糖原累积病(glycogen storage diseases,GSD)是由糖原代谢过程中酶的缺乏以及蛋白转运障碍所引起,以各组织中糖原累积为特点,尤以肝脏和肌肉为主的一组异质性的遗传性疾病.本文报告1例以间断便血为首发表现的1例GSD患儿资料.

Objective To investigate the long-term effects of GnRHa treatment on final height gain,gonadal function,and body mass index（BMI） in children with central precocious puberty （CPP） or early and fast puberty （EFP）,and to explore the influencing factors of height gain and early predictors.Methods Fifty patients with CPP and 44 patients with EFP who were treated with GnRHa for more

Background Hypospadias is a common congenital malformation of male external genitalia, which mainly manifests as an abnormal urethral opening on the ventral side of the penis. The etiology and clinical phenotype of hypospadias is highly heterogeneous, and its clinical diagnosis is challenging. Currently, over 70% of patients have an unknown etiology. Here, we performed a targeted analysis of gene mutations in 130 patients with hypospadias of unknown etiology to find the precise genetic cause. Methods We developed a targeted next-generation sequencing (NGS) panel, encompassing the exon coding regions of 105 genes involved in external genitalia and urogenital tract development and performed sequencing analysis on 130 children with hypospadias of unknown etiology. Results In total, 25 patients with hypospadias (19.2%) were found to have 20 mutations among the nine genes involved in external genitalia and urogenital tract development, including 16 reported and four novel mutation sites. Twenty-two patients (16.9%) had diagnostic variants. Multiple genetic mutations were identified in three of the 25 patients. Hypospadias combined with micropenis was the most common phenotype (68%) in 25 patients. Conclusions Higher frequency mutations were identified in SRD5A2 (52%) and AR (24%) in our patient cohort. Middle or posterior hypospadias with micropenis may be significant indicators of genetic variations. Polygenic inheritance may be a rare genetic cause of hypospadias.

Objective To explore the etiology and clinical characteristics of short stature. Method Clinical data of 2075 children with short stature treated from May 1995 to July 2017 were retrospectively analyzed. The etiology and morbidity of pathological short stature and normal variant short stature were analyzed. The clinical characteristics of growth hormone deficiency (GHD) , idiopathic short stature (ISS) , constitutional delay in growth (CDG) and familial short stature (FSS) were analyzed. The etiological differences between severe short stature [height standard deviation score (SDS) ≤-3] and general short stature (height SDS＞-3) were analyzed. Results Among 2075 children diagnosed with short stature, 1719 (82.84％) were pathological short stature, among which GHD (38.60％) and ISS (22.02％) were more common. Normal variant short stature was found in 356 children (17.16％) , with FSS and CDG accounting for 10.70％ and 6.46％ respectively. There were statistically significant differences in the sex ratio, age at initial diagnosis, height SDS, body mass index (BMI) , bone age and bone age delay among children with four common childhood short stature (GHD, ISS, CDG and FSS) (all P＜0.01) . Boys were more than girls in four kinds of childhood short stature. The height SDS was the lowest in GHD group and the highest in CDG group; BMI was highest in GHD group, but lower in CDG and ISS group. Bone age delay was highest in GHD group and lowest in CDG group. In severe short stature group, the rates of complete GHD, multiple pituitary hormone deficiency, small for gestational age infant, Turner syndrome, hypothyroidism and Russell-Silver syndrome were higher than those in general short stature group, but the rates of partial GHD, ISS, FSS and CDG was lower than those in general short stature group. Conclusion The etiology of short stature is complex. Analysis of the etiology and clinical features is helpful for clinical diagnosis and treatment.

