Objective To investigate the incidence of follicular stimulating hormone receptor (FSHR) gene C566T mutation in Chinese women with premature ovarian failure (POF) and to explore the etiologies of POF. Methods This case-control study was carried out between 73 Chinese women with idiopathic POF (POF group) and 35 controls (control group), including 25 normal females with a regular menstrual history and 10 normal post-menopause women. DNA was extracted from the peripheral blood of patients and controls. The exon 7 of FSHR gene was amplified by PCR. PCR products were subsequently digested by the enzyme BsmI and then subjected to electrophoresis on agarose gels and stained with ethidium bromide to determine the C566T mutation. DNA samples of random sampling were further analysed by sequencing the PCR products to confirm the mutation. Results BsmI digestion resulting in two fragments of 51 and 27 base pairs was noted for all 73 POF patients and 35 controls. PCR sequencing confirmed that the 566 allele of FSHR gene is C, demonstrating normal FSHR allele. Conclusions No FSHR gene C566T mutation is present in POF patients and controls. FSHR C566T mutation may be rare in Chinese women with POF.

Objective:To report the clinical features of KCNQ2-associated epilepsy and the novel mutations and unreported clinical phenotype of KCNQ2 gene, so as to provide help for treatment selection and prognosis evaluation.Methods:Among 979 patients with epilepsy and developmental delay who were admitted to the Department of Neurology,Children′s Hospital Affiliated to Capital Institute of Pediatrics from July 2015 to October 2019, a total of 13 patients were selected from 12 families with KCNQ2 gene mutation by whole exome sequencing technology. Suspected mutations were verified by Sanger sequencing on the probands and their parents to identify the source. The clinical phenotype and genotype were analyzed according to these results.Results:Among the 13 patients with epilepsy, the onset age of four cases were older than six months [two cases in infancy (epilepsy encephalopathy), one case in early childhood (epilepsy encephalopathy) and one case in adolescence (benign epilepsy)]. Eight cases were treated with oxcarbazepine, of whom five cases were seizure free, and two cases showed partial response (>50%). Two cases treated with topiramate were seizure free. Five novel mutations were found in this research, including c.379T>G(p.Y127D), c.1A>C(initial codon mutation), c.708G>C(p.W236C), c.1027G>T(p.A343S) and c.1649T>G(p.V550G).Conclusions:Although it was rare in clinical work, the variation of KCNQ2 gene existed in patients with childhood-onset epilepsy and adolescent-onset epilepsy. Meanwhile, five novel mutations of KCNQ2 gene were reported, which further expanded its gene spectrum. This research supported that oxcarbazepine was the efficient medicine for the KCNQ2-associated epilepsy. Genetic testing showed great help to the treatment of epilepsy.

Objective To compare the clinical outcomes of gonadotropin-releasing hormone (GnRH) antagonist (GnRH-ant) fixed protocol with GnRH agonist (GnRH-a) long protocol in infertile patients with normal ovarian reserve function in their first in vitro fertilization-embryo transfer (IVF-ET) cycle,and to explore the feasibility and advantage of GnRH antagonist protocol performed in normal responders.MethodsFrom January 2011 to June 2011,771 infertile women with normal ovarian reserve function underwent their first IVF or intracytoplasmic sperm injection (ICSI) cycles in Peking University Third Hospital,which were divided into 245 cycles in GnRH-ant fixed protocol group ( GnRH-ant group) and 526 cycles in GnRH-a long protocol group ( GnRH-a group).The data of general demographic,treatment and clinical outcome were compared between two groups.ResultsAge,infertile duration,body mass index (BMI),baseline serum follicle-stimulating hormone (FSH) and estradiol levels between two groups did not reached statistical difference (P ＞ 0.05 ).The level of estradiol was (12 289 ± 6856) pmol/L in GnRH-ant group and (14934±8007)pmol/L in GnRH-a group at day of hCG injection.The mean length of stimulation was ( 10.3 ± 1.2) days in GnRH-ant group and ( 12.8 ± 1.6) days in GnRH-a group.The dose of gonadotropin was (2013 ± 607 ) U in GnRH-ant group and (2646 ± 913 ) U in GnRH-a group.The number of ovum was 15 ± 7 in GnRH-ant group and 17 ± 8 in GnRh-a group.Those clinical parameter all reached statistical difference (P ＜0.05 ).The number of embryo was 7 ±4 in GnRH-ant group and 8 ± 5 in GnRH-a group,the rate of clinical pregnancy was 40.9％ (94/230) in GnRH-ant group and 45.6％ (216/474)in GnRH-a group,the rate of implantation was 26.1％ (128/490)in GnRH-ant group and 30.9％ (307/994) in GnRH-a group,the rate of continuing pregnancy was 38.7％ ( 89/230 ) in GnRH-ant group and 42.6％ (202/474) in GnRH-a group,those parameter did not reach statistical difference (P ＞ 0.05).The rate of moderate or severe ovarian hyperstimulation syndrome was 2.4％ ( 6/245 ) in GnRH-ant group and 4.2％ (22/526) in GnRH-a group,which did not show significant difference ( P ＞ 0.05 ).ConclusionIn the first IVF or ICSI cycle of the patients with normal ovarian reserve function,the fixed GnRH-ant protocol could get the same satisfied clinical outcome,and it is more economic,convenient and safer compared with low dose depot GnRH-a long protocol.

