Child ; Humans ; Leukemia, Myeloid, Acute ; complications ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; complications

ObjectiveTo observe the effects of hypoxia inducible factor-1 alpha (HIF-1α)-targeting small interfering RNA(siRNA) on the expression of HIF-1α and vascular endothelial growth factor (VEGF) in HaCaT ceils under hypoxic conditions. MethodsHaCaT cells were cultured and divided into four groups, normal control group (without any treatment), hypoxia group (cultured under hypoxic conditions for 24 hours),liposome control group (transfected with liposome followed by hypoxic culture for 24 hours), RNA interference group (transfected with HIF-1α-targeting siRNA/liposome complexes followed by hypoxic culture for 24 hours). Fluorescence real-time quantitative PCR was utilized to determine HIF-1oα and VEGF mRNA expression in HaCaT cells, and Western blot to detect HIF-1α and VEGF protein expression. ResultsNo significant difference was observed in the mRNA expression of HIF-1α between the hypoxia group and normal control group(0.907 ± 0.032 vs. 0.878 ± 0.034, F =1.108, P ＞ 0.05), while the expression levels of VEGF mRNA,HIF-1α and VEGF proteins were significantly higher in the hypoxia group than in the normal control group (0.935 ± 0.032 vs. 0.652 ± 0.053, 0.813 ± 0.047 vs. 0.236 ± 0.014, 0.791 ± 0.030 vs. 0.316 ± 0.013, all P ＜0.05). A significant decline was noted in the mRNA and protein expression levels of VEGF (0.230 ± 0.044 vs.0.978 ± 0.030, 0.213 ± 0.026 vs. 0.817 ± 0.049, both P ＜ 0.05) and HIF-1α(0.497 ± 0.033 vs. 0.806 ±0.040, 0.249 ± 0.028 vs. 0.833 ± 0.052, both P ＜ 0.05) in the RNA interference group than in the liposome control group. ConclusionsHypoxia may enhance the expression of HIF-1α and VEGF in HaCaT cells, and to inhibit the HIF-1α expression may suppress the expression of VEGF in HaCaT cells under hypoxia.

Objective:To evaluate the efficacy and safety of tirofiban,a platelet glycoproteinⅡb/Ⅲa Inhibitor,in percutaneous coronary intervention(PCI)of patients with acute non-ST segment elevation myocardial infarct(NSTEMI).Methods:A total of 114 patients with acute NSTEMI were enrolled in the trial from Sep.2005 to Jan.2007;they were randomly divided into 2 groups:tirofiban group(n=57)and placebo group(n=57).Patients in tirofiban group were given tirofiban for 24 h after PCI.All patients were routinely given heparin,aspirin and clopidogrel before CPI.The composite occurrence of death,myocardial infarction(MI),need for target vessel revascularization(TVR)after PCI,and the adverse effects(hemorrhage and thrombocypenia)were compared between the 2 groups.Results:One(1.8%)patient had angina pectoris and the other(1.8%)developed subacute thrombus in control group within 24 h after PCl;there was no such event in the tirofiban group.Two(3.6%)patients developed angina pectoris and 2(3.6%) developed subacute thrombus within 30 days after PCI in control group;one patient(1.8%)in birofiban group developed angina pectoris and one patient in birofiban group developed subacute thrombus.Each group had one case(1.8%)of upper digestive tract bleeding during hospitalization.No intracranial hemorrhage,skin/ mucosa hemorrhage,thrombocytopenia,or-death occurred in the 2 groups.Intravenous tirofiban treatment reduced the composite occurrence of death of NSTEMI patients after PCI(P