BACKGROUND:Dwarfism is a common severe growth disorder, but the etiology is unclear in the majority of cases. Recombinant human growth hormone may be a treatment option, but it has limited efficacy. The currently known laboratory assays do not meet the precision requirements for clinical diagnosis. Here, we have constructed a targeted next-generation sequencing (NGS) panel of selected genes that are suspected to be associated with dwarfism for genetic screening.METHODS:Genetic screening of 91 children with short stature of unknown etiology was performed with the help of the NGS panel. All the coding regions and exon-intron boundaries of 166 genes were included in the panel. To clarify the pathogenicity of these mutations, their clinical data were reviewed and analyzed.RESULTS:The assay identified p.A72G, p.I282V, and p.P491S variants of the PTPN11 gene and a p.I437T variant of the SOS1 gene in 4 cases with Noonan syndrome. A frameshift mutation (p.D2407fs) of the ACAN gene was identified in a case of idiopathic short stature with moderately advanced bone age. A p.R904C variant of the COL2A1 gene was found in a patient, who was accordingly diagnosed with Stickler syndrome. Severe short stature without limb deformity was associated with a p.G11A variant of HOXD13. In addition, we evaluated evidence that a p.D401N variant of the COMP gene may cause multiple epiphyseal dysplasia.CONCLUSIONS:Our findings suggest that syndromes, particularly Noonan syndrome, may be overlooked due to atypical clinical features. This gene panel has been verified to be effective for the rapid screening of genetic etiologies associated with short stature and for guiding precision medicine-based clinical management.

The clinical features and laboratory data of two patients with congenital lipoid adrenal hyperplasia (CLAH)were collected. The genomic DNA was extracted from the peripheral blood white cells in the two patients and their family members and the STAR gene was screened for mutations by PCR and Sanger sequencing. Patient 1 was a girl aged 2 years and 8 months,and she visited us because of continual cyanosis for more than two years. Physical examination showed no obvious pigmentation or clitoral hypertrophy,and Tanner stage was B1P1. Clinical examination revealed serum ACTH 1 284.1 pg/ml and 17α-hydoxyprogesterone(17-OHP)0.54 ng/ml, with Karyotype 46, XX. Genetic analysis showed compound heterozygous mutations of c.201_202delCT and c.229C>T in the STAR gene. Her father carried heterozygous c.201_202delCT mutation, and her mother showed heterozygous c.229C>T mutation. Patient 2 was a girl aged 22 years and referred to us because of dark skin for more than 21 years. Physical examination revealed generalized hyperpigmentation,with Tanner stage B5P2. Hormone examination showed ACTH>2 000 pg/ml and serum cortisol 0.77μg/dl. Karyotype analysis revealed 46,XX. Genetic analysis found compound heterozygous mutations of c.64+1G>C and c.707_708delinsCTT in the STAR gene,which descended from her father and mother respectively. Of note,c.64+1G>C is a novel splicing mutation of STAR gene.

PTPN11 is the most common mutation gene of RAS disease, which is located in the upstream of RAS/MAPK pathway and participates in signal transduction. Because the molecular mechanism of RAS's disease involves the same pathway, it may present a certain commonality in clinic, but the different genotypes with PTPN11 mutation may also express different phenotypes. Therefore, it is not easy to identify and diagnose this disease early in clinic. The present article aims to analyze the correlation between the clinical phenotype and genotype of 4 patients with RAS disease.

Objective To detect pathogenic genes of short stature with unknown etiology by a targeted next generation sequencing panel to analyze the correlation between genotypes and clinical phenotypes.Methods A total of 77 children diagnosed with unexplained short stature were enrolled for the study.These children were treated in Ruijin Hospital of Shanghai Jiao Tong University from 2007 to 2015.To search for genetic variation in 187 candidate genes which were associated with growth,we constructed a targeted next generation sequencing panel encompassing the coding regions of 187 genes.According to ACMG Guidelines,the sites of variation were determined.Sanger sequencing was used to verify the suspected pathogenic genes variation.The relationship between genotype and clinical phenotype was analyzed.Results Including 5 pathogenic variants,one likely pathogenic variant and one variant of uncertain significance,we identified 7 heterozygous variants of 7 cases in 77 cases of short stature with unknown etiology.A pathogenic variant p.D2407fs of ACAN gene was found in a case with advanced bone age.There were 3 reported pathogenic variants,including p.A72G,p.I282V and p.P491S of PTPN11 gene,which were diagnosed as Noonan syndrome.A case carrying known pathogenic variant COL2A1 (p.R904C) was diagnosed as Stickler syndrome.We still got one likely pathogenic variant COMP (p.D401 N),which could cause multiple epiphyseal dysplasia.There was a familial short stature of delayed bone age carrying a variant (p.S289Y) of uncertain significance,in which the genotype was in accordance with the clinical phenotype.Conclusion The ACAN gene defection is associated with the idiopathic short stature with advanced bone age.The likely pathogenic variant COMP(p.D401N) may cause multiple epiphyseal dysplasia.The newly-found heterozygous varians(p.S289Y) of GHSR gene may result in short stature,which needs further function verification.

