Objective: To study the effects of vitamin E and selenium on apoptosis and c-Myc expression in human leukemia cells. Methods: DNA gel electrophoresis and Northern blotting hybridization were used to detect the apoptosis and the expression of c-Myc gene, respectively. Human leukemia cell line HL-60 and K562 were cultured in vitro. Results: The apoptosis of HL-60 and K562 cells were induced after being exposed to vitamin E (100 ?mol/L) and selenium (8 ?mol/L) for 24 hours, respectively. In HL-60 cell line, c-Myc mRNA was down-regulated significantly by vitamin E(100 ?mol/L),but not selenium(8 ?mol/L).In contrast, the expression of c-Myc gene was repressed by selenium(8 ?mol/L) and not by vitamin E(100 ?mol/L) in K562 cell. Conclusion: Our observations suggest that c-Myc down-regulation and induction of apoptosis by selenium and vitamin E are important pathways in repressing leukemia cell proliferation.The results suggest there are different mechanisms of repressing leukemia cell proliferation for selenium and vitamin E.

Objective To explore the potential of breast cancer stem cells in differentiating into vascular endothelial cells and participating in the angiogenesis of breast tumor.Methods The expressions of mutant P53, CD31 ,VEGF and Her-2 amplification in tumor blood vessels were detected and the single cell suspension of breast cancer was prepared.CD44+/CD24-/low cells were selected and cultured in culture system.The expressions of CD31 and CD105 on endothelial cell surface were detected by EGM-2 endothelial cell culture medium.The ability of uptake of acetylated low-density lipoprotein was measured.The endothelial cells were cultured in vitro to observe their vascular structure.Results The expressions of CD31,VEGF and mutant P53 were found in paraffin sections of breast cancer tissue samples, and they were arranged along the vessel lumen or blood vessel sphere. Immunofluorescence showed that CD31 and DAPI signals were expressed intravascularly in breast cancer tissues. Four samples of Her-2 positive amplification and 9 cases of non-amplification were detected in the samples.The successful separation of breast cancer stem cells was carried out by immunomagnetic sorting.The percentage of CD44+ cells in the pre-sorting cells was (7.5±2.6)% and that of CD44+ cells was about (94.3±4.7)% after magnetic sorting (P<0.05).The ratio of CD24+ cells was (48.2±9.4)% before sorting,and that of CD24+ cells was (4.3±1.6)% after immunomagnetic bead sorting.The proportion of CD105+ cells in mammary gland cells was (4.5±0.9)% and the proportion of CD31+ cells was (6.2±1.3)%.After cultured for three generations,the percentages of CD105+ cells and CD31+ cells were (79.6±9.3)% and (84.1±10.7)%,respectively (P<0.05). DiL-Ac-LDL could be phagocytized by endothelial cells cultured in endothelial cell culture system.Vessel-like structure was found in the endothelial cell group while the control group did not have the tendency of vascular changes.Conclusion Breast cancer stem cell-derived endothelial cells differentiate into endothelial cells in vitro and have the ability of vascular formation,suggesting that breast cancer stem cells may be involved in the formation of tumor blood vessels.

Hepatitis B virus(HBV) infection is a severe health problem in the world.However,there is still not a satisfactory therapeutic strategy for the HBV infection.To search for new anti-HBV agents with higher efficacy and less side-effects,the inhibitory activities of traditional Chinese medicine Rheum palmatum L.ethanol extract(RPE) against HBV replication were investigated in this study.Quantitative real-time polymerase chain reaction(PCR) was employed to analyze the inhibitory activity of RPE against HBV-DNA replication in a stable HBV-producing cell line HepAD38; the expression levels of HBV surface antigen(HBsAg) and e antigen(HBeAg) were also determined by enzyme linked immunosorbent assay(ELISA) after RPE treatment.RPE could dose-dependently inhibit the production of HBV-DNA and HBsAg.The concentration of 50% inhibition(IC50) was calculated at 209.63,252.53 μg/mL,respectively.However,its inhibitory activity against HBeAg expression was slight even at high concentrations.RPE had a weak cytotoxic effect on HepAD38 cells(CC50 = 1 640 μg/mL) and the selectivity index(SI) was calculated at 7.82.Compared with two anthraquinone derivatives emodin and rhein,RPE showed higher ability of anti-HBV and weaker cytotoxicity.So Rheum palmatum L.might possess other functional agents which could effectively inhibit HBV-DNA replication and HBsAg expression.Further purification of the active agents,identification and modification of their structures to improve the efficacy and decrease the cytotoxicity are required.

