This study was aimed to investigate the growth inhibition effect of diimide G-quadruplex ligand on leukemia cells and to explore its molecular mechanisms. K562 leukemia cell lines were treated with various concentrations of the diimide G-quadruples ligand small molecule (0.1 - 10 micromol/L). Trypan blue exclusion assay was used to evaluate the proliferation inhibition. Cell apoptosis was observed using terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL). Telomerase activity was analyzed by telomere repeat amplification protocol. Gene expression was detected by microarray and confirmed by RT-PCR assay. The results showed that diimide small molecule inhibited the proliferation of K562 cells and induced apoptosis of these cells. After treating with diimide G-quadruplex ligand, telomerase activity of K562 cells was reduced and the transcriptional levels of some important genes were changed significantly. These genes were involved in cell apoptosis, cell signaling pathway and other key functions. In conclusion, the diimide G-quadruplex ligand is a small molecule that inhibits the proliferation and induces apoptosis in leukemia cells, and these functions may be related to telomerase inhibition and regulation of some important gene transcription.
Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; G-Quadruplexes ; Humans ; K562 Cells ; Leukemia ; genetics ; pathology ; Ligands ; Microarray Analysis ; Telomerase ; metabolism

Objective To study the expression of plasma oxidized low-density lipoprotein (oxLDL) in children with acute phase Kawasaki disease (KD), and investigate its value for early prediction of coronary artery lesions in KD. Methods Totally 80 children with KD were collected. Children were divided into four groups by the results of echocardiogram of coronary artery in different periods: CAL1 group (children with coronary artery lesions (CAL+) both in acute and sub-acute phase, 8 cases), CAL2 group (children with CAL+in acute phase but recovery normal (CAL-) in sub-acute phase, 10 cases), NCAL1 group (children with CAL-in acute phase but occur CAL+ in sub-acute phase, 10 cases) and NCAL2 group (children with CAL- both in acute and sub-acute phase, 52 cases). The serum samples (before the use of intravenous immunoglobulin) were collected in acute phase. Twenty healthy controls and twenty fever controls were enrolled into the study, and their serum samples were collected. OxLDL was measured by enzyme linked immunosorbent assay (ELISA). They were compared using ANOVA, pairwise comparison LSD-t test. And ROC curve analysis was used to determine the threshold. Results Compared with the control groups,plasma oxLDL levels were higher in children with KD, both CA+and CAL-[(15.0±3.3) mU/L, (12.3±3.5) mU/L vs (9.2±2.2) mU/L, (8.0±2.3) mU/L, F=20.435, P<0.05]. Plasma oxLDL levels were increased more significantly in children with CAL+ than children with CAL- in KD [(15.0 ±3.3) mU/L vs (12.3 ±3.5) mU/L, t=2.28, P=0.002]. There was significant difference in the concentration of oxLDL between the groups of Kawasaki disease (F=5.068, P=0.003). Plasma oxLDL levels were significantly higher in the NCAL1 group than those in the NCAL2 group [(14.5 ±3.8) mU/L vs (11.9±3.3) mU/L, t=2.29, P=0.02], but there were no statistically significant difference between the NCAL1 group and CAL1 or CAL2 group [(14.5±3.8) mU/L vs (15.9±3.9) mU/L, (14.5±3.8) mU/L vs (14.2±2.7) mU/L, t=0.73, 0.20;P=0.41, 0.84]. ROCs analysis indicated that oxLDL≥13.83 mU/L, could be the threshold for the prediction of coronary artery lesions with the sensitivity of 0.607 and a specificity of 0.75. Conclusion OxLDL plays an important role in coronary artery lesions in KD. The coronary endothelial dysfunction is earlier than coronary dilatation, and oxLDL is expected to become a reliable early predictor of coronary artery lesions in KD.

0.05).Conclusions SNP of 2350G→A in ACE gene is associated with MI,AA genotype is probably a genetic marker of MI in Han nationality.

Objective：This study was designed to establish and optimize the research methods for proteome,and to analyze the proteome components of human lung squamous carcinoma cell line NCI H520. Methods： A series of methods, including immobilized pH gradient two dimensional polyacrylamide gel electrophoresis(2DE), silver staining, PDQuest 2DE analysis software, peptide mass fingerprint based on matrix assisted laser desorption/ionization time of flying mass spectrometry (MALDI TOF MS) and SWISS PROT database searching, were used to separate and indentify the proteome of human lung squarmous carcinoma cell line NCI H520. Results： The good 2DE pattern including resolution and reproducibility was obtained. After silver staining, the 2DE image analysis by PDQuest 2DE software had detected average of 1146± 116 spots, and 851± 95 spots were matched. The average matching rate was 73.3％ . There had a good reproducibility of spot position in 2DE map, with average deviation in IEF direction of 1.52± 0.22 mm， while in SDS PAGE direction it was 1.97± 0.13 mm. Sixty spots were incised from silver staining gel randomly and digested in gel by TPCKtrypsin. Fifty four peptide mass fingerprints (PMF) maps were obtained by MALDI TOF MS. The typic peptide masses were searched in the SWISS PROT database by PeptIdent software. Forty four proteins were preliminarily identified. Some of them were cell cycle related proteins such as Cyclin H, some were signal transduction related proteins such as mitogen activated protein kinase, protein kinase C and receptor protein tyrosine kinase ERBB 2, some were oncogene related proteins such as Ras related protein RAB 36, etc. Conclusions： The main proteome research system including IPG 2DE, image analysis, MALDI TOF MS derived PMFs and database searching has been established. The data of NCI H520 obtained by above methods will be useful for the establishment of human lung squamous cell proteome database.

