雷珠单抗与视网膜激光光凝术治疗ROP疗效及ROP致病基因全外显子测序分析

发布时间：2018-05-08 13:28

本文选题：早产儿视网膜病 + 视网膜激光光凝术　；参考：《大连医科大学》2017年硕士论文

【摘要】：第一部分雷珠单抗与视网膜激光光凝术治疗ROP疗效分析目的:探讨298例早产儿视网膜病患儿首次采用雷珠单抗玻璃体腔注药术或视网膜激光光凝术两种治疗方法的治疗疗效。方法:本研究采用回顾性病例对照研究,对2007年10月至2016年1月收治在陆军总医院附属八一儿童医院的298例需要首次接受雷珠单抗玻璃体腔注药术或者视网膜激光光凝术治疗的早产儿视网膜病患儿临床资料进行分析,比较两不同治疗组中患儿临床特点是否存在差异,病变控制情况、治疗次数、病变最终治愈率、远期预后等是否存在不同,从而对首次采用的两种治疗方法的治疗疗效进行评价。结果:本研究中符合纳入标准需要首次接受雷珠单抗玻璃体腔注药术及视网膜激光光凝术治疗的早产儿视网膜病患儿为298例,其中首次选择的治疗方法为雷珠单抗注药术124例(41.61%)、激光术174例(58.39%)。根据病变分区不同,124例首次采用雷珠单抗注药术治疗人数中Ⅰ区病变人数有72例,占58.06%,Ⅱ区病变人数52例,占41.94%。174例首次采用视网膜激光术治疗人数中Ⅰ区病变人数有30例,占17.24%,Ⅱ区病变人数135例,占77.59%,两治疗组在病变的分区比较中存在差异性(χ~2=49.310,P0.001)。首次采用的两治疗组中APROP与非APROP构成比比较中无差异性(χ~2=0.154,P=0.695)。两治疗组患儿在出生体重、出生胎龄及其他多种临床特点的比较中均为P0.05,无显著性差异。在病变控制率的比较中两治疗组在首次治疗后无差异(χ~2=0.059,P=0.808),且两者分别在Ⅰ区和Ⅱ区不同病变区域中首次治疗后的病变控制率比较中也未见显著性差异(χ~2=1.436,P=0.231;χ~2=0.751,P=0.386)。不同病变类型比较中,两治疗组分别在急进型ROP与非急进型ROP首次治疗后病变控制率比较无统计学差异性(急进型ROPχ~2=0.475,P=0.491;非急进型ROPχ~2=2.534,P=0.111)。首次采用两种治疗方法后两组治疗次数的比较上也无显著差异(χ~2=0.818,P=0.664),且病变的最终治愈率上两者基本一致。通过检验分析发现胎膜早破、坏死性小肠结肠炎、外院转入ROP、病变分区、病变类型是否为APROP与病变控制情况相关,经Logistic回归分析发现胎膜早破、坏死性小肠结肠炎、病变类型是否为APROP是影响病变控制的独立危险因素(P0.05)。在激光术组随访人数较少的前提下,两治疗组远期预后比较无显著性差异(P0.05)。结论:1.本研究中纳入比较的两治疗组中患儿的临床基本特点一致,因而两治疗组在治疗疗效上具有可比性。2.注药术组中Ⅰ区病变所占比例高于激光术组中Ⅰ区病变所占比例,注药术组Ⅰ区病变多,而激光术组以Ⅱ区病变为主。3.两治疗组在首次治疗后的病变控制率、病变治疗次数及病变的最终治愈率的比较上并没有显著性差异,也说明两者在短期治疗效果上基本一致。4.通过随访发现两治疗组在首次治疗后远期治疗预后结果比较无统计学差异。5.在两治疗组临床治疗效果无差异的前提下,注药术治疗操作简单、无需麻醉及有创通气因而在临床治疗选择中更优于激光术组。第二部分ROP致病基因全外显子测序分析目的:通过实验筛选可能与早产儿视网膜病致病相关基因,分析其致病性,为下一步功能性验证实验做准备。方法:对10例早产儿视网膜病患儿血液样本进行全基因外显子测序,应用Phenolyzer系统预测与早产儿视网膜病相关基因,将两者进行比对,通过基因表型驱动分析方法,找到表型与基因的相关性,筛选可能与早产儿视网膜病发生相关的候选致病基因,并根据研究结果分析其致病性。结果:本研究数据显示测序质量及测序深度均满足研究要求,全基因外显子测序获得的基因总数为8211个,其中发生缺失突变的有687个,占8.37%,插入突变的有500个,占6.09%,单核苷酸突变(Single nucleotide variation,SNV)有7024个,占85.54%。缺失突变及插入突变主要以移码突变及非移码突变为主,而SNV主要以错义突变为主,突变的类型不同导致最终编码蛋白质功能亦不同。而通过Phenolyzer系统预测出与ROP疾病相关基因有50种,将全基因测序结果与Phenolyzer系统预测的基因进行表型驱动分析后发现LRP5基因可能为早产儿视网膜病致病基因。之后针对研究中基因突变类型、基因表达部位、人群中基因频率、是否存在致病性等进行分析,发现该基因突变为错义突变,可导致11号染色体1区3带第2次亚带中外显子23的第2900位密码子中间位置的碱基有G突变为T,最终导致第967位氨基酸由半胱氨酸变为苯丙氨酸,并且该基因突变为显性杂合突变,在三大人群中基因频率均0.01,提示基因突变具有研究意义,且通过SIFT、Polyphen2、Mutation Taster软件对该基因的致病性进行预测,提示该基因是具有致病性的。因而预测LRP5基因突变可能导致ROP的发生。结论:10例研究样本中1例样本中携带有LRP5基因突变,其可能为导致ROP疾病发生的致病基因,但仍需进一步的研究来验证该基因的功能。
[Abstract]:The first part of the treatment of ROP with reelzumumab and retinal laser photocoagulation in the treatment of 298 cases of premature infants with retinopathy of retinopathy of the first use of rezumumab glass cavity injection or retinal laser photocoagulation two treatment methods. Methods: This study used a retrospective case control study, from October 2007 to 2016. In January, the clinical data of 298 children with retinopathy of preterm infants, which were first accepted by lizumumab, or retinal laser photocoagulation, were treated in the children's Hospital Affiliated to the General Hospital of the army general hospital. The clinical characteristics of the children in the two different treatment groups were compared. The final cure rate and the long-term prognosis are different, and the therapeutic effect of the first two methods of treatment is evaluated. Results: in this study, 298 cases of premature infants with retinopathy of preterm infants were first accepted by the inclusion criteria of rezumumab and retinal laser photocoagulation for the first time. The selected treatment was 124 cases (41.61%) and 174 cases (58.39%) with laser surgery. According to the diseased area, 124 cases of the first use of lezumab injection were 72 cases, 58.06%, and 52 cases in the second region, accounting for the first time in the 41.94%.174 cases by retina laser treatment. There were 30, 17.24%, 135, 77.59%, and two in the two treatment group (x, P0.001). There was no difference in the ratio of APROP to non APROP in the first two treatment group (x ~2=0.154, P=0.695). The children in the two treatment group were born in birth weight, birth gestational age and a variety of other clinical cases. The comparison of the characteristics was P0.05 without significant difference. In the comparison of the rate of disease control, there was no difference between the two treatment groups after the first treatment (x ~2=0.059, P=0.808), and there was no significant difference (x ~2=1.436, P=0.231; Chi ~2=0.751, P=0.386) in the first treatment of the first treatment in the region I and the region II. In the comparison of different types of lesions, there was no significant difference in the control rate between the two treatment groups after the first treatment of the emergency ROP and the non emergency ROP (ROP Chi ~2=0.475, P=0.491; the non urgent ROP x ~2=2.534, P=0.111). There