地中海贫血铁过载GDF-15基因多态性及血清糖蛋白糖基化修饰的研究和意义
发布时间：2018-07-25 12:16
【摘要】：第一部分GDF-15基因多态性与地中海贫血铁过载发病风险相关性研究背景和目的GDF-15表达与地中海贫血铁过载发生、发展及治疗等方面有关,本部分研究GDF-15基因rsl058587,rsl059369和rs4808793单核苷酸多态性(SNP)位点在地中海贫血铁过载患者及健康人群中的基因分布及频率,探讨rsl058587,rsl059369和rs4808793位点基因多态性与地中海贫血铁过载遗传易感性的关系,从基因水平为地中海贫血铁过载的发病机制及治疗等方面研究提供实验基础。方法收集来自广西地区的地中海贫血铁过载患者122例,无铁过载患者162例,选取128例健康体检者作为对照组。采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术检测GDF-15基因rsl058587,rsl059369和rs4808793位点在病例组与对照组的基因分布频率,用DNA测序法测定对应基因序列,以Hardy-Weinberg平衡(HWE)定律检验纳入病例的群体代表性及可比性。SNP等位基因及基因型的分布差异用χ2检验统计,SNP多态性与地中海贫血铁过载发病风险的关系用logistic回归方法分析。结果三组在性别和年龄构成上无统计学差异(P0.05),具有可比性。经HWE定律检验,rs1059369和rs4808793两个位点P值均大于0.05,说明所选的样本具有群体代表性,而rs1058587位点P值小于0.001,未通过HWE平衡检验。rs1059369位点三组间的基因型分布差异有统计学意义(P0.05);对照组等位基因分布频率较铁过载组差异有统计学意义(P0.05),无铁过载组较对照组及铁过载组等位基因分布频率均无统计学差异(P0.05);铁过载患者AA及AT基因型频率较正常组和无铁过载组显著增加,地中海贫血铁过载的发病风险增加(OR=2.297,95%CI=1.158-3.1014,P=0.002);携带A基因型的个体患铁过载的风险增加(OR=2.259,95%CI=1.177-3.031,P=0.003);对显性模型及隐性模型进一步分析,等位基因A携带频率(AA+AT)在对照组、无铁过载组及铁过载组逐渐升高,有显著性差异(P=0.005),增加铁过载的发病风险(OR=2.553,95%CI=1.177-3.031,P=0.003)。rs4808793位点对照组与无铁过载组及铁过载组的基因型分布差异有统计学意义(P0.05);两个实验组基因型分布频率总体无统计学差异(P0.05),对照组等位基因分布频率较无铁过载组及铁过载组总体差异有统计学意义(P0.05),两个实验组等位基因分布频率总体无统计学差异(P0.05);铁过载患者CC及CG基因型频率较正常组和地无铁过载组显著增加,地中海贫血铁过载的发病风险增加(OR=1.295,95%CI=0.557-1.631,P=0.005);携带CC基因型的个体患铁过载的风险增加(OR=1.275,95%CI=0.658-1.589,P=0.016);对显性模型及隐性模型进一步分析,可看到等位基因C携带频率(CC+CG)在对照组、无铁过载组及铁过载组逐渐升高,有显著性差异(P=0.013),增加铁过载的发病风险(OR=1.159,95%CI=0.797-1.359,P=0.553)。结论GDF-15基因多态位点rs1058587存在GG、CG基因型,以CG型为主,而未发现CC基因型,极有可能是广西地区人群的突变型等位基因频率极低,检测标本量较小,不足以检测到突变纯合子个体。GDF-15基因多态位点rs1059369及rs4808793的多态性与地中海贫血及铁过载存在一定相关性,但rs4808793的多态性与地中海贫血铁过载相关性无统计学意义。第二部分GDF-15 SNP基因型与血清GDF-15、EPO、Hepc及铁代谢相关性的研究背景和目的GDF-15具有调节Hepc的作用,是铁代谢和红系生成中的调节因子。地中海贫血患者,尤其是重症患者血清中的GDF-15水平明显高于正常人群,这表明GDF-15与地中海贫血铁过载的发生发展密切相关,但GDF-15在地中海贫血铁过载中的机制及它对预后的影响仍不清楚,其循环水平及其基因型与地中海贫血铁过载的关系尚不明确,其与地中海贫血铁过载病变严重程度的关系对阐明铁过载的作用机制至关重要。因此本项目研究GDF-15在地中海贫血中的水平、与铁代谢的相关性,希望能发现GDF-15与铁过载的关系,为铁过载的诊断增加检测指标。方法疾病组:选取2016年8月至2017年2月广西医科大学血液内科及地贫之家就诊的284例地中海贫血患者,其中铁过载组122例,无铁过载组162例,男123例,女161例,年龄为0-60岁之间,地中海贫血铁过载入选病例诊断均符合张之南《血液病诊断及疗效标准》第三版及《铁过载诊断与治疗的中国专家共识》。地中海贫血铁过载组中依赖性输血患者68例,非依赖性输血患者54例,所有病例均来自广西地区,排除合并其它类型贫血、感染性疾病、肿瘤、肝脏及骨疾病等病史。对照组:同期广西医科大学第一附属医院的健康体检者128例,男性54例,女74例,均排除贫血、炎症、肿瘤等病史,纳入标准:血清铁蛋白、转铁蛋白水平处于正常参考区间,常规检查正常者。检测血清中GDF-15、EPO、Hepc、FER、Tr F水平并用WB验证。结果经检验,对照组、地中海贫血无铁过载组和地中海贫血铁过载组的血清Hb、GDF-15、EPO、Hepc、FER、Tr F、Hepc/FER均有显著性差异(P0.05)。地中海贫血铁过载组根据是否依赖输血分层分为依赖输血组和非依赖输血组,两组间的血清Hb、GDF-15、EPO、Hepc、FER、Tr F、Hepc/FER均有显著性差异,差异有统计学差异(P0.05)。分析GDF-15水平与Hb、EPO、Hepc、FER、Tr F等指标的相关性,发现所有研究对象血清中GDF-15水平与EPO、Hepc、FER等水平成分呈正相关(P0.001),而与Hb、Tr F水平、Hepc/FER比值呈负相关(P0.001)。GDF-15在地中海贫血患者中急剧升高,与GDF-15基因rs1059369和rs4808793位点基因型及等位基因分布频率的改变无相关性,与Hb水平呈负相关。结论1.地中海贫血患者GDF-15水平随着Hb水平的降低而升高,提示地中海贫血患者GDF-15水平升高与贫血及缺氧有关;2.地中海贫血患者GDF-15水平急剧升高与GDF-15基因多态位点rs1059369及rs4808793的多态性无相关性;3.较GDF-15、Hepc,Hepc/FER是一个更准确更优良的诊断铁过载的检测指标。第三部分地中海贫血铁过载血清糖蛋白糖谱差异研究及意义分析背景和目的铁过载可以引起多种脏器功能损害,涉及地中海贫血进展、疗效及预后。蛋白质糖基化修饰是一种常见的翻译后修饰,在免疫细胞粘附、信号传导、结构稳定、受体构成、细胞分化发育及免疫调控等方面均有重要作用。本部分研究应用凝集素集合芯片技术对地中海贫血铁过载患者血清样本进行分析,旨在研究其糖蛋白糖谱特征性变化,筛选地中海贫血铁过载诊断及活动分期相关特征性糖谱标志,为进一步诊疗研究提供思路及基础。方法实验主要包含以下步骤:(1)血清低丰度蛋白的收集:等体积混合4组(α地中海贫血铁过载组、α地中海贫血组无铁过载组、β地中海贫血铁过载组和β地中海贫血无铁过载组)患者血清,使用高丰度蛋白去除柱收集血清低丰度蛋白;(2)将收集的低丰度蛋白经除盐、超滤、测定浓度后处理为待标记缓冲液;(3)待测低丰度蛋白CY5标记;(4)凝集素芯片的点制及芯片的杂交;(5)芯片扫描、数据提取;(6)数据统计采用SPSS16.0,符合正态分布且方差齐的数据采用两独立样本的均数t检验,方差不齐者采用非参数秩和检验。(7)凝集素印迹技术检测AAL、LCA和WGA三种凝集素对血清糖蛋白的亲和能力,验证凝集素芯片结果的可靠性。结果本次研究中4组样品中共筛选出34种有统计学差异的凝集素阳性亲和信号,其中α地中海贫血筛选出19个差异性凝集素和β地中海贫血筛选出15个差异性凝集素。α地中海贫血铁过载组对AAL、LCA、ABL、AMA、MNA-M、DBA、GHA、GSL1、GSL1b4、HAL、GSL2、GNL、PHA-L、PSA、SSA和LAL凝集素的亲和力增强,而对RCA60、STL和WGA凝集素的亲和力减弱;β地中海贫血铁过载组对ABL、GSL2、LCA、STL、TL、AMA、PSA的亲和作用增强,而对Con A、GNL、NPL、IRA、HAL、HPL、DSA、MAL-II的亲和力减弱。凝集素印迹技术证明AAL、LCA和WGA三种凝集素对血清糖蛋白的亲和能力的结果与凝集素芯片结果一致。结论各种凝集素亲和信号变化趋势各有不同,表示地中海贫血铁过载发生过程中不同型别地中海贫血的血清糖蛋白糖链结构有复杂性与特异性的变化,这提示以不同型别血清特征性糖谱作为地中海贫血铁过载诊断或疾病分型的生物标志具备一定的实验基础。
