Animals ; Diaphragm ; physiopathology ; Electrophysiological Phenomena ; physiology ; Hypoxia ; physiopathology ; Male ; Rats ; Rats, Wistar ; Sleep Apnea Syndromes ; physiopathology

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 investigate the incidence of Keshan disease(KSD) in Shandong province and to supply evidence for scientific control of the disease.Methods According to the scheme of KSD monitoring in Shandong province,the KSD cases were searched in 7 counties,and representative 3 counties and 6 villages were selected as key monitoring sites.In each site,about 420 residents were checked by asking detailed disease history,physical examination,and electrocardiography (ECG) tracings.Suspected cardiac abnormalities were taken chest posteroanterior X-ray film.At the same time,residents hair and food samples were collected to detect selenium levels in internal and external environments,and residents' income and per capita share of grain and other basic information were collected.Results ①Two hundred and fifty-nine medical units were searched,1132 cases of myocardial disease were found out and 638 suspected cases were identified and 142 chronic KSD cases were diagnosed.②In the 6 monitoring sites,2538 residents were surveyed and 91 cases of KSD were detected(of which 77 cases of potential,14 cases of chronic),the detection rate was 3.58％.A total of 2127 residents were traced ECG and the incidence of abnormal ECG was 24.5％ (521/2127),of which ST-T changes,T-wave changes,sinus bradycardia,sinus arrhymia,sinus tachycardia,and atrial premature beats et al were more common.Of the 75 X-ray films,39 cases had enlarged heart(in which 21 of mild,13 of moderate,5 of severe).③A total of 406 hair and grain samples were collected,respectively,and the selenium content of hair was approaching the level of appropriate,and the selenium content of food was still low.The annual per capita incomes of each site were between 3000 - 5900 Yuan,and residents main staple food was flour.Conclusions KSD in Shandong province is in a relatively stable situation.But latent and chronic KSD cases can still be detected.The internal environment selenium levels are elevated since the local living standards and dietary nutrition are improved.

Objective To observe the electrocardiogram changes of threatened crowds in Keshan disease (KSD) endemic area in Shandong province. Methods In 2008,inhabitants from 21 villages of Zoucheng,Sishui,Tengzhou, Yishui, Pingyi, Wulian, Juxian and Qingzhou regions were selected as subjects undergoing electrocardiogram. No less than 100 people were chosen from each village and the examination rate was not lower than 85%. Results Among the 3378 inhabitants investigated,460 cases showed abnormal electrocardiogram and the total incidence of abnormal electrocardiogram was 13.62% (460/3378). The relatively high incidence was T-wave changes,QRS low voltage and ST-T changes,the detection rate being respectively 2.69% (91/3378), 1.92% (65/3378) and 1.72% (41/3378). The highest incidence of abnormal electrocardiogram (26.76%,55/213),the intermediate(21.50%,43/200) and the lowest(5.50%,12/218) was respectively found in Pingyi,Qingzhou and Sishui. Conclusions The threatened crowds in KSD endemic area in Shandong province are still in a state of high abnormal electrocardiogram detection,and electrocardiogram is of great value in the evaluation of KSD patients.

OBJECTIVETo maked a Nrf2 down-regulated cell line by over-expressing Keap1 in H460 cells to study the role of Nrf2 in drug resistance.

METHODSTransfecting H460 cells with mKeap1-pEGFP and screenig for Keap1 expressing clones by Western blotting with antibodies against Nrf2, HO-1, NQO1 and AKR1C. The cell line with Keap1 over-expression was further confirmed by real-time PCR. The cytotoxicity of H460-N5 to anti-cancer drugs was evaluated by MTS assay.

RESULTMTS assay results showed the enhanced cytotoxicity of anticancer drugs (Oxaliplatin, Doxorubicin and Etopside) to the H460 cell line with keap1 overexpression compared to the control cell line. In H460-N0 cells, the IC(50) values of Oxaliplation and Etopside were 93 micromol/L and 100 micromol/L respectively whereas the IC(50) values of the two drugs were 42 micromol/L and 30 micromol/L correspondingly in H460-N5 cells. A Nrf2 down-regulated cell line H460-N5 and a control cell line with GFP over-expression have been identified.Down-regulation of Nrf2 enhanced the cytotoxicity of Oxaliplatin, Doxorubicin and Etopside. The IC(50) value of Doxorubicin to H460-N0 cell was above 3 mg/L, but that to H460-N5 cell was about 2 mg/L.

CONCLUSIONA Nrf2 down-regulated cell line H460-N5 and a control cell line with GFP over-expression have been identified. Down-regulation of Nrf2 enhanced the cytotoxicity of Oxaliplatin, Doxorubicin and Etopside.

