BACKGROUND: Paraquat (PQ) is a world-wide used herbicide and also a type of common poison for suicide and accidental poisoning. Numerous studies have proved that the concentration of serum PQ plays an important role in prognosis. Spectrophotometry, including common spectrophotometry and second-derivative spectrophotometry, is commonly used for PQ detection in primary hospitals. So far, lack of systematic research on the reliability of the method and the correlation between clinical features of patients with PQ poisoning and the test results has restricted the clinical use of spectrophotometry. This study aimed to evaluate the reliability and value of spectrophotometry in detecting the concentration of serum PQ. METHODS: The wavelengths for detecting the concentration of serum PQ by common and second-derivative spectrophotometry were determined. Second-derivative spectrophotometry was applied to detect the concentration of serum PQ. The linear range and precision for detection of PQ concentration by this method were confirmed. The concentration of serum PQ shown by second-derivative spectrophotometry and HPLC were compared in 8 patients with PQ poisoning. Altogether 21 patients with acute poisoning 4 hours after PQ ingestion treated in the period of October 2008 to September 2010 were retrospectively reviewed. The patients were divided into higher and lower than 1.8 μg/mL groups based on their concentrations of serum PQ measured by second-derivative spectrophotometry on admission. The severity of clinical manifestations between the two groups were analyzed with Student's t test or Fisher's exact test. RESULTS: The absorption peak of 257 nm could not be found when common spectrophotometry was used to detect the PQ concentration in serum. The calibration curve in the 0.4–8.0 μg/mL range for PQ concentration shown by second-derivative spectrophotometry obeyed Beer's law with r=0.996. The average recovery rates of PQ were within a range of 95.0％ to 99.5％, relative standard deviation (RSD) was within 1.35％ to 5.41％ (n=6), and the lower detection limit was 0.05 μg/mL. The PQ concentrations in serum of 8 patients with PQ poisoning shown by second-derivative spectrophotometry were consistent with the quantitative determinations by HPLC (r=0.995, P<0.0001). The survival rate was 22.2％ in patients whose PQ concentration in serum was more than 1.8 μg/mL, and the incidences of acidosis, oliguria and pneumomediastinum in these patients were 55.6％, 55.6％and 77.8％, respectively. These clinical manifestations were different significantly from those of the patients whose PQ concentration in serum was less than 1.8 μg/mL (P<0.05). CONCLUSIONS: For common spectrophotometry, the wavelength at 257 nm was not suitable for detecting serum PQ as no absorbance was shown. Second-derivative spectrophotometry was reliable for detecting serum paraquat concentration. Serum PQ concentration detected by second-derivative spectrophotometry could be used to predict the severity of clinical manifestations of patients with PQ poisoning, and PQ content higher than 1.8 μg/mL 4 hours after ingestion could be an important predictive factor for poor prognosis.

To establish a cell model in vitro that stable expressing HBV by integrating HBA1.3 DNA into cell chromosome. The HBV1.3 full-length DNA was obtained by digested pGEM-HBV1.3 plasmid with HindIII and then was linked with PU-21 vector digested by HindIII. This was resulted in generation of a recombined plasmid named PU21-HBV plasmid. The recombined plasmid was introduced into HepG2 cells by electroporation. The transfected cells were screened with G418. The insertion and expression of HBV were identified by X-gal staining, RT-PCR and Southern blot. The result of PU21-HBV plasmid sequence demonstrated that HBV1.3 DNA was linked correctly with PU-21 vector. A series of positive cell colonies were obtained with G418 screening followed transfecting PU21-HBV plasmid into HepG2 cells. The results of Southern blot and RT-PCR exhibit that HBV1.3 DNA had successfully integrated into the chromosomes of HepG2 cells and had functional HBV gene transcription. HBV1.3 DNA was inserted into HepG2 genome and could stable transcript HBV RNA. The stable HBV expression cell line was constructed successfully. There are LoxP sites in the trapping vector PU21. With the Cre enzyme, interesting genes could be excganged into the LoxP sites. Therefore, double stable expression of interesting gene and HBV cell lines could be generated. The cell lines will be useful for further research some target gene function on replication of HBV.

Female ; Humans ; Laser Therapy ; methods ; Middle Aged ; Semicircular Canals ; surgery ; Vertigo ; etiology ; surgery

OBJECTIVETo study the effect of Shuangling Fuzheng anti-tumor preparation (SLAP) five groups on proliferation and c-myc gene expression of SGC-7901 cells in vitro.

METHODThe inhibitory effect of single SLAP (40 -640 microg x mL(-1)) and combined therapy with adriamycin (0.4, 4.0 microg x mL(-1) or cisplatin (0.1,1.0 microg x mL(-1) on human gastric carcinoma SGC-7901 cells proliferation were observed by MTT colorimetric analysis method. Technique of flow cytometry in vitro was used to measure the rate of positive sign of SLAP (80 - 320 microg x mL(-1)) on c-myc gene protein of SGC-7901 cells.

