Acetamides ; adverse effects ; Adult ; Alanine Transaminase ; blood ; Aspartate Aminotransferases ; blood ; Case-Control Studies ; Humans ; Liver ; drug effects ; metabolism ; pathology ; Liver Diseases ; epidemiology ; Middle Aged ; Occupational Exposure ; adverse effects ; Young Adult

AIMTo develop a method for separating salicylic acid drugs by aqueous and nonaqueous capillary electrophoresis with conductance detector.

METHODSFused-silica capillary (55 cm x 50 microns ID) was used. The effects of concentration and pH of the running buffer, running voltage and injection time were studied. Salicylic acid (SA), acetylsalicylic acid (ASA) and sulfosalicylic acid (SSA) can be separated in a 10 mmol.L-1 Tris-30 mmol.L-1 H3BO3 buffer (pH 8.0), the separation voltage and injection time were 24 kV and 10 s, respectively. But tailing peaks appeared. In order to improve the separation efficiency and the sensitivity, ethanol was used as nonaqueous solvent.

RESULTSHigh sensitivity and resolution for SA, ASA and SSA were obtained in ethanol media, and there was excellent linearity between peak area and concentration of the analytes in the concentration range of 0.05-100 mg.L-1, 5.0-250 mg.L-1 and 0.08-100 mg.L-1 for SA, ASA and SSA, respectively. All the correlation coefficients were over 0.995.

CONCLUSIONThe analysis of SA and ASA in aspirin tablets was tried and good results were obtained in ethanol media. There was higher sensitivity and separation efficiency than in aqueous media.

Aspirin ; analysis ; chemistry ; isolation & purification ; Benzenesulfonates ; Electrophoresis, Capillary ; methods ; Ethanol ; Salicylates ; isolation & purification ; Salicylic Acid ; isolation & purification ; Sensitivity and Specificity ; Tablets ; Water

Aim The purpose of this study was to compare the differential expression of 15-lipoxygenase isoenzymes in the pulmonary arteries between normoxia and hypoxia and to explore their roles in the formation of hypoxic pulmomary vasoconstriction. Method Eighteen SD rats were randomly divided into two groups(n=9):the normoxic control group breathing fresh gas and the hypoxic group breeding in animal hypoxic incubator.Immunohistochemical method,in situ hybridization and Western blot were employed to determine certain 15-lipoxygenase isoenzymes which involved in the process of hypoxic pulmonary vasoconstriction.Results ①In normoxic control group,the expression of 15-LO-1 protein was detected in the pulmonary arteries;but the expression of 15-LO-2 protein wasn’t detected.②The expression of 15-LO-1 protein in hypoxic group was much stronger than that in normoxic group (P

Carbon nanosheets load beta-cyclodextrin (β-CD-CNS) as a new modified electrode materials was reported for the electrochemical determination of sulfadiazine(SD). Carbon nanosheets(CNS) were prepared by a new method of ultrasonic electrolysis in which the β-CD was attached on CNS through ultrasonic dispersion method. The β-CD-CNS composite nanomaterials were immobilized onto glassy carbon electrodes with drops of coating method to construct an SD voltammetric sensor. The differential pulse stripping voltammetry (DPSV) was used to characterize the electrocatalytic behavior of the developed sensor. The Effects of some parameters on the response behavior of the sensor such as pH,modification amount,scanning rate,stirring speed,stirring time,deposition potential and time were investigated and optimized. The results indicated that the β-CD-CNS composite nanomaterials had excellent electroactivity for the SD in neutral solution and greatly improved the current response of SD. Under the optimal conditions, the SD had an irreversible characteristic oxidation peak around+0.87 V,and the oxidation peak current ip(μA) had a good linear relationship with the concentration C ( μmol/L) of the SD in concentration range of 0.05 μmol/L-13.5 μmol/L with correlation coefficients of 0.999. The detection limit was 12.2 nmol/L (S/N=3). The sensor was successfully applied for the trace SD determination in water and milk samples and the recoveries from the spiked samples were 80.0%-102% with RSD≤5.2%.

