Nitrites play multiple characteristic functions in invasion and metastasis of hepatic cancer cells, but the exact mechanism is not yet known. Cancer cells can maintain the malignant characteristics via clearance of excess mitochondria by mitophagy. The purpose of this article was to determine the roles of nitrite, reactive oxygen species (ROS) and hypoxia inducing factor 1 alpha (HIF-1 α) in mitophagy of hepatic cancer cells. After exposure of human hepatocellular carcinoma SMMC-7721 cells to a serial concentrations of sodium nitrite for 24 h under normal oxygen, the maximal cell vitality was increased by 16 mg x (-1) sodium nitrite. In addition, the potentials of migration and invasion for SMMC-7721 cells were increased significantly at the same time. Furthermore, sodium nitrite exposure displayed an increase of stress fibers, lamellipodum and perinuclear mitochondrial distribution by cell staining with Actin-Tracker Green and Mito-Tracker Red, which was reversed by N-acetylcysteine (NAC, a reactive oxygen scavenger). DCFH-DA staining with fluorescent microscopy showed that the intracellular level of ROS concentration was increased by the sodium nitrite treatment. LC3 immunostaining and Western blot results showed that sodium nitrite enhanced cell autophagy flux. Under the transmission electron microscopy (TEM), more autolysosomes formed after sodium nitrite treatment and NAC could prevent autophagosome degradation. RT-PCR results indicated that the expression levels of COX I and COXIV mRNA were decreased significantly after sodium nitrite treatment. Meanwhile, laser scanning confocal microscopy showed that sodium nitrite significantly reduced mitochondrial mass detected by Mito-Tracker Green staining. The expression levels of HIF-1α, Beclin-1 and Bnip3 (mitophagy marker molecular) increased remarkably after sodium nitrite treatment, which were reversed by NAC. Our results demonstrated that sodium nitrite (16 mg x L(-1)) increased the potentials of invasion and migration of hepatic cancer SMMC-7721 cells through induction of ROS and HIF-1α mediated mitophagy.
Acetylcysteine ; pharmacology ; Autophagy ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Movement ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Liver Neoplasms ; pathology ; Mitochondrial Degradation ; Neoplasm Invasiveness ; Nitrites ; metabolism ; Reactive Oxygen Species ; metabolism ; Sodium Nitrite ; pharmacology

Objective To research effects of water extraction of Salvia miltiorrhiza in sperm quality of mice. Methods Forty adult male Kunming mice were randomly divided into 5 groups, including low concentration group, medium concentration group, high concentration group, control group and positive control group in eight mice in each group. The low, medium and high concentration groups were given by gavage with 1 500, 3 000 and 6 000 mg/kg water extraction of salvia miltiorrhiza once a day respectively, while the control group was given gavage with 10 mL/kg normal saline daily, as well as the positive control group was injected with 100 mg/kg cyclophosphamide once; All mice were sacrificed after 30 days. We measured the weight gain, viscera coefficient and sperm quality in each mouse. Results There were no statistical difference in the weight of every week, increase of the weight, testis and epididymis coefficients among the control group, the low, medium and high concentration groups (P > 0.05), while the weight and weight increase were significantly higher than that in the positive control group (P < 0.05). Sperm concentration, viability, morphology, motility, PR, VSL, VAP of low, medium and high concentrations group were significantly improved than the control group (P < 0.05), and the sperm quality in the four groups were higher than that in the positive control group (P < 0.05). Conclusion Water extraction of salvia miltiorrhiza can obviously improve the sperm viability, morphology, and motility of mice.

Objective To evaluate the value of serum E2 levels during COH in predicting IVF-ET outcome. Method Data from 311 IVF-ET cycles received long protocol were collected and analyzed according to E 2 levels 5 days after stimulation:Group A (E2≤500 pmol/L), Group B (500 2 000 pmol/L). Results In groups with E2>1 000 pmol/L, the conditions of oocyte and embryo are higher than E2<1 000 pmol/L. With the increase of E2 levels, period cancellation rate increases. When E2> 2 000 pmol/L, the pregnancy rate become lowest. Conclusion E2 levels among 1 000~1 500 pmol/L during early COH predicts a better pregnancy outcome.

Objective To quantify the neurons in the temporal lobe white matter and find their distribution in the neurologically normal individuals.Methods The temporal lobe at the level of exterior geniculata body from brain autopsy samples of 14 neurologically normal individuals were made into large slice followed by quantitative analysis of neuron density,cell density,ratio and diameter of the neuronal nuclear and the distribution of white matter neurons using two-dimensional cell counting methods.Results With the depth of the white matter of the temporal lobe increasing,the neuron density decreased from 29.26 neurons/ mm~2 to 7.32 neurons/mm~2 and 0.00 neurons/mm~2,respectively;the cell density,neuron ratio and diameter of the neuronal nuclei all decreased.Conclusion There are neurons in the temporal lobe white matter of neurologically normal individuals,whose distribution of neurons is related to the depth of white matter.

Adolescent ; Antibodies, Antinuclear ; blood ; Child ; Histiocytic Necrotizing Lymphadenitis ; etiology ; Humans ; Lupus Erythematosus, Systemic ; complications ; Male ; Sjogren's Syndrome ; complications

OBJECTIVESTo investigate whether antisense Bmi-1 plasmid could inhibit the proliferation of Jurkat cells.

