Objective To investigate the role and intracellular signal transduction mechanism in the injury of renal tubular cells induced by postasphyxial-serum in neonate.Methods Human renal proximal tubular cell(HK-2 cell) was used as target cell. The experiment was designed as:control group, asphyxia group ,and pyrrolidine dithiocarbamate (PDTC)blocking group. The attacking concentration of serum was 20%, and the apoptosis rate of HK-2 cells was detected by flow cytometer.Results Compared with controls[(13.3?1.70)%],after being stimulated with postasphyxial-serum, the apoptosis rate of HK-2 cells of asphyxia group [(46.73?3.68)%] and PDTC blocking group [(31.19?2.79)%]were significantly increased(P

Objective To investigate the role of postasphyxial-serum of neonate in inducing injury of human renal proximal tubular cells(HK-2 cells).Methods HK-2 cells were used as target cell.The neonatal different concentration postasphyxial-serum of 1,3,7 days after asphyxia were used as attacking means.The experimental groups were divided into 15 groups:the 2.5%,5.0%,10.0%,(20.0%) attacking concertration groups of 1,3,7 day after asphyxia and control group of each concertration.The culture medium and concertration of the control group and the experimental groups were the same.The changes of morphology were observed under inverted microscope,the cell viability was measured by 3-(4,5-dimethyl-2-thiazoly1)-2,5-diphenyl-2H-tetrazolium bromide(MTT) method and the leakage rate of lactate dehydrogenase(LDH) was determined by biochemical methods.Results Compared with control group,the changes in morphology of HK-2 were most serious and obvious,the cell viability were obviously decreased(all P

Carcinoma, Non-Small-Cell Lung ; genetics ; Exons ; genetics ; Genes, erbB-1 ; genetics ; Humans ; Lung Neoplasms ; pathology

BACKGROUNDTo study apoptosis of peripheral blood cells of children with viral pneumonia, explore immunopathogenesis and the possibility of immunotherapy of patients with viral pneumonia.

METHODSFresh peripheral blood samples were collected from 28 patients with viral pneumonia and 24 healthy children were treated and run through the flow cytometry. The data were acquired using Cell Quest software and the percentage of live cells, viable apoptotic cells, non-viable apoptotic cells and dead cells of neutrophils and lymphocytes were counted. The patients with viral pneumonia were hospitalized at our hospital. The average age of patients was 1.3 years; 24 healthy children were served as control group (age 1.8 years, on average). T-test and variance analysis by SPSS FOR WINDOWS 10.0 software was used for statistical analysis.

RESULTSThe percentage of live neutrophils and lymphocytes in the acute stage and recovery stage in patients were significantly lower than that in control group (P < 0.01). The percentage of viable apoptotic neutrophils and lymphocytes in two stages in patients were significantly higher than that in control group (P < 0.05). Except for the percentage of live cells, non-viable apoptotic cells and dead lymphocytes, others had no difference between the patients and control groups.

CONCLUSIONApoptosis of neutrophils and lymphocytes of peripheral blood cells of children with viral pneumonia increased. Whereas the percentage of live cells decreased. Drugs that can accelerate apoptosis may be helpful in treatment of viral pneumonia.

Apoptosis ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Infant ; Lymphocytes ; pathology ; Male ; Neutrophils ; pathology ; Pneumonia, Viral ; pathology

Humans ; Immunoglobulin M ; Lung ; pathology ; Multiple Myeloma ; diagnosis ; pathology

OBJECTIVESalvia miltorrhiza Bunge (SMB) is a traditional Chinese herb, which is considered to promote blood flow and remove blood stasis. This study examined whether SMB can alleviate injury induced by hypoxia/reoxygenation (H/R) in human kidney proximal tubular cells-2 (HK-2 cells).

METHODSThere were 3 experimental groups: control, H/R injury and SMB-treated H/R injury. H/R injury of HK-2 cells was induced by first covering the cells with and then removing liquid paraffin wax. Different concentrations of compound SMB solution (0.05%, 0.10%, 0.15% or 0.20%) were administered to the SMB-treated H/R injury group before the hypoxic injury. After 4, 12 and 24 hrs of hypoxia and 4, 12, 24 and 48 hrs of reoxygenation, morphologic changes of HK-2 cells were observed under an inverted microscope. Cell viability was measured by the MTT method. Lactate dehydrogenase (LDH) activity in the culture supernatants was assayed using biochemical methods; TNF-alpha levels were determined by radioimmunoassay (RIA).

RESULTSThe number of HK-2 cells was significantly reduced in the H/R injury group after hypoxia, and reached a nadir 24 hrs after hypoxia treatment. Various concentrations of SMB-treated groups showed significantly greater number of HK-2 cells than the H/R injury group. SMB solution (0.10%) produced the best effect. The levels of LDH and TNF-alpha in the H/R injury group were significantly increased, and reached a peak between 24 hrs of hypoxia and 4 hrs of reoxygenation when compared to the control group. Pre-treating with 0.10% SMB resulted in significantly lower levels of LDH and TNF-alpha than in the untreated H/R injury group at various time points of H/R.

CONCLUSIONSSMB has protective effects against H/R injury of HK-2 cells, possibly through inhibition of inflammatory cytokines.