Objective To understand the epidemiological characteristics of common respiratory viruses among influenza-like illness (ILI) and severe acute respiratory infections (SARI) in Pudong New Area, Shanghai , so as to help estimate the disease burden and conduct the valuable control strategies . Methods Respiratory specimen ( throat swab or sputum ) from cases older than 65 years old with ILI/SARI were collected from outpatient and inpatient settings in four sentinel hospitals in Pudong New Area . Each specimen was tested by multiplex PCR for eight target viral etiologies , including influenza virus , human rhinoviruses ( HRV ) , human para-influenza virus ( PIV ) , adenoviruses ( ADV ) , respiratory syncytial virus ( RSV ) , human metapneumovirus ( hMPV ) , human coronavirus ( hCoV ) and human bocavirus ( hBoV) .Chi-square tests or Fisher's Exact Test were used to compare and analyze . Results From January 1st, 2014 to June 30st, 2016, a total of 363 elderly cases with ILI/SARI were enrolled, with 202 (55.65%) male and a median age of 70 years old.142(39.12%) patients were detected posi-tive for any one of the eight viruses.Influenza was the predominant virus (20.94%, 76/363), with the positive proportion of ( 29 .83%) among ILI cases and ( 12 .09%) among SARI cases .The Influenza infection presented two seasonal peaks in winter ( December to February ) and summer ( July to September ) . Conclusion Influenza is identified as the leading viral pathogen both among ILI and SARI cases older than 65 years old, and two seasonal epidemic peaks areobserved in Shanghai .Influenza vacci-nation strategy should be advocated for the elderly population in Shanghai .

Redox signal transduction, especially the oxidative modification of proein thiols, correlates with many diseases and becomes an expanding research area. However, there was rare method for quick and specific detection of protein thiols and their oxidative modification in living cells. In this article, we review the current chemical strategies for the detection and quantification of protein thiols and related cysteine oxidation. We also look into the future of the development of fluorescent probes for protein thiols and their potential application in the research of reactive cysteine proteomes and early detection of redox-related diseases.
Animals ; Cysteine ; metabolism ; Fluorescent Dyes ; Humans ; Nitrosation ; Oxidation-Reduction ; Proteins ; chemistry ; metabolism ; Reactive Nitrogen Species ; metabolism ; Reactive Oxygen Species ; metabolism ; Sulfenic Acids ; analysis ; Sulfhydryl Compounds ; analysis ; chemistry ; metabolism

AIMTo investigate the in vitro and in vivo stability of 9-nitrocamptothecin lactone form in rat plasma.

METHODSThe specific and accurate HPLC method was developed for quantifying 9-nitrocamptothecin lactone form and the total lactone and carboxylate forms simultaneously. By using of this method, the ratios of lactone form to the total in rat plasma at different time were determined in vitro and in vivo. The results were compared to determine which was the main factor influencing the stability of 9-nitrocamptothecin lactone form in rat plasma in vivo.

RESULTSThe stability of lactone form in rat plasma was much higher in vivo than that in vitro.

CONCLUSIONBlood cells help to increase the stability of 9-nitrocamptothecin lactone form. Clearance from blood in vivo is the primary factor which influences the plasma stability of 9-nitrocamptothecin lactone form. The kinetic process of 9-nitrocamptothecin lactone form and total drug in rats were both best fitted to a two-compartment model. However, the process of 9-nitrocamptothecin carboxylate form in vivo was best fitted to a one-compartment model.

Animals ; Antineoplastic Agents ; blood ; pharmacokinetics ; Area Under Curve ; Camptothecin ; analogs & derivatives ; blood ; pharmacokinetics ; Carboxylic Acids ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; methods ; Drug Stability ; Lactones ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley

BACKGROUNDChina is one of the countries with the highest incidence of H. pylori and more than 9090 isolates possessed the cagA gene. This study was to evaluate the biological activity of the H. pylori virulence factor cagA isolated from Chinese patients.

METHODScagA DNA fragments were amplified from the genomic DNA and subsequently cloned into the mammalian expression vector for cell transfection and DNA sequencing. cagA protein, phosphorylated-tyrosine cagA and the complex of cagA precipitated with SHP-2 were identified respectively by western blot in the crude cell lysate from conditionally immortalized gastric epithelial cells at 48 hours after transfection with cagA DNA. In addition, the ability of induction of scattering phenotype was examined after transient expression of cagA in AGS cells.

