The abuse of ketamine has gained popularity in recent years. It is important to develop rapid and accurate methods to determine ketamine and its metabolites in biological samples. The metabolites of ketamine are norketamine and dehydronorketamine in vivo. At present, there are blood, urine, hair and so on as specimens for detection, while the methods include GC, GC/MS, HPLC, LC/MS, HPCE etc. In this paper, these methods used for ketamine and its metabolites were reviewed in order to provide some preference for the study in relative fields.
Anesthetics, Dissociative/chemistry* ; Chromatography, High Pressure Liquid/methods* ; Forensic Medicine ; Gas Chromatography-Mass Spectrometry/methods* ; Hair/chemistry* ; Humans ; Ketamine/metabolism* ; Sensitivity and Specificity ; Substance Abuse Detection/methods*

OBJECTIVE: To improve the successful rate of operation and the livability in establishing acute myocardial ischemic rat model. METHODS: The successful rate of animal experiment is compared between traditional method and improved method. RESULTS: The successful rate of improved method was 90%, which was much higher than 30% in successful rate of traditional method. CONCLUSION The improved method may reduce the difficulty of operation remarkably and cut down the experiment expenditure, which is superior to traditional method.
Acute Disease ; Anesthesia, General/methods* ; Animals ; Coronary Vessels/surgery* ; Disease Models, Animal ; Intubation, Intratracheal/methods* ; Ketamine/chemistry* ; Male ; Myocardial Ischemia/pathology* ; Rats ; Rats, Sprague-Dawley

BACKGROUNDActivation of N-methyl-D-aspartate (NMDA) receptors and alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) receptors play an important role in the neurons death induced by ischemia. The mitigating effect of intravenous anesthetics on ischemic neuron injury is related to their influence on NMDA receptors. This study was performed to investigate the effect of ketamine-midazolam anesthesia on the NMDA and AMPA receptor subunits expression in the peri-infarction of ischemic rat brain and explore its potential mechanism of neuroprotection.

METHODSThirty Sprague Dawley (SD) rats were subjected to permanent middle cerebral artery occlusion under ketamine/atropine (100/0.05 mg/kg) or ketamine-midazolam/atropine (60/50/0.05 mg/kg) intraperitoneal anesthesia (n=15 each). Twenty-four hours after ischemia, five rats in each group were killed by injecting the above dosage of ketamine or ketamine-midazolam intraperitoneally and infarct size was measured. Twenty-four and 72 hours after ischemia, four rats in each group were killed by injecting the above dosage of ketamine or ketamine-midazolam intraperitoneally. After staining the brain tissue slices with toluidine blue, the survived neurons in the peri-infarction were observed. Also, the expression level of NMDA receptors 1 (NR1), NMDA receptors 2A (NR2A), NMDA receptors 2B (NR2B) and AMPA (GluR1 subunit) were determined by grayscale analysis in immunohistochemical stained slices.

RESULTSCompared with ketamine anesthesia, ketamine-midazolam anesthesia produced not only smaller infarct size [(24.1+/-4.6)% vs (38.4+/-4.2)%, P<0.05], but also higher neuron density (24 hours: 846+/-16 vs 756+/-24, P<0.05; 72 hours: 882+/-22 vs 785+/-18, P<0.05) and lower NR2A (24 hours: 123.0+/-4.9 vs 95.0+/-2.5, P<0.05; 72 hours: 77.8+/-4.1 vs 54.2+/-3.9, P<0.05) and NR2B (24 hours: 98.5+/-2.7 vs 76.3+/-2.4, P<0.05; 72 hours: 67.2 +/-7.5 vs 22.2+/-2.6, P<0.05) expression level in the peri-infarction following ischemia.

CONCLUSIONThe protective effects of ketamine-midazolam anesthesia on ischemic brain injury may related to decreasing NR2A and NR2B expression.

Anesthetics, Dissociative ; administration & dosage ; Animals ; Brain Chemistry ; drug effects ; Brain Infarction ; etiology ; metabolism ; pathology ; Brain Ischemia ; complications ; Immunohistochemistry ; Ketamine ; administration & dosage ; Male ; Midazolam ; administration & dosage ; Protein Subunits ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Receptors, AMPA ; biosynthesis ; Receptors, N-Methyl-D-Aspartate ; biosynthesis ; Time Factors