Objective To evaluate the impact of different doses of recombinant human B-type natriuretic peptid (rh-BNP) within the dosage of clinical rage on oxygen consumption during acute myocardial infarction (AMI) with heart failure (HF). Methods AMI-HF model of York pig was established by occluding coronary artery with balloon combined with in-jecting microthrombus. Then animals were randomized into rhBNP group and control group. Clinical dose of rhBNP ( 1.5μg/kg bolus followed by a continuous infusion with speed of 0.01, 0.02 and 0.03μg · kg-1 · min-1 for 60 minutes respectively in turn) was administrated in rhBNP group while equal volume of saline was given in the control group. Myocardial oxygen up-take (MOU) was measured by drawing blood from coronary artery and coronary sinus using a catheter. Coronary diameter was determined using quantitative coronary angiography. The observation points were at baseline (T0), instant after the mod-el establishment (T1), 60 min after continuous rhBNP infusion of 0.01, 0.02, 0.03μg·kg-1·min-1 (T2-T4) respectively. Re-sults Compared with the control group, pulmonary capillary wedge pressure, central venous pressure, heart rate, systolic blood pressure and MOU were significantly decreased after rhBNP administration. And cardiac output and coronary diame-ter were obviously increased with addition of rhBNP. There is a interaction of drug intervention and time. In rhBNP group, MOU was significantly decreased with drug administraion (T2-T4 vs T1,mL O2/L: 10.61 ± 0.35,9.85 ± 0.60,9.79 ± 0.31 vs 11.59 ± 0.37). Conclusion Intravenous administration of rhBNP in AMI-HF model could decrease MOU and PCWP while increase the cardiac output.

Objective To evaluate the application value of whole exome sequencing (WES) in diagnosis of NDDs (neuro-developmental disorders) children.Metheod WES was used for the diagnosis of 35 unexplained NDD children, which admitted to the outpatient and ward of Children′s hospital affiliated to Capital institute of pediatric from November 2015 to November 2016.These children′s clinical data was collected detailedly.Using bioinformatics software tools combining with patient′s phenotype, the candidate genetic/genomic variants of these patients were identified from WES data.The final pathogenicity of genetic/genomic variants was interpreted according to the guideline of the American College of Medical Genetics and Genomics (ACMG), meanwhile, the variants validation and co-separation analysis in the parents and their family members were performed by Sanger sequencing, real time-PCR and multiplex ligation-dependent probe amplification (MLPA).Results 14 pathogenic single nucleotide variants (SNVs) and three pathogenic copy number variations (CNVs) were detected in the 35 NDD children, the detection rate in this study is 48.6%.Among the 14 pathogenic SNVs, 11 of them are the definite NDD-related genes according to OMIM database (such as CHARGE syndrome, Wiedemann-Steiner syndrome, Cockayne syndrome, etc.), and six of them are de novo (6/11, 54.6%).Three pathogenic CNVs were identified from WES data, including two microduplications and one microdeletion.Meanwhile, a female child carrying a frame shift mutation in MECP2 was found and the germline mosaicism with low-frequency mutation of this site (8.4%) was confirmed by his father's sperm.Conclusions The diagnosis rate of WES in NDDs children is 48.6% in our small-sample study.In addition to pathogenic/likely pathogenic SNVs, CNVs can be detected successfully from WES data, which effectively improved the diagnosis yield in NDDs children.