Sucking mice, 2～3 day after birth, were intraperiton eally inoculated with 0.05mL F1M10 of Chen Strain Hantaan virus. At diff erent t ime point after inoculation, the brains were taken, routinely fixed and embedded in paraffin for preparing 5 μm serial sections. Traditional and confocal immun ochemical detection of viral antigens and heat shock protein 70 were performed t o exp lore the cerebral stress response after viral infection. The results showed that HSP70 immunoreactivities could be stably detected in the viral antigen positive neurons, but not in the viral antigen negative or uninfected control tissues. B y confocal microscopic examination, the HSP70 and viral antigens were colocolize d in the neuronal cytoplasm. Our result, comparable to our previous findings in human tissues and culture cells, indicated that Hantavirus infection can induce the expression of HSP70 in the infected cells, and HSP70 expression might be n ecessary but not sufficient to keep the cell survival.

Background Researches showed that the content of nitric oxide (NO) and nitric oxide synthetase (NOS) increases in blood,aqueous humor and tear of cataract patient.But the function of NO and NOS in cataract formation is still elusive.Objective The aim of this study was to explore the prevention and treatment effect of NOS inhibitor,1-nitro-arginine methyl ester (L-NAME),on galactose cataract.Methods Sixty clean three-week-old Wistar rats were equally and randomly divided into 3 groups.0.9％ Normal saline solution (30 ml/kg) was subcutaneously injected every day for 30 days in the rats of the control group,and 50％ of D-galactose solution (30 ml/kg) was used in the rats of the model and L-NAME group at the same way.L-NAME eye drops was simultaneously administered in the L-NAME group 3 times per day for 30 days.The eyes of the rats were examined under the slit lamp in 10,20 and 30 days,and the degree of lens opacification was scored.Lenses of the rats were obtained at the end of this experiment for the detect of NO,NOS contents.Flow cytometry was used to assay the caspase-3 level of rat lens.Repeated measurement two factor analysis of variance was used to analyze the difference of lens opacification scores in different groups and different time points,and one-way ANOVA was used to analyze the differences of NO,NOS and caspase-3 contents in lens among the groups.Results Lens opacification appeared in 10 days after injection of 50％ D-galactose solution in the rats of the model group and L-NAME group.Lens opacification score was higher among the different groups and different time points (Ftime =435.251,P =0.000 ;Fgroup =395.120,P=0.000).NO content in the lens was (0.45±0.15) μmol/g,(2.67 ± 0.47) μmol/g and (1.68±0.34) μmol/g in the control group,model group and L-NAME group,showing a significant difference (F=58.872,P=0.000).The NOS contents in the lens was (0.0160±0.0020) U/ml,(0.0370±0.0040) U/ml and (0.0270±0.0010) U/ml in the control group,model group and L-NAME group,showing a significant difference (F =66.174,P=0.000).Caspase-3 contents in the lens was (339.4 ± 37.9),(697.7 ± 46.5) and (650.7 ± 53.1),Showing a significant difference among them (F =100.005,P =0.000).Conclusions The increase of NO,NOS and caspase3 levels are associated with lens opacification.Topical administration of L-NAME eye drops can down-regulate NOS content in lens,reduce the NO formation and inhibit the apoptosis of lens epithelial cells.