was no significant difference in the comparison between the two groups after the first two treatments (x ~2=0.818, P=0). .664) and the final cure rate of the lesions were basically the same. Through the examination and analysis, it was found that premature rupture of the membranes, necrotizing enterocolitis, the external hospital was transferred to ROP, the lesion was divided, whether the type of APROP was related to the control of the disease, and the Logistic regression analysis found that the premature rupture of the membranes, necrotic enterocolitis, and the type of pathological changes were APROP. The independent risk factor (P0.05) for the control of rounded disease (P0.05). There was no significant difference in the long-term prognosis in the two treatment group (P0.05). Conclusion: the clinical basic characteristics of the children in the two treatment group were the same in the 1. study, so the two treatment group had the comparable.2. injection group in the therapeutic effect. The proportion of the lesion in the middle part I was higher than that in the laser group. The lesion in the group I was much more, and there was no significant difference in the rate of control, the number of treatment and the final cure rate of the lesion in the group of.3. two, which was the first treatment in the laser group. It also indicated that the two groups were treated in the short term. The curative effect was basically consistent with.4. through follow-up. It was found that there was no statistical difference between the two treatment groups after the first treatment and the prognosis of the long term treatment was no significant difference..5. was no difference in the effect of the clinical treatment in the two treatment group. The treatment operation was simple, no anesthesia and ventilation was needed, so second departments were better than the laser group in the clinical treatment. ROP pathogenic gene exon sequencing analysis objective: to screen the pathogenicity of retinopathy of premature infants by screening the genes related to retinopathy of prematurity, and to analyze its pathogenicity and prepare for the next functional verification experiment. Methods: the whole gene exon sequencing was carried out in 10 children with retinopathy of preterm infants, and the Phenolyzer system was used to predict and early. The genes related to retinopathy of birth were compared, and the correlation between phenotypes and genes was found by gene phenotypic driving analysis. The candidate genes associated with the occurrence of retinopathy in preterm infants were screened and the pathogenicity was analyzed according to the results. Results: the results of the study show that the quality of sequencing and the depth of sequencing are satisfied. The total number of gene exons sequenced is 8211, of which 687, 8.37%, 500, 6.09%, Single nucleotide variation, SNV, 8.37%, and 7024 of the single nucleotide mutations (variation, SNV), and the major 85.54%. deletion and the insertion mutation are mainly a code shift mutation and a non graft mutation, and SN V is mainly missense mutation, and the different types of mutation lead to the difference in the function of the final encoded protein, and 50 kinds of genes related to ROP disease are predicted by the Phenolyzer system. The LRP5 gene may be the pathogeny of the retinopathy of the premature infant after the analysis of the gene sequencing results and the gene predicted by the Phenolyzer system. Then the gene mutation type, the gene expression site, the gene frequency and the pathogenicity of the population were analyzed, and the mutation was found to be a missense mutation, which could lead to the G mutation of the G mutation of the base between the 2900th bits of the codon 23 of the second subbands of the 3 band and second subbands of the chromosome 1. Amino acids change from cysteine to phenylalanine, and the gene mutation is a dominant heterozygous mutation. The gene frequency of the gene is 0.01 in the three population, suggesting that the gene mutation is of research significance, and the pathogenicity of the gene is predicted by SIFT, Polyphen2, Mutation Taster software, suggesting that the gene is pathogenic. Therefore, the LRP5 base is predicted. Mutations may lead to the occurrence of ROP. Conclusion: in 1 samples of 10 cases, a mutation of LRP5 gene is carried in the sample, which may be a pathogenic gene that causes the occurrence of ROP disease, but further research is still needed to verify the function of the gene.

【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R774.1

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