Objective To investigate the effects of all trans-retinoic acid (ATRA) on the expressions of iron metabolism-related genes and their products in K562 cells and the possible relationship. Methods (1) The characteristics of K562 leukemic cell differentiation induced by ATRA was evaluated by Benzidine, Wright's, NSE and NBT staining.(2) The expression levels of cellular surface antigens (CD71 and CD 13) in K562 cells cultured with ATRA were measured by flow cytometry. (3) IRP/IRE binding activity was assessed by RNA/protein band-shift assay.(4) Ferritin was determined by radioimmunoassay.(5) The mRNA expression levels of H-Fn, TfR and IRP2 in K562 cells cultured with different concentrations of ATRA were delineated by RT-PCR method, confirmed by sequencing of RT-PCR products. Results K562 cells could be induced to differentiate into neutrophils by ATRA, confirmed by cytochemical staining. The expression of CD71 decreased while CD13 increased. The mRNA expression levels of TfR and IRP2 were decreased while mRNA expression level of H-Fn was increased in K562 cells cultured with ATRA, compared to that in control cells. Concomitantly,IRP binding activity was significantly decreased but the level of ferritin was significantly increased in K562 cells cultured with ATRA. Conclusions During the course of K562 cells induction and differentiation to myelocytes by ATRA, the expression level of iron metabolism-related genes and products were changed but the upstream-regulation mechanism still remains unclear.

OBJECTIVE: To approach efficacy of Dianning tablet in the treatment of refractoriness epilepsy.METHODS: 180 patients with refractoriness epilepsy were randomized into treatment group,control group Ⅰand control group Ⅱ(n=60).Treatment group were given Dianning tablet produced by our hospital.Control group Ⅰwere treated with carbamazepine tablet and clonazepam tablet while control group Ⅱ were treated with valproate sodium tablet and clonazepam tablet.The efficacies of three groups were compared after 6 months.RESULTS: Treatment group was better than control groups in total effective rate,seizure frequency and improvement of EEG (P

Objective To construct a eukaryotic vector which contains avian H5N1 influenza virus hemagglutinin (HA) antigen and the cholera toxin B subunit (CTB) and to investigate its expression in COS7 cells,and the ability to induce specific immune responses in vivo in different periods.Methods After cloned by polymerase chain reaction (PCR),CTB and HA genes were digested with BamH Ⅰ and connected into CTB-HA gene with T4 ligase.The connected gene was referred to as CH.After double digestion,CH gene was inserted into a eukaryotic recombinant plasmid pCI-neo.The pCI-CH plasmid was then transfected into COS7 cells.Western blot was used to detect the expression of HA antigen.After New Zealand white rabbits were immunized,the titer of HA antigen-specific antibody in serum and its specificity with other strains such as H1N1,H9N1,H3N2 and influenza B virus were determined by indirect enzyme-linked immunosorbent assay.Results The pCI-CH vector (DNA vaccine) was successfully constructed,which could be efficiently expressed in COS7 cells and induce specific antibodies against pCI-CH in rabbits.Cross reactions indicated that DNA vaccine pCI-CH specific antisera could not only react with H5N1 strain (P/N＞2.1),but also H1N1,H9N1 and H3N2 strains,but did not cross react with influenza B virus.Conclusion The newly constructed avian H5N1 influenza virus nucleic acid vaccine has good immunogenicity.