Objective To analyze the clinical and genetics characteristics in twin sisters with Cockayne syn-drome.Methods The identical twin sisters visited the Affiliated Children's Hospital of Capital Institute of Pediatrics in December 2016.The clinical presentations,course of treatment,blood biochemistry,metabolic screening and whole exon sanger sequencing were analyzed.Results These two patients were referred at 4 years and 5 months of age for growth failure and developmental delay.The younger sister manifested short stature(only 97 cm),low weight(14.0 kg)and little head circumference(43 cm),and the elder sister manifested short stature(only 98 cm),low weight(15.5 kg) and little head circumference(43 cm).They were born with out adverse event,and then they kept the head up at 8 months of age.They could sit at 10 months of age,but they had not acquired independent walking ability up till now. They spoke their first words at 2 year of age,and made little progress after that.They had a variety of abnormal clinical features including cognitive deficits,microcephaly,thin pointy nose,sunken eyes,small chin,photosensitive rash,hearing impairment,volitional tremor and hypermyotonia.They had been diagnosed as nephrotic syndrome at 4.5 years old,with little response to prednisone.The renal biopsy revealed minimal change nephropathy.Cerebrum and cerebellum atrophy was detected by magnetic resonance image scanning. Two heterozygous ERCC8 mutations in both patients,c.394_398delTTACA and large fragment deletion,were identified in the patient.The c.394_398delTTACA mutation originated from his father. The exon 4 deletion from his mother caused the defection of the protein. Conclusions Cockayne syndrome is a rare autosomal recessive disease. It is not only characterized by developmental delay,microcephaly, sunken eyes,photosensitive rash and auditory abnormalities,but also can be involved in nephrotic syndrome.Cockayne syndrome can be caused by compound heterozygous mutation,including c.394_398delTTACA and a large fragment deletion of exon 4 in ERCC8.

OBJECTIVETo explore the candidate disease causing gene for a case with floppy infant syndrome (FIS).

METHODSSingle nucleotide polymorphism array (SNP array) was used for analyzing the whole genome copy number mutations in the proband. Multiple PCR combined with denaturing high performance liquid chromatography (DHPLC) was employed to verify the suspected mutations in the proband and his family members.

RESULTSA large duplication arr [hg19] Xq13.1: 67 987 646-73 805 828, which spans approximately 5.818182 Mb and encompasses 66 known genes, was identified in the proband. The multiple PCR-DHPLC assay confirmed duplication of HDAC8, PHKA1, TAF1, DLG3, KIF4A, IGBP1, PJA1 and SLC16A2 genes in the proband. His mother and grandmother both had duplication of the above genes in one X chromosome, but his aunt had not.

CONCLUSIONThe large Xq13.1 duplication identified by the SNP array probably underlies the FIS in this family. For its high-throughput, high resolution and capacity of automation, SNP array has provided a first line method for the genetic testing for infants featuring developmental delay with unknown reason, mental retardation, autism, multiple malformation and FIS.

