2.Use of Green Fluorescent Protein to Study the Relationship between Legionella pneumophila and Its Protozoan Host
Shao-Song HUANG ; Run-Lin XU ; Yong-Jun LU ;
Microbiology 1992;0(02):-
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.
3.Effect of Polydatin on Epithelial-Mesenchymal Transition of Human Alveolar Epithelium A549 Cells Induced by TGF-β1.
Jun-chao YANG ; Lu XU ; Kang SONG ; Yuan WANG ; Run-di GAO ; Rui-lin CHEN ; Yu CAO
Chinese Journal of Integrated Traditional and Western Medicine 2016;36(4):466-470
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
4.Dicycloplatin differentially inhibits proliferation of human aortic smooth muscle and endothelial cells: potential for use in drug-eluting stents.
Lian-jun XU ; Run-lin GAO ; Chao WU ; Jue YE ; Li SONG ; Xin QIAN
Chinese Medical Journal 2012;125(24):4386-4392
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
5.Pharmacokinetics of gemcitabine in Chinese patients with non-small-cell lung cancer
Lin-Run WANG ; Ming-Zhu HUANG ; Nong XU ; Jian-Zhong SHENTU ; Jian LIU ; Jie CAI
Journal of Zhejiang University. Science. B 2005;6B(5):446-450
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.
6.Effects of gonadotropin-releasing hormone antagonists on the expression of vascular endothelial growth factor and its receptors in a rat model of ovarian hyperstimulation syndrome.
Xiao-mei TONG ; Song-ying ZHANG ; Tao SONG ; Wei-hai XU ; Xiao-na LIN ; Jing SHU ; Liu LIU
Chinese Medical Journal 2008;121(23):2434-2439
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
7.Outcome of conservative treatment and subsequent assisted reproductive technology for patients with early-stage endometrial adenocarcinoma and poor fertility potential.
Xiao-Mei TONG ; Hai-Yan ZHU ; Xiao-Na LIN ; Ling-Ying JIANG ; Wei-Hai XU ; Liu LIU ; Song-Ying ZHANG
Chinese Medical Journal 2012;125(19):3578-3580
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
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drug therapy
;
metabolism
;
Adult
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Antineoplastic Agents, Hormonal
;
Endometrial Neoplasms
;
drug therapy
;
metabolism
;
Female
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Follicle Stimulating Hormone
;
therapeutic use
;
Gonadotropins
;
therapeutic use
;
Humans
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Infertility
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Pregnancy
;
Progesterone
;
therapeutic use
;
Reproductive Techniques, Assisted
9.T wave and ST segment amplitude changes in the supine and standing electrocardiograms of children with orthostatic hypertension and their clinical significance.
Run-Mei ZOU ; Fang LI ; Ping LIN ; Yi XU ; Cheng WANG
Chinese Journal of Contemporary Pediatrics 2019;21(7):696-700
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
10.Observation on therapeutic effect of acupuncture at point "Xiajiquan" on muscular strength of finger after apoplexy.
Mei-Lin HU ; Ke-Xu ZHANG ; Run-Ping ZHANG
Chinese Acupuncture & Moxibustion 2007;27(11):813-814
OBJECTIVETo approach to the best therapy for recovery of fingers function after apoplexy. METHODS; One hundred and twenty-three cases were randomly divided into an observation group of 63 cases and a control group of 60 cases. The control group were treated by acupuncture at Quchi (LI 11), Shousanli ( LI 10), Waiguan (TE 5), etc., and the observation group by acupuncture at point "Xiajiquan" besides the acupoints for the control group. After treatment of 10 sessions, the therapeutic effects were assessed.
RESULTSThe cured and markedly effective rate was 74.6% in the observation group and 40.0% in the control group, the observation group being significantly better than the control group (P < 0.05).
CONCLUSIONPoint "Xiajiquan" has a better therapeutic effect on derivation of finger muscular strength.
Acupuncture Points ; Acupuncture Therapy ; methods ; Adult ; Aged ; Female ; Fingers ; physiopathology ; Humans ; Male ; Middle Aged ; Muscle Strength ; physiology ; Stroke ; physiopathology ; therapy