Animals ; SARS-CoV-2 ; Antibodies, Neutralizing ; Macaca mulatta ; COVID-19/prevention & control* ; Antibodies, Viral ; Vaccines

Objective To investigate the clinical and genetic characteristics of focal dermal hypoplasia (FDH) in children. Methods Clinical data, relevant examinations, histopathological features and genetic test results of a male newborn with FDH who was admitted to the neonatal ward of Beijing Children's Hospital of Capital Medical University were retrospectively analyzed. Reports of pediatric FDH patients with complete clinical data were retrieved from PubMed, Wanfang database and China National Knowledge Infrastructure from the establishment of these databases to March 2018 and characteristics of FDH was summarized. Results The case we reported here was a male neonate diagnosed with FDH in China, who was born with microcephaly, bilateral auricular cartilage dysplasia, tooth germ dysplasia and bipedal deformity and his skin, bone, gingiva, bilateral iris and pupils were all involved. Histopathological examination of the skin suggested dermal dysplasia. Genetic analysis showed a suspected chimeric nucleotide variation in PORCN gene (c.268 C>T), whereas no abnormalities were found in his parents and sister. A total of 60 cases (including the one we reported) of FDH diagnosed in childhood were reviewed, and 19 of them were confirmed in neonatal period. Fifty-seven of the 60 cases (95.0%) developed typical skin dysplasia and 56 cases (93.3%) with skeletal malformations, while other clinical manifestations vary. Histopathological examination suggested as dermis dysplasia, adipose tissue migration and reduction of appendage and collagen fibers. Among the 60 children, 19 (including four onset at neonatal period) underwent genetic testing and the results indicated PORCN gene mutation. Mutations in the four with neonatal-onset were c.956dupA, c.1061T>C, c.749C>T and c.268C>T. As the reported case was a boy, with only one X chromosome, the PORCN gene mutation could directly affected its function resulting in the abnormal phenotype. It was a de novo mutation as the same mutation was not detected in his parents. Conclusions FDH is a hereditary disease involving multiple systems with various clinical manifestations. Skin histopathological examination and genetic testing should be performed as soon as possible for early diagnosis and intervention. Accurate diagnosis is essential for genetic counseling, reproductive planning, prospective guidance and prognosis.

Objective: To summarize the clinical features of 7 rare cases of hemolytic disease of newborn (HDN), and to improve the understanding of rare HDN. Methods: Data of clinical information, laboratory findings, treatments and outcomes were collected and analyzed for four cases with HDN due to anti-M, two cases due to anti-Kidd, and one case due to anti-Duffy. All of them were admitted to the Department of Neonatology, Beijing Children's Hospital Affiliated to Capital Medial University from July 2007 to June 2017. Results: Among the four MN hemolytic babies, two were males and two were females. Jaundice was found in three cases. Two cases had hyperbilirubinemia, one of them had severe hyperbilirubinemia. All the four cases developed anemia, including severe anemia in three cases. Two cases of Kidd hemolytic disease and 1 case of Duffy hemolytic disease had jaundice and anemia, but did not reach the level of severe hyperbilirubinemia and severe anemia. MN hemolytic disease babies got negative results in direct antiglobulin test, whereas the Kidd and Duffy hemolytic disease babies had positive findings in direct antiglobulin test. None of the babies had blood transfusion, and they were discharged from the hospital. Conclusions Without maternal and fetal blood group incompatibility (ABO or Rh blood-group system), for early onset of jaundice, severe jaundice or anemia, antiglobulin test to mother and child earlier should be administered, and MN, Kidd, Duffy and other rare hemolytic disease of the newborn should be pay attention to.

Objective To study the application of the array comparative genomic hybridization (Array-CGH) for the detection of chromosomal disorders in newborns.Method The Array-CGH technique was used to analyze the whole genome of the patients who were suspected of chromosomal disease in neonatal ward of our hospital from January to December in 2014,and further verification in genomic unbalanced ectopia was carried out by FISH (fluorescence in situ hybridization,FISH).Result Among 514 patients,104 were found carrying chromosomal abnormalities with a detection rate of 20.2％.The most common chromosomal disease is the Down syndrome syndrome (24 cases),followed by the chubby Willy and Angel syndrome(17 cases),while the Wolf-Hirschhorn syndrome in 5 cases,Williams syndrome in 5 cases and the Criduchat syndrone in 5 cases.The results of FISH were consistent with Array-CGH.Conclusion The technique of Array-CGH can be used to scan the whole genome of children with unknown disease.As a high-throughput and rapid research method,this technique has important clinical significance in the screening of chromosomal diseases.

