OBJECTIVETo investigate the effect of adenosine A2A receptor knockout (A(2A)RKO) on relationship between continuous activation of phospho-c-Jun N-terminal kinase (P-JNK) and expression of nerve cell apoptosis in hippocampus CA1 domain of newborn mice after hypoxia/ischemia brain damage(HIBD) and its potential mechanism.

METHODSA(2A)RKO mice and adenosine A2A receptor wildtype (A(2A)RWT) littermates (n = 80) were divided into Sham operation group (S) and model group (M), 1, 3 and 7 day after HIBD, totally 8 groups. HIBD was developed with 7 day-old neonatal mice according classical Rice-Vannucci method. It was tested the effect of A(2A)RKO on short-term neurofunctional outcomes consisted of three developmental reflexes (righting, geotaxis and cliff aversion), the changes of brain pathology with hematoxylin-eosin (HE) staining and Nissl staining, the expressions of nerve cell apoptosis with terminal deoxynucleotidyl transferase mediated dUTP-biotin nick-end labeling(TUNEL) staining and P-JNK were observed by immunohistochemistry.

RESULTSThe neurological behavior injuries and brain histopathological damages and nerve apoptosis cells were aggravated in A(2A)RKO newborn mice after HIBD. The positive expressions of P-JNK were significantly higher in the ischemic hippocampus CA1 domain after HIBD than ones in group S respectively (P < 0.01), reaching to peak at 1 day and then began gradually decreasing. P-JNK expression in model knockout(MKO) at 1, 3 and 7 day increased greatly compared to those in the previous time point of corresponding model wildtype (MWT) (P < 0.01, P < 0.05, P > 0.05); there was a positive correlation between the expressions of P-JNK and nerve cell apoptosis after HIBD in newborn mice(r = 0.837, P < 0.01).

CONCLUSIONEarly continuous activation of P-JNK might be involved in the aggravated nerve apoptosis cells and brain damage induced by A(2A) RKO newborn mice after HIBD.

Animals ; Animals, Newborn ; Apoptosis ; Hypoxia-Ischemia, Brain ; metabolism ; pathology ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Mice ; Mice, Knockout ; Neurons ; drug effects ; metabolism ; pathology ; Receptor, Adenosine A2A ; genetics

A simple, reliable and sensitive liquid chromatography-isotope dilution mass spectrometry (LC-ID/MS) was developed and validated for quantification of olanzapine in human plasma. Plasma samples (50 microL) were extracted with tert-butyl methyl ether and isotope-labeled internal standard (olanzapine-D3) was used. The chromatographic separation was performed on XBridge Shield RP 18 (100 mm x 2.1 mm, 3.5 microm, Waters). An isocratic program was used at a flow rate of 0.4 m x min(-1) with mobile phase consisting of acetonitrile and ammonium buffer (pH 8). The protonated ions of analytes were detected in positive ionization by multiple reactions monitoring (MRM) mode. The plasma method, with a lower limit of quantification (LLOQ) of 0.1 ng x mL(-1), demonstrated good linearity over a range of 0.1 - 30 ng x mL(-1) of olanzapine. Specificity, linearity, accuracy, precision, recovery, matrix effect and stability were evaluated during method validation. The validated method was successfully applied to analyzing human plasma samples in bioavailability study.
Antipsychotic Agents ; blood ; pharmacokinetics ; Benzodiazepines ; blood ; pharmacokinetics ; Biological Availability ; Chromatography, Liquid ; methods ; Humans ; Indicator Dilution Techniques ; Isotope Labeling ; Reproducibility of Results ; Sensitivity and Specificity ; Tandem Mass Spectrometry ; methods

Objective To investigate the effects of different bolus and swallow patterns on esophageal manometry in patients with gastroesophageal reflux disease by high resolution manometry .Methods Patients with gastroesophageal reflux disease questionnaire score of more than 8 points and positive 24-hour pH monitoring were included in the study .All the patients were detected by liquid swallow ,solid swallow and continuous swallow .The parameters and comprehensive diagnosis were in accordance with the Chicago Standard .Results A total of 42 patients with gastroesophageal reflux disease were enrolled . Compared with the dynamic parameters of liquid swallow ,the residual pressure of upper esophageal sphincter [(11 .07 ± 3 .97 ,5 .29 ± 3 .36)mmHg] decreased ,the distal latency [(6 .28 ± 1 .87 ,8 .98 ± 2 .25)s] ,and lower esophageal sphincter relaxation time [(7 .79 ± 0 .98 ,10 .69 ± 13 .04)s] prolonged significantly (all P<0 .05) .In the comprehensive diagnosis of esophageal motility ,compared with liquid swallow (38 .1% ) , continuous liquid swallow showed a more sensitive positive diagnostic rate of ineffective esophageal motility (IEM) (63 .2% ) ,with a significant difference (P=0 .008) .Compared with that of liquid swallow ,the diagnostic rate (45 .2% ) of IEM by the solid swallow did not differ significantly (P=0 .581) . Among the ineffective contraction ,the rate of failed contraction (44.3% ) of solid swallow was higher than that of liquid swallow (22 .6% ) .Conclusion Solid swallow is more likely to induce severe esophageal hypomotility disorders than liquid swallow.Continuous swallow has more sensitivity in the diagnosis of IEM.Therefore.it can be used as a supplement to routine manometry in patients with eastroesophageal reflux disease.