OBJECTIVES: To investigate the mechanism by which circ_EPHB4 regulates temozolomide (TMZ) sensitivity of glioma cells through the miR-424-5p/Wnt3 signal axis. METHODS: We detected the expression levels of circ_EPHB4, miR-424-5p and Wnt3 mRNA in glioma specimens from 25 patients with primary glioma and 25 patients experiencing relapse following temozolomide-based chemotherapy and in TMZ-sensitive and -resistant glioma A172 and SHG44 cells with circ_EPHB4 knockdown using qRT-PCR, and Wnt3 protein expression level was detected with Western blotting. Cell viability, colony-forming ability, and apoptosis of the cells with circ_EPHB4 knockdown were assessed, and the targeted regulation relationship between circ_EPHB4, miR-424-5p, and Wnt3 was verified by dual luciferase reporter assay and RNA immunoprecipitation (RIP) experiments. The effect of circ_EPHB4 knockdown on tumorigenesis of glioma cells was evaluated in subcutaneous tumor-bearing nude mouse models. RESULTS: The expression of circ_EPHB4 was significantly increased in glioma tissues and cells as compared with normal neural tissues and astrocytes (P=0.014). In TMZ-resistant glioma cells, circ_EPHB4 knockdown resulted in an obvious reduction of IC50 value of TMZ, inhibited cell colony formation, and promoted cell apoptosis, and these effects were reversed by miR-424-5p knockdown. The expressions of miR-424-5p and circ_EPHB4 were negatively correlated in glioma tissues (P=0.011). MiR-424-5p knockdown also attenuated the effect of circ_EPHB4 knockdown on expressions of PCNA, P-gp, MRP1 and bax. CONCLUSIONS Circ_EPHB4 regulates Wnt3 expression through "sponge adsorption" of miR-424-5p, thereby modulating TMZ-resistant glioblastoma cell clonogenesis, apoptosis, and TMZ sensitivity, suggesting the potential of circ_EPHB4 as a therapeutic target for reversing drug resistance of gliomas.
MicroRNAs/genetics* ; Humans ; Temozolomide ; Glioma/genetics* ; Animals ; Mice, Nude ; Cell Line, Tumor ; Wnt3 Protein/metabolism* ; Mice ; Apoptosis ; RNA, Circular ; Drug Resistance, Neoplasm ; Brain Neoplasms/pathology* ; Signal Transduction

OBJECTIVES: To investigate the oncogenic role of circular RNA circ_EPHB4 in glioma and its molecular mechanism. METHODS: Microarray analysis was performed to identify the differentially expressed circRNAs in glioma tissues. The effects of circ_EPHB4 on glioma cell migration, invasion and epithelial-mesenchymal transition (EMT) in vitro and tumorigenicity in vivo were assessed using scratch wound healing assay, Transwell invasion assay and nude mouse models bearing subcutaneous tumors. RNA immunoprecipitation (RIP), RNA stability assays, and gene overexpression and silencing techniques were employed to validate the synergistic regulatory effect of circ_EPHB4 and the N6-methyladenosine (m⁶A) reader protein YTHDF3 on Wnt3 expression. RESULTS: Circ_EPHB4 was significantly overexpressed by 2.3 folds (|log2FC|=1.2, P<0.01) in glioma tissues compared to the adjacent tissues, and by 2.5 folds in glioma cell line U373 compared to normal cells (P<0.001). Overexpression of circ_EPHB4 significantly enhanced migration and invasion of glioma cells, and promoted the expressions of EMT markers N-cadherin and vimentin. In the tumor-bearing mouse models, the tumor volume in circ_EPHB4 overexpression group was significantly greater than that in the control group, and the lung metastatic foci increased by 4.2 folds. Overexpression of circ_EPHB4 promoted oncogenesis by upregulating Wnt3 expression, while YTHDF3 extended the half-life of Wnt3 mRNA in an m⁶A-dependent manner. Simultaneous knockdown of circ_EPHB4 and YTHDF3 resulted in an obvious reduction of Wnt3 mRNA expression by up to 47% compared to its level following knocking down either circ_EPHB4 or YTHDF3 alone. CONCLUSIONS Circ_EPHB4 and YTHDF3 promote glioma progression by jointly targeting the Wnt3 signaling pathway, which may provide a new therapeutic strategy for gliomas.
Glioma/genetics* ; Humans ; Animals ; Cell Line, Tumor ; RNA-Binding Proteins/genetics* ; RNA, Circular ; Epithelial-Mesenchymal Transition ; Mice, Nude ; Cell Movement ; Wnt3 Protein/genetics* ; Mice ; Disease Progression ; Adenosine/metabolism* ; Brain Neoplasms/metabolism* ; Gene Expression Regulation, Neoplastic

