Objective To observe the effects of Purpura decoction and its disassembled prescriptions on peripheral blood reticulated platelets (RP) and serum thrombopoietin (TPO) of idiopathic thrombocytopenic purpura (ITP) model mice, to explore the action mechanism and core structure of the prescription. Methods Totally 84 BalB/c mice were randomly divided into normal group, model group, prednisone group, Purpura decoction group, Buyi Qixue group, Buqi Huoxue group and Buxue Huoxue group. ITP mouse model was made by injecting intraperitoneally guinea pig-antimouse platelet serum. From the 5th day of modeling, mice were intragastrically administered. The normal group and model group were fed with normal saline 0.2 mL per mouse. Purpura decoction group was fed with Purpura decoction 0.43 mL per mouse. Buyi Qixue group was fed with Buyi Qixue decoction 0.36 mL per mouse. Buqi Huoxue group was fed with Buqi Huoxue decoction 0.27 mL per mouse. Buxue Huoxue group was fed with Buxue Huoxue decoction 0.3 mL per mouse. Prednisone group was fed with prednisone acetate suspension 0.2 mL per mouse. Ten days later, took the blood from the eyeball and isolated serum, the mouse peripheral platelet, RP and TPO were observed. Results After treatment, the platelet count of treatment groups was improved. Compared with before treatment, Purpura decoction group,Buqi Huoxue group and prednisone group had significant difference in platelet count (P<0.05). The RP of model group was significantly higher than the normal group (P<0.05), but RP absolute value was lower than the normal group (P<0.05). Compared with the model group, RP of treatment groups reduced (P<0.05), and RP was negatively correlated with platelet count (P<0.05). TPO level of the model group was higher than that of the normal group (P<0.01). TPO of treatment groups declined significantly, and had significant differences with that of the model group (P<0.05). Conclusion Core structure of Purpura decoction is qi tonifying and blood activating drug. The mechanism of treating ITP is related with lowering RP and TPO.

Objective To evaluate the role of μ-δ heterodimer in down-regulation of the expression of excitatory amino acid transporter 3 (EAAT3) in hippocampi caused by reinstatement of morphine-induced conditioned place preference (CPP) in rats.Methods Thirty-two healthy clean-grade male Sprague-Dawley rats,weighing 200-240 g,were assigned into 4 groups (n =8 each) using a random number table method:control group (group C),extinction group (group E),reinstatement group (group R) and reinstatement plus interference plasmid group (group RI).The model of morphine-induced CPP was established,and extinction of CPP was gradually induced by stopping administration.A small dose of morphine 5 mg/kg was intraperitoneally injected again to induce CPP reinstatement,and dwell time around the medicine box was recorded.μ-δ heterodimer interference plasmid 5 μl was injected into the lateral cerebral ventricle after successful establishment of CPP model in group RI.The content of glutamate (Glu) in hippocampi was measured using high-performance liquid chromatography.The EAAT3 expression in hippocampal CA1 and CA3 regions was detected using Western blot.Results Compared with group C,no significant change was found in the dwell time around the medicine box or content of Glu in hippocampi (P＞0.05),and the expression of EAAT3 in hippocampal CA1 and CA3 regions was significantly up-regulated in group E,and the dwell time around the medicine box was significantly prolonged,the content of Glu in hippocampi was increased (P＜0.05),and no significant change was found in the expression of EAAT3 in hippocampal CA1 and CA3 regions in group R (P＞0.05).Compared with group E,the dwell time around the medicine box was significantly prolonged,the content of Glu in hippocampi was increased,and the expression of EAAT3 in hippocampal CA1 and CA3 regions was down-regulated in group R (P＜0.05).Compared with group R,the dwell time around the medicine box was significantly shortened,the content of Glu in hippocampi was decreased,and the expression of EAAT3 in hippocampal CA 1 and CA3 regions was upregulated in group RI (P＜0.05).Conclusion μ-δ heterodimer is involved in down-regulation of EAAT3 expression in the hippocampus caused by reinstatement of morphine-induced CPP in rats.

Epithelial-mesenchymal transition (EMT) is involved in both physiological and pathological processes. EMT plays an essential role in the invasion, migration and metastasis of tumours. Autophagy has been shown to regulate EMT in a variety of cancers but not in head and neck squamous cell carcinoma (HNSCC). Herein, we investigated whether autophagy also regulates EMT in HNSCC. Analyses of clinical data from three public databases revealed that higher expression of fibronectin-1 (FN1) correlated with poorer prognosis and higher tumour pathological grade in HNSCC. Data from SCC-25 cells demonstrated that rapamycin and Earle's balanced salt solution (EBSS) promoted autophagy, leading to increased FN1 degradation, while 3-methyladenine (3-MA), bafilomycin A1 (Baf A1) and chloroquine (CQ) inhibited autophagy, leading to decreased FN1 degradation. On the other hand, autophagic flux was blocked in BECN1 mutant HNSCC Cal-27 cells, and rapamycin did not promote autophagy in Cal-27 cells; also in addition, FN1 degradation was inhibited. Further, we identified FN1 degradation through the lysosome-dependent degradation pathway using the proteasome inhibitor MG132. Data from immunoprecipitation assays also showed that p62/SQSTM1 participated as an autophagy adapter in the autophagy-lysosome pathway of FN1 degradation. Finally, data from immunoprecipitation assays demonstrated that the interaction between p62 and FN1 was abolished in p62 mutant MCF-7 and A2780 cell lines. These results indicate that autophagy significantly promotes the degradation of FN1. Collectively, our findings clearly suggest that FN1, as a marker of EMT, has adverse effects on HNSCC and elucidate the autophagy-lysosome degradation mechanism of FN1.
Autophagy ; Cell Line, Tumor ; Female ; Fibronectins ; Humans ; Lysosomes/metabolism* ; Ovarian Neoplasms ; Sequestosome-1 Protein/metabolism* ; Squamous Cell Carcinoma of Head and Neck