Protozoans,the natural host of the facultative intracellular pathogen Legionella species,play an important role in survival,proliferation,virulence and stress resistance of Legionella species. By repeating transformation and selection,a spontaneous mutant of plasmid over expressing green fluorescent protein was obtained. This mutant replicates and is maintained stably in Legionella cells. The colonies of L. pneumophila harbouring the mutated plasmid were intense green in colour even under the daylight. After feeding BF1 strain of Tetrahymena thermophila with transformed L. pneumophila,the intracellular dynamic of changing of bacterial shape,bacterial proliferation and lysis of the host cell due to the bacterial proliferation were observed clearly under fluorescent microscopy. Thus,the present paper provides a simple and intuitionistic strategy for investigating the ecological and cellular relationship between L. pneumophila and its host.

China ; Inventions

BACKGROUNDDicycloplatin is a relatively safe third generation platinum-complex anti-cancer drug. The present study focused on the effects of dicycloplatin on in vitro proliferation and apoptosis of human aortic smooth muscle cells (HASMC) and human aortic endothelial cells (HAEC).

METHODSProliferation of HASMC and HAEC, DNA content, and cellular levels of proliferation- and apoptosis-related proteins were assessed using the (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) assay, flow cytometry and Western blotting assays, respectively.

RESULTSDicycloplatin at 10 ng/ml significantly inhibited HASMC proliferation, however, 10 µg/ml were required to significantly inhibit HAEC proliferation. Cell cycle analysis showed that dicycloplatin was a non-specific inhibitor of the cell cycle. Although dicycloplatin significantly decreased proliferating cell nuclear antigen (PCNA) expression in HASMC at all concentrations tested, it did not significantly affect PCNA expression in HAEC; Bax and p53 protein expression was upregulated in dicycloplatin groups.

CONCLUSIONSDicycloplatin at nanogram concentrations significantly inhibits HASMC proliferation, although the effect is relatively weaker than that of sirolimus. In contrast, the effect of dicycloplatin on inhibition of HAEC proliferation is much less pronounced than that on HASMC. The latter characteristics point to the potential for use of dicycloplatin in drug-eluting stents.

Aorta ; cytology ; Blotting, Western ; Cell Proliferation ; drug effects ; Drug Combinations ; Drug-Eluting Stents ; Endothelial Cells ; cytology ; drug effects ; Flow Cytometry ; Glutamates ; pharmacology ; Humans ; Muscle, Smooth, Vascular ; cytology ; drug effects ; Organoplatinum Compounds ; pharmacology ; Sirolimus ; pharmacology

OBJECTIVETo explore the effect of polydatin on the growth of TGF-β₁induced humanalveolar epithelium A549 cells and the mechanism of polydatin for inhibiting the process of epithelial-mesenchymal transition (EMT).

METHODSA549 cells in vitro cultured were randomly divided into five groups, i.e., the blank group, the control group, the low dose polydatin group, the middle dose polydatin group, the high dose polydatin group. Common culture fluid was added in A549 cells of the blank group. Five ng/mLTGF-β₁contained culture fluid was added in A549 cells of the control group. 50, 100, and 150 μmol/mL of polydatin plus 5 ng/mL TGF-β₁contained culture fluid was added in A549 cells of low, middle, and high dosepolydatin groups, respectively. Morphological changes were observed and recorded at different time points. The optimal concentration of polydatin was determined by MTT method. Protein and mRNA expressions of E-cad epithelial cell marker) and Vimentin (mesenchymal cell marker) were detected by Western blot and Real-time PCR.

RESULTSUnder inverted phase contrast microscope, A549 cells turned from previous pebble shape to fusiform shape after intervened by polydatin and TGF-β1. The intercellular space was enlargedand the intercellular connection became loose. These phenomena were more obviously seen in the control group. A549 cells were more satiated in low, middle, and high dose polydatin groups than in the control group. The EMT inhibition was most obviously seen in the middle dose polydatin group at 48 h. Protein and mRNA expressions of E-cad showed an overall descending tendency after intervened by polydatin and TGF-β1 (P < 0.05). But compared with the control group, protein and mRNA expressions of E-cad were down-regulated in a lesser amplitude in each intervened group. Besides, the tendency was more obviously seen at 48 h than at 24 h. Protein and mRNA expressions of Vimentin showed an overall up-regulating tendency. But compared with the control group, protein and mRNA expressions of Vimentin were down-regulated in a lesser amplitude in each intervened group. Besides, the tendency was more obviously seen at 48 h than at 24 h (P < 0.05). CONCLUSIONS Polydatin could inhibit TGF-β1 induced EMT process of A549 cells time- and dose-dependently. It also played roles in inhibiting pulmonary fibrosis.

