Objective To investigate mechanisms underlying the prevention of experimental autoimmune encephalomyelitis (EAE) in mice by intraperitoneal infusion of myelin oligodendrocyte glycoprotein 35-55 (MOG35-55) (MOG i.p.).Methods C57BL/6 mice were immunized with MOG35-55 to establish the model of EAE and then were intraperitoneally injected daily with MOG35-55 or ovalbumin (OVA, served as control) from day 6 to day 16.EAE was evaluated daily using a general clinical scoring system and histological analysis.Numbers of lymphocytes in peripheral blood and central nervous system (CNS) were detected at different time points.Effects of MOG i.p.on the migration of MOG-T cells in vivo were analyzed by an adoptive transfer experiment.Maturation of splenic antigen-presenting cells (APCs) and migration of MOG-T cells in vitro were examined by fluorescence activated cell sorting (FACS) and a Transwell system, respectively.Results MOG i.p.protected the mice from development of EAE by blocking the lymphocyte recruitment to CNS.More effector T cells were trapped in the periphery of EAE and naive mice in adoptive transfer experiment after MOG i.p.treatment.MOG i.p.induced the maturation of splenic APCs and enhanced the expression of CD80, CD86 and major histocompatibility complex class Ⅱ (MHCⅡ) molecules.Mature APCs blocked the recruitment of effector T cells to CNS.Conclusion MOG i.p.protects mice from EAE by inducing the maturation of splenic APCs.

Objective This study aims to define the effects of hypothyroidism on c-fos and c-jun mRNA expression in rat testes to provide a theoretical basis for prevention and control of iodine deficiency disorders (IDD). Methods According to body weight (200 - 240 g), 20 Wistar male rats were divided into control group and hypothyroidism group (1 ml/100 g, 0.1% propylthiouracil by intragastric administration) by digital table. There were 10 male rats in each group and body weight was observed every 3 days. After 60 days, all rats were killed. The levels of thyroid hormones [total triiodothyronine (TT3), total thyroxine (TT4), and thyroid stimulating hormone (TSH)] were measured by radioimmunoassay. The mRNA expression levels of c-fos and c-jun in testes were detected by real-time quantitative PCR. Results Compared with control groups [(298.20 ± 12.15), (344.00 ± 13.73) g], the weights of hypothyroidism groups in 30 days [(239.00 ± 15.02) g] and in 60 days [(232.67 ± 17.86) g] were significant decreased (t=7.704, 11.380, all P<0.05). The levels of TT3 [(373.32 ± 101.31) ng/L] and TT4 [(4.00 ± 0.89) × 103 ng/L] in serum of hypothyroidism group were found to be significantly decreased, whereas the level of TSH [(5.77 ± 0.89) × 103 U/L] was increased in comparison with those of the control groups [(1 000.01 ± 273.53) ng/L, (44.33 ±7.84) × 103 ng/L, (1.87 ± 0.70) × 103 U/L, t = 5.262, 12.520, 8.413, all P< 0.05]. Compared with control group (1.00 ± 0.08, 1.01 ± 0.04), the c-fos and c-jun mRNA expression (0.67 ± 0.03, 0.75 ± 0.02) of hypothyroidism group was significant decreased (t = 12.382, 13.784, all P < 0.05). Conclusion Hypothyroidism may reduce mRNA expression levels of testicular c-fos and c-jun, and then damage the reproductive system in male rats.

OBJECTIVETo investigate the effect of microRNA-140 (miR-140) on the migration and invasion of colorectal cancer (CRC) cells and the possible mechanism.

METHODSmiR-140 mimics, miR-140 specific inhibitor or small interfering RNA (siRNA) against Smad3 were transfected into human CRC cell line RKO cells respectively, using Oligofectamine or Lipofectamine2000. Quantitative real-time PCR (real-time PCR) was used to measure the expression levels of miR-140 and Smad3 mRNA. Smad3 protein was analyzed by Western blot. The in vitro cell migrating and invasive abilities were determined by wound-healing and Transwell chamber assay after up-regulating or down-regulating miR-140 or knocking down Smad3.

RESULTSThe Western blot assays showed that the Smad3 protein level was significantly reduced after up-regulating miR-140 (0.04 ± 0.01), compared with that of (0.47 ± 0.02, P < 0.05) in the control group and that of (0.52 ± 0.06) in the negative control group (P < 0.05 for both). The results of real-time PCR indicated that no significant difference was found in the levels of Smad3 mRNA between miR-140 transfection and NC groups (1.11 ± 0.13 vs. 1.00 ± 0.06, P > 0.05). The wound-healing assay showed that the migrating ability was dramatically attenuated by miR-140 compared with that in the control and NC groups, whereas no significance was found when compared with that of the Smad3 siRNA transfected cells. The number of cells migrating through Transwell chamber without matrigel in the miR-140 group was (76.2 ± 4.4), remarkably lowered than that in the control (267.1 ± 4.9) and NC (336.1 ± 5.7) groups (P < 0.05 for both), but no significant difference between the miR-140 (76.2 ± 4.4) and Smad3 siRNA (83.5 ± 7.3) groups. Transwell chamber with matrigel assay showed that number of cells penetrating through the membrane was (109.5 ± 7.4) in the miR-140 group, significantly lower than that in the control (403.1 ± 5.1) and NC (392.6 ± 8.4) groups (P < 0.05 for both), while Smad3 siRNA transfection had a similar effect (138.8 ± 3.6)(P > 0.05). Down-regulation of miR-140 increased the level of smad3 protein expression, and partially reversed the inhibition of the cell migration and invasion mediated by miR-140. Co-transfection of miR-140 inhibitor and Smad3 siRNA had no significant effect on the Smad3 protein expression and the abilities of cell migration and invasion.

CONCLUSIONSmiR-140 regulates the Smad3 expression at the post-transcriptional level. miR-140 suppresses the migrating and invasive abilities of CRC cells, possibly through down-regulation of Smad3. The findings of this study suggest that miR-140 may have a unique potential as a possible biomarker candidate for diagnosis and therapy of tumor metastasis.

Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Colorectal Neoplasms ; metabolism ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; genetics ; Humans ; MicroRNAs ; Neoplasm Invasiveness ; RNA, Messenger ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Smad3 Protein ; genetics ; metabolism ; Transfection ; Up-Regulation