RESULTSGC-7901 cells proliferation were inhibited by single SLAP in dose of 40 - 640 microg x mL(-1) 24 h. Its inhibitory rate was increased with increase of dose. The inhibitory rate on SGC-7901 cells could be increased by SLAP in dose of 40 - 640 microg x mL(-1) plus adriamycin in dose of 0.4 and 4.0 microg x mL(-1) or plus cisplatin in dose of 0.1 and 1.0 microg x mL(-1). At the same time, SLAP (80 - 320 microg x mL(-1)) also could inhibite the expression of c-myc gene of SGC -7901 cells.

CONCLUSIONSingle SLAP had inhibiting effect on human gastric carcinoma cells proliferation with a dose-effect relationship and synergic effect while combined with adriamycin or cisplatin. To inhibit the expression of c-myc gene of human gastric carcinoma cells might be one of action mechanisms of SLAP, which inhibited tumor cells proliferation.

Antibiotics, Antineoplastic ; pharmacology ; Antineoplastic Agents ; pharmacology ; Antineoplastic Agents, Phytogenic ; administration & dosage ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; Dose-Response Relationship, Drug ; Doxorubicin ; pharmacology ; Drug Combinations ; Drug Synergism ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; Genes, myc ; Humans ; Plants, Medicinal ; chemistry ; Proto-Oncogene Proteins c-myc ; metabolism ; Stomach Neoplasms ; metabolism ; pathology

BACKGROUNDVisfatin, a visceral fat-derived adipocytokine, plays a significant physiological function in lipid metabolism. However, the precise function of visfatin and its regulation by thyroid hormones are still unknown. This study observed the plasma visfatin concentrations in subjects with hyperthyroidism and hypothyroidism in vivo, and investigated the possible regulation mechanism between visfatin and tri-iodothyronine (T3) in vitro as a further interpretation.

METHODSThe experiment in vivo included clinical subjects (57 patients with thyroid dysfunction and 29 euthyroid healthy volunteers) and an animal model (24 Wistar rats). All subjects were divided into hyperthyroidism, hypothyroidism and euthyroidism groups, with plasma thyroid hormones, thyrotropin, visfatin and triglyceride concentrations determined. Visfatin mRNA expression in visceral fat and liver of rats was detected by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR). The experiment in vitro studied 3T3-L1 cells and visfatin mRNA expression under nine different T3 concentrations (0, 0.1, 0.25, 0.5, 1, 5, 10, 20, 100 nmol/L) using quantitative real-time RT-PCR.

RESULTSClinical subjects and animal models showed elevated plasma visfatin concentrations in the hyperthyroidism group (20.466 ng/ml (15.263, 26.795 ng/ml) and (1209.164±165.292) ng/L) and hypothyroidism group (12.457 ng/ml (11.115, 15.454 ng/ml) and (1205.425±109.200) ng/L) compared to euthyroidism group (6.891 ng/ml (5.888, 8.803 ng/ml) and (926.650±54.002) ng/L, P<0.001). For animal models, visfatin mRNA expression in visceral fat in the hyperthyroidism and hypothyroidism groups increased about 3.33-fold and 1.98-fold compared to the euthyroidism group (P<0.001), which was positively correlated with plasma visfatin concentrations (r=0.713, P<0.001). However, no significant group difference (P>0.05) and correlation (r=0.121, P=0.572) was found in the liver. T3 induced a remarkable increase of visfatin mRNA expression in 3T3-L1 cells at low concentrations (0-0.5 nmol/L T3) followed by a sharp decrease at higher concentrations (0.5-100 nmol/L T3), with an inflection point at 0.5 nmol/L T3.

CONCLUSIONElevated circulating visfatin levels in subjects with hyperthyroidism and hypothyroidism are possibly due to an increase of visfatin mRNA expression in visceral fat, and a nonlinear regulation mechanism on visfatin mRNA expression under various T3 concentrations might be involved.

3T3-L1 Cells ; Adult ; Animals ; Female ; Humans ; Hyperthyroidism ; blood ; genetics ; metabolism ; Hypothyroidism ; blood ; genetics ; metabolism ; Male ; Mice ; Middle Aged ; Nicotinamide Phosphoribosyltransferase ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Triiodothyronine ; blood

OBJECTIVETo re-confirm and characterize the biophysical and pharmacological properties of endogenously expressed human acid-sensing ion channel 1a (hASIC1a) current in HEK293 cells with a modified perfusion methods.

METHODSWith cell floating method, which is separating the cultured cell from coverslip and putting the cell in front of perfusion tubing, whole cell patch clamp technique was used to record hASIC1a currents evoked by low pH external solution.

