PURPOSE: Gastric emptying has been evaluated by scintigraphy in spite of its limitations of time consumption, cost, and danger of radioisotope. Endoscopy is a simple technique, however, its validation for gastric emptying and quantification of food has not yet been investigated. The aim of our study was to assess endoscopic gastric emptying compared with scintigraphy and radiopaque markers (ROMs) studies. We also investigated the effect of a single dose of mosapride on gastric emptying. MATERIALS AND METHODS: Fifteen healthy volunteers underwent scintigraphy. Next day, subjects received a standard solid meal with ROMs and underwent endoscopy and simple abdomen X-ray after 3 hrs. After one week, the same procedure was repeated after ingestion of mosapride (5 mg for group 1, n = 8; 10 mg for group 2, n = 7) 15 min before the meal. Quantification of gastric residue by endoscopy was scored from 0 to 3, and the scores were added up. RESULTS: All subjects completed the study without any complication. The gastric emptying rate [T1/2 (min)] was in normal range (65.6 +/- 12.6 min). Endoscopic gastric emptying was correlated significantly with gastric clearance of ROMs (r = 0.627, p = 0.012). Endoscopic gastric emptying and gastric clearance of ROMs after administration of mosapride showed significant differences in the 10 mg group (p < 0.05). CONCLUSION: Endoscopy can evaluate gastric emptying safely and simply on an outpatient basis. A 10 mg dose of mosapride enhanced gastric emptying, assessed by both endoscopy and ROMs.
Adult ; Benzamides/*pharmacology ; Endoscopy/*methods ; Female ; Gastric Emptying/*drug effects ; Gastrointestinal Agents/*pharmacology ; Humans ; Male ; Middle Aged ; Morpholines/*pharmacology ; Radionuclide Imaging/*methods ; Stomach/radiography/radionuclide imaging ; Young Adult

Mosapride citrate (Mosapride) is a new prokinetic agent that enhances the gastrointestinal (GI) motility by stimulation of 5-HT4 receptors. This agent stimulates acetylcholine release from enteric cholinergic neurons in the GI wall. It was reported in several studies that mosapride selectively enhanced the upper, but not lower, GI motor activity. However, in these studies other 5-HT4 receptor agonists exerted stimulating effects on the motility of the colon. Moreover, it is well known that the receptors of 5-HT4 are also located in the colon. The purpose of this study was to estimate the effect of mosapride on the motility of the stomach, ileum and colon in the guinea pig and to investigate whether or not mosapride influenced the colonic motility. Mosapride significantly increased the amplitude of the contraction waves in the guinea pig stomach by electrical stimulation. In addition, it significantly increased the number of peaks, the area under the curve and the propagation velocity of the peristaltic contraction of the guinea pig ileum in a concentration dependent fashion. Mosapride also significantly shortened the transit time of the guinea pig colon. Accordingly, we concluded that mosapride exerted prokinetic effect on the entire GI tract of the guinea pig. Based on the possibility of similar results in humans, we suggest the potential use of mosapride for lower GI motor disorders such as constipation and upper GI motor disorders such as gastroesophageal reflex disease or gastroparesis.
Animals ; Benzamides/*pharmacology ; Colon/*drug effects ; Gastrointestinal Agents/*pharmacology ; Gastrointestinal Motility/*drug effects ; Guinea Pigs ; Ileum/*drug effects ; Morpholines/*pharmacology ; Stomach/*drug effects ; Support, Non-U.S. Gov't

OBJECTIVESTo explore the effects of insulin on the expression and the regulatory pathway of AQP9 in normal human liver cells.

METHODSNormal human liver cells L02 were cultured and treated with PI3K inhibitor LY294002, AKT inhibitor A-443654, MAPK inhibitors SB2030580 and insulin at different concentrations respectively. The AQP9 mRNA and protein expressions were detected with semi-quantitative RT-PCR and Western blot respectively.

RESULTSThe insulin (100 nmol/L approximately 500 nmol/L) treatment decreased the expression of AQP9 in normal human liver cells (P less than 0.05) concentration dependently, and the expression of AQP9 began to reduce from 3 hours of insulin stimulation (P less than 0.05), especially at insulin treatment for 12 hours (P less than 0.05); Incubated with the selective inhibitor of PI3K (LY294002) and AKT (A-443654), the inhibitory effects of insulin on AQP9 expression decreased (P less than 0.05); but it did not change significantly by blocking the MAPK signaling pathway.