Objective To compare selenium content in hair samples of children in Kashin-Beck Disease (KBD) areas and control areas,and to provide academic data for KBD control and prevention.Methods In 2011,Longnan,Qingyang,Dingxi City of KDB areas in Gansu Provines were selected as survey point,meanwhile,colleagues of Tianshui City and Lanzhou City were selected as control.Hair samples of children aged between 6 and 12 years were taken from each survey point to determine the hair selenium.After using the nitricperchloric mixed acid to digest the sample,the concentrations of selenium were determined by hydride generator atomic fluorescence spectrometry.Results Two hundred and fifty-four samples from KBD areas and 102 samples from control areas were collected.The hair selenium of KBD areas[(0.22 ± 0.07),(0.22 ± 0.00),(0.20 ± 0.07)mg/kg],were higher than the control areas[(0.32 ± 0.08),(0.42 ±0.11)mg/kg].After comparation of selenium content in every sampling point,we found the range of children's selenium content was relatively narrow in KBD areas (0.08-0.46 mg/kg).The minimum value and the maximum value in KBD areas were lower than that of control areas (0.15-0.51 mg/kg).A totle of 51.1％ (182/356) of children was seriously lack of selenium,the hair selenium content was ＜ 0.25 mg/kg.It was also found that Se content was not significantly different between boys and girls.Conclusions Children's selenium is deficient in KBD areas in Gansu.It is important to ensure salt supplement with selenium for a long period of time and through other ways to increase Selenium intake to control KBD.

1/2 cycle, three cases

Objective To investigate the changes in metabolic indexes and anesthesia administration during perioperation in patients undergoing orthotopic liver transplantation. Methods Eleven patients who underwent orthotopic liver transplantation were selected. Their pH, BE, HCO 3 -, K +, Ca 2+, blood glucose, PT, APTT and FIB were assessed at 60 min of pretransplantation, 20 min and 40 min of liver transplantation, 20 min and 40 min of new liver reperfusion. Results The pH, BE, HCO 3 -, K +, Ca 2+ during 20 min and 40 min of liver transplantation decreased more significantly than those during pretransplantation 60 min, PT and APTT extended markedly, the level of blood glucose did not change markedly; pH, BE, HCO 3-, K +, Ca 2+ and the level of blood glucose during new liver reperfusion 20 min and 40 min increased significantly compared to those during liver transplantation 20 min and 40 min; PT, APTT and FIB did not change markedly. Conclusion Metabolic indexes during liver transplantation change more markedly than those during pretransplantation or new liver reperfusion.

AIM: To establish the method for quality control of Gutongning Liniment(Herba Epimedii,Semen Strychni,Herba Ephedrae,etc.) METHODS: Herba Ephedrae,Radix Notoginseng,Ramulus Cinnamomi,Flos Caryophylli,Flos Daturae were identified by TLC,icariin content in Herba Epimedii and strychnine content in Semen Strychni were determined by HPLC. RESULTS: The identified characteristics of chromatography were distinct and the spots were clear.Icariin showed a good linear relationship at a range of 0.2-1.2 ?g,r=0.999 8.The(average) recovery was 10.3%,and RSD was 1.5%.Strychnine showed a good linear relationship at a range of 0.8-4.8 ?g,r=0.999 7.The average recovery was 99.17%,and RSD was 1.4%. CONCLUSION: The methods are accurate and reliable,and can be used to control the quality of this preparation effectively.

Manganese-oxidizing microorganisms are able to oxidize soluble Mn(II) into insoluble Mn oxides. Such microorganisms are very useful in treatment of Mn-contaminated water. In this research, a Mn(II)- oxidizing bacterium Bacillus sp. MK3-1 was isolated from Mn-contaminated soil. This bacterium has high MnCl2 resistance with a MIC of 20 mmol/L. The results showed that it is able to oxidize and remove more than 96% of Mn(II) in the culture medium. The immobilized solid-embedding Bacillus sp. MK3-1can re- moved 87.12% of manganese contaminated water. The final concentration of MnCl2 after the treatment reached the national discharge standard level. Scan electron microscope observation showed that the pro- duced Mn oxides located on the cell surfaces of Bacillus sp. MK3-1. Energy dispersive spectrdmeter analy- sis indicated that the content of manganese of cell surfaces of Bacillus sp. MK3-1 was 19.60% (W/W). At last we amplified a 903 bp multicopper oxidase gene mnxG encoding the putative Mn(II)-oxidizing protein.The product of mnxG showed 86% identity to the reported multicopper oxidase.