[Abstract]:Part 1 the correlation between GDF-15 gene polymorphism and the risk of iron overload in thalassemia and the purpose of GDF-15 expression is related to the occurrence, development and treatment of iron overload in thalassemia. This part studies the GDF-15 gene rsl058587, rsl059369 and rs4808793 single nucleotide polymorphisms (SNP) loci in the iron overload of thalassemia The genetic polymorphism of rsl058587, rsl059369 and rs4808793 loci and the genetic susceptibility to iron overload in thalassemia were investigated to provide experimental basis for the pathogenesis and treatment of iron overload in thalassemia. Methods collected from the Mediterranean region of Guangxi were collected. 122 patients with iron overload, 162 cases of iron free overload, and 128 healthy subjects were selected as the control group. Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) was used to detect the gene distribution frequency of GDF-15 gene rsl058587, rsl059369 and rs4808793 loci in the case group and the control group, and the DNA sequencing method was used to determine the corresponding base. Hardy-Weinberg equilibrium (HWE) law was used to test the distribution differences between the group representative and the comparable.SNP alleles and genotypes by the chi square test. The relationship between the SNP polymorphism and the risk of iron overload in thalassemia was analyzed by the logistic regression method. There was no statistical difference between the three groups in the sex and age composition. By HWE's law, the P values of two loci of rs1059369 and rs4808793 were both greater than 0.05, indicating that the selected samples were representative of the group, while the P value of the rs1058587 site was less than 0.001, and the genotype distribution difference between the three groups of the.Rs1059369 loci was statistically significant (P0.05) without HWE balance test (P0.05); the control group allele scores were statistically significant (P0.05). There was no significant difference in the frequency of distribution between the iron overload group and the iron overload group (P0.05). There was no statistical difference between the iron overload group and the control group and the iron overload group (P0.05). The frequency of AA and AT genotype in the iron overload patients was significantly higher than that in the normal group and the iron overload group, and the risk of iron overload in thalassemia was increased (OR=2.297,95%CI=1.158-3 .1014, P=0.002); the risk of iron overload with A genotype increased (OR=2.259,95%CI=1.177-3.031, P=0.003); further analysis of the dominant and recessive models, the A carrying frequency (AA+AT) of the allele (AA+AT) in the control group, the iron overload group and the iron overload group increased gradually (P=0.005), and increased the risk of iron overload (OR). =2.553,95%CI=1.177-3.031, P=0.003).Rs4808793 locus control group was significantly different from the iron overload group and iron overload group (P0.05), and there was no statistical difference in the genotype distribution frequency of the two experimental groups (P0.05). The frequency of allele distribution in the control group was more than that in the iron overload group and the iron overload group. P0.05, there was no significant difference in the frequency of allele distribution in the two experimental groups (P0.05); the frequency of CC and CG genotype in iron overload patients was significantly higher than that in the normal group and the iron free overload group, and the risk of iron overload in thalassemia increased (OR=1.295,95%CI= 