CONCLUSIONThe insulin treatment inhibited the expression of AQP9 and the PI3K/akt signal transduction pathway was involved in the mechanism.

Aquaporins ; metabolism ; Cells, Cultured ; Chromones ; pharmacology ; Hepatocytes ; drug effects ; metabolism ; Humans ; Insulin ; pharmacology ; Morpholines ; pharmacology ; Phosphatidylinositol 3-Kinase ; antagonists & inhibitors ; metabolism ; Protein-Serine-Threonine Kinases ; metabolism ; Signal Transduction

This study was purposed to investigate the reversal effect of glucosylceramide synthase (GCS) inhibitor D, L-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) hydrochloride, on multidrug resistance in K562/A02 cells and its mechanism. The IC(50) (the half maximal inhibitory concentration) of PDMP was measured by MTT method. Cell apoptosis and intracellular daunorubicin (DNR) concentration were detected by flow cytometry. The expression of GCS and mdr1 genes were assayed by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot. The results showed that the IC(50) of DNR in K562 and K562/A02 cells were 0.23 +/- 0.02 and 7.15 +/- 0.24 microg/ml respectively. When the concentration of PDMP was equal to or less than 20 micromol/L ( < / = 20 micromol/L), the obviously inhibitory effect on proliferation of K562 and K562/A02 cells was not observed, but both 20 micromol/L and 10 micromol/L PDMP could enhance the sensitivity of K562/A02 cells to DNR (p < 0.01) and the reversal multiples were 2.59 and 1.69 respectively. After treating with 20 micromol/L and 10 micromol/L PDMP for 48 hours, the concentration of DNR in K562/A02 cells increased (p < 0.05) and the apoptotic rate also was elevated (p < 0.01). The expressions of GCS and mdr1 genes were down-regulated at mRNA and protein levels after treating K562/A02 cells with 20 micromol/L PDMP for 48 hours. It is concluded that PDMP can enhance the sensitivity of K562/A02 cells to DNR by increasing cell apoptosis rate and accumulation concentration of DNR in cells, which may be related to down-regulated expressions of GCS and mdr1 genes.
Daunorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Enzyme Inhibitors ; pharmacology ; Humans ; Inhibitory Concentration 50 ; K562 Cells ; Morpholines ; pharmacology

OBJECTIVETo investigate the effects of LY294002, a phosphatidylinositol 3-kinase inhibition, on sperm motility in asthenozoospermia patients in vitro, and further analyze the possible molecular mechanism.

METHODSSperm aseptically obtained by masturbation and prepared by swim-up technique from 10 patients with asthenozoospermia and 10 healthy fertile men were incubated with different concentrations of LY294002. Measurements of motility were carried out at 10, 30 and 60 min in all specimens by CASA.

RESULTSThe sperm in asthenozoospermia patients treated with LY294002 showed a significant increase in sperm progressive motility, the percentage of motile cells, VSL and VAP.

CONCLUSIONLY294002 can enhance the motility of sperm in asthenozoospermia patients in vitro.

Asthenozoospermia ; drug therapy ; Cells, Cultured ; Chromones ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; Male ; Morpholines ; pharmacology ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; Sperm Motility ; drug effects ; Spermatozoa ; drug effects

OBJECTIVETo study the effect of endogeneous gangliosides (Gls) on integrin alpha2beta1-mediated adhesion of neuroblastoma cells to collagen (Col).

METHODSNeuroblastoma SK-N-SH cell line was cultured in the modified eagle's medium with the presence of 10 mum D-threo-1-phenyl-2-decanolamino-3-morphinolin-1-propanol (D-PDMP), an inhibitor of glucosylceramide synthase. Flow cytometry was used to detect the expression of integrin alpha2beta1 in the cell line. The effects of Mg2(+) and monoclonal antibodies to integrin alpha2beta1 on the adhesion of the cell line to immobilized Col were observed. The adhesion cell number was measured with the BCA method and presented with absorptance A570.