Objective To investigate the molecular mechanism of ALKBH5 reducing sepsis-induced myocardial dysfunction(SIMD).Methods The expression levels of ALKBH5 and TRAF1 in the blood of 50 SIMD patients and 50 healthy individuals were detected using reverse transcription fluorescence quantitative polymerase chain reaction(RT-qPCR),and the correlation between their expression levels was analyzed by person analysis;In vitro experiments,H9C2 myocardial cells were divided into 7 groups according to over expression of TARF1 and knockdown ALKBH5.The molecular mechanism of ALKBH5 targeting TRAF1 to regulate lipopolysaccharide(LPS)induced myocardial cell damage was studied through experiments such as CCK8,ELISA,and Western blot;In the in vivo experiment of rats,LPS induced rats were divided into 6 groups according to over expression of TARF1 and knockdown ALKBH5.Experimental methods such as colorimetry,ELISA,Western blot,HE staining,and immuno-histochemistry were used to study the mechanism of ALKBH5 targeting TRAF1 through NF-κB pathway in reduc-ing myocardial cell damage.Results The expression levels of ALKBH5 and TRAF 1 were downregulated in SIMD,and the Pearson analysis showed a positive correlation between them(P<0.001);In vitro experiments showed that overexpression of TRAF1 promotes cell proliferation,inhibits the expression of inflammatory factors and proteins involved in the NF-κB pathway,and knockdown ALKBH5 obtain the opposite resulst;In vivo experi-ments in rats showed that knockdown ALKBH5 promotes injury in cardiomyocytes,expression of inflammatory factors and NF-κB-related pathway proteins,and nuclear translocation of NF-κB p65 protein,but the overexpression of TRAF 1 yielded the opposite results.Conclusion ALKBH5 increases the stability of TRAF1 by reducing its meth-ylation,thereby inhibiting NF-κB pathway,thereby reducing SIMD.

Objective To investigate the material basis and antitumor mechanism of Marsdenia tenacissima (MT) on hepatocellular carcinoma (HCC) by bioinformatics, network pharmacology and molecular docking technology. Methods Active ingredients of MT were collected by literature search and screened by Swiss ADME website, which targets were predicted by Swiss Target Prediction. The chip data of HCC (GSE147888) were downloaded from the NCBI Gene Expression Omnibus (GEO) database. Differentially expressed genes were screened by R software. HCC-related targets were collected from the Genecards and OMIM databases. The Venny online tool was used to obtain the intersection of the herbal medicine targets and the disease targets. Subsequently, drug-target network and protein–protein interaction (PPI) network were constructed by Cytoscape software and String platform. GO enrichment analysis and KEGG pathway analysis were performed to analysis the functions and pathways enriched by key genes. The expression of key genes in HCC and its effect on survival were analyzed by the GEPIA database. The Human Protein Atlas (HPA) was used to analyze the immunohistochemical expression of key genes in HCC. Finally, molecular docking was carried out to investigate interactions between the top five targets and their related active compounds. Results A total of 50 active components were screened and 12 common targets were identified related to MT and HCC. Scutellarein-4-Methylether, Tenasogenin, Sinapic Acid, Dresgenin and Kaempferol were considered as the critical components. JUN, MMP9 and PTGS2 were recognized as key therapeutic targets. The GO analyses demonstrated that key targets mainly involved in the process of gene silencing and inflammatory response. KEGG analysis suggested that key targets were enriched in TNF signaling pathway and IL-17 signaling pathway. Survival analysis by the GEPIA showed significant differences in the expression of ESR1, MMP1, MMP9, JUN, and PPARG between high and low risk groups. Immunohistochemical results showed that ESR1 and MMP9 were differentially expressed in normal and hepatocellular carcinoma tissues. The molecular docking results verified that the drug active ingredient could be stably bound to the target protein. Conclusion This study reflected the multi-component, multi-target and multi-pathway characteristics of the MT in the treatment of HCC, which could provide a scientific basis for the clinical application of MT in HCC.

Objective To understand the ability and level of emergency rescue at general hospitals in Tianjin city. Methods Such actions as formulating plans and examination forms, establishing assessment indicators and evaluation criteria, and simulation exercises were performed to evaluate the capacity of 28 general hospitals in terms of their organizational structure, emergency response, event reporting, and summary assessment. Results The emergency response assessment system consisted of 4 level-1 indicators, 19 level-2 indicators and 58 level-3 indicators. 28 hospitals were found high in their overall emergency response capacity, but some were found with setbacks. For example, the " organizational structure" scored the highest in 4 first-level indicators, up to 88. 91％ , while " incident report" scored the lowest, down to 67. 99％ . Among level-2 indicators, professional emergency professional procedures and initial reporting scored the lowest. Conclusions In order to further improve the ability of medical institutions to respond to emergency events, the hospitals are recommended to enhance their backup resources for emergency response, their staff′s awareness of first aid knowledge and first aid skills, as well as their timeliness of initial reports and the completeness of progress reports.