OBJECTIVETo examine the effect of trastuzumab on cell proliferation, colony formation and changes of HER-2 proteins in human breast cancer cell line SKBR3 and human ovarian cancer cell line SKOV3 cells which overexpress p185 HER-2 but shed high or low HER-2 extracellular domain (ECD) levels.

METHODSSKBR3 cells and SKOV3 cells were treated with or without trastuzumab. Cell number and the rate of colony formation were calculated. Western blot analysis was used to detect p185 HER-2, HER-2 ECD and phospho-HER-2. Two-site ELISA assay was used for the detection of HER-2 ECD.

RESULTSTrastuzumab inhibited cell proliferation, colony formation, and decreased or eliminated the levels of two uncharacterized phospho-proteins (molar weight about 90 000 and 40 000) in SKBR3 cells shedding high level of HER-2 ECD expression. These responses were not observed in SKOV3 cells shedding low level of HER-2 ECD expression. But total p185, phospho-p185 and phospho-p95 proteins did not appear to change in SKBR3 and SKOV3 cells after treatment with trastuzumab. Trastuzumab reacts not only with proteolytic cleavage HER-2 ECD containing HER-2 ECD I , II , III and IV subdomains of p185 HER-2 extracellular domain, but also with the secreted autoinhibitor p68/ECD III a specifying 340 residues, identical to subdomains I and II from the extracellular domain of p185 HER-2, followed by a unique C-terminal sequence of 79 aa encoded by intron 8, which suggested that there may be a trastuzumab binding site on p68/ECD III a protein. Comparing with HER-2 ECD levels of the same number of SKBR3 cells, there was no significant decrease of HER-2 ECD shedding level after treatment with or without trastuzumab for 4 days in serum-free medium.

CONCLUSIONAntitumor effects of trastuzumab may be related to the two uncharacterized phospho-p90 and/or phospho-p40 proteins. There is probably a trastuzumab epitope on p68/ECD III a. The decrease of HER-2 ECD levels may be positively correlated with the number of SKBR3 cells.

Antibodies, Monoclonal ; pharmacology ; Antibodies, Monoclonal, Humanized ; Antineoplastic Agents ; pharmacology ; Blotting, Western ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Ovarian Neoplasms ; metabolism ; pathology ; Phosphorylation ; Receptor, ErbB-2 ; metabolism ; Trastuzumab

Objective To conduct a comprehensive performance evaluation of fully automated coagulation analyzer in testing coagulation factors , protein S ( PS) , protein C ( PC) , anti-thrombin Ⅲ( AT-Ⅲ) , and von Willebrand factor antigen (vWF:Ag).Methods According to the Clinical and Laboratory Standards Institute (CLSI) EP5-A2, EP9-A2, and WS/T 406-2012 specifications, coagulation factors (FⅡ, FⅤ, FⅦ, FⅧ, FⅨ, FⅩ, FⅪ, and FⅫ) , PS, PC, AT-Ⅲ, and vWF:Ag were detected in the apparatus to evaluate the accu-racy , within-run and between-run imprecisions , linear range , carryover rate , method comparisons , and reference range of the ACL TOP 700 coagulation analyzer .Results The instrument had high accuracy and precision , a good linear range, and low carryover rate (0-2.63%, <3%).The biases of the 12 College of American Pa-thologists ( CAP) controls compared with the target value were all less than 15%, excepting the low value of PC . The within-run imprecisions of 8 coagulation factors were 1.7%-4.4%, and the between-run imprecisions were 2.2%-7.5%.The within-run imprecisions of 3 thrombophilia screen tests and vWF:Ag were 1.0%-7.0%and 2.2%-3.1%, and he between-run imprecisions were 1.5%-10.5% and 2.1%-4.1%, respectively . The carryover rates of the 12 items ranged from 0 to 2.63%.Results of liner verification test showed the correla-tion coefficients of PS , PC, AT-Ⅲ, and vWF:Ag were 0.982-0.988 .Results of method comprisons showed the correlation coefficients of ACL TOP 700 and other coagulation analyzer were more than 0.975 , and the recommen-ded reference ranges of all the 12 items were appropriate for our laboratory .Conclusions ACL TOP 700 coagu-lation analyzer has a good performance in accuracy , imprecision , carryover rate , linear range , and method com-parison .It is suitable for detection of clinical specimens .