RESULTSUsing cell floating method, the amplitude of hASIC1a currents activated by pH 5.0 in HEK293 cells is twice as large as that by the conventional method where the cells remain attached to coverslip. The time to reach peak at two different recording conditions is (21+/-5) ms and (270+/-25) ms, respectively. Inactivation time constants are (496+/-23) ms and (2284+/-120) ms, respectively. The cell floating method significantly increases the amiloride potency of block on hASIC1a [IC50 is (3.4+/-1.1) micromol/L and (2.4+/- 0.9) micromol/L, respectively]. Both recording methods have similar pH activation EC50 (6.6+/-0.6, 6.6+/-0.7, respectively).

CONCLUSIONASICs channel activation requires fast exchange of extracellular solution with the different pH values. With cell floating method, the presence of hASIC1a current was re-confirmed and the biophysical and pharmacological properties of hASIC1a channel in HEK293 cells were precisely characterized. This method could be used to study all ASICs and other ligand-gated channels that require fast extracellular solution exchange.

Acid Sensing Ion Channels ; Amiloride ; pharmacology ; Biophysics ; instrumentation ; methods ; Cell Culture Techniques ; instrumentation ; methods ; Cell Line ; Cell Membrane ; chemistry ; drug effects ; metabolism ; Culture Media ; chemistry ; pharmacology ; Extracellular Fluid ; chemistry ; metabolism ; Humans ; Hydrogen-Ion Concentration ; drug effects ; Membrane Potentials ; drug effects ; physiology ; Nerve Tissue Proteins ; chemistry ; drug effects ; metabolism ; Neuropharmacology ; instrumentation ; methods ; Patch-Clamp Techniques ; instrumentation ; methods ; Perfusion ; instrumentation ; methods ; Sodium Channel Blockers ; pharmacology ; Sodium Channels ; chemistry ; drug effects ; metabolism ; Time Factors

OBJECTIVETo detect the polymorphisms of Radix Plygoni Multiflori in chongqing by means of a new marker system SRAP.

METHODDifferent shaples of Radix Plygoni Multiflori from major production areas were collected. The SRAP was used to asses divergence among 16 populations. The data were analyzed using unweighted pairgroup method, based on arithmetic averages (UPGMA) bootstrap analysis. Cluster analyses was performed by using DPSv3.01 software, the alkaloid was extracted from P. ternate with chlorolform.

RESULT104 combinations generated 250 polymorphie bands, the cluster analysis indicated that 16 materials could be distinguished into two main groups and one special type, Nei&Li similarity coefficient ranged from 0.23-0.99, and the average distance is 0. 44.

CONCLUSIONThe results of the study showed a potential application of SRAP fingerprinting for identification of Radix Plygoni Multiflori.

Cluster Analysis ; DNA, Plant ; genetics ; Genetic Markers ; Genetic Variation ; Nucleic Acid Amplification Techniques ; methods ; Phylogeny ; Plant Roots ; genetics ; Plants, Medicinal ; classification ; genetics ; Polygonum ; classification ; genetics

OBJECTIVETo screen the differential expression gene profile in nasal mucosa of seasonal allergic rhinitis (SAR) and SAR with asthma, oligonucleotide microarray (Affymetrix HG-U133-plus2) was employed to analyze the changes of gene expressions with GeneSpring software.

METHODSInferior turbinate mucosa was obtained from five SAR patients and four SAR with asthma patients. Total RNA was extracted from the nasal mucosal biopsies and pooled into one SAR control pool and one SAR with asthma patient pool, and biotin-labeled cRNA probes were hybridized with Affymetrix HG-U133-plus2 array. The hybridization results were confirmed by RT-PCR analysis. The analysis of differential expression profiles were performed by GeneSpring software 7.3.

RESULTSOut of 47,000 analysed transcripts, 1,900 genes were differentially expressed at least 2-fold in which 849 genes were up-regulated and 1,051 genes were down-regulated in nasal mucosa of SAR with asthma patients compared with that in SAR patients. These genes were involved in cell metabolism, gene transcription, cell proliferation, signal transduction, immune response, enzyme activity, transmembrane receptor activity, cytoskeletal protein binding, and many other aspects. Pathway analysis displayed 161 groups, of which including more than 20 genes were as follow: cytokine-cytokine receptor interaction, focal adhesion, cell adhesion molecules (CAMs), regulation of actin cytoskeleton, cell communication, gap junction, MAPK signaling pathway, calcium signaling pathway, leukocyte transendothelial migration, and purine metabolism.

CONCLUSIONSThe data suggested that multigentic expression and regulation changes were involved in the development of SAR and SAR complicated with asthma, whose molecular mechanisms might be elucidated by identification of these differential genes.

