Paper 1:A Case Of Orf Paper 2:Effect Of MicroRNA-181b-5p Inhibitors On The Proliferation,Cell Cycle,Apoptosis,Migration And Invasion In Cutaneous Squamous Cell Carcinoma Cell Line SCC12

Objective: Discuss the clinical features and prevention of orf.Method: Analysis clinical characteristics,treatment and literature reviews of one orf patient.Results: The woman patient was 40 years old.The patient presented with nodules on her right thumb?the back of right hand and forearm with itching for two weeks,maculopapule on trunk one day.Lesions were dark red nodules,which tops were evenness and had hilar depression in the center surrounded with a white ring.Histopathology showed:The epidermal showed pseudoepitheliomayous hyperplasia.The papillary dermis is markedly edematous.Massive capillary proliferation and dilation presented in the dermis.The dermal showed diffuse infiltration of inflammatory cells,which consisted of lymphocytes,histiocytes,eosinophils.Diagnosis: Orf.Conlustions:(1)Occasional contact sick sheep in the non-pastoral area may also has the risk of orf infection,we should pay attention.(2)Direct transmission from sick sheep and its pollution are the main reasons of human orf infection,and therefore individual protection is very important when contact with sick sheep.Therefore,sick sheep must be isolated and its contaminants be given strict disinfection.Objective:To investigate the effect of micro RNA-181b-5p inhibitors on the biological behavior of cutaneous squamous cell carcinoma cell line SCC12 by observing the role of micro RNA-181b-5p inhibitors in cell proliferation,cell cycle,apoptosis,migration and invasion.Methods:1.QRT-PCR was performed on SCC12 and Ha Ca T to determine mi R-181b-5p expression;2.SCC12 was transfected with GFP,to determine transfection condition and efficiency by inverted fluorescence microscope and FCM(flow cytometry).3.SCC12 cells were randomly divided into three groups : Experimental group(transfected with mi R-181b-5p inhibitors),control group(transfected with micro RNA inhibitor N.C)and blank group.4.We used q RT-PCR to analyze the expression of mi R-181b-5p in different cell groups and evaluate the transfection effect about 48 hours.5.CCK-8 method and cell cycle was used to observe the changes of proliferation activities of SCC12 cells after transfect of mi R-181b-5p.6.Flow cytometry was used to analyze the changes of apoptosis of SCC12 cells after transfect of mi R-181b-5p.7.Transwell method was used to observe the changes of migration and invasion activities of SCC12 cells after transfect of mi R-181b-5p.Results:1.QRT-PCR showed that mi R-181b-5p expression in SCC12 sign ificantly increased to about 1.82 fold of that in Ha Ca T(P<0.05).2.Cells after transfection in good condition,and transfection efficiency can reach(95.6±0.4)%.3.The results showed that,compared with the control group and blank group,experimental group effectively down regulated the expression level of mi R-181b-5p in SCC12 cells(P<0.05).4.After the inhibition of mi R-181b-5p,the ability of cell proliferation of SCC12 cells can be inhibited(P<0.05).5.Analysis of cell cycle demonstrated a marked blockage of the G1 stage of the SCC12 cells treated with mi R-181b-5p(P<0.05).6.Apoptosis rate of the SCC12 cells has significantly increased after transfection with mi R-181b-5p inhibitors compared to that of correspongding negative control and blank(P<0.05).7.After the inhibition of mi R-181b-5p,the migration abilities of cells no obvious change(P>0.05).8.After the inhibition of mi R-181b-5p,the invasion abilities of cells no obvious change(P>0.05).Conclusions:1.Mi R-181b-5p expression in SCC12 cells significantly increased than that in Ha Ca T cell.2.Transfection with mi R-181b-5p inhibitors significantly affected SCC12 cell line proliferation,apoptosis and cell cycle.Mi R-181b-5p may become a new target in the diagnosis and treatment of cutaneous Squamous Cell Carcinoma.