RESULTSThere was a high expression of integrin alpha2beta1 in the SK-N-SH cell line without D-PDMP treatment. Endogenous Gls in the cells were almost depleted after 6-day exposure to D-PDMP, but the integrin alpha2beta1 expression was not significantly changed. 1 mmoL/L Mg2(+) treatment increased significantly the number of adhesion cells in the SK-N-SH cell line. The adhesion to Col of the SK-N-SH cells exposed to D-PDMP which Gls was depleted was significantly reduced compared with the control SK-N-SH cells treated with 1 mmoL/L Mg2(+) (A570: 0.33 +/- 0.016 vs 0.57 +/- 0.033; P < 0.01). After endogeneous Gls was added into the Gls-depleted SK-N-SH cells, the adhesion of the cells was restored (A570: 0.52 +/- 0.035). The adhesion of SK-N-SH cells was significantly blocked by anti-alpha2 and anti-beta1 monoclonal antibodies, with A570 of 0.31 +/- 0.018 and 0.36 +/- 0.021 respectively.

CONCLUSIONSEndogenous tumor Gls increases neuroblastoma cell adhesion to Col by regulating the function of integrin alpha2beta1, but has no effects on the integrin expression. It is suggested that tumor Gls may play a role in migration, invasion and metastasis of tumor cells.

Antibodies, Monoclonal ; immunology ; Cell Adhesion ; Cell Line, Tumor ; Collagen ; physiology ; Gangliosides ; physiology ; Humans ; Integrin alpha2beta1 ; physiology ; Magnesium ; pharmacology ; Morpholines ; pharmacology ; Neuroblastoma ; pathology

This study was aimed to investigate the effects of LY294002, a specific inhibitor of phosphatidylinositol 3-kinase, on growth and apoptosis of MCL Jeko-1 cell line and its mechanism. The proliferation inhibitory rate of Jeko-1 cells treated by different doses of LY294002 was assayed by MTT method; the level of apoptosis of Jeko-1 cells was detected by flow cytometry; the expression level of apoptosis-related protein Cyclin D1, Bcl-2, procaspase-3 and PI3K/Akt signaling pathway protein phosphorylated-Akt (p-Akt), phosphorylated-TOR (p-mTOR), phosphorylated-P70S6K (p-P70S6K) phosphorylated-Akt (p-Akt) in Jeko-1 cells were determined by Western blot. The results showed that the growth of Jeko-1 cell line was inhibited by LY294002. The apoptosis rates of Jeko-1 cells treated with 0, 5, 10 and 20 µmol/L of LY294002 for 24 hours were (3.25 ± 1.27)%, (11.34 ± 2.35)%, (22.81 ± 2.74)%, (43.61 ± 3.48)% respectively, the difference between them was statistically significant (P < 0.01). Phosphorylation levels of PI3K/Akt signaling pathway protein p-Akt, p-mTOR, p-P70S6K decreased, the expression of apoptosis-related protein cyclin D1, Bcl-2, procaspase-3 was down-regulated.It is concluded that the LY294002 can inhibit Jeko-1 cell proliferation, which may be realized through down-regulating the phosphorylation level of p-Akt, p-mTOR, p-P70S6K, inhibiting the P13k/Akt signaling pathway, and promoting the cell apoptosis.
Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chromones ; pharmacology ; Gene Expression Regulation, Leukemic ; Humans ; Morpholines ; pharmacology ; Phosphorylation ; Signal Transduction ; drug effects

OBJECTIVETo explore the mechanisms involved in Staphylococcus aureus (S. aureus) invading human monocytic U937 cells.

METHODSS. aureus were added to U937 cells at multiplicity of infections (MOI) of 20:1 for 0, 15, 30, 60, and 90 minutes, respectively. Cell apoptosis was analyzed with Hoechst 33258 staining and Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) flow cytometry analysis. Akt and nuclear factor-kappaB (NF-kappaB) activities were detected by Western blotting.

RESULTSInfection of U937 cells with S. aureus induced rapid cell death in a time-dependent manner, and the cells displayed characteristic features of apoptosis. S. aureus-induced apoptosis was associated with a prominent downregulation of activated (phosphorylated) Akt and NF-kappaB. The inhibition of phosphorylated Akt by LY294002 led to the inhibition of NF-kappaB in a dose-dependent manner. Inhibition of Akt with LY294002 caused further increase in apoptosis of U937 cells.

CONCLUSIONSS. aureus can stimulate the apoptosis of U937 cells. S. aureus induces apoptosis of U937 cells by inhibiting Akt-regulated NF-kappaB.

