ABSTRACT
Objective To evaluate the release of leucine-enkephalin (L-EK) in the cerebrospinal fluid induced by intrathecal human embryonic kidney (HEK293) cells modified with human preproenkephalin (hPPE)gene and the analgesic efficacy of L-EK for bone cancer pain.Methods Forty CIBP female Sprague-Dawley rats were randomized into transplantation (CIBP+ hPPE/HEK293,n =20) and control (CIBP + HEK293,n =20)groups using a random number table.At 1 day before inoculation of cancer cells (T1,baseline) and 8,15,21,25,32 and 35 days after inoculation (T2-7),thermal paw withdrawal latency (TWL) was measured,and the number of licking/biting the claw on the transplantated side and degree of hindlimb limping during free activities were recorded.After observation at T4,10 rats were chosen from each group and sacrificed and the cerebrospinal fluid of rats was collected in an ice bath for detection of hPPE expression using radioimmunoassay.Results Compared with control group,TWL was significantly prolonged,the concentration of L-EK in the cerebrospinal fluid was increased,and the number of licking/biting the claw on the transplantated side and degree of hindlimb limping during free activities were decreased at T4-7 in transplantation group.Conclusion Intrathecal HEK293 cells modified with hPPE gene can continuously secrete L-EK and mitigate bone cancer pain.
ABSTRACT
ObjectiveTo construct human embryonic kidney cells (HEK293) modified with human preproenkephalin (hPPE) gene.MethodshPPE gene fragments were obtained from recombinant plasmid pcDNA3.1( + )/hPPE by using restriction endonuelease Hind Ⅲ and Not Ⅰ.Homologous recombination of lentivirus and hPPE gene was produced by using recombinant DNA technology.HEK293 cells were then transfected with the recombinant lentivirus vectors.The expression of hPPE gene in HEK293 cells was detected by Western blot.ResultsThe results of DNA sequencing indicated that the positive clone of recombinant lentivirus was completely consistent with sequencing of hPPE in Genebank.The titer of the concentrated virus was 2.07 × 108 TU/ml.GFP fluorescence was not seen in HEK293 cells transfected with the lentiviral vector under fluorescence microscope.A strong fluorescence was seen in HEK293 cells transfected with Ubc-GFP-L.V.empty viral vector.Positive expression of hPPE was demonstrated in HEK293 cells transfected with lentiviral vector by Western blot.Conclusion HEK293 cells modified with hPPE gene were successfully constructed and the target gene hPPE was stably expressed in HEK293 cells.
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OBJECTIVE To study the prevention strategy of community-acquired pneumonia(CAP) for retired old cadres of PLA.METHODS We adopted the first and second prevention strategy of disease to deduce the frequency for CAP.RESULTS The old cadres were divided into common prevention group and important prevention group by evaluating their body condition.The common prevention group was taken the first prevention strategy:healthy education,inhabit surroundings improvement and proper exercises.The second prevention was carried out to personnel who have chronic disease of respiratory organs which included eliminating the risk factors and making efforts to find, diagnose and treat infection as early as possible.CONCLUSIONS The first and second prevention is an effective working mode which improves the level of CAP prevention.
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Objective: To observe the effect of chronic psychological stress on change of function of learning and memory, and CD4, and CD8 in mice. Methods: 50 BALB/c mice were randomly divided into control group and immobilization stress group (n=25 in each group). The mice mode of stress was made by restraint for 6 hours per day. Suspend tail test was performed on change of behaviors in mice. RIA was employed to measure levels of neuropeptide Y in the brain. Results: Compared with the control group, the mean escape latency of the mice in the chronic restraint groups was longer than that of the control mice (P