Cadherins ; metabolism ; Cell Line ; Epithelial Cells ; drug effects ; Epithelial-Mesenchymal Transition ; drug effects ; Glucosides ; pharmacology ; Humans ; Stilbenes ; pharmacology ; Transforming Growth Factor beta1 ; pharmacology ; Vimentin ; metabolism

To determine the pharmacokinetics of gemcitabine (2',2'-difluorodeoxycytidine) in Chinese non-small-cell lung cancer (NSCLC) patients. Six study subjects were administered gemcitabine at a fixed dose rate of 10 mg/m2 per min (1200 mg/m2,two hours infusion) and carboplatin, and plasma gemcitabine concentrations were measured by ion-pair reversed-phase high-performance liquid chromatography (HPLC). 3P97 Pharmaceutical Kinetics Software was used for the calculation of pharmacokinetic parameters. The obtained mean parameters, elimnation half life (t1/2) (10.67±3.38 min), area under the curve hematologic toxicology result showed that the regimen was effective on and tolerated by the patients.

Adult ; Enchondromatosis ; pathology ; Female ; Granulosa Cell Tumor ; pathology ; Humans ; Ovarian Neoplasms ; pathology

Conservative treatment with high doses of progestin is an alternative to standard hysterectomy for young patients with early-stage endometrial adenocarcinoma who desire to preserve their fertility. Here we report a patient with well-differentiated early-stage endometrial adenocarcinoma and poor fertility potential who failed to become pregnant in two in vitro fertilization-embryo transfer cycles and suffered a relapse after conservative treatment. This case illustrates that assessment of fertility potential is critical at the time of initial evaluation and conservative treatment planning for patients with endometrial adenocarcinoma.
Adenocarcinoma ; drug therapy ; metabolism ; Adult ; Antineoplastic Agents, Hormonal ; Endometrial Neoplasms ; drug therapy ; metabolism ; Female ; Follicle Stimulating Hormone ; therapeutic use ; Gonadotropins ; therapeutic use ; Humans ; Infertility ; Pregnancy ; Progesterone ; therapeutic use ; Reproductive Techniques, Assisted

BACKGROUNDOvarian hyperstimulation syndrome (OHSS) is one of the most life-threatening complications of assisted reproduction treatments. Gonadotropin-releasing hormone antagonists (GnRHanta) are thought to be effective in preventing this complication, and some clinical trials have found lower incidences of OHSS in patients treated with GnRHanta. Our aim was to investigate the effects of GnRHanta on vascular permeability and the expression of vascular endothelial growth factor (VEGF) and its receptors in a rat model of OHSS.

METHODSAn immature early OHSS rat model was established. Three ovarian stimulation protocols were used: pregnant mare serum gonadotropin/human chorionic gonadotropin (hCG) alone, with a GnRHanta, or with a gonadotropin-releasing hormone agonists (GnRHa). Blood and tissue samples were collected at 48 hours after hCG administration. Vascular permeability was evaluated by measuring the Evans-Blue content of extravasated peritoneal fluids. The expression of VEGF and its receptors, including flt-1 and KDR, were detected by reverse transcriptase-polymerase chain reaction and Western blotting.

RESULTSTreatment with both a GnRHanta and a GnRHa resulted in significant reductions in serum estradiol and peritoneal vascular permeability, as well as decreased ovarian expression of VEGF and its two receptors. However, GnRHanta treatment caused a greater reduction in serum estradiol concentrations, and in VEGF receptor mRNA expression than GnRHa. There were no significant reductions in the expression of VEGF or its receptors in extra-ovarian tissues, including the liver, lungs and peritoneum.

CONCLUSIONOur results reveal that GnRHanta are more potent than GnRHa in preventing early OHSS through down-regulation of the expression of VEGF and its receptors in hyperstimulated ovaries.

