Objective To explore the effect of repetitive transcranial magnetic stimulation (rTMS) on behaviors and hippocampal glucocorticoid receptor (GR) protein expression in chronic stress depression model rats and the possible antidepressant mechanism of rTMS. Method Seventy-five male Sprague-Dawley rats were randomly divided into the blank control group (n=15) and the stress-induced group (n=60). Singly housing and chronic unpredictable mild stress (CUMS) were used to induce the depression model in stress-induced group. Forty-five CUMS rats were selected and ran?domly divided into rTMS group (receiving 10 Hz rTMS intervention for 3 weeks), sham group (receiving pseudo rTMS treatments for 3 weeks) and depression group (with no further treatment). Body weight measurements and performance in the sucrose consumption and forced swimming test (FST) were evaluated before modeling, after modeling and after inter?vention. The GR protein and GR mRNA expression level in the hippocampus were examined after intervention. Results Compared with control group, the body weight growth rate and the sugar water preference were significantly lower in stress-induced group (P<0.01), and the immobility time of FST was significantly longer (P<0.01). After the 3-week rTMS intervention, the body weight growth rate and the sugar water preference in rTMS group, which were insignificantly differ?ent from control group (P>0.05), were higher than those in sham group and depression group (P<0.01). The immobility times of FST in rTMS group and control group were shorter than sham group and depression group (P<0.01). Compared with rTMS group and control group, GR and GR mRNA expression levels in the hippocampus were significantly reduced in sham group and depression group (P<0.01). Conclusion rTMS can improve depression behavior of CUMS rats, which may be associated with upregulation of GR expression in the hippocampus.

Objective To observe the effect of repetitive transcranial magnetic stimulation (rTMS) on behavior in response to chronic but unpredictable mild stress and explore potential neuroendocrine mechanisms.Methods Forty adult SD male rats were randomly divided into a control group (n =8) and a model preparation group (n=32).The control group was given normal care while a model of depression was induced in the model preparation group through giving an unpredictable mild stimulus (CUMS).The depressive rats were randomly divided into a model group,an rTMS group and a sham rTMS group (8 cases in each group).The rTMS group and sham rTMS groups accepted the rTMS or sham stimulation for 3 weeks.The changes in behavior in each group were quantified using body weight,sucrose consumption and an open field test before and after stimulation.Enzyme-linked immunosorbent assays (Elisas) were conducted to detect plasma adrenocorticotropic hormone (ACTH) and corticosterone (CORT) levels.Reverse-transcription polymerase chain reactions (RT-PCRs) were carried out to allow the detection of mRNA expression in hypothalamus related to levels of adrenocorticotropic hormone releasing hormone (CRH).Results After the modeling there were significant differences between the model preparation group and the control group in terms of weight increase,sucrose consumption and open field test results.After rTMS the rate of weight increase,sucrose consumption and the scores in the open field test of the rTMS group had increased significantly more than in the control group.Elisas showed significantly higher plasma ACTH and CORT levels in the model group as well.The average expression of CRH mRNA in the model group was significantly higher than in either of the other two groups.Conclusions rTMS can relieve depression-like behavior induced by chronic stress,at least in rats.This may be related to a downgrading of the hyperactive functioning of the hypothalamus-pituitary-adrenal axis.

Objective: To investigate the expression of ciRS-7 in esophageal squamous cell carcinoma (ESCC) and its effect on the cellular proliferation, migration and invasion. Methods: The cancer tissues and paired adjacent normal tissues from 60 ESCC patients treated in the Fourth Hospital of Hebei Medical University between May, 2016 andApril, 2017 were selected for this study. The expressions of ciRS-7 were detected by qRT-PCR. After over-expressing or silencing of ciRS-7, the proliferation of ESCC cell line TE1 was measured by CCK-8 assay; and the migration and invasion were tested by wound healing assay and Transwell invasion assay，respectively. Finally, the effect was validated via animal experiment. Results: CiRS-7 was highly expressed in ESCC tissues (P<0.05), and its expression level was closely related to pathological grade and lymph node metastasis (P<0.05). Over-expression of ciRS-7 significantly increased the proliferation, migration and invasion (all P<0.05) of TE1 cells; while silencing of ciRS-7 remarkably suppressed the proliferation, migration and invasion (all P<0.05). Conclusion: CiRS-7 was up-regulated in ESCC and could enhance ESCC cell proliferation, migration and invasion, suggesting that ciRS-7 could be used as a potential target for the diagnosis and treatment of ESCC.

