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Objective: The aim of this study was to explore the roles of FOXN2 (Fork head Box N2) in mediating the proliferation and invasion of hepatocellular carcinoma (HCC) cells., Patients and Methods: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to determine expression of FOXN2 in HCC tissues and cells. Transfection of plasmid containing FOXN2 was used to exogenously overexpress FOXN2 in vitro. Cell Counting Kit-8 (CCK-8) assay and transwell assay were applied to detect the proliferation and invasion of HCC cells, respectively., Results: FOXN2 expression decreased significantly in both HCC tissues and cells (p<0.05). Upregulation of FOXN2 significantly inhibited the proliferation and invasion of HCC cells (p<0.05)., Conclusions: FOXN2 acts as a regulator in the progression of HCC. Our findings suggest that FOXN2 may be a novel therapeutic monitoring and prognosis biomarker in HCC.

Objective: To investigate the role of circ100284 in osteosarcoma (OS) and the underlying mechanism., Patients and Methods: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to detect the level of circ100284 in OS tissues and cells, and to examine the association between its level and clinicopathological features such as tumor size, tumor stage, and survival time. In addition, circ100284 was knocked out in MG63 and U2OS cells to observe the effect of circ100284 on cell viability, migration, cycle, and apoptosis by Cell Counting Kit-8 (CCK-8), transwell assay, and flow cytometry assay. Correlations of circ100284 with lysine-specific histone demethylase 1A (LSD1) and enhancer of zeste homolog 2 (EZH2) target proteins were analyzed by RNA co-precipitation experiments. Furthermore, the chromatin immunoprecipitation assay (ChIP)-qPCR assay was performed to analyze the relationship between circ100284 and its target protein and target gene., Results: Circ100284 had a high level in OS tissues. The high expression of circ100284 was positively correlated with tumor size, pathological stage, and lung metastasis, and negatively correlated with patient survival time. Knocking down circ100284 in OS cells damaged the cell viability and invasiveness, blocked cell cycle, and promoted cell apoptosis. Further experiments showed that circ100284 could epigenetically inhibit cell proliferation by negatively regulating Phosphatase and tensin homolog (PTEN) and epithelial membrane protein 1 (EMP1) in OS., Conclusions: Circ100284 promotes the progression of OS cells by downregulating PTEN and EMP1.

Objective: Mesenchymal stem cells (MSCs) induce allograft immune tolerance, but low efficacy severely limits their wide application. In this work, Netrin-1 was used to maintain MSC function in an IR environment to study its role in the immune tolerance induction of the allograft., Materials and Methods: The experiments were divided into three groups: the control group, the IR group and the Netrin-1 group (Netrin-1 was added to MSC medium and then cultured for 48 h). After digestion, MSCs were mixed with TLR4 and TLR3 antibodies (BD), incubated for 20 min, and washed with Phosphate-Buffered Saline (PBS) three times. The mean fluorescence intensity (MFI) of TLR4 and TLR3 was detected by flow cytometry. Isolated lymphocytes were divided into four groups: the control group (no treatment), the MSC group (lymphocytes were co-cultured with MSCs in the control group), the rejection group (lymphocytes were co-cultured with MSCs in the IR group), and the Netrin-1 group (MSCs in the IR group) was stimulated by Netrin-1 for 48h., Results: Our study found that compared with control mice, toll-like receptor (TLR3) expression in bone marrow MSCs decreased as the expression of TLR4 increased, the secretion of transforming growth factor-β (TGF-β) and interleukin-10 (IL-10) was reduced, while the secretion of IL-6 significantly increased in immune rejection (IR) mice. MSCs in IR mice promoted T-cell proliferation and reduced the ratio of Treg cells. Netrin-1 inhibited the pro-rejection effect of these MSCs, further inhibited T-cell proliferation and facilitated an increase in the ratio of Treg cells. The animal experiment results showed that MSC transplantation in the rejection group would shorten the mean survival time of the skin graft and induce the infiltration of lymphocytes. Netrin-1 prolonged the mean survival time of the skin graft by enhancing MSC function. The immunohistochemistry results showed that, compared with the rejection group, the T cell number in the skin graft significantly decreased in the Netrin-1 group., Conclusions: MSC can be divided into immune-tolerant and pro-rejection types in organ transplantation and Netrin-1 can induce the transformation of MSC from the pro-rejection to immune-tolerant type and markedly prolong the skin graft survival time.

