HA117’s Contribution To The Etiology Of HD

SECTION ONE THE EXPRESSION AND POTENTIAL ROLES OF DPF3 AND ITS RELATED GENES,SMARCD3,MEF2 IN COLON TISSUES FROM PATIENTS WITH HDObjective To detect the expression of DPF3,Smarcd3 and MEF2 in the proximal anastomosis(control) and stenotic segments of colon tissues from patients with HD and analyze the potential roles of DPF3,Smarcd3 and MEF2 underlying the development of HD.Methods All the resected colon tissues were obtained randomly from the patients with HD in the Children’s Hospital of Chonqing Medical University between June 2012 and April 2014. Quantitative RT-PCR and Western blotting were used for the detection of the DPF3,Smarcd3 and MEF2 m RNAs and proteins expression files in different portions of colon tissues respectively. Immune histochemical method and optical microscope digital radiography were used to analyze the protein stainings. Graph Pad Prism 5(Graph Pad Software, Inc, USA) and SPSS statistical software(version 17.0) were used for spreadsheets and statistical analysis respectively.Results The expression of DPF3 m RNA and protein in proximal anastomosis and stenotic segments was 1.76±1.30 、 1.07±0.93 and 2.31±0.62、1.95±0.57 respectively. The expression of Smarcd3 m RNA and protein in proximal anastomosis and stenotic segments was 0.68±0. 62、0.38±0.42 and 0.85±0.43、0.61±0.34 respectively. The expression of MEF2 m RNA and potein in proximal anastomosis and stenotic segments was:1.01±0. 63 、 0.68±0.39 and 2.29±0.38 、 1.83±0.76 respectively.Besides,both m RNA and protein of DPF3,Smarcd3 and MEF2 in stenotic segments were remarkably less than in proximal anastomosis,sharing the same trend.Positive staining of DPF3,Smarcd3 and MEF2 was observed in proximal anatomosis,whereas no significant staining existed in stenotic segment.Conclusions DPF3,Smarcd3 and MEF2 may be involved in the process of HD.SECTION TWO HA117 REGULATED THE EXPRESSION OF NEURO-RELATED GENES IN 293 T CELL LINEObjective To study the underlying mechanism of HA117 regulating the expression of neuro-related genes,DPF3,Smarcd3 and MEF2 in 293 T cell line.Methods Over-expression lenti-virus of HA117 and lenti-virus vector were used to transfect 293 T cell line. Quantitative RT-PCR and western blotting were used to detect the expression files of HA117 and neuro-related genes in 293 T,293T/Vector and 293T/HA117. Optical microscope digital radiography were used to analyze the fluorescence of lenti-virus. Graph Pad Prism 5(Graph Pad Software, Inc, USA) and SPSS statistical software(version 17.0) were used for spreadsheets and statistical analysis respectively.Results The expression of HA117 RNA in 293 T,293T/Vector and 293T/HA117 were 0.020±0.003、0.027±0.031、1.116±0.372;the expression of DPF3 m RNA in 293 T 、 293T/Vector and 293T/HA117 were 1.653±0.187、1.527±0.185、0.984±0.276;the expression of Smarcd3 m RNA in 293T、293T/Vector and 293T/HA117 were 1.653±0.156、1.013±0.161、0.532±0.134;the expression of MEF2 m RNA in 293 T,293T/Vector and 293T/HA117 were 0.821±0.258、1.677±0.779、0.875±0.384. Expression files of DPF3,Smarcd3 and MEF2 m RNA in 293T/HA117 were remarkably less than in 293 T,sharing the opposite trend with HA117.Conclusions The expression of HA117 in 293T/HA117 significantly increased than that in 293T(P<0.05),suggesting that over-expression lenti-virus of HA117 transfected 293 T efficiently.However,the expression of neuro-related genes in 293T/HA117 decreased apparently than that in 293T(P<0.05),indicating that HA117 could down-regulate the expression of neuro-related genes,DPF3,Smarcd3 and MEF2 in 293 T cell line,which may be the underlying mechanism of HA117 being involved in the etiology of HD.