目的 分析混合性性腺发育不良(mixed gonadal dysgenesis,MGD)伴外生殖器畸形患儿的临床特征及其与分子遗传学的相关性.方法 收集2002年1月至2014年12月因外生殖器畸形就诊于上海交通大学医学院附属瑞金医院儿内科5例患儿病例资料,其染色体为45,X/46,XY嵌合体或包含45,X/46,XY的其他嵌合体.分析其临床特征并进行相关辅助检查,采用多重连接依赖式探针扩增技术(multiplex ligation-dependent probe amplification,MLPA)检测外周血DNA中Y染色体微缺失情况及性发育过程相关基因拷贝数变化.结果 5例MGD患儿临床表型不一,其中3例为男性抚养,2例女性抚养.4例存在不同片段及数量的Y染色体微缺失,5例均未检测到WNT4、NR5A1、SOX9、Cxorf21的拷贝数异常.结论 MGD临床表型谱广泛,外生殖器畸形严重程度不一.临床表型和外周血染色体核型嵌合比例之间无明显关联.Y染色体微缺失在MGD中的发生率很高,微缺失的范围大小可能和外生殖器的男性化程度正相关.

目的 探讨46,XY外生殖器畸形的临床及分子遗传学特征,并分析二者相关性.方法 2004年1月至2014年12月因外生殖器畸形就诊于上海交通大学医学院附属瑞金医院儿内科的染色体为46,XY的患儿77例,收集患儿的临床资料,分析总结其临床特征及实验室检查特点,采用候选基因分析策略,选择15个候选基因,利用依托Illumina-Miseq测序平台的二代测序(next generation sequencing,NGS)技术对候选基因的全外显子测序筛查突变.结果 77例46,XY外生殖器畸形患儿临床表型多样.NGS检测到18例患儿存在候选基因有氨基酸改变的突变位点或多态性位点,其中9例患儿存在6种5α-还原酶2型(steroid-5-alpha-reductase,alpha polypeptide 2 gene,SRD5A2)基因变异,包括一处以往未见报道突变位点.在4例患儿中检测到4种雄激素受体(androgen receptor,AR)基因的变异,包含1处新发现突变.2例患儿检测到MAMLD1基因的1处纯合多态性位点.1例患儿中存在INSL3基因的复合杂合改变.1例患儿检测到MID1基因纯合突变.另有1例患儿检测NR5A1基因多态性纯合改变.结论 46,XY外生殖器畸形临床表现多样,以严重型尿道下裂为主.外生殖器发育过程中相关基因的突变在尿道下裂患儿中的检出率较高(14/64,21.8％).这其中以SRD5A2和AR的突变为最常见,MAMLD1、INSL3、MID1和NR5A1作为尿道下裂的候选基因,尽管阳性检出率不高,但是亦不可忽视.

Objective: To explore the clinical manifestations and molecular features of 46, XX male syndrome. Method: The clinical and molecular data of five 46, XX male syndrome cases treated in the Department of Pediatrics of Shanghai Ruijin Hospital form August 2010 to August 2014 were retrospectively analyzed. Result: The five patients were all sociopsychologically males and came to hospital respectively for short stature, ambiguous genitalia or gynecomastia. They were all below the normal male's average height, and their karyotype was all 46, XX. One case in five was verified as sex determining region of Y chromosome (SRY gene) positive revealed no abnormality in their external genitalia. He had short stature since childhood, whose SRY gene fragments were shown by FISH transferred to the ends of X chromosome. Three cases in four were SRY gene negative with ambiguous genitalia of cryptorchidism and testicular dysplasia to different degrees. The copy number variations of SOX9 gene was found in one case, the loss of heterozygosity area in DHH gene of one case. Another SRY gene negative patient who had normal male external genitalia, came to the hospital due to puberty gynecomastia, that of SOX9 gene and its upstream gene both increased. Conclusion: The main clinical characteristics of 46, XX male syndrome are male phenotype, 46, XX karyotype, gonad of testis or ovotestis and no uterus. In addition, short stature, ambiguous genitalia or gynecomastia can be one reason for hospital visits. SRY gene translocation, SOX9 gene and its upstream gene copy number increase all can lead to 46, XX male syndrome. The cause of some may play an important role in 46, XX male syndrome, but has not yet been determined.