Objective To investigate the effects of iodine excess on mitochondrial superoxide production and mitoehondrial membrane potential(△ψ)changes in Fisher rat thyroid cell line(FRTL)cells.Methods FRTL cells were treated with 10-4mol/L potassium iodine(KI),10 U/L thyrotropin(TSH),10-4 mol/L KI+10 U/L TSH respectively for 24 h.Effects on cell proliferation were assayed by methyl thiazolyl tetrazolium(MTT)colorimetric method.Changes of mitochondrial superoxide production and △ψ were measured by live cell imaging and spectrofluorometer using MitoSOX and rhodamine 123(rh123)respectively.Results Absorbance(A)in the KI group (0.794±0.144)showed a significant decline compared to the control group(1.000 ±0.183,P<0.05),whereas a significant elevation was observed in the TSH group(1.215±0.156,P<0.05).No significant differences was found between the KI+TSH group(1.025±0.254)and the control group(P>0.05),but the former was marked higher than the KI group(P<0.05).Compared to the control group(9.74±3.24).MitoSOX mean fluorescence intensity (MFI)in the KI and KI+TSH groups(18.16±6.57,13.33±2.92)were significantly increased(all P<0.05),which was a significant decline in the TSH group(6.64±2.15,P<0.05).MitoSOX MFI in the KI+TSH group was lower than the KI group(P<0.05).Rh123 MFI in the KI and KI+TSH groups(210 593±31 328,295 525±34 243)showed significant decline than the control group(407 824±37 198,all P<0.05).Compared with the KI group.the KI+TSH group pronouncedly attenuated the reduction of Rh 123 MFI(P<0.05).No significant differences of Rh 123 MFI were found between the TSH group(411 187 ± 72 852) and the control group(P > 0.05). Conclusion Iodine excess (10-4 mol/L KI) may lead to peroxide damage on the mitochondria of FRTL cells, and cell proliferation is inhibited. Combining treatment with 10 U/L TSH may attenuate mitochondrial peroxide damage and inhibition of cell proliferation caused by iodine excess.

Objective To determine the influence of protein fusion on the biological characteris- tics of hymidine kinase(TK)and human immunodeficiency virus(HIV)Tat recombinant protein. Methods By utilizing polymerase chain reaction(PCR)technique,different fragments containing two,four or six glycines(Gly)were inserted between the HIV Tat gene and TK,and cloned into PBK vector.After testified by sequencing,the vectors were transfected into E coli.After induced by iso- propyl thiogalactose(IPTG),bacilli were collected and destructed by ultrasonic,the fusion proteins were determined by monoclonal antibody against HIV protein.HepG2 cells were incubated in DMEM supplement with 10?g/mL HIV-Gly(n)-TK(n=0,2,4,6)fusion protein,TK-HIV Tat and only HIV Tat.HepG2 cells in different groups were detected by immunofluorescence assay 24 hours after transduction with HIV Tat monoclonal antibody.The rate of apoptosis after cells were incubated with gencilovir(10?g/mL)for 3 days was determined by cell flow cytometry,while survival cell ratio was recorded by trypan blue.The data were analyzed by statistics(t-test).Results The Tat-Gly(n)-TK (n= 0,2,4,6)recombinant genes were constructed and inserted into PBK vectors,which were expressed in E coli and then purified.Cells in different groups,which were incubated with Tat-Gly (n)-TK(n=0,2,4,6)fusion proteins,Tat-TK fusion protein,TK-Tat fusion proteins or only Tat proteins respectively,were detected by immunofluorescence assay.The intensities of fluorescence in different groups were almost same,but the ratios of cell survival or apoptosis were different.The highest ratio of cells apoptosis(14.77%)was in the group that cellular culture medium was mixed with Tat-Gly(4)-TK fusion protein,followed by the groups containing 6,2 glycines or no TK gene in genes(4.30%,12.69% and 1.03%,respectively).There were significant differences between each 2 groups among the all groups(t-test,P

Objective To study the effect of trachea intubation and mechanical ventilation on the prognosis and discharge rate of patients with successful cardiac-pulmonary resuscitation.Method The clinical data of 389 patients,who were admitted from January 2005 to February 2007,were retrospectively analyzed.The relation between trachea intubation time and discharge rate was studied.According to the time from cardiac arrest to finishing trachea intubation,patients were divided into group A (within 3 minutes,n=209) and group B (over 3 minutes,n=143);according to the time from reaching emergency medicine department to finishing trachea intubation,the rest patients were divided into group C (within 5 minutes,n=9) and group D (over 5 minutes, n=38) minutes.The discharge rate was calculated between groups.The software of SPSS 11.0 was used for statistical analysis.Results The successful rate was 9.75% (389/3988),and 59 patients were discharged, with discharge rate 1.48% (59/3988).The discharge rate of group A was 19.62% (41/209),and was significantly higher than that of group B 6.99 % (10/143) (P

Two high protease producing strains, whic h are desi gnated Aspergillus oryzae AS100 and AS102, were screened and applied as main fermentation strains of swine blood meal. A Saccharomyces cerevisiae Y113 and a Bacillus Asp007 were also used as assistant fermentation strains. The enzyme production abilities we re studied,and a series of parameters of the fermentation technology were deter mined. Through swine blood meal fermentation, a high-protein fermented feed wa s produced, which is strong soy flavor and was rich in protein(69%), free amino acid, vitamin such as VitD 3 and niacin and organic elements such as Fe. It is a kind of high-protein feed for animals and it could be used as feedstuff addi tive.

