ABSTRACT
OBJECTIVE@#To select specific DNA aptamer for determining ketamine by FluMag-SELEX.@*METHODS@#Based on magnetic beads with tosyl surface modification as solid carrier and ketamine as target, a random ssDNA library with total length of 78 bp in vitro was compounded. After 13 rounds screening, DNA cloning and sequencing were done. Primary and secondary, structures were analyzed. The affinity, specificity and Kd values of selected aptamer were measured by monitoring the fluorescence intensity.@*RESULTS@#Two ssDNA aptamers (Apt#4 and Apt#8) were successfully selected with high and specific abilities to bind ketamine as target with Kd value of 0.59 and 0.66 μmol/L. The prediction of secondary structure was main stem-loop and G-tetramer. The stem was the basis of stability of aptamer's structure. And loop and G-tetramer was the key of specific binding of ketamine.@*CONCLUSION@#FluMag-SELEX can greatly improve the selection efficiency of the aptamer, obtain the ketamine-binding DNA aptamer, and develop a new method for rapid detection of ketamine.
Subject(s)
Aptamers, Nucleotide/metabolism , DNA , DNA, Single-Stranded/genetics , In Vitro Techniques , Ketamine/metabolism , Oligonucleotides , SELEX Aptamer Technique/methodsABSTRACT
OBJECTIVE@#To explore the effect of electroacupuncture on heroin seeking behavior and FosB expression in relevant brain regions.@*METHODS@#Rat model of heroin relapse behaviors was developed with progressive fixed ratio program,and model rats were randomly divided into 3 groups: a restraint group, a needle retention group, and a electroacupuncture group. The heroin seeking behavior was elicited by a small dose of heroin. FosB expression in relevnt brain region was assessed with immunohistochemical technique.@*RESULTS@#Tests on reinstatement of drug seeking behavior induced by heroin priming showed that compared with the restraint group, active pokes in the electroacupuncture group decreased significantly(P<0.05). Compared with the restraint group, the expression of FosB positive nuclei in Acd, Pcg and CeA of rats brain both in the electroacupuncture group and the needle retention group (P<0.05) decreased significantly. In LC, the expression of FosB positive nuclei in the needle retention group decreased significantly compared with the restraint group (P<0.05).@*CONCLUSION@#Continuous acupuncture and needle retention attentuate the reinstatement of heroin-seeking behaviors induced by heroin priming, and the inhibitory effect may be mediated partially by the expression of FosB in relevant regions which are involved in the process of heroin addiction.
Subject(s)
Animals , Male , Rats , Amygdala , Metabolism , Behavior, Animal , Brain , Metabolism , Electroacupuncture , Methods , Heroin Dependence , Metabolism , Psychology , Therapeutics , Nucleus Accumbens , Metabolism , Proto-Oncogene Proteins c-fos , Random Allocation , Rats, Sprague-DawleyABSTRACT
OBJECTIVE@#To develope an easy to use, rapid and accurate test for detecting buprenorphine based on the principle of competitive immunoassay.@*METHODS@#Monoclonal antibody against buprenorphine was conjugated with colloidal gold and dispensed on the glass fiber. The complete antigen Buprenorphine-BSA and the goat anti-mouse IgG polyclonal antibody were separately sprayed on the nitrocellulose membrane as the test line (T line) and the control line (C line). The rapid test kit was the final assembled product of test strip with the plastic cover.@*RESULTS@#A total of 100 urine samples were tested for buprenorphine by immunochromatographic and GC/ MS methods. The accuracy was 99.0%. It is found the test kit can only detect by cross reaction with other 45 kind drugs.@*CONCLUSION@#Rapid test kit can detect buprenorphine in the samples in 5 minutes. The cut-off value of the test is 100 ng/mL.