OBJECTIVETo examine the association between CD133 expression and invasion of gastric cancer, and to elucidate whether CD133 can promote the invasion and metastasis of gastric cancer via epithelial-mesenchymal transition (EMT).

METHODSThe CD133(+) and CD133(-) KATO-III( cells were sorted by magnetic activated cell sorting (MACS). The invasion ability was detected by Transwell method. RT-PCR and Western blot were used to detect the expression of EMT-related factors in KATO-III( cells before and after CD133 was knocked out by siRNA method. The expressions of CD133 and EMT-related proteins of cancer and adjacent normal tissues in 50 patients with gastric cancer were detected by Western blot, and correlations among protein expressions were also analyzed.

RESULTSAs compared to CD133(-) cells, the number of broken-membrane cells was significantly higher (67.7±10.5 vs. 13.3±6.8, P=0.001) and the invasion ability was stronger (P<0.05) in CD133(+) cells, while the mRNA expression levels of Snail and N-cadherin were significantly higher in CD133(+) cells (0.311±0.015 vs. 0.223±0.016, P=0.040; 0.581±0.020 vs. 0.270±0.018,P=0.004), and the protein expression levels of Snail and N-cadherin were significantly higher in CD133(+) cells as well (0.513±0.015 vs. 0.179±0.023, P=0.030; 0.538±0.028 vs. 0.202±0.032, P=0.020), but E-cadherin mRNA and protein levels were significantly lower in CD133(+) cells (0.231±0.009 vs. 0.460±0.015, P=0.040; 0.426±0.030 vs. 0.748±0.027, P=0.040). After CD133 knock-out, the expressions of Snail and N-cadherin were down-regulated (P<0.05) and the expression of E-cadherin was up-regulated (P<0.05). As compared to normal mucosal tissues, the protein expression levels of Snail, N-cadherin and CD133 in gastric cancer tissues were significantly higher(0.635±0.119 vs. 0.485±0.116, P=0.029; 0.599±0.114 vs. 0.259±0.108, P=0.020; 0.754±0.154 vs. 0.329±0.134, P=0.001), while the protein expression of E-cadherin in gastric cancer tissues was lower (0.378±0.123 vs. 0.752±0.156, P=0.003). The protein expressions of Snail and N-cadherin were positively correlated with CD133 expression (r=0.278, P=0.048; r=0.406, P=0.003) and the protein expression of E-cadherin was negatively correlated with CD133 expression (r=-0.504, P=0.000).

CONCLUSIONCD133(+) cells in primary lesion of gastric cancer have relatively higher invasion ability, which may promote the metastasis of gastric cancer via up-regulation of EMT-related factors.

AC133 Antigen ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; metabolism ; Cell Line, Tumor ; Epithelial-Mesenchymal Transition ; Female ; Glycoproteins ; metabolism ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Peptides ; metabolism ; Stomach Neoplasms ; metabolism ; pathology

OBJECTIVETo investigate the molecular pathology in families with long QT syndrome (LQTS) including Jervell-Longe-Nielsen syndrome (JLNS) and Romano-ward syndrome (RWS) and Brugada syndrome (BS) in Chinese population.

METHODSPolymerase chain reaction and DNA sequencing were used to screen for KCNQ1, KCNH2, KCNE1, and SCN5A mutation.

RESULTSWe identified a novel mutation N1774S in the SCN5A gene of the BS family, a novel mutation G314S in a RWS family which had also been found in Europe, North America, and Japan, and a single nucleotide polymorphisms (SNPs) G643S in the KCNQ1 of the JLNS family. In this JLNS family, another heterozygous novel mutation in exon 2a was found in KCNQ1 of the patients.

CONCLUSIONNew mutations were found in our experiment, which expand the spectrum of KCNQ1 and SCN5A mutations that cause LQTS and BS.

