Efficacy Analysis Of Ranibizumab And Retinal Laser Photocoagulation For Treatment Of ROP And Analysis Of Whole-exome Sequencing Of ROP Pathogenic Gene

Part 1 Efficacy analysis of Ranibizumab and retinal laser photocoagulation for treatment of ROPObjective: To explore efficacy analysis of Ranibizumab and retinal laser photocoagulation for treatment of ROP.Methods: In this study,a retrospective case-control study was conducted on 298 children with retinopathy of prematurity who underwent Ranibizumab intravitreal injection or retinal laser photocoagulation admitted to Bayi children hospital from October 2008 to January 2016.Clinical data were analyzed to compare two treatment groups whether there were differences in clinical features,the disease control,the times of treatment and cure rate,then the efficacy of the two treatment groups was evaluated.Results: In this study,298 cases with retinopathy of prematurity who underwent Ranibizumab intravitreal injection or retinal laser photocoagulation were enrolled in the study.There were 124 cases(41.61%)with the treatment of Ranibizumab intravitreal injection and 174 cases in the laser group(58.39%).According to the different zones of the lesions,the number of patients in zone ?was 72 cases(58.06%),and the number of patients in the zone ? was 52 cases(41.94%)in the Ranibizumab intravitreal injection group.In the laser group,there were 30 cases(17.24%)in zone?and 135 cases in zone?,accounting for 77.59%.There were significant differences in the zones of lesions between the two treatment groups(?~2=49.310,P<0.001).There was no difference in the composition ratio of APROP and non-APROP between the two treatment groups(?~2=0.154,P=0.695).There was no significant difference between the two groups in the birth weight,birth gestational age and other clinical features,because of P>0.05.The control rate of ocular lesions after the first treatment of two groups has no significant difference(?~2=0.059,P=0.808).There was no significant difference between the two groups in the control rate of lesions in zone? and ?(?~2=1.436,P=0.231;?~2=0.751,P=0.386).In the two groups,we found that the rate of disease control with APROP was no statistical difference in the two treatment groups(?~2=0.475,P=0.491),and the same to non-APROP(?~2=2.534,P=0.111).There was no significant difference in the times of treatments between the two treatment groups(?~2=0.818,P=0.664),and the finally cure rate of the two treatments is basically the same.We found that premature rupture of membranes,necrotizing enterocolitis,transfer of patients with ROP into the outer courtyard,the zone of the lesion and whether the lesion type was APROP or non-APROP were related with disease control by test(P<0.05).Logistic regression analysis showed that premature rupture of membranes,necrotizing enterocolitis,and whether the lesion type was APROP or non-APROP were the independent risk factors of disease control(P<0.05).There was no significant difference in the long-term prognosis of the two groups(P>0.05).Conclusions:1.In this study,the clinical basic characteristics of children in the two treatment groups were similar,and efficacy of treatment in the two groups was comparable.2.In the Ranibizumab intravitreal injection group the proportion of zone?was higher than laser surgery group.Most of the lesions with Ranibizumab intravitreal injection were located in zone?.Most of lesions were located in zone?in laser surgery group.3.There was no significant difference in the rate of disease control,the times of treatment and the final cure rate of the lesion in the two treatment groups,which indicated that the two treatments were basically the same in the efficacy of short-term treatment.4.There was no significant difference between the two treatment groups in the prognosis of long-term treatment after follow-up.5.Because of simple surgical operation,without anesthesia and invasive ventilation and other advantages in clinical treatment,Ranibizumab injection surgery was better than laser surgery.Part 2 Analysis of whole-exome sequencing of ROP pathogenic geneObjective: By experiments we screen out pathogenic genes associated with ROP possiblely,and carry out the pathogenicity of ROP by analytical studies,preparing for the next functional validation experiments.Methods: A total of 10 infants with ROP were enrolled in this study.Collect blood samples and get on the whole-exome sequencing.Phenolyzer system was used to predict the gene related with the pathogenesis of ROP.The result of two tests was compared by w KGGSeq,finding out the correlation between the phenotype and the gene.From the results of whole-exome sequencing,we can Screen out the candidate pathogenic genes of ROP.The pathogenicity was analyzed for candidate genes with ROP.Results: The data show that the sequencing quality and sequencing depth of whole-exome sequencing meet the research requirements.The total number of genes related to ROP disease was fifty predicted by Phenolyzer system.The total number of genes was 8211 screened out by whole-exome sequencing.The number of deletation mutation was 687,accounting for 8.37%.The number of insert mutation was 500,accounting for 6.09% and the number of SNV was 7024,accounting for 85.54%.Deletion mutations and insertion mutations are mainly based on frameshift mutations and non frameshift mutations,while SNVs were predominantly missense mutations.Comparing the result of predicted gene by Phenolyzer system with the result of whole-exome sequencing,LRP5 gene was found out.The gene was suspected as the pathogenic genes of ROP.By the analysis of gene mutation type,position of gene expression,gene frequency in the population and whether it was pathogenic,we found out that the gene was a missense mutation located in 11q13.2 Exon23.The base G was changed into T situated in Codon 2900,eventually leading to the amino acid at position 967 from cysteine to phenylalanine.The mutation of gene was dominant heterozygous mutation and pathogenic.The gene frequencies in the three genomic databases were less than 0.01,suggesting that mutation of the gene had research significance.And through the software of SIFT,Polyphen2,Mutation Taster,we can predict pathogenicity of gene.LRP5 gene mutations may lead to the occurrence of ROP.Conclutions: One of the 10 samples of the study was carrying a LRP5 gene mutation which may be a pathogenic gene that leads to ROP but the further studies were needed to verify the function of the gene.