METHODSThe antisense plasmid was constructed by PCR amplification of a 171 bp segment spanning Bmi-1 start codon and zinc finger structure and the PCR product was subsequently inserted reversely to plasmid pLNCX2. The final construct was confirmed through restriction enzyme digestion. G418 was added into the medium after the plasmid was successfully introduced into Jurkat cells by using lipofectin-mediated DNA transfection. The proliferation of Jurkat cells were determined by MTT and colony formation assays. Cell cycle was determined by flow cytometry. The p16 expression of Jurkat cells was studied by immunofluorescent histochemistry.

RESULTSThe growth rate of antisense Bmi-1 transfected Jurkat cells was significantly lower than that of the controls, and the colony forming capacity of the transfected cells decreased significantly (P < 0.01), the colony numbers being (90.7 +/- 9.07)/10(3) cells, (83.3 +/- 6.11)/10(3) cells and (56.0 +/- 5.56)/10(3) cells for control cells, empty plasmid transfected Jurkat cells and antisense Bmi-1 transfected Jurkat cells, respectively. The percentage of G, phase cells was increased and the p16 expression of antisense Bmi-1 transfected cells was significantly upregulated than that of control cells.

CONCLUSIONAntisense Bmi-1 can inhibit the growth and upregulate the expression of p16 of Jurkat cells in vitro.

Cell Cycle ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Humans ; Jurkat Cells ; Nuclear Proteins ; genetics ; Oligonucleotides, Antisense ; genetics ; Plasmids ; genetics ; Polycomb Repressive Complex 1 ; Proto-Oncogene Proteins ; genetics ; Repressor Proteins ; genetics ; Transfection

Eight compounds from six Chrysanthemum morifolium cv. 'Hangju' were determined and multivariate statistics, including principle component analysis (PCA) and partial least squares discriminate analysis (PLS-DA) were used to investigate the potential damaging effect of sulfr-fumigating process. Meanwhile, S, Mn, Fe, Cu, Pb were also detected by using ICP-MS and ICP-OES. In this study, dramatic chemical changes were found that the contents of flavonoid aglycones remarkably increased while those of glycosides and hydroxycinnamoylquinic acids were significantly reduced. The PCA score plots showed six samples were clearly classified into the non-fumigated and sulfur-fumigated types. And according to VIP > 1, the most important chemical markers were luteolin, apigenin and luteolin-7-O-glucoside which indicated that the extracted three components might have a marked effect on the discrimination among different group samples. And then, it was found that the residue of sulfur of SHJ were significantly higher than NHJ (P < 0.05). The established approach was applied to rapidly discriminate sulfur-fumigated HJ with combining the quantitative chemical analysis and multivariate statistical analysis, and then the result will provide some evidence to evaluat the quality of HJ and control its processing.
Chemistry, Pharmaceutical ; methods ; Chrysanthemum ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Flowers ; chemistry ; Fumigation ; methods ; Sulfur ; chemistry

Child ; Humans ; Male ; Mental Disorders ; diagnosis

OBJECTIVETo explore the effects of advanced oxidation protein products (AOPP) on expressions of stromal cell-derived factor-1alpha (SDF-1alpha) in ECV304 cells and the signal pathway that mediated the effects.

METHODSAOPP-BSA was made from bovine serum albumin (BSA) and sodium hypochlorite. After treated with AOPP-BSA of different concentrations (50, 100, 200 micromol/L), the expressions of SDF-1alpha mRNA in ECV304 cells were measured by reverse transcription-polymerase chain reaction (RT-PCR) and the expressions of SDF-1alpha protein and the levels of phosphorylated extracellular signal-regulated kinase (ERK) in ECV304 cells were analyzed by Western blot. In inhibition test, U0126, the special inhibitor of ERK of different concentrations (0.1, 1, 10 rmol/L) were added into ECV304 cells culture media for 1 hour, then the cells were treated with AOPP-BSA for 24 hours, at last the protein levels in supernatant were detected by enzyme-linked immunosorbent assay (ELISA).

RESULTSAOPP-BSA obviously promoted the expressions of SDF-1alpha mRNA and increased the levels of SDF-1beta protein of ECV304 cells in dose-dependent manner (all P < 0.01), after 15 minutes treated with 200 micromol/L AOPP-BSA, the levels of phosphorylated ERK of ECV304 cells increased significantly (P < 0.01). When the ERK pathway was blocked by U0126, the promoting effects of AOPP-BSA on expressions of SDF-la protein in ECV304 cells were significantly inhibited in dose-dependent manner (P < 0.05).

CONCLUSIONAOPP induced the expression of SDF-la of ECV304 cells, ERK signal pathway is an important pathway that mediated the effects.

Advanced Oxidation Protein Products ; pharmacology ; Cell Line ; Chemokine CXCL12 ; metabolism ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Humans ; MAP Kinase Signaling System ; Oxidative Stress ; Phosphorylation ; RNA, Messenger ; genetics

Adenocarcinoma ; diagnosis ; metabolism ; pathology ; secondary ; Brain ; metabolism ; pathology ; Brain Neoplasms ; diagnosis ; metabolism ; pathology ; secondary ; Carcinoembryonic Antigen ; metabolism ; Diagnosis, Differential ; Female ; Humans ; Keratin-7 ; metabolism ; Lung Neoplasms ; pathology ; Magnetic Resonance Imaging ; Middle Aged ; Nuclear Proteins ; metabolism ; Thyroid Nuclear Factor 1 ; Transcription Factors ; metabolism