Objectives To investigate the closing time of patent foramen ovale in newborns and infants in order to provide appropriate follow-up time points.Methods From September 1,2010 to April 30,2011,131 of l 202 full-term infants with patent foramen ovale were finished follow up at 3,6 and 12 months of age in Beijing Children's Hospital,Capital Medical University.If the foramen ovale was not closed at 12 months of age,the patients were followed up until two years of age.The closing time and the effects of complicated patent ductus arteriosus (PDA) were analyzed statistically using two independent t test,Chi-square and trend Chi square tests.Results Of the 131 full-term infants with patent foramen ovale,72 were males,and 59 were females.The foramen ovale size in neonatal period was not statistically different between males and females [(2.94 ±0.86) vs (2.95 ± 0.92) mm,t=0.641,P=0.964].The foramen ovale closing rate at 3 months was 21.4％ (28/131),67.9％ (89/131) at 6 months,and 95.4％ (125/131) at 12 months.The rate of foramen ovale closing decreased with larger foramen ovale at 3,6 and 12 months of age (x2trend were 42.930,101.050 and 63.260,all P＜0.05).Six patients with patent foramen ovale at 12 months of age were followed up until 2 years of age:two cases with foramen ovale ＜5.0 mm in the neonatal period were closed,one of two cases with foramen ovale ≥5.0 but ＜6.0 mm was closed,and one of two cases with foramen ovale ≥ 6.0 mm was closed.Of the 131 cases,121 were simple patent foramen ovale,and l0 were complicated with PDA.There were no significant difference in neonatal foramen ovale size between children with simple patent foramen ovale and those with PDA [(2.95 ±0.88) vs (2.82±0.83) mm,t=0.782,P=0.649].The closing rates in the simple patent foramen ovale group at 3,6,and 12 months of age were 21.5％ (26/121),57.9％ (55/95) and 87.5％ (35/40),respectively,and showed no significant difference from those with PDA (2/10,6/8 and 1/2,x2=0.012,0.946 and 1.536,all P＞0.05).In the simple patent foramen ovale group,the closing rate at 3 months was less than that at 6 months and 12 months 0x2 were 10.410 and 62.515,both P＜0.01).There was no difference in the closing rate in patients with PDA at 3,6 and 12 months (x2=5.748,P＞0.05).Conclusions Asymptomatic patent foramen ovale with a foramen ovale ＜5.0 mm may not require follow-up.But patients with a foramen ovale ≥ 5.0 mm,even asymptomatic,should be followed up using thoracic echocardiography at 2 years,and further follow-up is required if unclosed.

OBJECTIVETo determine the disease-causing mutation in a newborn with hereditary spherocytosis.

METHODSGenomic DNA was extracted from peripheral blood samples of the patient and her parents. Next-generation sequencing was used to analyze the related genes. Suspected pathogenic mutation was verified with polymerase chain reaction and Sanger sequencing.

RESULTSAn insertional mutation g.834_833insC was identified in the coding region of ankyrin-1 (ANK1) gene, which has caused a frame shift, resulting premature termination of protein translation.

CONCLUSIONThe hereditary spherocytosis in the neonate was probably due to the g.834_833insC mutation of the ANK1 gene.