Objective: To investigate the effect of miR-124 on the invasion and migration of esophageal cancer KYSE170 cells by regulating autophagy. Methods: miR-124 mimic was transfected into esophageal cancer KYSE170 cells. Transwell assay was used to detect the change of invasion and migration ability of cells. Dual luciferase reporter gene assay was used to verify the targeted regulation of BECN1 (Beclin1) by miR-124, and Western blotting was used to analyze the expressions of BECN1, P62 and LC3 protein. siRNA targeting BECN1 was transfeted into KYSE170 cells, and then the cell invasion and migration ability was calculated by Transwell assay. The expressions of BECN1, P62 and LC3 protein were detected by Western blotting. miR-124 mimic and BECN1 over-expression plasmid were co-transfected into KYSE170 cells, and then Transwell assay was used to detect the changes of cell invasion and migration ability, and Western blotting to examine the expression levels of autophagy-related gene. Results: The invasion and migration ability of KYSE170 cells were significantly inhibited after transfection with miR-124 mimic (All P<0.05). The expression of autophagyrelated protein P62 was increased, and the expression of BECN1 and LC3 was significantly decreased (All P<0.01); in addition, the activity of luciferase reporter gene was also significantly reduced (P<0.01). Silencing BECN1 expression inhibited the invasion and migration of esophageal cancer KYSE170 cells (P<0.01). However, after co-transfection with BECN1 over-expression plasmids, the effects of miR-124 mimic on the autophagy, invasion and migration of esophageal carcinoma KYSE170 cells were significantly weakened (P<0.01), it was also accompanied with lower P62 expression, and higher LC3 expression (P<0.01). Conclusion: miR-124 mimic can inhibit the invasion and migration of esophageal carcinoma cells. The mechanism may be related to the autophagy-related gene BECN1 expression.

Objective: To evaluate the expression of melanoma antigen A family（MAGE-As）in the peripheral blood of patients with esophageal carcinoma (EC), and to analyze its correlations to the clinicopathological features and the prognosis of EC patients. Methods: mRNA expression of MAGE-As in peripheral blood from 153 EC patients and 30 healthy donors was detected using multiplex semi-nested PCR. In addition, restriction endonuclease treatment was used to determine the expression of MAGE-As family members, including MAGE-A1, A2, A3, A4 and A6. Results: The positive expression of MAGE-As was observed in 30 of 153 EC patients (19.61%) in peripheral blood. The positive expression rate of MAGE-A1, A2, A3, A4, A6 was 10.46% (16/153), 16.34%(25/153), 9.8% (15/153), 11.11% (17/153) and 18.30%(28/153), respectively. Additionally, the expression of MAGE-As was positively associated with clinical stage, lymphatic metastasis and distant metastasis (all P<0.05). The positive expressions of MAGE-As and its sub-type genes were all associated with low 5-year overall survival of ES patients (all P<0.05). Expression of MAGE-As, tumor volume, lymphatic metastasis and distant metastasis can be used as independent prognostic factors for the survival of EC patients (all P<0.01). Conclusion: The expression of MAGE-As in peripheral blood of EC patients was associated with the prognosis of EC, and may be used as an important indicator for the prognosis of esophageal carcinoma.

[Abstract] Objective: To investigate the regulating effects of miR-92b on the expression of EZH2 (enhancer of zeste homolog 2) gene and the proliferation and invasion abilities of esophageal cancer (EC) cells. Methods: Fifteen cases of esophageal cancer tissues that preserved in the research center of the Fourth HospitalAffiliated to Heibei Medical University from January 2016 to January 2017 were selected for this study. The bioinformatics tool was used to predict the possible miRNAs that might target EZH2. The mimics of predicted miRNAs were transfected into human esophageal carcinoma cell lines Eca109, respectively. Then the regulation effect of miRNAs on EZH2 gene expression was validated by real-time PCR, Western blotting and dual luciferase reporter experiment. In the meanwhile, EZH2 over-expression plasmids were co-transfected into esophageal carcinoma Eca109 cells, and the effects of miRNAs and EZH2 expression changes on the proliferation, apoptosis , invasion and migration of esophageal carcinoma cells were detected by CCK-8 method, Flow Cytometry, Transwell Invasion and migration assay, respectively. Results: Bioinformatics analysis showed that miR-92b, let7a and miR-25 could combine with potential binding sites at 3’-terminal non-translation region of EZH2 gene. Real-time PCR results showed that only miR-92b was able to regulate the expression of EZH2, and miR-92b was negatively correlated to EZH2 in esophageal cancer (P<0.01). Compared with mimic-NC, the expression of EZH2 mRNA, protein and luciferase activity in Eca109 cells after miR-92b mimic transfection was significantly down-regulated (both P<0.01). However, miR-92b mimic transfection had no effect on the apoptosis of Eca109 cells. Moreover, the proliferation, invasion and migration of Eca109 cells were significantly inhibited after transfection with miR-92b-mimic (P<0.01). In addition, after co-transfection with EZH2 over-expression plasmids, the effects of miR-92b-mimic on the proliferation, invasion and migration of Eca109 cells were significantly weakened (P<0.01). Conclusion: miR-92b can inhibit the proliferation,invasionandmigrationofesophagealcarcinomacells,anditsmechanismmayberelatedtoitstargetregulationofEZH2. ··