OBJECTIVETo explore the role of CD(8)(+)CD(28)(-) T regulatory cells (Tr) in the immunological pathogenesis of acute infection with Epstein-Barr virus in children.

METHODSThe present study enrolled 25 children with infectious mononucleosis (IM) and 25 age-matched healthy children. Flow cytometric analysis was performed to detect the percentage of CD(3)(+), CD(3)(+)CD(4)(+), CD(3)(+)CD(8)(+), CD(8)(+)CD(28)(+) by determining the ratio of positive cells in lymphocytes. Reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time PCR were used to analyze IL-6, IL-10, IFN-gamma expression in CD(8)(+)CD(28)(-) Tr cells and ILT-3, ILT-4 expression in monocytes/macrophages.

RESULTSThe proportions of CD(8)(+)CD(28)(-)T cells in children with acute-phase IM was significantly higher than those in the controls (P < 0.01). The expression level of IL-6, IL-10, IFN-gamma, ILT-3, ILT-4 mRNA significantly increased compared to those of the controls (P < 0.01).

CONCLUSIONThe CD(28) expressed on CD(8)(+) T cells in vivo is gradually lost with age and CD(8)(+)CD(28)(-) cells increase up 50% to adult. EBV can directly infect B cells, trigger CD(8)(+) CTL response and destroy the target cells to cause serious immunopathological lesion. Therefore we speculate that the expansion of CD(8)(+)CD(28)(-) Tr cells in children with IM may be an adaptive immune response to avoid serious inflammation and autoimmune reactions.

CD28 Antigens ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Case-Control Studies ; Child ; Child, Preschool ; Cytokines ; immunology ; Epstein-Barr Virus Infections ; immunology ; Female ; Flow Cytometry ; Herpesvirus 4, Human ; Humans ; Infectious Mononucleosis ; immunology ; virology ; Male ; Membrane Glycoproteins ; metabolism ; Receptors, Cell Surface ; metabolism ; Receptors, Immunologic ; metabolism ; T-Lymphocytes, Regulatory ; immunology

OBJECTIVETo establish a new method for screening active ingredients of Chinese herbs by determining different bio-thermodynamic effects of 3 genosides on splenic lymphocyte of mice.

METHODSUsing a thermal bioactivity monitoring system, the maximum heat output (mHO), average metabolic heat (MH) and constant of decrease rate (DR) of lymphocyte were determined based on the growth metabolic power-time curve, and the outcomes were verified by MIT.

RESULTSThe mHO and MH increased and the DR decreased after lymphocytes being exposed to the 3 genosides in different concentrations, arranged upon their potency as genoside Rg3 > genoside Rg2 > genoside Rg1 (merely insignificant effect). MTT showed the same results.

CONCLUSIONHeat activity monitoring system could precisely display the different bio-thermal dynamic effects of 3 genosides on splenic lymphocyte.

Animals ; Cell Survival ; drug effects ; Cells, Cultured ; Energy Metabolism ; drug effects ; Ginsenosides ; pharmacology ; Lymphocytes ; cytology ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Spleen ; cytology ; Thermodynamics

Objective To observe the clinical efficacy of acupoint iontophoresis with Sinomenine hydrochloride in treating knee osteoarthritis (KOA).Method Sixty KOA patients were randomized into a treatment group and a control group, 30 cases each. The treatment group was intervened by iontophoresis with Sinomenine hydrochloride into specific acupoints via a direct current iontophoresis appliance; the control group was intervened by oral administration of Meloxicam tablets. The Lequesne index, Visual Analogue Scale (VAS) and knee joint space were observed before and after the intervention.Result The Lequesne and VAS scores were significantly changed after the treatment in the two groups (P<0.01,P<0.05). After the intervention, there was a significant difference in the Lequesne score between the two groups (P<0.01). After the treatment, the knee joint space didn't show significant intra-group or between- group differences (P>0.05).Conclusion Acupoint iontophoresis with Sinomenine hydrochloride is an effecttive method in treating KOA.