Objective To explore the effects of thiamine and riboflavin on H2O2-induced DNA oxidative damage in human umbilical vein endothelial cell line ECV304.Methods ECV304 cells were incubated with 10,100,500,1000 mg/L of thiamine or 20,100,300,500 nmol/L of riboflavin for 24 h,and then oxidative damage of cells were induced by 25 mol/L H2O2 for 30 min.DNA damage was detected with single cell gel electrophoresis(SCGE)assay.ECV304 cells incubated without H2O2,thiamine and riboflavin were served as negative controls,and those incubated with H2O2 and without thiamine and riboflavin were served as positive controls.Results H2O2 induced DNA damage,and the indices of percent of DNA damage cells,percent of tail DNA,tail length and Olive tail moment were increased.The indices of cells pretreated with 10,100,500 mg/L of thiamine or 20,100,300 nmol/L riboflavin were significantly decreased(P0.05).Conclusion Proper supplementation of thiamine and riboflavin may decrease H2O2-induced DNA oxidative damage,while excess thiamine and riboflavin supplementation may be harmful to DNA and enhance the susceptibility to H2O2 potentially.

AlM:To evaluate the clinical efficacy of intravitreal injection of ranibizumab combined with macular grid photocoagulation for diabetic macular edema ( DME) .METHODS:Totally 60 eyes ( 60 patients ) with DME were randomly divided into 2 groups: 30 eyes of simple injection group underwent intravitreal injection of ranibizumab, and 30 eyes of combined treatment group underwent intravitreal injection of ranibizumab and macular grid photocoagulation 1wk later. The best corrected visual acuity ( BCVA ) , central macular thickness ( CMT ) measured by optical coherence tomography ( OCT ) and postoperative complications were observed.
RESULTS:ln simple injection group, the BCVA after operation were separately 0. 390 ± 0. 075 (4wk), 0. 367 ± 0. 088 (8wk) and 0. 319 ± 0. 064 (12wk), the CMT after operation were separately 221. 63 ± 112. 34μm (4wk), 337. 73±99. 56μm (8wk) and 432. 92 ± 100. 46μm (12wk), which were much better than pre-operation. But during follow-up, the BCVA presented down trend and the CMT was on the rise slowly. ln combined treatment group, the BCVA after operation were separately 0. 385 ± 0. 036 (4wk), 0.382±0.079 (8wk) and 0.377±0.097 (12wk),the CMT after operation were separately 249. 77 ± 106. 55μm (4wk), 270. 40 ± 92. 88μm (8wk) and 275. 84 ± 97. 34μm (12wk ), which were satisfactory and steady during follow-up, better than simple injection group (P<0. 05).CONCLUSlON:lntravitreal injection of ranibizumab can effectively improve visual acuity and decrease central foveal thickness for patients with DME, combining with macular grid photocoagulation can ensure therapeutic effects steady and permanent.

Amygdala ; drug effects ; metabolism ; Animals ; Conditioning (Psychology) ; Morphine ; adverse effects ; Rats ; Receptors, Opioid, kappa ; metabolism

OBJECTIVETo investigate the effects of transcriptional inhibitors 5, 6-dichloro-1-b-D-ribofuranosylbenzimidazole (DRB) and alpha-Amanitin on the localization of Nrf2 in the nucleus.

METHODSA549 cells were treated with DRB (50 mg/L) or alpha-Amanitin (2.5 mg/L)for 1 h and 6 h in serum-free medium, respectively. The expressions of Nrf2, HO-1, NQO1 and AKR1C were detected by Western blotting analysis. The localization of Nrf2 was determined by laser scanning confocal microscopy after cells were treated with either DRB or agr:-Amanitin for 1 h.

RESULTSThe expressions of Nrf2 and Nrf2-ARE gene batteries HO-1, AKR1C and NQO1 were decreased after 6 h treated with either DRB or alpha-Amanitin. The expression of SC35 was up-regulated but RNA Pol II was down-regulated; Y12 and NPC did not significantly change. The localization of Nrf2 in the cell nucleus did not change significantly.

CONCLUSIONDRB and alpha-Amanitin can down-regulate the expression of Nrf2 and its targeting proteins HO-1, AKR1C and NQO1, but may have no effect on the localization of Nrf2.

