【作者】 李婕；

【导师】 梁雁；

【作者基本信息】 华中科技大学 ， 儿科学， 2013， 硕士

【摘要】 目的：研究我国Turner综合征（Turnersyndrome,先天性卵巢发育不良综合征）GHR外显子3缺失多态性分布频率，探讨Turner综合征患儿中rhGH治疗疗效与该多态性的相关性。方法：对31例rhGH治疗满一年及9例治疗满2年的Turner综合征患儿进行回顾性研究分析。用△Ht-SDS、△GV及△Ht为衡量rhGH治疗疗效的指标。PCR方法检测GHR外显子3缺失多态性，用Hardy-Weinberg遗传平衡方法检验各基因型的理论值，用SPSS12.0软件（X2检验、方差分析、t检验、pearson相关分析及线性回归分析）对结果进行统计分析。结果：（1）31例Turner综合征患儿中fl/fl-GHR、fl/d3-GHR、d3/d3-GHR分布频率分别为41.9%、45.2%及12.9%，与德国、巴西分布频率无差异（p＞0.05），韩国分布频率有显著差异（p＜0.05）。（2）rhGH治疗1年治疗前与治疗后，GV分别为（2.94±0.89cm/年，7.36±1.34cm/年）；Ht-SDS分别为（-3.47±1.33，-2.78±1.24）均有显著差异（p＜0.05）。（3）9例治疗两年Turner综合征患儿，治疗前、治疗第一年及治疗第二年GV分别为（3.39±0.56cm/年，7.98±0.73cm/年，7.07±0.98cm/年）,三者之间均有显著差异（p＜0.05），治疗前、治疗第一年及治疗第二年Ht-SDS分别为（3.48±1.51，2.48±1.25，1.01±1.02），治疗前Ht-SDS与治疗第一年Ht-SDS无显著差异（p＞0.05），治疗第二年Ht-SDS与治疗前及第一年Ht-SDS均有显著差异（p＜0.05）。（3）不同基因型患儿（fl/fl-GHR，fl/d3-GHR，d3/d3-GHR）rhGH治疗1年后，第一年△GV、△Ht-SDS、△Ht及rhGH剂量均无显著差异p＞0.05）。（4）不同基因型患儿（fl/fl-GHR，fl/d3-GHR+d3/d3-GHR）rhGH治疗2年后，第二年△GV（3.56±0.51cm/年，3.92±1.01cm/年）、两年△Ht及两年△GV平均均无显著性差异（p＞0.05）。（5）根据染色体核型将31例患儿分为45，X核型组、嵌合型核型组及结构畸变型核型组，生长激素治疗1年后，三组患儿△Ht-SDS分别为（0.54±0.35SDS，0.86±0.44SDS，0.56±2.44SDS），嵌合型核型组△Ht-SDS与45，X核型组有显著差异（p＜0.05），其余两组△Ht-SDS无显著差异（p＞0.05），△GV（4.42±1.31cm/年，4.39±1.22cm/年，4.53±1.41cm/年）及△H（t7.24±1.07cm，7.55±1.29cm，7.18±2.44cm）三组之间无显著差异（p＜0.05）。(6)生长激素治疗一年，△Ht-SDS与初治GV、Ht-SDS、BMI及治疗前GV的单因素相关分析均有相关性（p＜0.05），治疗时BA、GHR基因型及rhGH剂量单因素相关分析无相关性（p＞0.05）；治疗一年△GV与上述参数均无相关性（p＞0.05）；多因素线性回归分析△Ht-SDS与初治时CA、治疗前GV相关（p＜0.05），回归系数分别为-0.096和0.217，△Ht-SDS与治疗时Ht-SDS及BMI无相关性（p＞0.05）。（7）生长激素治疗两年后△Ht-SDS及△GV平均与治疗时CA、BA、Ht-SDS、BMI、rhGH剂治疗前GV单因素相关分析均无相关性（p＞0.05）。结论：本研究中31例Turner综合征患儿GHR外显子3缺失多态性的频率分布与部分国外报道有显著差异，生长激素治疗能明显改善Turner综合征患儿身高，我国Turner综合征患儿GHR基因型与生长激素治疗一年及两年后的疗效无明显相关性，染色体核型可能与治疗疗效相关，生长激素治疗一年，初治时年龄与生长激素治疗疗效呈负相关，未治疗前身高增长速率与生长激素治疗疗效呈正相关。目的：对4临床疑诊Noonan综合征患儿进行临床特征分析及PTPN11基因诊断。方法：总结4例疑诊Noonan综合征患儿的临床表现并行染色体核型分析，选择PTPN11基因为目的基因，用PCR扩增PTPN11基因的热点突变外显子（2、3、4、7、8、12、13），PCR产物经ABI3730测序仪进行测序获得峰图，DNABaser序列汇编软件分析测序结果发现突变位点，最后在人类基因组突变数据库（HGMD）中查看突变位点是否为致病突变。结果：4例患儿，均有身材矮小；睑裂下斜、眼距宽等特殊面容；双肘外翻、先天性心脏病等特点，染色体核型均正常。2例PTPN11基因测序正常，2例患儿在PTPN11基因上存在致病突变，患儿APTPN11外显子13第1510位碱基由A被G替代（1510A→G），导致第504位氨基酸由甲硫氨酸变为缬氨酸（met504val）。患儿B，PTPN11外显子3第236位碱基A被G替代（236A→G），导致第79位氨基酸由谷氨酰胺变为精氨酸（Gln79Arg）。两个突变均为已报道的Noonan综合征的致病突变。结论：Noonan综合征临床症状与Turner综合征、GHARGE综合征及Costello综合征不易鉴别，PTPN11基因是Noonan综合征最常见致病基因，对高度怀疑Noonan综合征上的患儿可行PTPN11等相关基因检测，由于Noonan综合征遗传异质性较强，基因检测阴性结果并不能排除Noonan综合征。更多还原

