目的 比较宫内发育迟缓(intrauterine growth restriction,IUGR)新生儿与适于胎龄儿(appropriate for gestational age,AGA)代谢产物水平的差异,探讨新生儿代谢产物的变化与IUGR的联系,以及新生儿组织和器官损害的可能的代谢机制,为临床干预提供理论依据.方法 研究纳入2015年12月至2019年12月至江西省儿童医院新生儿科住院的320例I-UGR新生儿与302例同期住院的适于胎龄儿,完善血串联质谱检查,比较两组新生儿代谢产物浓度的差异.结果 与对照组相比,氨基酸代谢中缬氨酸、瓜氨酸、苯丙氨酸、脯氨酸浓度明显增加,差异有统计学意义;肉碱代谢中游离肉碱、乙酰肉碱、丙酰肉碱、丁酰肉碱、已酰肉碱、辛酰肉碱、月桂烯酰肉碱、肉豆蔻酰肉碱、肉豆蔻烯酰肉碱、棕榈酰肉碱、棕榈烯酰肉碱、十八碳酰肉碱与十八碳烯酰肉碱浓度明显增加,差异有统计学意义.结论 在IUGR新生儿中存在氨基酸和肉碱、酰基肉碱的代谢异常.

Objective: To explore the application value of mass spectrometry (MS) combined with next generation sequencing (NGS) in diagnosing neonatal inherited metabolic diseases (IMD). Methods: The clinical information, metabolites in blood and urine, and gene sequencing results of 19 neonates with IMD coming from the Department of Neonatology of Jiangxi Provincial Children's Hospital from March 2017 to September 2019 were analyzed retrospectively. The metabolites in blood were detected by liquid chromatography tandem mass spectrometry and urine were detected by gas chromatography-mass spectrometry respectively.Meanwhile, the whole bloods were dectected by neonatal genetic disease panel based on NGS. Results: Twelve neonates had the same results between MS and NGS among the 19, 2 had different results from MS to NGS, and 4 had no disease indication by MS but were diagnosed by NGS whose clinical phenotype were partially consistent with NGS results. One of them who did not carry out MS was considered as the diagnosis of IMD because of the detection of gene, and was followed up on this basis. Conclusion: MS could diagnose IMD relatively quickly to guide clinical treatment, and while NGS could verify the results of MS detection. Combination of MS and NGS would understand the cause of disease on genetic level, so as to guide further treatment and genetic consultation.

2020年2月29日是第十三个国际罕见病日,目前世界上有超过7000种罕见病,其共同特征如下:⑴对人体危害大;⑵对症治疗的药物少导致药物难寻;⑶几乎没有有效的根治方法.由于罕见病的特殊性,加之儿童疾病诊断的复杂性,儿科疑难罕见病异戊酸血症(IVA)的诊治过程可谓"难上加难",大多数罕见病进行正确的遗传咨询和产前诊断可有效预防.近年来基因组学、代谢组学的发展为诊治IVA提供了广阔前景.

OBJECTIVE:To detect potential mutation of TCOF1 gene in a Chinese family affected with Treacher-Collins syndrome.METHODS:Clinical data of the patient was collected. The analysis included history taking, clinical examination and genetic testing. All coding regions of the TCOF1 gene were subjected to PCR amplification and Sanger sequencing.RESULTS:A novel mutation c.2261ins G (p.E95X) of the TCOF1 gene was discovered in the patient. The same mutation was not found in his parents and 100 healthy controls.CONCLUSION:The c.2261insG (p.E95X) mutation of the TCOF1 gene probably underlies the disease in the patient. Genetic testing can facilitate diagnosis and genetic counseling for families affected with TCS.

Objective:To study the expression and interaction between miR-194 and PTPN12 in the process of age-related atrophy of thymus for clarifying the regulatory mechanism in the process of this disease.Methods:C57BL/6 mouse were divided into 4 groups as 1 month,6 months,10 months and 19 months old and each group has 6 cases.Thymus tissue was removed and thymic stromal cells were isolated.And thymus epithelial cells were washed out by CD45 antibody and LS column after anesthesia.Fluorescence quantitative real-time PCR and Western blot were used to detect the changes of miR-194 and PTPN12 gene expression in thymus epithelial cells with aging.miR-194 and PTPN12 luciferase reporter vectors were transfected into HEK293 cells,and the auto fluorescence values were detected at 24 h and 48 h,respectively in vitro.Results:The expression level of miR-194 decreased (P<0.05),while the expression level of PTPN12 mRNA increased (P<0.05) as the age increased.And the correlation between miR-194 and PTPN12 mRNA expression was found to be negative(P<0.05).In vitro,luciferase reporter gene results show that miR-194 has a direct effect on the 3'UTR region of PTPN12 gene and had the highest binding efficiency in 48 h.Conclusion:PTPN12 is one of the target genes of miR-194,which is involved in the aging process of thymus and is an important factor regulating the function of thymic ep-ithelial cells.