Objective To analyze the clinical characteristics of two patients with 3β-hydroxysteroid dehydrogenase deficiency and to explore their molecular genetic defects.Methods The clinical features and laboratory data of two patients were collected.The exons of HSD3B2 gene were amplified by PCR and sequenced by Sanger sequencing.Results Patient 1, aged 5 yrs old, was raised as a girl with 46, XY karyotype, presented with hyperpigmentation, female infant vulva, clitoral hypertrophy, and bilateral cryptorchidism;Patient 2, aged 11 yrs old, was raised as a girl at birth but as a boy after 1 yr old for known 46, XY karyotype, presented with hyperpigmentation, micropenis and severe hypospadias.Both patients had markedly elevated adrenocorticotropin and decreased cortisol.Two homozygous missense mutations in HSD3B2 gene were identified:conversions of codon Pro155 toLeu(p.P155L)inpatient1,andcodonAla82toThr(p.A82T)inpatient2,bothofwhichwerereportedforthe first time in China.Conclusion The patients with 3β-hydroxysteroid dehydrogenase deficiency in 46,XY karyotype mainly present with male pseudohermaphroditism and adrenocortical deficiency, and the diagnosis should rely on the steroids detection and HSD3B2 gene screening.

Background The correlation between gestational hypertension-preeclampsia (GH-PE) and placenta previa (PP) is controversial. Specifically, it is unknown whether placenta previa has any effect on the various types of preeclampsia (PE), and the role PP with concurrent placenta accreta (PA) play in the occurrence of GH-PE are not well understood. Objective The aim of this study was to identify the effects of PP on GH, mild and severe preeclampsia (MPE and SPE), and early- and late-onset preeclampsia (EPE and LPE). Another aim of the study was to determine if concurrent PA impacts the relationship between PP and GH-PE. Methods A retrospective single-center study of 1,058 patients having singleton pregnancies with PP was performed, and 2,116 pregnant women were randomly included as controls. These cases were collected from a tertiary hospital and met the inclusion criteria for the study. Clinical information, including PP and the gestational age at the onset of GH-PE were collected. Binary and multiple logistic regression analyses were conducted after the confounding variables were controlled to assess the effects of PP on different types of GH-PE. Results There were 155 patients with GH-PE in the two groups. The incidences of GH-PE in the PP group and the control group were 2.5% (26/1058) and 6.1% (129/2116), respectively (P = 0.000). Binary and multiple regression analyses were conducted after controlling for confounding variables. Compared to the control group, in the PP group, the risk of GH-PE was reduced significantly by 78% (AOR: 0.216; 95% CI: 0.135–0.345); the risks of GH and PE were reduced by 55% (AOR: 0.451; 95% CI: 0.233–0.873) and 86% (AOR: 0.141; 95% CI: 0.073–0.271), respectively; the risks of MPE and SPE were reduced by 73% (AOR: 0.269; 95% CI: 0.087–0828) and 88% (AOR: 0.123; 95% CI: 0.055–0.279), respectively; and the risks of EPE and LPE were reduced by 95% (AOR: 0.047; 95% CI: 0.012–0.190) and 67% (AOR: 0.330; 95% CI: 0.153–0.715), respectively. The incidence of concurrent PA in women with PP was 5.86%; PP with PA did not significantly further reduce the incidence of GH-PE compared with PP without PA (1.64% vs. 2.51%, P>0.05). Binary logistic regression analyses were conducted after controlling for confounding variables, compared with the non-PP + GH-PE group, and the AOR of FGR in the non-PP + non-GH-PE group was 0.206 (0.124–0.342). Compared with the PP + GH-PE group, the AOR of FGR in the PP + non-GH-PE group was 0.430 (0.123–1.500). Conclusion PP is not only associated with a significant reduction in the incidence of GH-PE, but also is associated with a reduction in incidence of various types of PE. Concurrent PA and PP do not show association with a reduction in incidence of GH-PE.