【Abstract】 Object:To study the distribution of growth hormone receptor (GHR) Exon3polymorphism in Chinese Turner syndrome patient, to explore the relationship between the polymorphism and the Recombinant human growth hormone (GH) treatment efficacyMaterial and methods:This was a retrospective study of31Turner syndrome, GH had been administered to the31patient at least one year, nine of which has been treated for two years, the change of height standard deviation (ΔHt-SDS), the change of growth rate (ΔGV) and height change (ΔHt) as indicators to measure GH treatment efficacy, Genomic DNA was isolated from peripheral blood lymphocytes, The GHR Exon3polymorphism was detected by PCR assay, Hardy-Weinberg genetic equilibrium method to test the theoretical value of each genotype, the data was analysed by the SPSS12.0software(t test, Chi-square test, pearson correlation analysis and multiple linear regression analysis)Results:(1) the distribution frequency of the fl/fl-GHR, fl/d3-GHR d3/d3-GHR in this study were41.9%,45.2%and12.9%, respectively, there was no difference.(2) Before the growth hormone therapy and after1year of treatment, the growth rate (GV,2.94±0.89vs7.36±1.34) and Height SD Score (Ht-SDS,-3.47±1.33vs-2.78±1.24) of the31Turner syndrome children are significantly different (p<0.05)(3) In the nine Turner syndrome patient who were treated for two years, the growth velocity of the year before treatment, the first year treatment and the second year treatment was (3.39±0.56vs7.98±0.73vs7.07±0.98)respectively, There were statistical significance between the them(p<0.05). The Ht-SDS of the time begin to treatment, the first year and the second year treatment was (3.48±1.51vs2.48±1.25vs1.01±1.02) respectively, there was no significant difference between the first year treatment and the time begin to treatment(p>0.05), there was significant difference between the second year treatment and the first year as well as the time begin to treatment(p<0.05).(4)There were no difference in Δ GV;ΔHt;ΔHt-SDS and rhGH dose between the three group(fl/fl-GHR vs fl/d3-GHR vsd3/d3-GHR) at the first year treatment(p>0.05).(5) After two years treatment,there were no difference the second year ΔGV; total two year ΔHt and the two year ΔGV between the fl/fl-GHR group and(fl/d3-GHR+d3/d3-GHR) group(p>0.05).(6) According to the karyotype,the31patient were devided into three group:45x karyotype group, chimeric karyotype group and structural aberrations karyotype group, the ΔHt-SDS of the three group were0.54±0.35SDS、0.86±0.44SDS and0.56±2.44SDS,the ΔHt-SDS was significant difference between the45x karyotype group and chimeric karyotype group(p<0.05),there were no significant difference between the other two group. there were no difference in ΔGV and ΔHt between the three group.(7)In the patient who were treated for one year, according the pearson analysis, the ΔHt-SDS was correlation with start age, Ht-SDS, BMI and GV before treatment (p<0.05), ΔHt-SDS wasn’t correlation with BA, GHR genotype,rhGH dose (p>0.05). there was no correlation between the ΔGV and the parameters. according to Linear regression, the ΔHt-SDS was correlation with start age, GV(p<0.05), regression coefficient was-0.096and0.217,respectively.(9) In the patient who were treated for two year, according to the pearson analysis, Δ Ht-SDS wasn’t correlatGVo,vsBMI,vs BA, vs growth hormone dose were greater than0.05, The p value of pearson analysis in ΔGV vs on with GVo,, BMI, BA, and rhGH dose(p>0.05)Conclusion:In this study, the distribution of GHR exon3deletion polymorphism frequency in Chinese Turner syndrome are different from foreign Turner syndrome, which indicated that the GHR genotype distribution may have ethnic variations, There was no significant correlation between the GHR Exon3polymorphism and the effect of GH treatment in Chinese Turner syndrome, Karyotype may be associated with the efficacy of GH therapy, The efficacy of growth hormone was negatively correlated with initial treatment age and was positively correlated with untreated growth velocity. Objective:analysis the clinical characteristics and detect the PTPN11gene in4clinically suspected Noonan syndrome patients.Methods:Summary clinical manifestations of4cases who were suspected Noonan syndrome, Analysis of the karyotype, we select the PTPN11gene for the purpose gene,PTPN11gene hotspot mutations in exons (2,3,4,7,8,12,13) was amplified by PCR, the PCR product was sequenced by ABI3730sequence analyzer to obtain the peak of FIG. DNA Baser sequence assembly software analysis sequencing results showed that the mutation site, and finally in the human Genome mutation Database (HGMD) View the mutation site is the causative mutation.Results:Four infants have short stature, Down-slanting palpebral fissures, ptosis, orbital hypertelorism, Cubitus valgus and congenital heart disease, chromosome karyotypes were normal,2patients PTPN11gene sequencing were normal,2patients in the presence of disease-causing mutations in the PTPN11gene, in patient A, The A→G transition at position1510in PTPN11Exon13, predicting an met504val substitution. in patient B, The A→G transition at position236in PTPN11Exon3, predicting an Gln79Arg substitution, Two mutations are reported pathogenic mutations of the Noonan syndrome.Conclusion:The clinical feature of Turner syndrome,GHARGE syndrome, Costello syndrome similar to Noonan syndrome, PTPN11gene is the most common disease-causing gene in Noonan syndrome, in children who was suspected with Noonan syndrome can carry on the PTPN11gene testing, Noonan syndrome is a genetically heterogeneous disease, negative result does not rule out Noonan syndrome.更多还原