0.557-1.631, P=0.005); the risk of iron overload carrying CC genotype was the risk of iron overload. Increase (OR=1.275,95%CI=0.658-1.589, P=0.016); to further analyze the dominant and recessive models, we can see that the allele C carrying frequency (CC+CG) in the control group, the iron overload group and the iron overload group gradually increase, there is a significant difference (P=0.013), increasing the risk of iron overload (OR=1.159,95%CI=0.797-1.359, P=0.553). Conclusion GDF-15 gene Polymorphic loci rs1058587 exists GG, CG genotype, CG type, and no CC genotypes. It is very likely that the frequency of mutant alleles in Guangxi population is very low and the sample size is small. It is not enough to detect the polymorphism of.GDF-15 gene polymorphisms of rs1059369 and rs4808793 in the mutant homozygote and thalassemia and iron overload. There is a certain correlation, but there is no significant correlation between the polymorphism of rs4808793 and the iron overload of thalassemia. The background and purpose of the study on the correlation between the second part of the GDF-15 SNP genotype and serum GDF-15, EPO, Hepc and iron metabolism is the regulatory factor in the regulation of Hepc. It is a regulatory factor in the formation of iron metabolites and red lines. The serum levels of GDF-15 in patients, especially in severe patients, are significantly higher than those in the normal population, which indicates that GDF-15 is closely related to the development of iron overload in thalassemia, but the mechanism of GDF-15 in the iron overload of thalassemia and its impact on the prognosis are still unclear. The relationship between the circulation level and its genotype and the iron overload of thalassemia It is not clear that the relationship between the severity of thalassemia and the severity of iron overload is essential to elucidate the mechanism of iron overload. Therefore, this project studies the level of GDF-15 in thalassemia and the correlation with iron metabolism. We hope to find the relationship between GDF-15 and iron overload, and increase the detection index for the diagnosis of iron overload. 284 cases of thalassemia were selected from the Department of Hematology of Guangxi Medical University from August 2016 to February 2017, including 122 cases of iron overload group, 162 cases of iron free overload group, 123 men, 161 women and 0-60 years old. The cases of thalassemia iron overloading cases were all conformed to Zhang Zhinan's diagnostic and therapeutic standards for hematological diseases and the standard of curative effect. The Chinese expert consensus of the three edition and < iron overload diagnosis and treatment >. 68 patients with dependent blood transfusion in the thalassemia iron overload group and 54 cases of non dependent blood transfusion, all cases were from Guangxi, excluding other types of anemia, infectious diseases, tumor, liver and bone disease. Control group: the same period of Guangxi Medical University. 128 cases of health checkup in a Affiliated Hospital, 54 men and 74 women, were all excluded from anemia, inflammation, tumor and other diseases. The serum ferritin, transferrin level was in normal reference range, normal subjects were examined. The serum levels of GDF-15, EPO, Hepc, FER, Tr F were tested by WB. The results were tested, the control group, thalassemia was no There were significant differences in serum Hb, GDF-15, EPO, Hepc, FER, Tr F and Hepc/FER (P0.05) in the iron overload group and the iron overload group of thalassemia. There were significant differences between the two groups of serum Hb, GDF-15, EPO, EPO. Statistical difference (P0.05). The correlation between GDF-15 level and Hb, EPO, Hepc, FER, Tr F and other