Objective To investigate the clinical characteristics and molecular pathological mechanism of McCune-Albright syndrome ( MAS) in order to provide a guidance for the precision medicine of MAS. Method The clinical data and genetic findings of 41 patients with MAS were analyzed retrospectively. Results (1) MAS girls had the phenotype of peripheral precocious puberty with premature sexual development and high estradiol, low LH and FSH, and the increased volume of uterus and ovary. ( 2 ) In 41 MAS cases, there were 17 cases with GNAS1 gene mutation, and the total positive rate was 41. 5%, of which the classic triad was 66. 7%, two signs 56. 3%, and 12. 5% in only one classic sign. GNAS1 gene mutation was found in 78. 6% of patients with polyostotic fibrous dysplasia of bone, while only 55. 0% in patients with cafe au lait skin spots. Children with precocious puberty and fibrous dysplasia of bone is an important basis for clinical diagnosis of MAS, but cafe au lait skin spots does not seem to be the specifical manifestation of MAS. Conclusion Clinically MAS was lack of typical clinical manifestations, and the most important clinical weight factor for the diagnosis of MAS was peripheral precocious puberty with fibrous dysplasia of bone. GNAS1 gene screening may be helpful to improve the clinical accurate diagnosis of MAS.

Background: The mini-pubertal period of almost six hours in neonatal male rats is thought to be an important stage in sexual development. Objectives: The aim of this study was to investigate the effect of hormone inhibition during mini-puberty on testicular function in male rats. Materials and Methods: We measured serum testosterone (T), luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels in male rats at different time points after birth by enzyme linked immunosorbent assay (ELISA) and established a “no mini-puberty” model by ether inhalation. The mRNA expression levels of testicular androgen receptor (AR), insulin-like growth factor 3 (INSL3), anti-Mullerian hormone (AMH), and ghrelin were determined by real-time polymerase chain reaction (PCR) assays on postnatal days 45 and 75. Testicular tissue biopsies were stained with hematoxylin and eosin (H & E) and the structure, number and maturity of testis cells (including spermatogenic, Sertoli, and Leydig cells) were observed under microscopy at the same time. Results: Serum T and LH levels peaked at two hours after birth, while FSH peaked at hour 0, and bilateral testicular weight peaked at four hours after birth. The rats that underwent ether inhalation five minutes after birth had markedly reduced serum hormone levels. The mini-puberty model group revealed visible morphologic alterations in the tests on postnatal day 45. Then, on postnatal day 75, the mRNA expression level of AMH significantly decreased (P < 0.05) in the same group. Conclusions: The inhibition of mini-puberty period in male rats was demonstrated to have an effect on their testicular function to some extent.

Objective To detect the relationship between the molecular defects and their phenotypes in children with growth hormone insensitivity syndrome (GHIS).Methods 21 patients defined as GHIS were enrolled in the study.4 candidate genes (GHR,IGFALS,JAK2,and STAT5B) were analyzed by genomic DNA sequence screening and clinical relevance analysis.Results The statistical descriptions of the patients were showed as an average height standard deviation (SDS)-4.33 ± 1.91 (-9.17 to-2.21),average serum peak values of GH (22.67 ±20.98) tg/L (11.33 to 104.21 μg/L),basal serum insulin-like growth factor-Ⅰ SDS-2.65 ± 0.53 (-3.57 to -1.79),insulin-like growth factor-binding protein 3 SDS-1.77 ± 1.64 (-4.13 to 0.96).Bone age of backward difference (chronological age-bone age) (43.10 ± 19.54) months (6 to 82 months).One of two children with severe growth failure and mid-face hypoplasia was found to a homozygote for G to A gene mutation in the intron 6 splice donor consensus sequences (IVS6 ds+ 1 G-A) in the GHR gene,causing its functional defect.3 cases with mild dwarf were found gene variations as novel finding:c.1097T＞C c.1098C＞T p.V366A pathogenic variant,c.1229C＞T p.S410L and nt1843707 A→G of 5' UTR region in the IGFALS gene.JAK2 and STAT5b genes mutations were not found.Conclusion Molecular pathology of GHIS is considered as involving the defects of GHR and its signal pathway.The mutation of intron 6 splice donor sequences in GHR gene has been reported which affect the function of GHR.The 3 novel type base variants in IGFALS gene,causing non severe dwarfism,might be suspected with pathogenic roles of GHIS.