20-Hydroxysteroid Dehydrogenases ; genetics ; metabolism ; Alpha-Amanitin ; pharmacology ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Dichlororibofuranosylbenzimidazole ; pharmacology ; Heme Oxygenase-1 ; genetics ; metabolism ; Humans ; Lung Neoplasms ; metabolism ; pathology ; NF-E2-Related Factor 2 ; genetics ; metabolism ; Nucleic Acid Synthesis Inhibitors ; pharmacology

Objective To observed hepatolenticular degeneration gene (ATP7B gene) expressed in Me32aT22/2L cell and detected whether A TP7B could transport redundant copper and reduce copper-induced cell apoptosis, which will make the basement for future gene therapy. Methods Piasmid pRc/CMV-WD containing A TP7B cDNA was transfected into Me32aT22/2L cell using lipofectamin2000 transfection methods. Intracellular distribution of ATP7B was detected by immunofluorescence histochemistry method and copper transportation function was studied using high-concentration copper incubation experiments, meanwhile, cell apoptosis induced by high-concentration copper incubation was observed. Results Expression of A TB7B gene could be detected and located around nuclei within the Me32aT22/2L cell. After incubation of high- concentration copper solution after 24, 48 and 72 hours, in empty vector group intracellular copper content was (600.60±69.71) ng/mg, (890.72±65.74) ng/mg and (1189.20±85.71) ng/mg respectively, however, intracellular copper content was (351.33±49.86) ng/mg, (427.38±30.95) ng/mg and (539.10±34.91) ng/mg in A TP7B group. Comparison of two groups has a statistical significance (P<0.01). Meanwhile, cell apoptotic rate was markedly decreased in A TPTB group compared with empty vector group (P<0.01). Conclusion Exogenous A TP7B gene could be expressed in Me32aT22/2L cell and A TPTB could transport intracellular redundant copper, subsequently reduced copper-induced cell apoptosis.

Child ; Child, Preschool ; Female ; Growth Disorders ; blood ; diagnosis ; Human Growth Hormone ; deficiency ; Humans ; Insulin-Like Growth Factor Binding Protein 3 ; blood ; Insulin-Like Growth Factor I ; analysis ; Male

OBJECTIVETo study the difference of plasma actual bicarbonate between the children with growth hormone deficiency (GHD) and idopathatic short stature (ISS) and to value the plasma bicarbonate standard deviation scores (SDS) in diagnosis of GHD.

METHODSForty-seven short stature children were divided into two groups (GHD and ISS) according to the peak GH response to provocative test. Plasma actual bicarbonate concentrations anion gap (AG), base excess and electrolytes were measured in 47 children with short stature before GH provocative tests.

RESULTSThe mean plasma actual bicarbonate concentrations, bicarbonate SDS were (22.60+/-1.29)mmol/L and -0.27+/-0.98 respectively in GHD children, which were significantly lower than those of ISS children (P<0.01), whereas AG was higher than that of ISS children [(11.73+/-4.52 vs 7.87+/-1.70) mmol/L], P<0.01. Seventy-two percent of patients with bicarbonate SDS

CONCLUSIONPlasma actual bicarbonate concentrations and bicarbonate SDS are lower in patients with GHD than those in patients with idopathatic short stature. Evaluation of plasma bicarbonate SDS of short stature children can predict the probability of GHD, especially when bicarbonate SDS is less than 1 s.

Acid-Base Equilibrium ; Adolescent ; Bicarbonates ; blood ; Child ; Child, Preschool ; Female ; Growth Disorders ; metabolism ; Human Growth Hormone ; deficiency ; Humans ; Male

This study was purposed to investigate the inhibition of hTERT antisense oligodeoxynucleotide (ASODN) on the proliferation and telomerase activity in HL-60 cells and to explore the relativity between the telomerase activity and the expression of hTERT gene in HL-60 cells. After treated by hTERT ASODN the expression of hTERT was detected by RT-PCR, the morphological changes of HL-60 cells was observed with inverted microscopy, the cell proliferation was measured by MTT method, and the telomerase activity was determined with TRAP-ELISA and TRAP-PAGE. The results showed that after sealing hTERT gene with ASODN for 72 hours, the expression of hTERT gene was significantly inhibited, the cell growth was repressed and the ability of proliferation decreased, and the effect was specific in sequence and dependent in dose and time. OD(450-690) values were 2.648 +/- 0.42, 1.504 +/- 0.47, 1.223 +/- 0.39, 0.944 +/- 0.16 respectively, as the cells were treated with 0, 10, 20, 30 micromol/L ASODN for 72 hours. The difference was significant as compared 10, 20, 30 micromol/L groups with 0 micromol/L ASODN group respectively (P < 0.05), but the difference was no significant when compared 20 micromol/L SODN group (2.376 +/- 0.65) with untreated group (2.648 +/- 0.42) (P > 0.05). TRAP-PAGE detection revealed that comparing ASODN groups with SODN groups the telomerase image bands were decreased and least was found in groups of 30 +/- mol/L. It is concluded that the hTERT ASODN may inhibit the proliferation and down-regulate the telomerase activity in HL-60 cells by sealing the expression of hTERT gene.
Cell Proliferation ; drug effects ; HL-60 Cells ; Humans ; Oligonucleotides, Antisense ; biosynthesis ; genetics ; Telomerase ; biosynthesis ; genetics ; metabolism ; pharmacology ; Transfection

OBJECTIVETo recognize the clinical features of the bronchiolitis obliterans.