RESULTSThe C-terminal half of cagA contained only one repeated sequence and three tandem five-amino-acid motifs glutamic acid-proline-isoleucine-tyrosine-alanine (EPIYA). Moreover, the amino acid sequence of D2 region in repeated sequence was aspartic acid-phenylanaline-aspartic acid (D-F-D) which was significantly distinguished from the three repeated sequences and aspartic acid-aspartic adid-leucine (D-D-L) in the western standard strain NCTC11637. Western blot revealed that cagA became phosphorylated in tyrosine site and bound with SHP-2 after transient expression of cagA DNA in gastric epithelial cells. Transient expression of cagA in AGS cells showed that cagA was able to induce the elongation phenotype although to a lesser extent than western strains.

CONCLUSIONScagA perturbs cell signaling pathways by binding with SHP-2. However, significant difference exists in amino acid sequence and biological function of cagA in Chinese compared with those of western countries.

Amino Acid Sequence ; Antigens, Bacterial ; chemistry ; physiology ; Bacterial Proteins ; chemistry ; physiology ; Blotting, Western ; Cells, Cultured ; Gastric Mucosa ; Humans ; Intracellular Signaling Peptides and Proteins ; Molecular Sequence Data ; Phenotype ; Phosphorylation ; Protein Tyrosine Phosphatase, Non-Receptor Type 11 ; Protein Tyrosine Phosphatases ; metabolism ; Repetitive Sequences, Amino Acid ; Signal Transduction

OBJECTIVETo investigate clinical effects and advantages of Wiltse paraspinal approach to thoracolumbar burst fractures.

METHODSFrom June 2008 to June 2010, the data of 53 patients with thoracolumbar burst fractures with no obviously nerve injury were retrospectively analyzed, including 43 males and 10 females with an average age of 41 years ( ranged, 19 to 62 years). For segmental distribution, 6 cases were T11, 11 cases were T12, 22 cases were L1 and 14 cases were L2. Among them, 28 cases were treated with Wiltse paraspinal approach, and 25 cases with postmiddle approach. The operation time, blood loss, postoperative drainage,VAS score of back, Cobb angle of injured cord, changes of collapse of vertebral and median sagittal diameter of injured vertebral were observed.

RESULTSCompared with two methods, there were advantages in improving operation time, blood loss, postoperative drainage and VAS score of back, but there were no significant differences in improving Cobb angle of injuried cord, changes of collapse of vertebral and median sagittal diameter of injuried vertebral.

CONCLUSIONWiltse paraspinal approach to thoracolumbar burst fractures can achieve the same reduction with postmiddle approach,and has advantages of minimally invasive, less blood, simple operation and rapid recovery, and worth popularizing.

Adult ; Female ; Follow-Up Studies ; Humans ; Lumbar Vertebrae ; diagnostic imaging ; injuries ; surgery ; Male ; Middle Aged ; Spinal Fractures ; diagnostic imaging ; surgery ; Thoracic Vertebrae ; diagnostic imaging ; injuries ; surgery ; Tomography, X-Ray Computed ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the clinical significance of 24-hour blood pressure monitoring (ABPM) for evaluating the treatment outcome of nerve-mediated syncope (NMS) in children.

METHODSTwenty-eight children with NMS confirmed by a head-up tilt table test (HUTT) (12 males and 16 females, aged 6-13 years) and with a chief complaint of unexplained syncope or pre-syncope between February 2010 and August 2012, were included in the study. These children received health education combined with therapy using oral rehydration salts solution and were then reexamined for clinical symptoms as well as HUTT and ABPM results.

RESULTSOf 28 NMS cases, 22 were vasodepressive type, 5 were mixed type, and 1 was cardioinhibitory type. The follow-up showed that 27 (96%) of all cases had improved clinical symptoms, and 18 (64%) had improved HUTT results. The ABPM follow-up revealed no significant changes in 24-hour mean systolic pressure, 24-hour mean diastolic pressure, daytime mean systolic pressure, daytime mean diastolic pressure, nighttime mean systolic pressure, nighttime mean diastolic pressure, day-night difference of systolic pressure, and day-night difference of diastolic pressure after treatment (P>0.05). The percentage of children with a dipper blood pressure pattern increased from 29% (8/28) before treatment to 50% (14/28) after treatment; the percentage of children with a non-dipper blood pressure pattern decreased from 71% (20/28) before treatment to 50% (14/28) after treatment.

CONCLUSIONSAs an effective, objective and non-invasive monitoring means, ABPM is of some clinical significance for evaluating the treatment outcome of NMS in children.

Adolescent ; Blood Pressure Monitoring, Ambulatory ; Child ; Female ; Health Education ; Humans ; Male ; Rehydration Solutions ; administration & dosage ; Syncope ; physiopathology ; therapy ; Tilt-Table Test ; Treatment Outcome