indexes was analyzed. It was found that the level of GDF-15 in the serum of all the subjects was positively correlated with EPO, Hepc, FER and other levels. There was no correlation between the change of genotype and allele distribution frequency of rs1059369 and rs4808793 loci and negative correlation with the level of Hb. Conclusion the level of GDF-15 in 1. thalassemia patients increased with the decrease of Hb level, suggesting that the level of GDF-15 in thalassemia patients was associated with anemia and anoxia, and 2. thalassemia patients had a sharp GDF-15 level. No correlation between the polymorphism of GDF-15 gene polymorphic loci rs1059369 and rs4808793; 3. compared with GDF-15, Hepc, Hepc/FER is a more accurate and better diagnostic indicator of iron overload. The third part of thalassemia iron overload serum glycoprotein sugar spectrum differences and significance analysis background and purpose iron overload can cause a variety of viscera. Functional impairment involves the progression, efficacy and prognosis of thalassemia. Glycosylation is a common post-translational modification, which plays an important role in immune cell adhesion, signal transduction, structural stability, receptor composition, cell differentiation and development, and immunoregulation. The serum samples of patients with anemia of iron overload were analyzed in order to study the characteristic changes of glycoprotein sugar spectrum, screening the diagnosis of thalassemia iron overload and the characteristic glycan markers related to activity stages, and provide ideas and basis for further diagnosis and treatment. The method experiment mainly includes the following steps: (1) the collection of serum low abundance protein: equal volume The mixed 4 groups (alpha thalassemia iron overload group, alpha thalassemia free iron overload group, beta thalassemia iron overload group and beta thalassemia free iron overload group) were used to collect serum low abundance protein with high abundance protein removal column, and (2) the low abundance protein was collected by desalination, ultrafiltration, and the concentration was treated as a pending buffer. Liquid; (3) to test low abundance protein CY5 markers; (4) dot and chip hybridization of agglutinin chips; (5) chip scanning, data extraction; (6) data statistics using SPSS16.0, data consistent with normal distribution and variance of data using two independent samples of the average number of t tests, square difference using non parametric rank sum test. (7) lectin Blotting Technology to detect AAL, LC The affinity of A and WGA three lectins to serum glycoproteins was proved to be reliable. Results in this study, 4 groups of samples were screened out of 34 different agglutinin positive affinity signals, of which alpha thalassemia screened 19 differential agglutinin and beta thalassemia to screen 15 differential coagulabilities. AAL, LCA, ABL, AMA, MNA-M, DBA, GHA, GSL1, GSL1b4, HAL, GSL2, GNL, HAL, and agglutinin affinity. The affinity of HPL, DSA and MAL-II was weakened. Lectin blot technique showed that the affinity of three lectins to serum glycoproteins of AAL, LCA and WGA was consistent with the results of lectin chip. Conclusion the changes of various agglutinin affinity signals are different, indicating the blood of different types of thalassemia in the process of iron overload in thalassemia. The sugar chain structure of the glycoprotein has a complex and specific change, which suggests that the specific glycemic spectrum of different types of sera has a certain experimental basis as a biomarker for the diagnosis of thalassemia iron overload or the classification of disease.
【学位授予单位】：广西医科大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R556.61
