Objective:To explore the relationship between ABCB1 or ABCG2 gene polymorphisms and therapeutic effects of gefitinib in non-small cell lung cancer (NSCLC) patients, and establish a prediction model of efficacy.Methods:A total of 176 NSCLC patients with epidermal growth factor receptor (EGFR) -sensitive mutation treated with gefitinib admitted to Department of Pulmonary Disease of Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine of Hebei Province from December 2018 to December 2020 were employed as subjects, and all patients were detected ABCB1 and ABCG2 gene polymorphisms. Patients were divided into remission group and non-remission group according to curative effect after 3 months of gefitinib treatment. The clinical data, ABCB1 and ABCG2 gene polymorphisms, levels of serum carcinoembryonic antigen (CEA) and carbohydrate antigen 125 (CA125) were compared between the two groups. The related factors of failure to remission after treatment were analyzed by multivariate logistic regression analysis. Combined with ABCB1 and ABCG2 gene polymorphisms, the prediction model for gefitinib efficacy was constructed and the nomogram was drawn.Results:During the follow-up period, 5 patients were lost to follow-up and 7 patients withdrew from the trial due to intolerable adverse effects, finally 108 patients were employed as remission group, and 56 patients were employed as non-remission group. The numbers of GG, GT and TT at ABCB1 rs2032582 in the remission group were 49, 50 and 9, and those in the non-remission group were 12, 35 and 9, with a statistically significant difference ( χ2=9.56, P=0.008). The numbers of GG, GA and AA at ABCG2 rs2231137 in the remission group were 13, 72 and 23, and those in the non-remission group were 11, 42 and 3, with a statistically significant difference ( χ2=7.74, P=0.021). Before treatment, the levels of serum CEA in the remission group and the non-remission group were (34.28±5.11) ng/ml and (37.88±7.05) ng/ml, with a statistically significant difference ( t=3.74, P<0.001). The levels of CA125 of the two groups were (27.24±6.50) U/ml and (33.31±6.09) U/ml, with a statistically significant difference ( t=-5.79, P<0.001). Multivariate logistic regression analysis showed that TT at rs2032582 of ABCB1 gene ( OR=12.99, 95% CI: 3.17-53.23, P<0.001), GG at rs2231137 of ABCG2 gene ( OR=7.75, 95% CI: 1.36-44.07, P=0.021) and GA ( OR=6.94, 95% CI: 1.47-32.84, P=0.015), CA125 ( OR=1.18, 95% CI: 1.10-1.28, P<0.001) were independent risk factors of failure to remission in NSCLC patients with EGFR sensitive mutation after treatment. The consistency index (C-index) of nomogram for predicting failure to remission was 0.92 (95% CI: 0.86-0.94) . Conclusion:ABCB1 rs2032582 and ABCG2 rs2231137 polymorphisms are related to therapeutic effects of gefitinib in the NSCLC patients, the nomogram based on the two genes combined with serum CA125 can predict efficacy of gefitinib.

Objective: To study the effect of plumbagin on epithelial-mesenchymal transition (EMT) and underlying mechanisms in human tongue squamous cell carcinoma (TSCC) cells. Methods: Methyl thiazolyl tetrazolium assay was apllied to examine the proliferation inhibition effect and half maximal inhibitory concentration (IC₅₀) of plumbagin (0.1, 1.0, 5.0, 10.0, 20.0 μmol/L) in 12, 24, 48 h in TSCC cells. Transwell assay was used to count the number of transmembrane cells and scratch test was performed to examine cells mobility. The flow cytometry was applied to measure intracellular reactive oxygen species (ROS) level in control group, plumbagin group (1.0 μmol/L, 24 h) and glutathione (GSH)+plumbagin group. The expression of E-cadherin, vimentin, Slug, p38 mitogen activated protein kinases (p38MAPK) and phospho-p38MAPK (p-p38MAPK) proteins were determined by Western blotting. The expression of E-cadherin, vimentin and Slug were detected by Western blotting in control group, plumbagin group, activator combined group (p38MAPK activator+plumbagin) and inhibitor combined group (p38MAPK inhibitor+plumbagin). Results: After the treatment of plumbagin for 12, 24, and 48 h, the IC₅₀ of TSCC cells were 10.3, 3.1, 1.5 μmol/L. After treated by 1.0 μmol/L plumbagin for 24 h, the number of transmembrane cells were significantly reduced ([50±13], P<0.05) in comparison to control group (204±6), as well as the cells mobility ([18.2±2.3]%, P<0.05) in comparison to control group ([49.3±1.2]%). Compared to control group (2.32±0.52), the ROS level was increased in plumbagin group (902.20±10.69), while compared to plumbagin group, the ROS level was reduced in GSH combined group (2.18±0.15). In comparison to control group, the expression of E-cadherin was up-regulated (P<0.05), and vimentin, Slug, p-p38MAPK/p38MAPK were down-regulated in plumbagin group (P<0.05). In comparison to plumbagin group, the expression of E-cadherin was down-regulated (P<0.05), and vimentin, Slug, p-p38MAPK/p38MAPK were up-regulated in GSH combined group (P<0.05). Treatment of cells with p38MAPK activator could decrease the expression of E-cadherin significantly (P<0.05) and increase the expression of vimentin (P<0.05) and Slug (P<0.05) in comparison to plumbagin group. Treatment of cells with p38MAPK inhibitor could increase the expression of E-cadherin significantly (P<0.05) and decrease the expression of vimentin (P<0.05) and Slug (P<0.05) in comparison to plumbagin group. Conclusions Plumbagin inhibits EMT via ROS/p38MAPK-mediated pathway in human TSCC cells.

