Study On The Expression Of CircRNA In The Plasma Of Patients With Hand-arm Vibration Disease And Its Diagnostic Value

Objective:In this study,the expression profiles of plasma circ RNAs in HAVD,vibration-exposed controls were identified using Arraystar Human circ RNA microarray to find circ RNAs that were associated with vascular injury and significantly different from HAVD.We also analysed the correlation between the circ RNA and HAVD,and explored diagnostic value of plasma circ RNA in HAVD.Methods:1.Research object:According to the inclusion and exclusion criteria,workers who underwent occupational physical examination from June 2021 to January 2023（microarray testing phase:4 cases each of HAVD group,VEC group and NVEC group;validation phase:23 cases of HAVD group,20 cases of VEC group and 20 cases of NVEC group）were selected as study subjects in a sports equipment factory company in Zhongshan City.2.Circ RNA microarray and data analysis:The circ RNA expression profiles of 12plasma samples were detected using Arraystar Human Circ RNA microarray.t-test was applied to screen circ RNAs associated with vascular injury in the database.Circ RNAs with differences were shown by plotting trend scatter plots and volcano plots.Followed by construction of Cytoscape using circ RNA/mi RNA/m RNA interaction network map.3.Real-time fluorescence quantitative PCR and data analysis:The expression levels of hsa＿circ＿0003258,hsa＿circ＿0004566 and hsa＿circ＿0080319 were verified in 63 plasma samples using quantitative real-time fluorescence PCR.Spearman correlation analysis was used to analyse the years of exposure to HTV for workers in grinding positions,frequency weighted acceleration(denoted by‘a_hw’),hand autonomic symptoms（finger numbness,finger tingling,VWF,finger coldness）in relation to hsa＿circ＿0003258,hsa＿circ＿0004566,hsa＿circ＿0080319 expression levels.Ridge regression analysis of the relationship between age,height,weight,years of exposure to HTV,smoking,alcohol,a_hw,hsa＿circ＿0003258,hsa＿circ＿0004566,hsa＿circ＿0080319 on HAVD.Receiver operating characteristic curve（ROC curve）to assess the sensitivity and specificity of hsa＿circ＿0003258,hsa＿circ＿0004566,hsa＿circ＿0080319 for the diagnosis of HAVD and to find cut-off values.Results:1.Results of circ RNA microarray:In this study,a total of 12 study subjects were analysed by Arraystar Human circ RNA microarray analysis,in which a total of 10circ RNAs were differentially expressed,contain six upregulated circ RNA and four downregulated circ RNA.We find three circ RNAs（hsa＿circ＿0003258,hsa＿circ＿0004566,hsa＿circ＿0080319）were finally screened to construct the reciprocal relationship map.2.Expression of circ RNA:q RT-PCR experiments showed that hsa＿circ＿0003258 and hsa＿circ＿0080319 were highly expressed in the VEC and HAVD groups,with statistically significant differences（P<0.05）.hsa＿circ＿0004566 was highly expressed in the VEC group,with statistically significant differences（P<0.05）.3.Analysis of relevant factors of circ RNA:hsa＿circ＿0003258 and hsa＿circ＿0080319were positively correlated with years of exposure to HTV,a_hw,finger numbness,finger tingling,VWF,and finger chill（P<0.05）,and hsa＿circ＿0004566 was positively correlated with years of vibration,a_hw,finger numbness,and finger tingling（P<0.05）.4.Ridge Regression Analysis of HAVD:Ridge regression analysis showed that age,height,weight,smoking,alcohol consumption,years of exposure to HTV,and hsa＿circ＿0004566 did not affect HAVDS（P>0.05）.ahw,circ＿0003258,and circ＿0080319produced a significant positive relationship on HAVDS（P<0.05）.5.Analysis of the diagnostic value of HAVD:ROC analysis showed a statistically significant AUC of 0.82 for plasma hsa＿circ＿0080319（P<0.001）,standard error of 0.05,95%CI of 0.71-0.93,sensitivity of 0.83,specificity of 0.78,and cut-off value of 3.87.hsa＿circ＿0003258,hsa＿circ＿0004566 were not statistically significant in the diagnosis of HAVD（P>0.05）.Conclusion:1.Hsa＿circ＿0003258 was highly expressed in the VEC and HAVD groups;Hsa＿circ＿0004566 was highly expressed in the VEC group;Hsa＿circ＿0080319 was highly expressed in the VEC and HAVD groups and shows an upward trend.2.Hsa＿circ＿0003258,hsa＿circ＿0080319 were positively correlated with years of vibration,a_hw,finger numbness,finger tingling,VWF,finger chills,hsa＿circ＿0004566years of vibration,a_hw,finger numbness,finger tingling.3.a_hw,hsa＿circ＿0003258,hsa＿circ＿0080319 produced a significant positive effect relationship on HAVD.4.Hsa＿circ＿0080319 has potential as a diagnostic marker for HAVD.