本文编号：2143793
[Abstract]:Part 1 the correlation between GDF-15 gene polymorphism and the risk of iron overload in thalassemia and the purpose of GDF-15 expression is related to the occurrence, development and treatment of iron overload in thalassemia. This part studies the GDF-15 gene rsl058587, rsl059369 and rs4808793 single nucleotide polymorphisms (SNP) loci in the iron overload of thalassemia The genetic polymorphism of rsl058587, rsl059369 and rs4808793 loci and the genetic susceptibility to iron overload in thalassemia were investigated to provide experimental basis for the pathogenesis and treatment of iron overload in thalassemia. Methods collected from the Mediterranean region of Guangxi were collected. 122 patients with iron overload, 162 cases of iron free overload, and 128 healthy subjects were selected as the control group. Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) was used to detect the gene distribution frequency of GDF-15 gene rsl058587, rsl059369 and rs4808793 loci in the case group and the control group, and the DNA sequencing method was used to determine the corresponding base. Hardy-Weinberg equilibrium (HWE) law was used to test the distribution differences between the group representative and the comparable.SNP alleles and genotypes by the chi square test. The relationship between the SNP polymorphism and the risk of iron overload in thalassemia was analyzed by the logistic regression method. There was no statistical difference between the three groups in the sex and age composition. By HWE's law, the P values of two loci of rs1059369 and rs4808793 were both greater than 0.05, indicating that the selected samples were representative of the group, while the P value of the rs1058587 site was less than 0.001, and the genotype distribution difference between the three groups of the.Rs1059369 loci was statistically significant (P0.05) without HWE balance test (P0.05); the control group allele scores were statistically significant (P0.05). There was no significant difference in the frequency of distribution between the iron overload group and the iron overload group (P0.05). There was no statistical difference between the iron overload group and the control group and the iron overload group (P0.05). The frequency of AA and AT genotype in the iron overload patients was significantly higher than that in the normal group and the iron overload group, and the risk of iron overload in thalassemia was increased (OR=2.297,95%CI=1.158-3 .1014, P=0.002); the risk of iron overload with A genotype increased (OR=2.259,95%CI=1.177-3.031, P=0.003); further analysis of the dominant and recessive models, the A carrying frequency (AA+AT) of the allele (AA+AT) in the control group, the iron overload group and the iron overload group increased gradually (P=0.005), and increased the risk of iron overload (OR). =2.553,95%CI=1.177-3.031, P=0.003).Rs4808793 locus control group was significantly different from the iron overload group and iron overload group (P0.05), and there was no statistical difference in the genotype distribution frequency of the two experimental groups (P0.05). The frequency of allele distribution in the control group was more than that in the iron overload group and the iron overload group. P0.05, there was no significant difference in the frequency of allele distribution in the two experimental groups (P0.05); the frequency of CC and CG genotype in iron overload patients was significantly higher than that in the normal group and the iron free overload group, and the risk of iron overload in thalassemia increased (OR=1.295,95%CI= 0.557-1.631, P=0.005); the risk of iron overload carrying CC genotype was the risk of