To establish a pig kidney cell line LLC-PK1/BCRP in which human breast cancer resistance protein was highly expressed, the expression vector pcDNA3.1(+)-BCRP which contained BCRP gene was constructed and transfected into LLC-PKI cells via liposomes. After selecting with G418, population doubling time, flow cytometry and Western blotting analysis were used to evaluate the cell line. MTT assays were employed to determine the drug resistance index of mitoxantrone and doxorubicin. Invert fluorescent microscope was used to observe the efflux of fluorescence dye Hoechst 33342 by BCRP, furthermore, the BCRP's inhibitor GF120918 was applied to reverse the efflux of Hoechst 33342. The experiment results showed that the expression of BCRP protein increased in LLC-PK1/BCRP cell. The population doubling time of LLC-PK1/BCRP cell was a little longer than that of the parental cell LLC-PK1. The resistance indexes to mitoxantrone and doxorubicin were 51.95 and 6.09 times, respectively, higher than LLC-PK1 cell. The efflux of Hoechst 33342 was significantly enhanced and could be reversed by GF120918. So a LLC-PK1/BCRP cell line was established, which highly expressed BCRP protein successfully. This cell line could be a valuable model to further investigate the biological profile of BCRP and select the substrate and inhibitor of BCRP.

It is meaningful to prompt the research-aimed experiments in the laboratory edu-cation of undergraduate students majoring in pharmacy,which means under the guidance of tutors, students collect information,design the experiment procedure and conduct the whole experiment independently. Under this novel mode of laboratory education, students’ abilities of indepen-dent-thinking and comprehensive-experimental conduction are largely improved. Meanwhile,the sense of team-work is enhanced. To sum up,the research-aimed experiments is significantly ben-eficial to training undergraduate students for future scientific researches.

ObjectiveTo investigate the effect of intermittent theta burst stimulation (iTBS) of the multi-target cerebral cortex after stroke on functional recovery of the upper limb of the hemiplegic side. MethodsFrom November, 2019 to August, 2020, 40 stroke patients in Gansu Provine Hospital Rehabilitation Center were included and randomly divided into single-target stimulation group (n = 20) and multiple-target stimulation group (n = 20). Both groups underwent basic neurorehabilitation drug therapy and conventional rehabilitation exercises. The single-target stimulation group received repetitive transcranial magnetic stimulation (rTMS) (iTBS mode) only in the primary motor cortex (M1) of the affected side. The multi-target stimulation group received rTMS (iTBS mode) in the cerebellar cortex of the healthy brain and M1 of the affected side, once a day, six days a week, for four weeks. Before and after treatment, the scores of Fugl-Meyer Assessment-Upper Extremities (FMA-UE), Action Research Arm Test (ARAT) and modified Barthel Index (MBI), and the latency and amplitude of somatosensory-evoked potentials N20 were compared. ResultsNo serious adverse reaction occurred during treatment. After treatment, the scores of FMA-UE, MBI and ARAT, and the amplitude and latency of N20 improved in both groups (|t| > 3.478, |Z| > 2.243, P < 0.05); and the scores of FMA-UE and ARAT, and the amplitude of N20 were better in the multiple-target stimulation group than in the single-target stimulation group (t > 2.939, Z = -2.697, P < 0.01). ConclusionMulti-target stimulation is superior to single-target stimulation for improving upper limb motor function and N20 amplitude in the hemiplegics after stroke.