OBJECTIVE To investigate the effect of Huangqin decoction （HQD）-based self-assembled nanoparticles （SAN） co-loaded with terbinafine （TBF） （TBF-HQD-SAN NPs） on the transdermal absorption of TBF. METHODS High-speed centrifugation combined with dialysis was used to separate HQD-SAN， and TBF-HQD-SAN NPs were obtained by loading TBF using the ultrasound magnetic stirring method； the particle size distribution， Zeta potential and polydispersity index （PDI） of the nanoparticle were characterized， and the encapsulation efficiency （EE） and drug loading （DL） of TBF were determined； using in vitro and in vivo transdermal experiments， the differences in transdermal performance between TBF-HQD-SAN NPs and TBF raw materials， as well as TBF and HQD-SAN physical mixture （TBF-HQD-SAN PM）， were compared and analyzed. RESULTS TBF- HQD-SAN NPs were spherical with a particle size of （177.60±2.57） nm， a PDI of 0.197 4±0.007 9， and a Zeta potential of （－14.63±0.85） mV. The EE and DL of TBF were （99.49±0.71）% and （3.22±0.10）% ， respectively. In vitro transdermal experiments， compared with TBF raw materials， the steady-state permeation rate （Jss） and skin retention of TBF-HQD-SAN NPs increased by 3.34 times and 27.56 times， respectively （P＜0.05）； compared with TBF-HQD-SAN PM， its Jss and skinretention were increased by 2.04 times and 7.44 times， respectively （P＜0.05）. In vivo transdermal experiments 69号） showed that， the area under the drug-time curve and the maximum concentration of TBF-HQD-SAN NPs increased by 2.13 times and 2.06 times respectively compared to TBF raw materials， and increased by 1.59 times and 1.65 times respectively compared to TBF-HQD-SAN PM （P＜0.05）. CONCLUSIONS TBF-HQD-SAN NPs can significantly enhance the in vitro and in vivo transdermal absorption efficiency and skin retention of TBF.

Objective:To evaluate the role of spinal mammlian target of rapamycin (mTOR)/ribosomal S6 kinase 1 (S6K1)/glioma associated oncogene homolog 1 (Gli1) signaling pathway in chronic morphine tolerance in mice.Methods:Healthy male Kunming mice, aged 8-10 weeks, weighing 23-25 g, were used in the study.The experiment was performed in two parts.Experiment I Fifty mice were randomly assigned into 2 groups: normal saline group (group S, n=10) and morphine group (group M, n=40). In M and S groups, morphine and normal saline 10 mg/kg were injected subcutaneously, respectively, twice a day for 7 consecutive days.The thermal pain threshold (TPT) was measured and the maximum analgesic effect percentage (MPE) was calculated at 1 day before administration and 30 min after the last administration every day.Ten mice in each group were randomly selected and sacrificed after measurement of TPT at 1, 3, 5 and 7 days after administration in group M and after the last measurement of TPT in group S, and the lumbar segment (L 4-6) of the spinal cord was removed.Experiment Ⅱ Forty mice were randomly divided into 4 groups ( n=10 each): KU-0063794+ morphine group (group KU+ M), dimethyl sulfoxide (DMSO)+ morphine group (group DM+ M), morphine+ KU-0063794 group (group M+ KU) and morphine + DMSO group (group M+ DM). Morphine 10 mg/kg was injected subcutaneously twice a day for 7 consecutive days in 4 groups.At 1-3 days of morphine injection, mTOR specific inhibitor KU-0063794 200μl (1 μg/μl) and 10% DMSO 200 μl was injected intraperitoneally in KU+ M group and DM+ M group at 30 min before administration twice a day.At 5-7 days of morphine injection, KU-0063794 200μl (1 μg/μl) or 10% DMSO 200 μl was injected intraperitoneally in group M+ KU or group M+ DM at 30min before administration, respectively, twice a day.TPT was measured and MPE was calculated at 1 day before morphine injection and at 30 min after the last administration every day.The animals were sacrificed after the last measurement of TPT, and the lumbar segment (L 4-6) of the spinal cord was removed for determination of the expression of spinal mTOR, phosphorylated mTOR (p-mTOR), S6K1, phosphorylated S6K1 (p-S6K1) and Gli1 (using Western blot). Results:Experiment Ⅰ Compared with group S, MPE was significantly increased at each time point after administration at 3, 5 and 7 days after administration, expression of spinal p-mTOR, p-S6K1 and Gli1 was significantly down-regulated ( P<0.05), and no significant change was found in mTOR and S6K1 in group M ( P>0.05). Experiment Ⅱ Compared with group DM+ M, MPE was significantly decreased at 3-7 days after morphine injection, expression of p-mTOR, p-S6K1 and Gli1 in spinal cord was down-regulated ( P<0.05), and no significant change was found in expression of mTOR and S6K1 in group KU+ M ( P>0.05). Compared with group M+ DM, MPE was significantly increased at 6-7 days after morphine injection, expression of p-mTOR, p-S6K1 and Gli1 in spinal cord was down-regulated ( P<0.05), and no significant change was found in mTOR and S6K1 in group M+ KU ( P>0.05). Conclution:Spinal mTOR/S6K1/Gli1 signaling pathway is involved in the development and maintenance of chronic morphine tolerance in mice.