iron overload. Increase (OR=1.275,95%CI=0.658-1.589, P=0.016); to further analyze the dominant and recessive models, we can see that the allele C carrying frequency (CC+CG) in the control group, the iron overload group and the iron overload group gradually increase, there is a significant difference (P=0.013), increasing the risk of iron overload (OR=1.159,95%CI=0.797-1.359, P=0.553). Conclusion GDF-15 gene Polymorphic loci rs1058587 exists GG, CG genotype, CG type, and no CC genotypes. It is very likely that the frequency of mutant alleles in Guangxi population is very low and the sample size is small. It is not enough to detect the polymorphism of.GDF-15 gene polymorphisms of rs1059369 and rs4808793 in the mutant homozygote and thalassemia and iron overload. There is a certain correlation, but there is no significant correlation between the polymorphism of rs4808793 and the iron overload of thalassemia. The background and purpose of the study on the correlation between the second part of the GDF-15 SNP genotype and serum GDF-15, EPO, Hepc and iron metabolism is the regulatory factor in the regulation of Hepc. It is a regulatory factor in the formation of iron metabolites and red lines. The serum levels of GDF-15 in patients, especially in severe patients, are significantly higher than those in the normal population, which indicates that GDF-15 is closely related to the development of iron overload in thalassemia, but the mechanism of GDF-15 in the iron overload of thalassemia and its impact on the prognosis are still unclear. The relationship between the circulation level and its genotype and the iron overload of thalassemia It is not clear that the relationship between the severity of thalassemia and the severity of iron overload is essential to elucidate the mechanism of iron overload. Therefore, this project studies the level of GDF-15 in thalassemia and the correlation with iron metabolism. We hope to find the relationship between GDF-15 and iron overload, and increase the detection index for the diagnosis of iron overload. 284 cases of thalassemia were selected from the Department of Hematology of Guangxi Medical University from August 2016 to February 2017, including 122 cases of iron overload group, 162 cases of iron free overload group, 123 men, 161 women and 0-60 years old. The cases of thalassemia iron overloading cases were all conformed to Zhang Zhinan's diagnostic and therapeutic standards for hematological diseases and the standard of curative effect. The Chinese expert consensus of the three edition and < iron overload diagnosis and treatment >. 68 patients with dependent blood transfusion in the thalassemia iron overload group and 54 cases of non dependent blood transfusion, all cases were from Guangxi, excluding other types of anemia, infectious diseases, tumor, liver and bone disease. Control group: the same period of Guangxi Medical University. 128 cases of health checkup in a Affiliated Hospital, 54 men and 74 women, were all excluded from anemia, inflammation, tumor and other diseases. The serum ferritin, transferrin level was in normal reference range, normal subjects were examined. The serum levels of GDF-15, EPO, Hepc, FER, Tr F were tested by WB. The results were tested, the control group, thalassemia was no There were significant differences in serum Hb, GDF-15, EPO, Hepc, FER, Tr F and Hepc/FER (P0.05) in the iron overload group and the iron overload group of thalassemia. There were significant differences between the two groups of serum Hb, GDF-15, EPO, EPO. Statistical difference (P0.05). The correlation between GDF-15 level and Hb, EPO, Hepc, FER, Tr F and other indexes was analyzed. It was found that the level of GDF-15 in the serum