METHODClinical manifestation, chest X-ray, computed tomography (CT) and pulmonary function of 4 cases with bronchiolitis obliterans were retrospectively analyzed.

RESULTTwo cases were after Stevens-Johnson syndrome (SJS), the other 2 were after severe pneumonia, including one suffered from adenovirus pneumonia. Cough, tachypnea and wheezing persisted in all the 4 patients. The symptoms lasted for at least 6 weeks, in one case for over one year. Crackles and wheezing were present in all the 4 cases. Hyperinflation was seen in chest radiographs in all cases. On pulmonary CT/high-resolution CT (HRCT), patchy opacity and bronchial wall thickening were seen in each patient. Areas of air trapping were seen in three cases. Bronchiectasis was seen in 2 cases, atelectasis and mosaic perfusion were seen respectively in one case. PO(2) was low in all the four cases. Wheezing was not responsive to beta(2) agonist and other bronchodilating therapy. Prednisone was used at a dose of 1 mg/(kg.d) in 3 cases. Two cases were followed up for 3 months. The clinical condition of one case was improved, whose wheezing and bronchiolar constriction disappeared, cough and dyspnea were also relieved. However, the condition of one patient was not improved, although the wheezing disappeared. The HRCT of these two cases showed no improvement.

CONCLUSIONClinical symptoms of BO were cough, tachypnea, and wheezing after acute lung injury. Crackles and wheezing were the most common signs in the BO. Chest radiographs showed hyperinflation. Pulmonary CT showed bronchial wall thickening, bronchiectasis, atelectasis, and mosaic perfusion. Pulmonary function tests suggested obstruction of small airway.

Bronchiolitis Obliterans ; etiology ; pathology ; physiopathology ; Child ; Child, Preschool ; Humans ; Infant ; Male ; Pneumonia ; complications ; Pneumonia, Viral ; complications ; Prognosis ; Respiratory Function Tests ; Stevens-Johnson Syndrome ; complications ; Tomography, X-Ray Computed

OBJECTIVETo investigate the effect of Luteolin alone or combination with chemotherapentic drugs on the cytoxicity of cancer cells.

METHODSCultured A549, Hela, MCF-7, AGS, MGC-803, Caco2 and HepG2 cells were treated with Luteolin or the combination of Luteolin with other chemotherapeutic agents (Bexarotene, Cisplatin and Bleomycin). Cell viability was measured by MTS assay and IC(50) was calculated.

RESULTSThe IC(50) of Bexarotene to Hela cells was 2 micromol/L, but with the combination of 5 micromol/L of Luteolin that reduced to 0.2 micromol/L. However, the combination of Bexarotene and Luteolin did not show significant benefit in MGC-803, HepG2 cells, Caco2 and MCF-7 cells. The IC(50) of Cisplatin to Hela cells was over 30 micromol/L,but it decreased to 3 micromol/L in the presence of 5 micromol/L Luteolin; Luteolin also sensitized Cisplatin in MGC-803, HepG2 and A549 cells studied. The IC(50) of Bleomycin to Hela cells was over 100 micromol/L, but it was about 1 micromol/L in the presence of 5 micromol/L Luteolin. A549 cells were resistant to Bleomycin with an IC(50) of 100 micromol/L, 10 micromol/L Luteolin greatly enhanced the cytotoxicity of Bleomycin to the cells with the IC(50) of about 10 micromol/L. The inhibitions of MGC-803, HepG2, A549 and AGS cells didn't change by combination of Luteolin.

CONCLUSIONLow concentration of Luteolin has little toxic effect on the cancer cell lines tested in the study, but it can sensitize chemotherapeutic drugs in various cancer cell lines.

Antineoplastic Agents ; pharmacology ; Breast Neoplasms ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Synergism ; Humans ; Lung Neoplasms ; pathology ; Luteolin ; pharmacology ; Neoplasms ; pathology