目的 探讨FLT3-ITD在AML患儿的临床意义.方法 收集我院2013年-2015年收治的45例AML患儿骨髓样本,提取全基因组DNA,采用PCR扩增产物克隆测序法检测FLT3-ITD基因突变情况,解析序列,整理并分析其插入的重复序列,同时分析FLT3-ITD阳性患儿的临床特征及对预后的影响.结果 45例初诊AML患儿中,FLT3-ITD突变阳性患儿4例(8.9%),表现为杂合突变和纯合突变,解析出4条插入序列,插入长度范围为18-99bp.阳性患儿伴染色体正常核型1例,异常核型3例,突变型患儿白细胞及骨髓原始细胞比例高于野生型患儿,差异有统计学意义,但在性别、年龄、血红蛋白、血小板无显著差异.结论 FLT3-ITD突变检测有助于急性白血病基因分型,是儿童AML预后不良的一个危险因素,可作为预后的分子标志物.

目的 检测环氧化酶-2(COX-2)基因启动子区CRE位点甲基化状况,探讨其在子宫内膜异位症(EMs)发生中的作用.方法 收集南昌大学第一附属医院40例子宫内膜异位症患者的子宫内膜作为病例组,同期40例正常子宫内膜作为对照组,采用焦磷酸盐测序技术检测病例组和对照组子宫内膜组织中COX-2基因启动子区CRE位点甲基化频率;采用实时定量PCR及免疫组化技术分别检测病例组和对照组子宫内膜组织中COX-2 mRNA及蛋白的表达;分析COX-2基因启动子区CRE位点甲基化频率与COX-2表达水平的相关性.结果 对照组内膜组织COX-2基因启动子区CRE位点的甲基化频率(40.81％±6.10％)显著高于病例组(14.74％±2.84％),P＜0.05;而对照组COX-2 mRNA表达水平(0.98±0.37)及蛋白表达水平(1.00±0.73)显著低于病例组的mRNA表达水平(2.63±0.44)及蛋白表达水平(2.74±1.18),P均＜0.05.随着COX-2基因启动子区CRE位点甲基化频率升高,COX-2基因表达水平下降,二者呈负相关(P＜0.05).结论 COX-2基因启动子区CRE位点去甲基化与EMs患者在位内膜COX-2高表达相关,在EMs的发生、发展中具有重要作用,提示子宫内膜异位症是一种表现遗传性疾病.

患儿 女 ,9个月 ,系第3胎第3产 ,足月顺产 ,无窒息史.3个月会抬头 ,7个月会坐 ,现不会爬.无反复发热、腹泻和其他慢性疾病史.无多饮多尿 ,无夜间哭闹.体检 :身高 65 .6 cm,体重7.0kg,均低于同龄同性别儿童的第三个百分位数.毛发偏多 ,前囟2.5cm × 2.5cm,塌鼻梁 ,有颈蹼 ,盾状胸不明显 ,轻度肋外翻 ,心肺听诊无特殊 ,腹部平软 ,肝脾肋下未触及 ,神经系统无阳性发现 ,四肢较粗短.

目的：探讨串联质谱（MS／MS）技术在遗传代谢病（IEM）筛查工作中的应用价值。方法利用 MS／MS 技术对4135例可疑 IEM 患儿的血液样本进行氨基酸、酰基肉碱谱检测筛查。结果4135例患儿筛查发现阳性病例102例，阳性率为2．5％。其中有机酸代谢病20例，占19．6％（20／102）；氨基酸代谢病30例，占29．4％（30／102）；脂肪酸氧化障碍疾病52例，占51．0％（52／102）。结论MS／MS 技术在 IEM 的筛查诊断中高效、准确，是 IEM 筛查的有效工具。