CD146 Antigen ; metabolism ; Humans ; Pericytes ; cytology ; metabolism ; Receptor, Platelet-Derived Growth Factor beta ; metabolism

Objective To learn the current capacity building of emergency response teams in Tianjin for the purpose of goals setting.Methods Health Emergency Capacity Questionnaire was issued to 89 secondary and above medical institutions and 19 CDCs in Tianjin.The form covered such items as basic institutional information, workforce makeup, emergency preparations, detection and early warning, emergency response, and summary/assessment.Data collected in the questionnaire were subject to descriptive and correlation analysis.Results Tianjin has scored an initial success in emergency medicine as evidenced in its emergency response mechanisms in place, elevated capacity in emergency medical rescue and disposal, and enhanced competence of emergency teams.Rooms of improvement however include insufficient professionalism and independence of health emergency, inadequate emergency commanding and decision making system functions, insufficient laboratory test capacity at district/county levels, and insufficient social involvement in health emergency.Conclusions Top-down design should be emphasized, health emergency response should be enhanced in terms of management and response planning system, while capacity building of the teams and long-term primary care emergency mechanism deserve higher attention.

Objective To observe effect of cognitive level and self-care ability of protective motivation intervention based on the quantitative evaluation in elderly patients with osteoporosis. Methods A total of 116 cases patients with senile osteoporosis from May 2013 to April 2015 were divided into the observation group and the control group. The control group implemented routine care, the observation group was given protective motivation intervention based on quantification protection motivation assessment strategies. Then cognitive level, self-care ability, quality of life were compared between two groups. Results Disease related knowledge cognition level:osteoporosis risk factors, sports knowledge, calcium score in the observation group were significantly higher than those in the control group (t=8.428, 10.104, 6.775, P < 0.05). Self-care ability: self-care ability, self-care responsibility, self-care knowledge scores in the observation group were significantly higher than those in the control group(t=2.230-5.162, P<0.05). Life quality:physical function, role-physical, bodily pain, general health, vitality, social function, role-emotional, mental health ect scores in the observation group were significantly higher than those in the control group (t=6.804-10.488, P < 0.05). Conclusions Protective motivation intervention based on the quantitative evaluation is helpful to enhance the cognitive level of disease knowledge and improve the self-care ability, as well as improve the quality of life.

Objective To observe the clinical efficacy of Zhishanghan Zhufeng Formula in treating knee osteoarthritis.Methods Totally 90 patients were randomly divided into treatment group and control group, 45 cases in each group. The treatment group was treated with Zhishanghan Zhufeng Formula for 30 days. The control group was treated with routine medicine (diclofenac sodium sustained release tablets and glucosamine sulfate tablets) for 30 days. The clinical efficacy and the changes of VAS, WOMAC score and knee temperature before and after treatment of two groups were observed.Results TCM clinical symptoms were significantly improved, and the effects in the treatment group were more evident compared with the control group (P<0.05). After treatment, VAS and WOMAC score of the two groups decreased obviously, while the knee temperature increased, and the treatment group was superior to the control group, with significant significance (P<0.05).Conclusion Zhishanghan Zhufeng Formula has effective clinical efficacy for knee osteoarthritis.

FXYD6, FXYD domain containing ion transport regulator 6, has been reported to affect the activity of Na(+)/K(+)-ATPase and be associated with mental diseases. Here, we demonstrate that FXYD6 is up-regulated in hepatocellular carcinoma (HCC) and enhances the migration and proliferation of HCC cells. Up-regulation of FXYD6 not only positively correlates with the increase of Na(+)/K(+)-ATPase but also coordinates with the activation of its downstream Src-ERK signaling pathway. More importantly, blocking FXYD6 by its functional antibody significantly inhibits the growth potential of the xenografted HCC tumors in mice, indicating that FXYD6 represents a potential therapeutic target toward HCC. Altogether, our results establish a critical role of FXYD6 in HCC progression and suggest that the therapy targeting FXYD6 can benefit the clinical treatment toward HCC patients.
Animals ; Antibodies, Monoclonal ; pharmacology ; Antineoplastic Agents ; pharmacology ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Female ; HEK293 Cells ; Humans ; Ion Channels ; antagonists & inhibitors ; metabolism ; Liver Neoplasms ; drug therapy ; metabolism ; Mice, Inbred BALB C ; Mice, Nude ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Tumor Burden ; drug effects ; Xenograft Model Antitumor Assays