OBJECTIVES: To learn which inhibition of nitric oxide synthase with L-nitro arginine methylester(L-NAME) induces a preeclampsia-like syndrome in pregnant rabbits and high dose of L-arginine reverse the adverse changes induced by nitric oxide synthesis inhibition in pregnant rabbits. MTERIAL AND METHOD: Twenty Newzealand rabbits with 22-days of gestation were injected subcutaneously with 400mg of L-NAME for 7days and 100mg/kg L-arginine was also given intravenously 10 of 20 L-NAME injected pregnant rabbits. They are compared with the control group in which same volume of saline was subcutaneously injected to 5 rabbits with same condition. They were anesthesized by ketamine 50 mg/kg and roupum 2 mg/kg intramuscularly. Cutdown of femoral artery was performed and 22 gauge angioneedle was inserted. On manometer,three way catheter was connected, zeroed with saline, and blood pressure was read. Blood samples were taken from the vein of ear and checked for count of blood cells and bood chemistry (BUN/Cr, GOT/GPT, LDH, Uric acid). Urine protein was measured with nelaton catheterized urine. We injected drugs for 7 days begining on 22 days after mating and performed cesarian section to deliver fetus. To observe their effects on organs, lung, liver, placenta and kidney were taken and fixed with formalin. The sliced kidney tissue in thickness of 1 mm, was fixed with glutaraldehyde for electron microscopy and stored at 4degree C. Special staining was done for closed observation of pattern changes. For statistical analysis, mean+/-SEM was used. The control and experimental groups were compared by unpaired t-test and the differences were significant if probability level is less than 0.05(<0.05). RESULT: Mean blood pressure(MAP) in the experimental group I was significantly high compared to the control group(P<0.05). There was no significant differences in MAP between experimental group II and control group. Urine Protein, BUN, Cr, GOT/GPT, LDH, platelet count in the experimental group I was significantly high(p<0.05) compared to the control group. There was no significant difference between experimental group II and control group. In light microscopic examination, lung, liver, kidney, placenta showed specific finding in experimental group I. Misconstructive of glomerulus in the experimental kidney was well preserved under EM examination. Interstitium of kidney was widened by increase of mesangial matrix. Mild effacement of foot process and cytoplasm of proximal tubule containing electron dense myelin figure like structure were observed. CONCLUSION: Long term injection of L-arginine analogue produced preeclampsia like syndrome and pathologic changes of organ system in pregnant rabbits. Concurrent high dose of L-arginine reversed such chages.
Arginine* ; Blood Cells ; Blood Pressure ; Catheters ; Chemistry ; Cytoplasm ; Ear ; Femoral Artery ; Fetus ; Foot ; Formaldehyde ; Glutaral ; Ketamine ; Kidney ; Liver ; Lung ; Microscopy, Electron ; Models, Animal* ; Myelin Sheath ; NG-Nitroarginine Methyl Ester ; Nitric Oxide Synthase* ; Nitric Oxide* ; Placenta ; Platelet Count ; Pre-Eclampsia* ; Pregnancy ; Rabbits ; Veins

OBJECTIVETo study the effects of ketamine combined with penehyclidine hydrochloride on the learning and memory abilities and the expression of synaptophysin in the hippocampus CA3 region in the brain of neonatal rats.

METHODSEighty seven-day-old Sprague-Dawly rats were randomly intraperitoneally injected with 50 mg/kg of ketamine (K group), 2 mg/kg of penehyclidine hydrochloride (P group), 50 mg/kg of ketamine plus 2 mg/kg penehyclidine hydrochloride (PK group) or normal saline (control group). The rats were trained and tested in a Morris water maze 14 days after administration. The immunhistochemical method was used to ascertain the expression of synaptophysin in the hippocampus CA3 region 24 hrs, 14 days and 28 days after administration.

RESULTSIn the Morris water maze training, the rats in the PK group performed worst, followed by the K group. The rats from the P and NS groups performed well. Compared with the NS group, the expression of synaptophysin in the K and the PK groups decreased significantly 24 hrs and 14 days after administration (p<0.05). The PK group had lower synaptophysin expression than the K group 24 hrs and 14 days after administration (p<0.05). Up to 28 days after administration, the synaptophysin expression increased in all of the four groups and there were no significant differences between groups.

CONCLUSIONSKetamine combined with penehyclidine hydrochloride may inhibit more significantly learning and memory abilities and the synaptophysin expression in the hippocampus CA3 region than ketamine alone in neonatal rats. Penehyclidine hydrochloride alone has no effect on learning and memory abilities and the synaptophysin expression. The synaptophysin expression may increase to a normal level by training and with increasing age.

Animals ; Animals, Newborn ; Cholinergic Antagonists ; pharmacology ; Drug Therapy, Combination ; Excitatory Amino Acid Antagonists ; pharmacology ; Hippocampus ; chemistry ; drug effects ; Ketamine ; pharmacology ; Maze Learning ; drug effects ; Memory ; drug effects ; Quinuclidines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; physiology ; Synaptophysin ; analysis