Objective: To investigate the iodine nutritional status of children and pregnant women in Hulunbuir, Inner Mongolia after reduction of salt iodine content, and to provide theoretical bases for scientific iodine supplementation. Methods: In May to October 2018, according to "Inner Mongolia Iodine Deficiency Disorders Surveillance Project (2016)", in 14 banners (cities, districts) of Hulunbuir, each banner (city, district) was divided into 5 sampling areas according to the location of east, west, south, north and middle, and 40 non-boarding children aged 8-10 years old (age matched, half male and half female) and 20 pregnant women were selected. Salt samples and urine samples were collected to detect salt and urinary iodine levels. Salt iodine was detected based on the "General Test Method in Salt Industry-Determination of Iodine" (GB/T 13025.7-2012), and urinary iodine was detected based on the "Method for Determination of Iodine in Urine by As³⁺-Ce⁴⁺ Catalytic Spectrophotometry" (WS/T 107-2009), the iodine nutritional status was determined according to the standards of urinary iodine recommended by WHO/UNICEF/ICCIDD. At the same time, the goiter condition of children was examined by B-ultrasound. Results: A total of 4 018 salt samples from homes of children and pregnant women were collected, the median of salt iodine was 22.61 mg/kg, the iodized salt coverage rate was 94.50% (3 797/4 018), the qualified rate of iodized salt was 96.92% (3 680/3 797), and the consumption rate of qualified iodized salt was 91.59% (3 680/4 018). A total of 2 790 urine samples from children were collected, the median of urinary iodine was 179.15 μg/L; and 1 228 urine samples from pregnant women were collected, the median of urinary iodine was 156.88 μg/L. There were 9 banners (cities, districts) where children were at the iodine appropriate level, 4 banners (cities, districts) were higher than the iodine appropriate level and 1 banner was at iodine excessive level. There were 4 banners (cities, districts) where pregnant women were at the iodine deficiency level, 8 banners (cities, districts) were at the iodine appropriate level and 2 banners (cities) were higher than the iodine appropriate level. A total of 2 629 children were examined thyroid gland, and the goiter rate was 0.99% (26/2 629). Conclusions After reduction of salt iodine content, the iodine nutrition of children and pregnant women in Hulunbuir is generally at an appropriate level. In some banners (cities, districts), children and pregnant women are at iodine deficiency level, iodine over appropriate level or iodine excessive level. Iodine nutrition monitoring measures of children and pregnant women should be strengthened.

Infantile spasm (IS) is a common epileptic encephalopathy in infancy, characterized by typical epileptic spasm, developmental delay and hypsarrhythmia on interictal electroencephalogram (EEG). Adrenocorticotropic hormone (ACTH) is the first-line treatment medicine for IS. Although ACTH has shown good response to IS and has been widely used, the regime is not identical and the mechanism is still unclear. This paper focuses on the clinical application of ACTH for IS and the anticonvulsant mechanisms of ACTH, in order to provide clinical and theoretical basis for ACTH application.

Objective To analyze the clinical manifestation and genetic testing in a patient with Adams-Oliver syndrome (AOS) and summarize clinical and genetic characteristics of the dedicator of cytokinesis (DOCK) 6 gene related AOS through reviewing related references.Methods Information of the proband who was hospitalized in Affiliated Children Hospital of Capital Institute of Pediatrics in October 2016 and her family members as well as their DNA samples were collected.The gene sequencing was performed using next generation sequencing technology.Using "Adams-Oliver syndrome"and "DOCK6" as key words,the relevant articles were searched from the Pubmed,China National Knowledge Internet and Wanfang databases and reports of 19 cases were reviewed.Results The proband is an eight months old girl.She presented with severe developmental delay,terminal transverse limb defects and visual loss after birth,and then suffered from tonic seizures and myoclonic seizures at two months old.By physical examination she was found to have esotropia and visual loss.The distal phalanx and nail of the right second-fourth fingers were absent,while the phalangette of the left second-fourth fingers and bilateral distal phalanges of toes were short with small nails attachment.Thyroid function test showed hypothyroidism.The ocular fundus examination showed the residual vitreous artery in the left eye and the retinal pigment degeneration in the right eye.CT scan showed multiple bilateral periventricular calcification and cranial magnetic resonance imaging showed bilateral periventricular lesion.Two heterozygous mutations were identified in DOCK6 gene:one was a known pathogenic mutation (p.L1064Vfs*60),and the other was a novel splice site mutation (c.873+ 1G＞A).By analyzing this case and reported 19 cases,the common performances of DOCK6 gene related AOS included terminal transverse limb defects (20/20),aplasia cutis congenita (18/20),ocular abnormalities (13/20),seizures (12/20),mental retardation (12/20),microcephaly (10/20),cardiovascular malformations (10/20),intrauterine growth retardation (6/20).The mutation of the DOCK6 gene was found to be dominated by frameshift mutation and splice site mutation.Conclusions If either terminal transverse limb defects or aplasia cutis congenita was detected in a patient,AOS should be under consideration.In addition,autosomal recessive inheritance,nervous system and eyes involvement will further indicate DOCK6 gene related AOS.