.90) ％ and paraquat group (89.68 ± 4.26) ％ (P＜0.05).Trypan blue staining assay showed that the death rate of ceils in the combination group (53.58 ± 1.57) ％ was significantly higher than the 5-Aza-2'-dc group (7.44 ± 2.31) ％ and paraquat group (12.90 ± 1.21) ％ (P＜0.05) Conclusion 5-Aza-2'-dc promotes V79 cells damage caused by paraquat.

Objective To study the reactive oxygen level and the expression of anti-apoptotic protein Bcl-2 and pro-apoptotic protein Bax after treatment of DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine (5-Aza-2'-dc) and paraquat in V79 cells.Methods Cultured V79 cells were divided into 5-Aza-2'-dc treatment group (group A),paraquat treatment group (group B),5-Aza-2'-dc and paraquat treatment group (group C,V79 cells were pretreated with 5-Aza-2'-dc for 12h followed by exposure to paraquat for 12h) and control group (group D).Reactive oxygen level in V79 cells was measured by DCFH-DA flow cytometry and expression of Bcl-2 and Bax was detected by Western blot.Results Reactive oxygen levels and expression levels of Bcl-2 and Bax in V79 cells were significantly different (P＜0.05) in 5-Aza-2'-dc and paraquat treatment group (group C),compared with 5-Aza-2'-dc treatment group (group A),paraquat treatment group (group B) and control group (group D).Expression levels of Bcl-2 and the ratio of Bcl-2 and Bax were lower while reactive oxygen levels and expression levels of Bax were higher in group C than in groups A,B and D.Conclusion 5-Aza-2'-dc regulates DNA methylation by the imbalancing the reactive oxygen metabolism and apoptosis,thus up-regulating the toxic effect of paraquat on V79 cells.

BACKGROUND: Cardiac stem cells can differentiate into cardiomyocytes in vitro under induction of pilose antlerpolypeptides, which provides a new therapeutic idea for myocardial injury.OBJECTIVE: To explore the effect of pilose antler polypeptides on the apoptosis rate and membrane stability ofmitochondria in cardiac stem cells.METHODS: We chose healthy male Wistar rats born 2 days to extract cardiac stem cells. The culture dish was used asthe experimental unit, and extracted cells were divided into the following four groups (n=12). Blank control group: Thesame amount of buffer was added for induction; 5-azacytidine group: induced with 5-azacytidine (3 μmol/L); pilose antlerpolypeptides group: induced with pilose antler polypeptides (800 mg/L); combined group: induced with pilose antlerpolypeptides (800 mg/L) and 5-azacytidine (3 μmol/L). After 48 hours induction, apoptosis rate of cardiac stem cells ineach group was detected with flow cytometry. The membrane potential of mitochondria in cardiac stem cells wasdetected with immunofluorescence. The expression level of Nkx 2.5, GATA4, ATF-2, and MEF-2C in cardiac stem cellswas detected using western blot assay.RESULTS AND CONCLUSION: There were small, round and bright cells after 2 weeks culture, and cell colonies ofcardiac stem cells formed. The apoptosis rate of cardiac stem cells in the 5-azacytidine group increased significantly (P <0.05) and its membrane potential decreased significantly compared with the blank control group (P < 0.05). Theapoptosis rate of cardiac stem cells in the pilose antler polypeptides group and combined group decreased significantlycompared with the 5-azacytidine group (P < 0.05, P < 0.01), and their membrane potential increased significantly (P <0.05, P < 0.01). The results of western blot showed that the expression level of Nkx2.5 increased significantly in thepilose antler polypeptides group and combined group compared with the 5-azacytidine group (P < 0.05), while theexpression levels of GATA4 and ATF-2 in each group were low and there were no significant differences among groups(P > 0.05). The expression level of MEF-2C in each group was at a middle level, and there were no significantdifferences among groups (P > 0.05). To conclude, our experimental findings indicate that pilose antler polypeptidescould decrease the apoptosis rate and improve membrane stability of mitochondria in cardiac stem cells. Themechanism may be related to the increased expression of Nkx 2.5, but whether the mechanism is related to GATA4,ATF-2 and MEF-2C needs to be further studied.