Objective To explore on FK506 promoting proliferation of Schwann cells in vitro and NGF of Schwaun cells secreted highly by itself. Methods Purified Schwann cells divide into six groups:group A (control group) DMEM/F12 contained 10% calf bloodserum; group B contained 0.1 ng/ml FK506; group C contained 0.5 ng/ml FK506; group D contained FK506:1.0 ng/ml;group E contained FK506:5.0 ng/ml; group F contained FK506:10 ng/ml. Morphology of Schwann cells were oboyrved by invert microscope and evaluated Schwann cells in immunocytochemistry staining with anti-S-100. The best concentration of FK506 who promoted proliferation of Schwann cells by MTT. Cell cycle of Sehwarm cells were determined by flow cytometry. The level of NGF in the conditioned media was determined by an enzyme-linked immunoadsordcnt assay after 72 h. Results Group C was the best concentration which promoting proliferation of Schwann cells among 5 groups, when the concentration 1.0 ng/ml FKS06 to promote Schwann cell proliferation gradually weakened. Detected by flow cytometry showed that: containing 10% fetal DMEM/F12 bovine serum for 24 h,72 h and 48 h after Schwann cells in S phase percentage were 27.8%,39.3% and 58.4% in the 0.5 ng / ml FK506 for 24 h,72 h and 48 h after Schwann cells S percentage period were 54.2% ,60.3% and 94.6%. S phase of the latter than the former in 24 h,72 h and 48 h, respectively higher: 26.4% and 21% and 36.2%. FK506 detected by ELISA promote Schwann cell proliferation after the expression of NGF in the experimental study found: 0.5 ng/ml FK506 for 72 h after the Schwann cells secreted by the NGF as high as 0.188 ng/ml, rcspectiveIy. Conclusion FKS06 can promote proliferation of Schwann cells at early time in vitro and Schwann cells' good situation is highly kept to secrete NGF.

A rapid method was developed for the determination of 5 common fatty acids, palmitic acid, stearic acid, oleic acid, linoleic acid and linolenic acid in industrial oleic acid based on ultra_performance convergence chromatography_mass spectrometry ( UPC_MS) . The sample was dissolved by n_hexane, followed by clean_up of extract using 0. 22 μm organic phase filter. The fatty acids were separated in 3 min on the column of Acquity UPC2 BEH 2_EP by gradient elution with carbon dioxide and methanol/acetonitrile (1∶1, V/V) system, and finally detected by MS detector in ESI- mode. Through the optimization of UPC2_MS condition, the reasonable linearity was achieved for all the analytes over the range of 0. 5-100 mg/L with the correlation coefficients ( R2 ) greater than 0. 9985. The recoveries for five fatty acids at three spiked levels were in the range from 89 . 3% to 106 . 67% with relative standard deviations of 0 . 8%-3 . 0%. The limits of detection for target compounds in the method ranged from 0. 07 mg/L to 0. 26 mg/L. The real sample analysis showed that this method was simple,fast and had a good separation effect. There was no need of derivatization for fatty acid samples. This work would provide a fast and effective detection method for UPC2 technology in oil related research field.

Aim To observe the improvement of visual-auditory cognitive functions in the human entering high altitude by taking in tea polyphenols.Methods Thirty eight males living at 3 700 m high altitudes for 90 days constantly were randomly divided into two groups: ①group Ⅰ(placebo,40 mg/day); ②group Ⅱ(TP,300 mg/day).Cognitive functions were measured by integrated visual and auditory continuous performance test and the difference between groups was evaluated by the comparisons of post-treatment to pre-treatment.Results Compared with pre-treatment,PruA was significantly higher after taking in TP(P

