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 miR-155 in rats exposed to PM2.5 and its signifi-cance.Methods Thirty-two 12-weeks-old healthy male SD rats were randomly divided into PM2.5-primary expo-sure group (P1),PM2.5-three times exposure group (P3), saline-control group and blank-control group (O), with 8 rats in each group. The pathological changes of lung tissue of each group were observed by HE staining and the expressions of miR-155 in lung tissue of each group were measured by RT-PCR. The expressions of TNF-α, IL-6 and IL-1β in serum of each group were measured by ELISA.One-way ANOVA and LSD-t test were used for statistical analysis,and Pearson′s correlation analysis was used to evaluate the relationship between the levels of miR-155 and TNF-α,IL-6 and IL-1β. Results The expression of inflammatory cytokines TNF-α,IL-6 and IL-1β in P1 and P3 increased when compared with that in saline control group and blank-control group, and the amount of expression increased with the increase of the number of exposure and dose;the difference between groups was statistically sig-nificant(P<0.01).At the same time,with the increase of the dose and number of PM2.5,the inflammatory damage of lung tissue was enhanced.The expression of miR-155 in lung tissue also increased with the increase of the dose and number of PM 2.5,and it had significantly positive correlation with serum inflammatory cytokines(P<0.01),and had statistical significance. Conclusions The lung tissue of rats exposed to PM2.5 can produce inflammatory injury, and the damage is enhanced with the increase of exposure times and dose.The expression of miR-155 is positively correlated with inflammatory injury in rats.Consequently,miR-155 may participate in PM2.5-induced inflammatory lung injury in rats.

Objective:To explore the protective role and possible mechanism of neural stem cells combined with edaravone in cortical neurons after oxygen-glucose deprivation/reperfusion (OGD-R).Methods:(1) Neural stem cells from brain tissues of SD fetal rats aged 14-16 d were cultured in vitro, and identified with Nestin, glial fibrillary acidic protein (GFAP), and microtubule-associated protein 2 (MAP2) immunofluorescent staining. Expressions of neuron-specific nuclear protein (NeuN) and β-Tubulin were detected by immunofluorescent staining in primary cortical neurons from SD rats born within 24 h. (2) Primary cortical neurons were divided into normal group (normal culture), OGD-R model group (re-oxygenated culture for 24 h after hypoxia for 1.5 h), OGD-R+neural stem cells group (re-oxygenated co-culture with cortical neurons and neural stem cells for 24 h after hypoxia for 1.5 h), OGD-R+edaravone group (re-oxygenated culture for 24 h after hypoxia for 1.5 h; 100 μmol/L edaravone before hypoxia), OGD-R+neural stem cells+edaravone group (re-oxygenated co-culture with cortical neurons and neural stem cells for 24 h after hypoxia for 1.5 h; 100 μmol/L edaravone before hypoxia); 24 h after each treatment, neuron proliferation in each group was detected by cell counting Kit 8 (CCK8), apoptosis rate was detected by flow cytometry, contents of interleukin-1β (IL-1β), and tumor necrosis factor α (TNF-α) in neuronal supernatant were detected by enzyme-linked immunosorbent assay (ELISA), and real-time quantitative fluorescent polymerase chain reaction (qRT-PCR) and Western blotting were used to detect the mRNA and protein expressions of Notch1, Hes1 and Hes5, respectively. Results:(1) Immunofluorescent staining results showed that neural stem cells were positive for Nestin, GFAP and MAP2, and cortical neurons were positive for NeuN and β-Tubulin; all of them were successfully identified. (2) Compared with normal group, OGD-R model group, OGD-R+neural stem cell group and OGD-R+edaravone group had decreased neuron viability, increased apoptosis, increased supernatant IL-1β and TNF-α contents, and increased Notch1 mRNA and protein expressions, with significant differences ( P<0.05). Compared with OGD-R model group, OGD-R+neural stem cells+edaravone group had increased neuron viability, decreased apoptosis, decreased supernatant IL-1β and TNF-α contents, and decreased mRNA and protein expressions of Hes1 and Hes5, with significant differences ( P<0.05). Compared with the OGD-R model group, OGD-R+edaravone group and OGD-R+neural stem cell+edaravone group had significantly decreased Notch1 mRNA and protein expressions ( P<0.05). Conclusion:Combination of edaravone and neural stem cell therapy can reverse the neuronal damage caused by OGD-R, whose mechanism may be by inhibiting the expressions of inflammatory factors and key signaling molecules in Notch signaling pathway, such as Notch1, Hes1, and Hes5.

Objective:To investigate the association between congenital hypothyroidism (CH) and the adverse outcomes during hospitalization in very low birth weight infants (VLBWI).Methods:This prospective, multicenter observational cohort study was conducted based on the data from the Sino-northern Neonatal Network (SNN). Data of 5 818 VLBWI with birth weight <1 500 g and gestational age between 24-<37 weeks that were admitted to the 37 neonatal intensive care units from January 1 st, 2019 to December 31 st, 2022 were collected and analyzed. Thyroid function was first screened at 7 to 10 days after birth, followed by weekly tests within the first 4 weeks, and retested at 36 weeks of corrected gestational age or before discharge. The VLBWI were assigned to the CH group or non-CH group. Chi-square test, Fisher exact probability method, Wilcoxon rank sum test, univariate and multivariate Logistic regression were used to analyze the relationship between CH and poor prognosis during hospitalization in VLBWI. Results:A total of 5 818 eligible VLBWI were enrolled, with 2 982 (51.3%) males and the gestational age of 30 (29, 31) weeks. The incidence of CH was 5.5% (319 VLBWI). Among the CH group, only 121 VLBWI (37.9%) were diagnosed at the first screening. Univariate Logistic regression analysis showed that CH was associated with increased incidence of extrauterine growth retardation (EUGR) ( OR=1.31(1.04-1.64), P<0.05) and retinopathy of prematurity (ROP) of stage Ⅲ and above ( OR=1.74(1.11-2.75), P<0.05). However, multivariate Logistic regression analysis showed no significant correlation between CH and EUGR, moderate to severe bronchopulmonary dysplasia, grade Ⅲ to Ⅳ intraventricular hemorrhage, neonatal necrotizing enterocolitis in stage Ⅱ or above, and ROP in stage Ⅲ or above ( OR=1.04 (0.81-1.33), 0.79 (0.54-1.15), 1.15 (0.58-2.26), 1.43 (0.81-2.53), 1.12 (0.70-1.80), all P>0.05). Conclusion:There is no significant correlation between CH and in-hospital adverse outcomes, possibly due to timely diagnosis and active replacement therapy.

[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. ··

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.