Antineoplastic Agents ; pharmacology ; Antioxidants ; metabolism ; pharmacology ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Cell Line, Tumor ; Down-Regulation ; Drug Resistance, Neoplasm ; genetics ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Kelch-Like ECH-Associated Protein 1 ; Lung Neoplasms ; metabolism ; pathology ; NF-E2-Related Factor 2 ; genetics ; metabolism ; physiology ; Response Elements ; physiology ; Signal Transduction ; physiology ; Transfection

OBJECTIVETo study the oxidative stress induced by consumption of mercury-contaminated rice in rats, and to assess the possible public health risk of mercury contamination in Wanshan mining area.

METHODSSprague Dawley rats were fed the mercury-contaminated rice produced from Wanshan area for 90 days. The antioxidant status and the free radicals in rat serum were evaluated.

RESULTSHigh mercury accumulation in organs of rats fed the mercury-contaminated rice confirmed the server pollution of mercury in Wanshan mining area. The intensity of electron spin resonance (ESR) signal increased by 87.38% in rats fed the rice from Wanshan compared with that in the control rats fed the rice from Shanghai, suggesting that chronic dietary consumption of rice from mercury mining area could induce an aggravation of free radicals. Feeding the mercury-contaminated rice was associated with significant decreases in the antioxidant enzymatic activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) and concentration of serum nitric oxide (NO), but it had no effect on serum nitric oxide synthase (NOS) activity. Feeding the mercury-contaminated rice raised the level of serum malonyldialdehyde (MDA), indicating the occurrence of oxidative stress.

CONCLUSIONThe long-term dietary consumption of mercury-contaminated rice induces the aggravation of free radicals and exerts oxidative stress.

Animals ; Brain ; metabolism ; China ; Environmental Pollutants ; analysis ; pharmacokinetics ; toxicity ; Food Contamination ; analysis ; Free Radicals ; blood ; Glutathione Peroxidase ; blood ; Industrial Waste ; adverse effects ; Kidney ; metabolism ; Liver ; metabolism ; Malondialdehyde ; blood ; Mercury ; analysis ; pharmacokinetics ; toxicity ; Methylmercury Compounds ; analysis ; pharmacokinetics ; toxicity ; Nitric Oxide ; blood ; Nitric Oxide Synthase ; blood ; Oryza ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; blood

A full-length cDNA encoding a MYB-related regulatory gene was isolated from a cDNA library prepared from mRNAs of the red line callus of S. medusa by TD-PCR. The cDNA, designated SmP, is 969 nucleotides long and has an open reading frame of 771 bp with a deduced amino acid sequence of 256 residues. The putative protein of SmP has two typical conversed R2R3-Myb DNA-binding domains in N-terminal and displays a rather high degree of similarity to OsMYB from rice and LBMI from tobacco, showing 73% and 70% identity within the DNA-binding domains. However, the C-terminal domain of the SmP protein does not show obvious similarity to any other known protein sequence. It is rich in hydrophilic amino acids, especially in serine residues (18.38%), partly organized in homopolymeric stretches, a feature often found in activation domain of transcription factors.
Amino Acid Sequence ; Cloning, Molecular ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; classification ; genetics ; metabolism ; Polymerase Chain Reaction ; Saussurea ; classification ; genetics ; metabolism ; Sequence Homology, Amino Acid ; Transcription Factors ; chemistry ; genetics ; metabolism

OBJECTIVETo investigate the effect of tBHQ and sulforaphane on the protein expression in Nrf2-ARE signaling pathway of Caco2 cells.

METHODSHuman colorectal carcinoma Caco2 cells were treated with 20 micromol/L tBHQ and 5 micromol/L sulforaphane (SFN) respectively. Real time PCR, Western blotting and immunoflourescence staining (IF) were performed to measure the target gene expression.

RESULTSNrf2, AKR1C1 and NQO1 protein expressions were increased time-dependently in Caco2 cells after treatment with tBHQ and SFN. Time-course experiments showed that tBHQ and SFN increased the accumulation of Nrf2, and concomitantly increased the protein levels of AKR1C1 and NQO1. Real-time PCR and Western blotting showed that tBHQ and SFN significantly increased the expression of Nrf2 at 8h after the treatment, and AKR1C1 and NQO1 at 16 h. Confocal microscopy technique showed that Nrf2 accumulated in the nucleus at 6-8 h after treatment with tBHQ. After 1 h treatment with tBHQ the nuclear Nrf2 maintained at elevated level for at least 4 h with tBHQ withdrawn.

CONCLUSIONtBHQ and SFN induced nuclear accumulation of Nrf2 and activated Nrf2-dependent regulation of ARE-mediated gene expression in Caco2 cells. In addition, the results provide experimental evidence for choosing the dose and frequency of the inducer in cancer chemoprevention study and in developing inhibitors of Nrf2-ARE signaling pathway.