Apoptosis ; Chromones ; pharmacology ; Humans ; Morpholines ; pharmacology ; NF-kappa B ; physiology ; Phosphatidylinositol 3-Kinases ; physiology ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; antagonists & inhibitors ; physiology ; Staphylococcus aureus ; pathogenicity ; U937 Cells

Activation of cannabinoid CB1 receptors (CB1Rs) regulates a variety of physiological functions in the vertebrate retina through modulating various types of ion channels. The aim of the present study was to investigate the effects of this receptor on cell excitability of rat retinal ganglion cells (RGCs) in retinal slices using whole-cell patch-clamp techniques. The results showed that under current-clamped condition perfusing WIN55212-2 (WIN, 5 μmol/L), a CB1R agonist, did not significantly change the spontaneous firing frequency and resting membrane potential of RGCs. In the presence of cocktail synaptic blockers, including excitatory postsynaptic receptor blockers CNQX and D-APV, and inhibitory receptor blockers bicuculline and strychnine, perfusion of WIN (5 μmol/L) hardly changed the frequencies of evoked action potentials by a series of positive current injection (from +10 to +100 pA). Phase-plane plot analysis showed that both average threshold voltage for triggering action potential and delay time to reach threshold voltage were not affected by WIN. However, WIN significantly decreased +dV/dtmax and -dV/dtmax of action potentials, suggestive of reduced rising and descending velocities of action potentials. The effects of WIN were reversed by co-application of SR141716, a CB1R selective antagonist. Moreover, WIN did not influence resting membrane potential of RGCs with synaptic inputs being blocked. These results suggest that activation of CB1Rs may regulate intrinsic excitability of rat RGCs through modulating evoked action potentials.
Action Potentials ; Animals ; Benzoxazines ; pharmacology ; Evoked Potentials ; In Vitro Techniques ; Membrane Potentials ; Morpholines ; pharmacology ; Naphthalenes ; pharmacology ; Patch-Clamp Techniques ; Piperidines ; pharmacology ; Pyrazoles ; pharmacology ; Rats ; Receptor, Cannabinoid, CB1 ; physiology ; Retinal Ganglion Cells ; physiology

The present study aimed to investigate whether cannabinoids could modulate the response mediated by ATP receptor (P2X purinoceptor). Whole-cell patch-clamp recording was performed on cultured rat trigeminal ganglionic (TG) neurons. The majority of TG neurons were sensitive to ATP (67/75, 89.33%). Extracellular pretreatment with WIN55212-2, a cannabinoid receptor 1 (CB1 receptor) agonist, reduced ATP-activated current (I(ATP)) significantly. This inhibitory effect was concentration-dependent and was blocked by AM281, a specific CB1 receptor antagonist. Pretreatment with WIN55212-2 at 1×10(-13), 1×10(-12), 1×10(-11), 1×10(-10), 1×10(-9) and 1×10(-8) mol/L reduced I(ATP) (induced by 1×10(-4) mol/L ATP) by (8.14±3.14)%, (20.11±2.72)%, (46.62±3.51)%, (72.16±5.64)%, (80.21±2.80)% and (80.59±3.55)%, respectively. The concentration-response curves for I(ATP) pretreated with and without WIN55212-2 showed that WIN55212-2 shifted the curve downward, and decreased the maximal amplitude of I(ATP) by (58.02±4.21)%. But the threshold value and EC(50) (1.15×10(-4) mol/L vs 1.27×10(-4) mol/L) remained unchanged. The inhibition of I(ATP) by WIN55212-2 was reversed by AM281, suggesting that the inhibition was mediated via the CB1 receptor. Pretreatment with forskolin [an agonist of adenylyl cyclase (AC)] or 8-Br-cAMP reversed the inhibition of I(ATP) by WIN55212-2. These results suggest that the inhibitory effect of cannabinoids on I(ATP) is mediated via the CB1 receptors, that lead to inhibition of the AC-cAMP-PKA signaling pathway.
Adenosine Triphosphate ; physiology ; Animals ; Benzoxazines ; pharmacology ; Cannabinoids ; pharmacology ; Morpholines ; pharmacology ; Naphthalenes ; pharmacology ; Neurons ; drug effects ; physiology ; Patch-Clamp Techniques ; Pyrazoles ; pharmacology ; Rats ; Receptor, Cannabinoid, CB1 ; agonists ; antagonists & inhibitors ; Signal Transduction ; Trigeminal Ganglion ; drug effects ; physiology