of all the subjects was positively correlated with EPO, Hepc, FER and other levels. There was no correlation between the change of genotype and allele distribution frequency of rs1059369 and rs4808793 loci and negative correlation with the level of Hb. Conclusion the level of GDF-15 in 1. thalassemia patients increased with the decrease of Hb level, suggesting that the level of GDF-15 in thalassemia patients was associated with anemia and anoxia, and 2. thalassemia patients had a sharp GDF-15 level. No correlation between the polymorphism of GDF-15 gene polymorphic loci rs1059369 and rs4808793; 3. compared with GDF-15, Hepc, Hepc/FER is a more accurate and better diagnostic indicator of iron overload. The third part of thalassemia iron overload serum glycoprotein sugar spectrum differences and significance analysis background and purpose iron overload can cause a variety of viscera. Functional impairment involves the progression, efficacy and prognosis of thalassemia. Glycosylation is a common post-translational modification, which plays an important role in immune cell adhesion, signal transduction, structural stability, receptor composition, cell differentiation and development, and immunoregulation. The serum samples of patients with anemia of iron overload were analyzed in order to study the characteristic changes of glycoprotein sugar spectrum, screening the diagnosis of thalassemia iron overload and the characteristic glycan markers related to activity stages, and provide ideas and basis for further diagnosis and treatment. The method experiment mainly includes the following steps: (1) the collection of serum low abundance protein: equal volume The mixed 4 groups (alpha thalassemia iron overload group, alpha thalassemia free iron overload group, beta thalassemia iron overload group and beta thalassemia free iron overload group) were used to collect serum low abundance protein with high abundance protein removal column, and (2) the low abundance protein was collected by desalination, ultrafiltration, and the concentration was treated as a pending buffer. Liquid; (3) to test low abundance protein CY5 markers; (4) dot and chip hybridization of agglutinin chips; (5) chip scanning, data extraction; (6) data statistics using SPSS16.0, data consistent with normal distribution and variance of data using two independent samples of the average number of t tests, square difference using non parametric rank sum test. (7) lectin Blotting Technology to detect AAL, LC The affinity of A and WGA three lectins to serum glycoproteins was proved to be reliable. Results in this study, 4 groups of samples were screened out of 34 different agglutinin positive affinity signals, of which alpha thalassemia screened 19 differential agglutinin and beta thalassemia to screen 15 differential coagulabilities. AAL, LCA, ABL, AMA, MNA-M, DBA, GHA, GSL1, GSL1b4, HAL, GSL2, GNL, HAL, and agglutinin affinity. The affinity of HPL, DSA and MAL-II was weakened. Lectin blot technique showed that the affinity of three lectins to serum glycoproteins of AAL, LCA and WGA was consistent with the results of lectin chip. Conclusion the changes of various agglutinin affinity signals are different, indicating the blood of different types of thalassemia in the process of iron overload in thalassemia. The sugar chain structure of the glycoprotein has a complex and specific change, which suggests that the specific glycemic spectrum of different types of sera has a certain experimental basis as a biomarker for the diagnosis of thalassemia iron overload or the classification of disease.
【学位授予单位】：广西医科大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R556.61
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