AIM: To investigate whether aorta-derived CD_(105)~+ cells show characteristics of mesenchymal stem cells, and if dexamethason enhances this kind of CD_(105)~+ cells to differentiate into adipocytes. METHODS: The distribution of CD105 in aorta was assessed by immunohistochemistry. The aorta wall cells were isolated and immunophenotypes were identified by FACS. CD_(105)~+ cells were sorted using MACS CD105 micromagnetic beads. The differentiation of CD_(105)~+ cells into adipocytes and osteoblasts was induced under different conditions and indicated by staining of Oil red O, detecting of alkaline phosphatase activity, calcium accumulation stained with silver nitrate and transmission electron microscope analysis, respectively. RESULTS: The endothelial cells, a part of medial smooth muscle cells and adventital fibroblasts were CD105 positive. The isolated aortic arch cells were positive for CD105, CD106, CD44, CD29, and negative for CD45, CD11a, CD11b and HLADR. The CD_(105)~+ cells differentiated into adipocytes contained Oil-Red-O-positive lipid droplets, the osteocytes with calcium deposition and alkaline phosphatase activity. Ultrastructurally, it was observed that some needle-shaped crystal calcium deposition similar to bone spicules was inside the cytoplasm of induced osteocytes. When the dexamethason was absent in the adipogenic medium, there were no adipocytes with lipid droplets. CONCLUSION: The results demonstrate that CD_(105)~+ cells show characters of MSCs reside in aortic wall, and is able to differentiate into adipocytes and osteocytes in vitro. Dexamethasone enhances aorta-derived CD_(105)~+ with characters of MSCs to differentiate into adipocytes. These suggeste that MSCs might be related with atherosclerosis. [

AIM: To explore the mechanism underlying inducible nitric oxide (NO) caused injury of endothelial cells during inflammation. METHODS: The activity of iso-enzymes of NO synthase (NOS), NO level and iNOS expression were examined using NADPH method, Griess reaction and RT-PCR, respectively. Furthermore, the lactate dehydrogenase (LDH) release rate, malondialdehyde (MDA) content were also measured. RESULTS:Co-administration of cytokines (TNF-? 5?10 5 U/L, IL-1? 2?10 5 U/L, INF-? 2?10 5 U/L) and LPS (10 mg/L) caused an obvious increase in NOS activity, NO levels (about two-fold) and a significant injury of the cells. At the same time, a significant increase in iNOS mRNA was also detected. Wheareas, treatment of the cells separately with cytokines or LPS for 24 h had no significant effect on NOS activity and NO level in cell lysates, however, it caused a significant increase in LDH release and MDA content. Also, the effect of cytokines and LPS on cell viability was concentration-and time-dependent. L-NMMA, a inhibitor of NOS, can suppress inducible NO production and protect cells against NO induced injury. CONCLUSION:Co-administration of cytokines (TNF-?, IL-1? and INF-?) and LPS significant activated iNOS and NO production which, in turn, induced oxidative reaction in endothelial cells.

AIM: To explore the effect of aspirin on inducible nitric oxide synthesis and gene expression under inflammation in endothelial cells. METHODS: Using NADPH, Griess methods and RT-PCR, the activity of isozymes of NO synthase (NOS), nitric oxide (NO) level, and iNOS mRNA expression were examined respectively. Also, the lactate dehydrogenase (LDH) release rate, malondialdehyde (MDA) content and cell viability were measured. RESULTS: Aspirin (3 mmol/L) reduced inducible NO production and NOS activity(P

AIM: To explore the regulatory effects of ferritin expression and intracellular iron change on aspirin resistance to oxidative damage in endothelial cells. METHODS: Using ELISA to measure the levels of ferritin expression under different aspirin concentrations, in the presence of iron cheltor desferioxamine and add to FeCl 3. Then using RNA-protein bandshift assay and RT-PCR to examine the activation of IRP and the expression of IRP 2 mRNA onaspirin induced ferritin formation. RESULTS: Aspirin at low concentration (0.1mmol/L) induced significant increase in ferritin expression in a concentration-dependent fashion up to 25% over basal levels. Aspirin induced cytoprotection from H 2O 2 damage increased significantly following ferritin formation in endothelial cells.However, in the presence of iron chelator desferrioxamine, aspirin enhanced ferritin synthesis was abrogated with a 3 fold increase in the activity of IRP and significant increase in IRP 2 mRNA level. In contrast, FeCl 3 and aspirin both increased the level of induced ferritin synthesis with significant decrease in IRP activity and IRP 2 mRNA level. CONCLUSION: The effect of aspirin induced ferritin synthesis on resistance to oxidative damage in endothelium was operated through down-regulating IRP activation and IRP 2 mRNA level.