Adolescent ; Adult ; Asthma ; complications ; genetics ; Female ; Gene Expression Profiling ; Gene Expression Regulation ; Humans ; Nasal Mucosa ; metabolism ; Oligonucleotide Array Sequence Analysis ; Rhinitis, Allergic, Seasonal ; complications ; genetics ; Young Adult

OBJECTIVETo investigate the correlation between fetal chromosomal abnormalities and the characteristic features of prenatal ultrasound findings.

METHODSA total of 510 cases were underwent chromosome examination by amniotic fluid or cord blood analysis to identify fetal chromosomal abnormalities. The correlation between the abnormalities and the characteristics of the prenatal ultrasound findings was analyzed.

RESULTSFifty-three cases of abnormal karyotypes were detected with a positivity rate of 10.2%. Of these cases, 32 cases had chromosome number abnormalities, including 15 with 21-trisomy, 11 with 18-trisomy, 2 with 13-trisomy, 2 with 45, XO monomer and 2 with 92, XXXX tetraploid. Chromosome structural abnormalities were found in 21 cases, including 4 with translocation, 3 with insertion, 6 with inversion, 4 with deletion and 4 with derivation. Prenatal ultrasound showed obvious structural abnormalities in 22 cases (41.5%), structural malformation with ultrasonographic soft markers in 18 cases (34.0%), and separate ultrasonographic soft markers in 8 cases (15.1%).

CONCLUSIONPrenatal ultrasound fetal abnormalities and chromosome abnormalities are closely related. Prenatal ultrasound of fetal chromosomal abnormalities usually presents with a variety of significant structural abnormalities. A greater number of malformations is associated with a greater risk of chromosomal abnormalities and increased occurrence of ultrasonographic soft markers.

Adult ; Chromosome Aberrations ; Chromosomes, Human, Pair 18 ; Down Syndrome ; diagnosis ; Female ; Fetal Diseases ; diagnosis ; diagnostic imaging ; Humans ; Pregnancy ; Trisomy ; diagnosis ; Ultrasonography, Prenatal ; methods

BACKGROUNDOur previous study has confirmed that one bout of exhaustion (Ex) can cause hippocampus neurocyte damage, excessive apoptosis, and dysfunction. Its initial reason is intracellular calcium overload in hippocampus triggered by N-methyl-D-aspartic acid receptor (NMDAR) over-activation. NMDAR activation can be suppressed by γ-aminobutyric acid (A) receptor (GABAAR). Whether GABAAR can prevent intense exercise-induced hippocampus apoptosis, damage, or dysfunction will be studied in this study.

METHODSAccording to dose test, rats were randomly divided into control (Con), Ex, muscimol (MUS, 0.1 mg/kg) and bicuculline (BIC, 0.5 mg/kg) groups, then all rats underwent once swimming Ex except ones in Con group only underwent training. Intracellular free calcium concentration ([Ca2+]i) was measured by Fura-2-acetoxymethyl ester; glial librillary acidic protein (GFAP) and synaptophysin (SYP) immunofluorescence were also performed; apoptosis were displayed by dUTP nick end labeling (TUNEL) stain; endoplasmic reticulum stress-induced apoptosis pathway was detected by Western blotting analysis; Morris water maze was used to detect learning ability and spatial memory.

RESULTSThe appropriate dose was 0.1 mg/kg for MUS and 0.5 mg/kg for BIC. Ex group showed significantly increased [Ca2+]i and astrogliosis; TUNEL positive cells and levels of GFAP, B cell lymphoma-2 (Bcl-2) associated X protein (Bax), caspase-3, caspase-12 cleavage, CCAAT/enhancer binding protein homologous protein (CHOP), and p-Jun amino-terminal kinase (p-JNK) in Ex group also raised significantly compared to Con group, while SYP, synapse plasticity, and Bcl-2 levels in Ex group were significantly lower than those in Con group. These indexes were back to normal in MUS group. BIC group had the highest levels of [Ca2+]i, astrogliosis, TUNEL positive cell, GFAP, Bax, caspase-3, caspase-12 cleavage, CHOP, and p-JNK, it also gained the lowest SYP, synapse plasticity, and Bcl-2 levels among all groups. Water maze test showed that Ex group had longer escape latency (EL) and less quadrant dwell time than Con group; all indexes between MUS and Con groups had no significant differences; BIC had the longest EL and least quadrant dwell time among all groups.

CONCLUSIONSActivation of GABAA R could prevent intense exercise-induced synapses damage, excessive apoptosis, and dysfunction of hippocampus.

Animals ; Apoptosis ; physiology ; Body Weight ; physiology ; Endoplasmic Reticulum Stress ; physiology ; Hippocampus ; metabolism ; Male ; Physical Exertion ; physiology ; Rats ; Rats, Sprague-Dawley ; Receptors, GABA ; genetics ; metabolism ; Synapses ; pathology