Objective To study the clinical feature and genetic mutations of alkaptonuria(AKU).Methods One patient was diagnosed with AKU disease via gas chromatography mass spectrometry (GC/MS) in the investigated family.All exon of homogentisate 1,2 dioxygenase(HGD) gene were amplified in the family by means of polymerase chain reaction(PCR) and followed by using direct DNA sequencing and Polyphen software was used to predict protein function.Results The infant only had red-brown urine,with no skin,joints,or viscera lesion,and GC/MS suggested AKU.Two heterozygous mutations of AKU were identified c.34A ＞ C(p.N12H) in exon 2 c.240A ＞ T(p.Q80H) in exon 4 and c.910A ＞ G(p.K304E) in exon 12 from the proband.The heterozygous change c.34A ＞ C(p.N12H) in exon 2 was found in the proband's mother and sister with normal phenotype.The proband's father had the heterozygous change c.910A ＞ G(p.K304E) in exon 12.Conclusions The proband is hetero-zygous compound AKU disease patient carrying on one allele with the c.34A ＞ C(p.N12H) mutation inherited from his mother and the other allele with the c.910A ＞ G(p.K304E) from his father.The parents and his sister are heterozygous carrier with normal phenotype.The p.N12H and p.K304E mutations are novel mutations not reported around world yet,which has the earliest onset age of AKU with the only symptom of dark urine.

目的:探讨甲硫氨酸腺苷转移酶2A(MAT2A)在胃癌组织及癌旁组织中的表达及意义.方法:分别应用real-time PCR及Western blot技术检测48例胃癌组织和30例癌旁正常胃组织中MAT2A mRNA与蛋白的表达.结果:real-time PCR与Western blot结果显示,胃癌组织中MAT2A mRNA相对表达量明显高于癌旁正常胃组织(3.99±2.52 vs.1.17±1.04);MAT2A蛋白相对表达量明显高于癌旁正常胃组织(0.62±0.05 vs.0.20±0.02),差异均有统计学意义(均P＜0.05).结论:MAT2A在胃癌组织中表达上调,提示其在胃癌发病机制中起了重要的作用,其作用机制值得进一步研究.

目的 探讨miR-181a在胸腺增龄性萎缩过程中的表达,及其与靶基因ZFP3612的相互作用关系,阐明胸腺增龄性萎缩过程分子调节机制.方法 C57BL/6小鼠分为1月龄组、9月龄组、15月龄组,每组各6只.取出胸腺并分离细胞,经CD45抗体染色,LS柱吸附洗脱筛选出胸腺细胞.实时荧光定量PCR法检测胸腺细胞中miR-181a与靶基因ZFP3612随年龄增长的表达变化趋势,并进行相关性分析.体外应用HEK293细胞转染荧光素酶报告载体,48 h后检测自发荧光值.结果 随年龄增长,miR-181a呈现表达下调趋势(P＜0.05),而靶基因ZFP3612则呈现出表达上升趋势(P＜0.05).对miR-181a与ZFP3612表达量进行相关性分析,发现两者呈负相关性(P ＝0.04).体外细胞转染荧光素酶报告基因证实,miR-181a与ZFP3612 3'UTR区域结合(P＜0.05).结论 miR-181a与靶基因ZFP3612与胸腺增龄性萎缩过程密切相关,是调节胸腺细胞功能的重要因子.

目的 研究儿童急性淋巴细胞白血病(acute lymphoblastic leukemia,ALL)细胞与分子遗传学特征及临床意义.方法 应用常规细胞遗传学、实时定量PCR、荧光原位杂交等技术,对178例儿童ALL细胞与分子遗传学特征进行分析,并探讨其临床意义.结果 178例儿童ALL遗传学异常者92例占51.69％,其中融合基因TEL-AML1/t(12;21) (p13;q22)阳性35例(38.04％),BCR-ABL/t (9;22) (q34;q11)阳性9例(9.78％);E2A/PBX1/t(1;19) (q23; p13)阳性9例(9.78％);MLL-AF4/t(4;11) (q21;q23)阳性3例(3.26％;),HOX11L2/t(5;14) (q35;q32)阳性7例(7.61％);SIL-TAL1(ip32-)阳性5例(5.43％)、E2A-HLF/t (17;19) (q22;p13)阳性1例(1.09％).复杂核型异常4例(4.35％),包括:t (8; 14) (q24; q32),der (15)t (1; 15) (p11; q11);t(6; 10) (p25; p11),11p+;t(2;14) (p11;q32),9p-;t(9; 11) (q21; p15).高超二倍体(48～65) 17例(18.48％),近三倍体(65～78)2例(2.17％),其它有2p-/5p-/6p+/9p-/9q-/10p-/11p+/17p+/+5/+8等.结论 儿童ALL具有其特有的细胞与分子遗传学特征,临床上将常规细胞遗传学、分子生物学、荧光原位杂交等技术等相结合,将对白血病的诊断、治疗选择和预后判断及微小残留病检测等具有十分重要的意义.