Calcium ; metabolism ; Cell Hypoxia ; Cells, Cultured ; Cytoprotection ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Kidney Tubules, Proximal ; drug effects ; pathology ; L-Lactate Dehydrogenase ; metabolism ; Plant Extracts ; pharmacology ; Salvia miltiorrhiza ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVETo investigate the mutations in exon 19 of epidermal growth factor receptor (EGFR) gene in non-small cell lung cancer from Chinese patients.

METHODSGenomic DNA was extracted from 72 lung cancer tissues. Then the exon 19 of EGFR gene was amplified by nested PCR and sequenced.

RESULTSIn 13 tumor tissues, multi-nucleotide in-frame deletion mutations at the exon 19 of EGFR gene, had been detected. There were 4 mutation types. The mutation rate was 18.1%. The mutations were all heterozygous. There was association of the exon 19 mutation of EGFR gene with adenocarcinoma, female patients and non-smokers.

CONCLUSIONThere were multi-nucleotide in-frame deletion mutations in exon 19 of EGFR gene. Mutations of the exon 19 of EGFR gene were higher in female, non-smoking and adenocarcinoma patients.

Adenocarcinoma ; genetics ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; genetics ; DNA Mutational Analysis ; Exons ; genetics ; Female ; Genes, erbB-1 ; genetics ; Humans ; Male ; Middle Aged ; Mutation ; Polymerase Chain Reaction ; Sex Factors ; Smoking

Porous carbon nanoparticles ( NPC) were prepared by ZnCl2 activation and carbonization using citrus waste as carbon source. A sample pretreatment method with NPC as dispersive solid phase extraction (d-SPE ) absorbent was established for the determination of organophosphorus pesticides in fruits and vegetables by gas chromatography. The NPC was characterized by scanning electron microscopy (SEM), X-ray diffraction ( XRD), FT-IR spectra, Raman spectroscopy, Brunauer, Emmett and Teller surface area(BET). Those results showed that the NPC was an amorphous porous carbon material with pore size in the range of 0-15 nm. Its specific surface area and pore volume were 1243 m2 / g and 1. 28 cm3 / g, respectively. The analysis conditions, including the amount and clean up time of adsorbent, were optimized by analysis of 14 kinds of oranophosphorus pesticides in fruits and vegetables with gas chromatography-flame photometric determination(GC-FPD). Moreover, the comparison for NPC with commercial materials of PSA, C18 and GCB was investigated in this study. The results indicated that the purification time was only 2 min using 0. 01 g NPC. The cost of NPC was about 25% of C18 , 21% of PSA and 16% of GCB. Because of the porous structure of NPC, the purification efficiency was significantly higher than the three commercial materials mentioned above. Under the optimum conditions, the calibration curves of the 14 organophosphorus pesticides were linear in the range of 0. 02-1. 00 mg / L with good correlation coefficients (R2>0. 99) and detection limits (S / N=3) of 0. 63-5. 30 μg / kg. The recoveries of the pesticides at three spiked levels ranged from 71. 3% to 114. 7%with the relative standard deviations (RSDs) of 0. 9% -12. 9% . The method is simple, rapid, sensitive, and low cost, and can satisfy the requirements of detection of organophosphorus pesticide residues in fruits and vegetables, displaying a good application prospect.

BACKGROUND: Calcium calmodulin-dependent kinase Ⅱ (CaMKⅡ) can be more active in patients with left ventricular hypertrophy (LVH), which in turn causes phosphorylation of ryanodine receptors, resulting in inactivation and the instability of intracellular calcium homeostasis. The present study aimed to determine the effect of CaMKⅡ–ryanodine receptor pathway signaling in rabbits with left ventricular hypertrophy and triggered ventricular arrhythmia. METHODS: Forty New Zealand rabbits were randomized into four groups (10 per group): sham group, LVH group, KN-93 group (LVH+KN-93), and ryanodine group (LVH+ryanodine). Rabbits in the LVH, KN-93, and ryanodine groups were used to establish a left ventricular hypertrophy model by the coarctation of the abdominal aorta, while those in the sham group did not undergo the coarctation. After eight weeks, action potentials (APs) were recorded simultaneously in the endocardium and epicardium, and a transmural electrocardiogram (ECG) was also recorded in the rabbit left ventricular wedge model. Drugs were administered to the animals in the KN-93 and ryanodine groups, and the frequency of triggered APs and ventricular tachycardia was recorded after the rabbits were given isoprenaline (1 mol/L) and high-frequency stimulation. RESULTS: The frequency (animals/group) of triggered APs was 0/10 in the sham group, 10/10 in the LVH group, 4/10 in the KN-93 group, and 1/10 in the ryanodine group. The frequencies of ventricular tachycardia were 0/10, 9/10, 3/10, and 1/10, respectively. The frequencies of polymorphic ventricular tachycardia or ventricular fibrillation were 0/10, 7/10, 2/10, and 1/10, respectively. The frequencies of triggered ventricular arrhythmias in the KN-93 and ryanodine groups were much lower than those in the LVH group (P<0.05). CONCLUSIONS: KN-93 and ryanodine can effectively reduce the occurrence of triggered ventricular arrhythmia in rabbits with LVH. The CaMKⅡ–ryanodine signaling pathway can be used as a new means of treating ventricular arrhythmia.