Anticarcinogenic Agents ; pharmacology ; Antioxidants ; metabolism ; pharmacology ; Caco-2 Cells ; Calcium-Transporting ATPases ; antagonists & inhibitors ; Humans ; Hydroquinones ; pharmacology ; Isothiocyanates ; NF-E2-Related Factor 2 ; genetics ; metabolism ; physiology ; Oxidative Stress ; genetics ; physiology ; Response Elements ; physiology ; Signal Transduction ; drug effects ; Thiocyanates ; pharmacology

BACKGROUNDEvaluation of fetal central nervous system (CNS) agenesis by ultrasonography (US) is frequently limited, but magnetic resonance imaging (MRI) has its own advantages and is gaining popularity in displaying suspected fetal anomalies. The purpose of this study was to explore the value of MRI in detecting fetal CNS agenesis.

METHODSThirty-four women (aged from 22 to 35 years, average 27 years) with complicated pregnancies (16 - 39 weeks of gestation, average 30 weeks) were examined with a 1.5 T superconductive MR unit within 24 hours after ultrasonography. Half-Fourier acquisition single-shot turbo spin-echo (HASTE) T(2)-weighted imaging (T(2)WI) sequence were performed in all patients, and fast low angle shot (FLASH) T(1)-weighted imaging (T(1)WI) sequence were applied sequentially in seven of them. Comparison of the results was made between the MRI and US findings as well as autopsy or postnatal follow-up MRI findings.

RESULTSThe gyrus, sulcus, corpus callosum, thalamus, cerebellum, brainstem, and spinal cord of fetus were shown more clearly on T(2)-weighted MR images than on T(1)-weighted MR images. MRI corrected the diagnosis of US in 10 cases (10/34, 29.41%) and the diagnosis was missed only in 1 case (1/34, 2.94%).

CONCLUSIONMRI has advantages to US in detecting fetal CNS anomalies and is a supplement to US in complicated pregnancies.

Adult ; Central Nervous System ; abnormalities ; diagnostic imaging ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Pregnancy ; Ultrasonography, Prenatal

OBJECTIVETracheobronchomalacia is one of the common respiratory tract dysplasia in children. Its symptoms are nonspecific, and routine methods are unreliable in the assessment of tracheobronchomalacia in children. In addition, many physicians are confused about its clinical characteristics, so tracheobronchomalacia is often underdiagnosed. The purpose of this study was to explore the clinical features of tracheobronchomalacia in children and to investigate the diagnostic value of flexible bronchoscopy for children with tracheobronchomalacia.

METHODFor diagnosis and treatment, 229 children out of 4725 patients hospitalized in the division of respiratory disorders were examined by Olympus BF3c-20 flexible bronchoscopy or by Olympus BF-P20 flexible bronchoscopy under general anesthesia with propofol, in Chongqing Children's Hosptial from April 2004 to April 2006. Fifty-three cases were confirmed to have tracheobronchomalacia by bronchoscopy, patients' data including airway lesion, age, sex, clinical characteristics, aided examinations, treatment, final outcomes, were collected and analyzed.

RESULTS(1) Of the 53 children with tracheobronchomalacia, 31 were not suspected for this diagnosis prior to bronchoscopy, who were instead misdiagnosed as refractory pneumonia, difficult-to-control asthma, bronchial foreign body, bronchopulmonary dysplasia and pulmonary atelectasis of unknown origin or bronchiolitis. (2) In the 53 children with tracheobronchomalacia aged one month to eight years, 41 were infants, 6 were younger than two years, 4 were younger than 3 years and the rest 2 cases were older than 3 years. The risk of tracheobronchomalacia related inversely with ages. Ten cases were girls and 43 were boys. (3) Eleven cases had tracheomalacia, 24 bronchomalacia, 18 tracheobronchomalacia; 12 cases had malacia on left lung, 11 on right lung, 19 on both sides; 21 children were mild cases, 25 moderate cases, 7 severe cases. (4) In the 53 children with tracheobronchomalacia, 28 had recurrent or prolonged wheezing, 16 chronic cough, 5 recurrent respiratory infections, 2 atelectasis of unknown origin, and 2 dyspnea.

CONCLUSIONSThe infants and toddlers seem to be predisposed more to the bronchomalacia than the older children. Clinical features of children with airway malacia were variable and atypical, expiratory stridor and cough are the most commonly reported symptoms. Flexible bronchoscopy should be regarded as a "golden standard" method for diagnosing TBM.

Bronchoscopy ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Tracheobronchomalacia ; diagnosis ; pathology