Objective:To establish the rat model of gestational diabetes ( GDM ) in-duced by administering Streptozotocin( STZ) intraperitoneally with low dosage in single injec-tion to investigate the proper method for duplicating reliable GDM model in pregnant rats. Method:Twenty virgin Sprague-Dowlgy( SD) rats of comparable pre-weight were used for the experiments. The control group and the study group comprised 10 rats respectively. The study group was made diabetes by a single i. p injection of STZ (25mg/kg body wt) dissolved in 0. 1 M citrate buffer,PH 4. 2 on the first day of gestation,while the control group was injected with the buffer only at the same gestation age. Fasting and non-fasting blood glucose was determined on day 4,day 11 and day 18 of pregnancy. An oral glucose tolerance test(OGTT) was per-formed on the 18th day in each group to evaluate the degree of insulin resistance. On the 21st day the rats were sacrificed. The blood serums were determined for blood insulin and the pan-creas was removed for examination for morphological and ultra structural changes. Results:Non-fasting blood glucose in the study group was much higher than that in the control group. The level of blood glucose and insulin during OGTT in the study group was higher than that in the control group(P<0. 05). ISI in the study group was significantly higher(P<0. 001),but InsI was much lower than that in the control group(P<0. 001). The mild ?-cell damage in the study group was observed by means of the H-E staining. Conclusion:A single i. p injection of STZ may produce signs similar to those of GDM,such as higher levels in blood glucose and insulin, higher insulin resistance,lower insulin secretion and the mild ?-cell damage. Consequently,a single and low dose STZ could duplicate the rats model for GDM.

Objective To explore the causes of ambiguous genitalia.Methods Clinical data of 106 cases with ambiguous genitalia from Ruijin Hospital of Shanghai Jiaotong University School of Medicine were retrospectively analyzed.DNA fragments of related genes from parts of patients were amplified by means of polymerase chain reaction (PCR) and were directly sequenced to detect gene mutations.Results (1)The 106 ambiguous genitalia patients presented a variety of clinical phenotypes.Karyotype of 42 cases(39.6％)were 46,XX,while 62 cases(58.5％)were 46,XY and 2 cases(1.9％)were abnormal.(2)Forty(95.2％)patients with 46,XX were diagnosed with congenital adrenal hyperplasia(CAH) ;one case(2.4％) was adrenal cortical tumor and one case (2.4％) was 46,XX [sex determining region of Y choromosome (SRY) positive] male syndrome.(3) Fifty-three cases (85.5 ％) out of 46,XY karyotype were directly sequenced with steroid-5-alpha-reductase,alpha polypeptide 2 gene (SRD5A2),androgen receptor gene (AR) and steroidogenic factor-1 gene(SF-1).Sequencing analysis of SRD5A2 revealed 8 patients with compound heterozygous or homozygous mutations.A patient carried a novel missense mutation of SF-1 and another patient had a mutation of AR.(4) One abnormal karyotype was 46,XX/46,XY and the other was 46,XX/46,XY/46,X.+ may.ish (DYZ3 +) (DXZ1-).Conclusions (1) CAH is the most common cause of genital ambiguity in 46,XX patients but some rare causes such as adrenal cortical tumors or SRY positive should not be ignored.(2) To find the causes of 46,XY genital ambiguity,direct DNA sequencing analysis of candidate genes would be the better choice because of the complicate pathogenesis.(3)Abnormal karyotype also can lead to ambiguous genitalia.