Animals ; Blotting, Western ; Chorionic Gonadotropin ; pharmacology ; therapeutic use ; Disease Models, Animal ; Female ; Gene Expression ; drug effects ; Gonadotropin-Releasing Hormone ; analogs & derivatives ; antagonists & inhibitors ; pharmacology ; therapeutic use ; Ovarian Hyperstimulation Syndrome ; drug therapy ; genetics ; metabolism ; Rats ; Rats, Wistar ; Receptors, Vascular Endothelial Growth Factor ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVE: To examine the changes in T wave and ST segment amplitude in the supine and standing electrocardiograms (ECG) of children with orthostatic hypertension (OHT) and to determine their clinical significance. METHODS: A total of 49 children with OHT were included in the OHT group. Forty-three age- and sex-matched healthy children were included in the control group. Heart rate and T wave and ST segment amplitude were measured in both groups. T wave amplitude and ST segment amplitude in supine ECG were compared with those in standing ECG within each group. The differences in supine vs standing T wave amplitude and ST segment amplitude were compared between the OHT and control groups. RESULTS: In the control group, T wave amplitude in leads aVR, V, and V-V were significantly lower in standing ECG than in supine ECG (P<0.05); ST segment amplitude in leads V and V were significantly higher in standing ECG than in supine ECG (P<0.05). In the OHT group, T wave amplitude in leads II, aVR, aVF, and V-V were significantly lower in standing ECG than in supine ECG (P<0.05); ST segment amplitude in lead II was significantly higher in standing ECG than in supine ECG (P<0.05). The differences in T wave amplitude in lead II and V between supine and standing ECG were significantly higher in the OHT group than in the control group (P<0.05). Difference in T wave amplitude in lead V6 between supine and standing ECG was a significant diagnostic marker for OHT (P<0.05). This marker had 72.10% sensitivity and 57.10% specificity for the diagnosis of OHT at the optimal cut-off value of 0.105 mV. CONCLUSIONS Difference in T wave amplitude in lead V between supine and standing ECG has certain diagnostic value for OHT.
Child ; Electrocardiography ; Heart Rate ; Humans ; Hypertension ; Sensitivity and Specificity ; Standing Position

OBJECTIVECardiotonic steroids (CTS) can bind to Na+, K+ -ATPase to activate complex intracellular signaling cascades regulating the proliferation and apoptosis of cells. The aim of this study was to investigate the effects of ouabain at different concentrations on growth regulation in various kinds of leukemia cell lines and explore the pathogenesis of leukemia, the functions of Na+, K+ -ATPase as a signal transduction conductor and its effects on cell growth.

METHODSUsing the MTT assay, the survival rates of leukemia cell lines were observed 6, 12 and 24 hrs after treatment with 1 or 10 nmol/L ouabain. The expression of Na+, K+ -ATPase alpha1 subunit of leukemia cells was detected by Western blot.

RESULTSThe MTT results showed that ouabain at 1 nmol/L or 10 nmol/L induced proliferation of lymphocytic leukemia B95 and Jhhan cell lines, as well as megakaryocytic leukemia M07e and Meg01 cell lines. Ouabain at 1 nmol/L or 10 nmol/L increased the expression of Na+, K+ -ATPase alpha1 subunit. There were significant differences in the proliferation and the expression of Na+, K+ -ATPase alpha1 subunit of the leukemia cell lines between the ouabain treatment and the blank control groups 24 hrs after ouabain treatment (P<0.05). The proliferation effect of leukemia cell lines was in a direct proportion with the ouabain concentration and incubation time.

CONCLUSIONSNa+, K+ -ATPase plays an important role in signal transductions. Through binding to ouabain, Na+, K+ -ATPase may regulate proliferation of leukemia cell lines of different origins. Ouabain at 1 nmol/L or 10 nmol/L may induce proliferation of lymphocytic leukemia cell lines (B95, Jhhan) and megakaryocytic leukemia cell lines (M07e, Meg01), and the proliferation effect was in a direct proportion with the concentration and incubation time of ouabain.

Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Leukemia ; pathology ; Ouabain ; pharmacology ; Signal Transduction ; Sodium-Potassium-Exchanging ATPase ; analysis ; genetics ; physiology