Adolescent ; Adult ; Base Sequence ; ERG1 Potassium Channel ; Ether-A-Go-Go Potassium Channels ; genetics ; Female ; Humans ; Jervell-Lange Nielsen Syndrome ; genetics ; KCNQ1 Potassium Channel ; genetics ; Long QT Syndrome ; congenital ; genetics ; Male ; Middle Aged ; Molecular Sequence Data ; Muscle Proteins ; genetics ; Mutation ; NAV1.5 Voltage-Gated Sodium Channel ; Pedigree ; Potassium Channels, Voltage-Gated ; genetics ; Romano-Ward Syndrome ; genetics ; Sodium Channels ; genetics

OBJECTIVETo identify the mutation of a Chinese family with inherited long QT syndrome(LQTS).

METHODSThe disease-causing gene was tentatively determined in light of the clinical manifestations and electrophysiological properties, and then polymerase chain reaction and DNA sequencing were used for screening and identifying mutation.

RESULTSA missense mutation G940A(G314S) in the KCNQ1 gene was identified, which was the 'hot spot' of long QT syndrome mutation.

CONCLUSIONThe mutation that is involved with long QT syndrome in Chinese patients is the same as that in the European, American and Japanese patients.

China ; DNA Mutational Analysis ; Family Health ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; KCNQ1 Potassium Channel ; genetics ; Long QT Syndrome ; diagnosis ; genetics ; Male ; Mutation, Missense ; Pedigree ; Polymerase Chain Reaction

OBJECTIVETo investigate the changes in proliferation, invasiveness, clone sphere formation and chemosensitivity of human gastric cancer cell lines of KATO-III CD133(+) cells transfected with small interfering RNA (siRNA) against CD133 gene.

METHODSCD133(+) cells of KATO-III cell lines were isolated by magnetic activated cell sorting (MACS). CD133 siRNA was designed and synthesized, and then transfected into KATO-III CD133(+) cells. Cell fluorescence counting under confocal laser scanning microscope was used to determine the transfection efficiency after transfection with the CD133 FITC-siRNA. The knock-down effect of the CD133 gene and expression of epithelial-mesenchymal transition (EMT)-related factors were detected by RT-PCR and Western blotting. Cell counting kit-8 assay (CCK-8), transwell chamber and colony sphere forming assay were performed to measure the variation of cell proliferative, invasive, colony formation viability and chemosensitivity to 5-FU after the above-mentioned treatment.

RESULTSThe transfection efficiency was (87.7±8.1)%. The CD133 mRNA and protein expression levels in the interference group were lower than those in negative control group. Twenty-four, 48 and 72 hours after transfection, cells proliferation activity was significantly inhibited in the interference group compared with negative control group, (all P<0.01). Seventy-two hours after transfection, compared with negative control group, cells proliferation activity was reduced by (52.1±8.0)%. The invasive cell number reduced (41.7±6.0 vs. 130.3±11.0, P<0.05) and clone formation rate decreased significantly [(24.3±4.3)% vs. (45.1±6.4)%, P<0.01] in the interference group. EMT-related gene E-cadherin protein expression increased, while the Snail and N-cadherin protein expression reduced in the interference group (all P<0.01). The cells sensitivity to 5-FU was significantly enhanced in the interference group, and the cell inhibition rate of 5-Fu was (62.4±3.3)%, higher than that in negative control group [(21.5±2.2)%, P<0.01].

CONCLUSIONSThe expression of CD133 gene plays an important role in cell proliferation, invasiveness, colony formation and resistance to chemotherapy of KATO-III CD133(+) gastric cancer cells. It suggests that CD133 can be used as one of surface markers for detection of gastric cancer stem cells. Inhibition of CD133 expression may be a promising way for gastric cancer biotherapy.

AC133 Antigen ; Antigens, CD ; genetics ; metabolism ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Fluorouracil ; pharmacology ; Glycoproteins ; genetics ; metabolism ; Humans ; Peptides ; genetics ; metabolism ; RNA Interference ; RNA, Small Interfering ; genetics ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Transfection