OBJECTIVE：To s tudy the intestinal absorption differences of 6 kinds of active constituents of Polygonum orientale （kaempferol，isokaempferol，vitexin，protocatechuic acid ，kaempferol-3-O-β-D-glucoside and quercetin ）in normal and myocardial ischemia（MI）model rats. METHODS ：UPLC-MS/MS method was adopted to determine the contents of 6 active components in the intestinal circulatory perfusion fluid. Totally male SD 80 rats were divided into normal group and model group ，with 40 rats in each group. Model group was given isoproterenol hydrochloride （50 mg/kg） subcutaneously to induce MI model；normal group was given constant volume of normalsaline， once a day ， for consecutive 2 days. 24 h after successful molding ，normal group and model group received in-situ intestinal circulatory perfusion experiment. The effects of different concentration s of P. orientale extract（5.0，10.0， 20.0 mg/mL），different intestinal segments （duodenum，jejunum，ileum，colon），P-glycoprotein（P-gp）inhibitors（verapamil） and bile on the intestinal absorption of each constituent were explored. RESULTS ：The linear ranges of concentrations of kaempferol， isokaempferol， vitexin， protocatechuic acid ， kaempferol-3-O-β-D-glucoside and quercetin were 3.15-50.40， 3.21-51.31，1.63-52.43，1.60-50.94，1.31-20.97，8.07-129.25 µg/mL（r＞0.999）. The lower limits of quantification were 7.86， 8.45，6.52，4.00，3.28，16.14 ng/mL，respectively. RSDs of precision ，matrix effect and stability tests were all lower than 11％； the accuracy were 85.64％-107.65％，which were in line with the requirements of biological sample quantification analysis. Except for there was no statistical significance in the absorption of kaempferol absorption in duodenum of model group at different concentrations，absorption of other five constituents in duodenum of normal and model rats increased with the increase of the concentration of active constituents ，and absorption of medium- and/or high- concentration active constituents （except quercetin ）in model group was significantly lower than normal group （P＜0.05）. In normal group ，the absorption of kaempferol was more in jejunum，ileum and colon ，isokaempferol was more in ileum ，vitexin and protocatechuic acid were more in jejunum and ileum ， kaempferin-3-O- β-D-glucoside was more in duodenum ，jejunum and colon ，quercetin was more in colon ；in the model group ，the absorption of Polygonum orientale in jejunum and colon was more ，the absorption of isokaempferol in 4 intestinal segments was little different ，vitexin was mainly absorbed in ileum ，protocatechuic acid and kaempferol- 3-O-β-D-glucoside was mainly absorbed in jejunum ，quercetin was mainly absorbed in duodenum and ileum ；in the same intestine ，the absorption of constituents in the model group was less than normal group. After adding verapamil ，absorption of all constituents in the normal group increased ，but the difference was not statistically significant （P＞0.05）；absorption of kaempferol ，isokaempferol，vitexin，protocatechuic acid and kaempferol- 3-O-β-D-glucoside were all increased significantly in model group （P＜0.05），while there was no statistical significance in the increase of quercetin （P＞0.05）. After the bile flowed into the duodenum ，absorption of protocatechuic acid was increased significantly in normal group （P＜0.05）；absorption of other active constituents were increased significantly in model group，except for isokaempferol and quercetin （P＜0.05）. CONCLUSIONS ：Six active constituents of P. orientale were absorbed in the whole intestine of normal and MI model rats ，and the absorption of above constituents may be enhanced more significantly by P-gp inhibitor and bile under pathological condition.

The cochlear auditory epithelium contains two types of sound receptors, inner hair cells (IHCs) and outer hair cells (OHCs). Mouse models for labelling juvenile and adult IHCs or OHCs exist; however, labelling for embryonic and perinatal IHCs or OHCs are lacking. Here, we generated a new knock-in Fgf8^P2A-3×GFP/+ (Fgf8^GFP/+) strain, in which the expression of a series of three GFP fragments is controlled by endogenous Fgf8 cis-regulatory elements. After confirming that GFP expression accurately reflects the expression of Fgf8, we successfully obtained both embryonic and neonatal IHCs with high purity, highlighting the power of Fgf8^GFP/+. Furthermore, our fate-mapping analysis revealed, unexpectedly, that IHCs are also derived from inner ear progenitors expressing Insm1, which is currently regarded as an OHC marker. Thus, besides serving as a highly favorable tool for sorting early IHCs, Fgf8^GFP/+ will facilitate the isolation of pure early OHCs by excluding IHCs from the entire hair cell pool.
Animals ; Mice ; Hair Cells, Auditory, Inner ; Cochlea/metabolism* ; Hair Cells, Auditory, Outer/metabolism* ; Disease Models, Animal ; Fibroblast Growth Factor 8/metabolism*