Objective: The aim of this study was to investigate the role of long non-coding RNA (lncRNA) TCL6 in early abortion and to explore its underlying mechanism., Patients and Methods: The expression levels of lncRNA-TCL6 and epidermal growth factor receptor (EGFR) in placental tissues of normal pregnancy, threatened abortion pregnancy, spontaneous abortion pregnancy, and induced abortion pregnancy were detected by quantitative Real-Time Polymerase Chain Reaction (qRT-PCR), respectively. Trophoblast cells were transfected with siRNA to knock-down lncRNA-TCL6. Cell viability of trophoblast cells was detected by cell counting kit-8 (CCK-8) assay. The protein expression levels of EGFR, extracellular regulated protein kinases (ERK) and protein kinase B (AKT) in trophoblast cells after lncRNA-TCL6 knockdown were detected by Western blot. Rescue experiments were performed to investigate the relationship between EGFR and lncRNA-TCL6., Results: The expression of lncRNA-TCL6 in placenta tissues of threatened abortion pregnancy was significantly higher than that of normal pregnancy. Meanwhile, the expression of lncRNA-TCL6 in placenta tissues of spontaneous abortion pregnancy was also markedly higher than induced abortion pregnancy. However, the expression of EGFR showed an opposite trend. After knockdown of lncRNA-TCL6 in trophoblast cells, the protein expression levels of EGFR, ERK, and AKT were significantly increased when compared with those of the control group. CCK-8 assay indicated that cell viability was remarkably increased after knockdown of lncRNA-TCL6, which could be reversed by EGFR knockdown., Conclusions: Compared with normal pregnancy, lncRNA-TCL6 was highly expressed in placental tissues of threatened abortion pregnancy. Moreover, the expression of lncRNA-TCL6 in placenta tissues of spontaneous abortion pregnancy was significantly higher than induced abortion pregnancy. Knockdown of lncRNA-TCL6 promoted the proliferation of trophoblast cells and inhibited the abortion via the EGFR signaling pathway.

Objective: The use of adipose-derived stem cells (ADSCs) to cure the optic nerve injury was never shown previously. Here, we implanted purified ADSCs into optic nerve injury of rats., Materials and Methods: Male Sprague Dawley (SD) rats were used in this study. The vision degeneration was detected by Flash-visual evoked potential (F-VEP) assay. The expression of Macrophage-1 (Mac-1), myeloid differentiation factor 88 (MyD88), and nuclear transcription factor-κB (NF-κB) were studied by Western blot. The expression of interleukin (IL)-6 and tumor necrosis factor (TNF)-α in the optical nerve lysates were assessed by enzyme-linked immunosorbent assay (ELISA)., Results: We found out that ADSC implantation inhibits the amplitude decrease and latency increase of the P1 wave caused by the optic nerve injury. The expression of the inflammation associated proteins of the toll-like receptor 4 (TLR4) signaling pathway, including Mac-1, MyD88, NF-κB, IL-6, and TNF-α, were inhibited in the ADSC therapy group compared to the control group., Conclusions: Our results indicated that ADSC implantation can inhibit the inflammation after the optic nerve injury and improve the functional vision impairment. These findings suggested ADSC implantation as a translational therapy method for optic nerve injury in clinics.

Objective: Oxidative stress caused by reactive oxygen species (ROS) plays an important role in the pathogenesis of endometriosis. The gene AT-rich interactive domain 1A (ARID1A), is frequently down regulated and inactivated in endometriosis. This report is focused on the molecular mechanism of the correlation between oxidative stress and ARID1A gene expression in endometrial cell oxidative damage model., Patients and Methods: In this study, the ARID1A gene expression level and its promoter methylation level were detected in 30 endometriosis and normal tissues. The primary endometrial cell was co-cultured with H2O2. Then, MDA and Gpx level were used to test the ROS level, RT-PCR was employed to detect the expression level of ARID1A. At last, the ARID1A gene promoter methylation level was detected by methylation-specific PCR (MSP). Finally, the expression level of DNMT1 was detected by both RT-PCR and Western blot., Results: The expression level of ARID1A gene was down regulated in endometriosis compared with normal tissues. The low expression level of ARID1A gene was associated with its promoter hyper-methylation. In H2O2 simulated endometrial cells, ARID1A gene expression level was decreased. Finally, ROS regulated ARID1A gene expression by changing the methylation level of ARID1A gene promoter. Finally, both the mRNA level and protein level of DNMT1 increased in H2O2 simulated endometrial cells., Conclusions: In endometriosis, the down-regulated ofARID1A gene was highly correlated with its promoter hyper-methylation. ROS decreased the expression level of ARID1A gene via regulating methylation of its promoter which contributing to the understanding of the pathogenesis of endometriosis. The possible mechanism of ARID1A gene promoter hyper-methylation is ROS up-regulated DNMT1gene expression.