Amino Acid Sequence ; Ankyrins ; genetics ; Base Sequence ; Female ; Humans ; Infant, Newborn ; Infant, Newborn, Diseases ; diagnosis ; genetics ; Molecular Sequence Data ; Mutation ; Spherocytosis, Hereditary ; diagnosis ; genetics

ObjectiveTo analyze the clinical manifestation of hemolytic disease of the newborn (HDN) due to anti-M and Rhesus system.MethodsClinical information was collected and analyzed for three cases with HDN due to anti-M and 64 with Rhesus hemolytic disease, who were admitted to Department of Neonatology, Beijing Children's Hospital Affiliated to Capital Medical University from February 2011 to January 2015, as well as another 28 cases of HDN due to anti-M with complete information retrieved from literature in Wanfang and China National Knowledge lnfrastructure (CNKI) Database from 1992 to 2014.Chi-square test was performed for statistical analysis.ResultsTwo out of the 64 Rh hemolytic babies gave up therapy due to kernicterus and another two out of the 31 MN hemolytic babies, obtained from literature, died 24 h after birth because of anemia or edema, while the rest survived. Although more babies were the first child of the family in HDN due to anti-M than those of Rh hemolytic disease [26%(8/31) vs 9%(6/64),χ2=4.487, P=0.034], but lower incidence of jaundice [81%(25/31) vs 98%(63/64),χ2=9.686,P=0.002], less proportion of presentation of jaundice within 24 h after birth [29% (9/31) vs 64%(41/64),χ2=10.279,P=0.001] and lower positive rate of direct antiglobulin test [39%(12/31) vs 100%(64/64), Fisher exact test,P=0.000] were shown in HDN due to anti-M. No significant difference was found in the incidences of hyperbilirubinemia [58%(18/31) vs 66%(42/64),χ2=0.513], severe hyperbilirubinemia [23%(7/31) vs 36%(23/64),χ2=1.724], anemia [81%(25/31) vs 89%(57/64),χ2=1.253] and severe anemia [29%(9/31) vs 34%(22/64),χ2=0.271] between HDN due to anti-M and Rh hemolytic babies (allP>0.05).ConclusionsHDN due to anti-M and Rhesus hemolytic disease can cause severe pathological jaundice and/or anemia in newborns. Indirect antiglobulin test should be offered when direct antiglobulin test is negative which is helpful in the diagnosis of HDN due to anti-M.

Objective Design and synthesize short hairpin RNA (shRNA) expression vector of RNA for specific silencing of heparanase (HPA) gene,screened plasmid which silence effects is the best.Observe the function of ceil invasion after inhibiting the expression of HPA in cervical carcinoma cell lines (HeLa).Methods The genomic sequence of HPA gene was retrieved from GenBank database.Designed four pairs of specific oligonucleotide sequences and a negative control according to the shRNA design principles.They were inserted into the vector pYr-1.1,vectors,and transfected into HeLa cells via lipofectamine.Reverse transcription (RT)-PCR and immunofluorescence were employed to detect the expression of HPA gene in the transfected cells at the mRNA and protein levels,respectively.The plasmid were screened and transfected into HeLa cells,then transwell small room stromal invasion experiment were employed to observe the cervical carcinoma cell invasion.Results RT-PCR results of transfected HeLa cells shown that the mRNA amplification multiples were 0.54 ±0.05 in the HPA-592 group,0.89 ±0.18 in HPA-995 group,0.82 ±0.22 in the HPA-1351 group,0.91 ±0.47 in HPA-1658 group.While,they were 1.31 ±0.72 and 1.09 ±0.16 in negative control and blank control group,respectively.Green fluorescence was visible in the cytoplasm,which indicated that the HPA protein was expressed in the cytoplasm,of them the weakest green fluorescence in the HPA-592 group.The relative numbers of invasive cells among the HeLa cells were as follows:182 ±6 in the blank control group,258 ± 17 in the negative control group,and 44 ± 4 in the HPA-592-specific interference group(P ＜ 0.01).Conclusion Successfully screened shRNA vector targeting human HPA,efficiently inhibit expression of HPA gene when transfected into HeLa cells,and significantly reduced the invasion capacity of cervical carcinoma cells.

Objective To study the efficacy and safety of Gonadotropin-releasing hormone agonists (GnRH-a) combined with laparoscope conservative surgery in treatment of moderate or severe endometriosis.Methods From Jan.2007 to Jan.2010,68 patients with moderate or severe undergoing treatment in Renmin Hospital of Wuhan University were enrolled in this retrospective study.Three groups were classified,which were 25 patients in GnRH-a group,subcutaneous injection Leuprorelin on the second day of menstruation,every 4 weeks for 3 months.Twenty-three patients in Marvelon group,orally one marvelon tablet on the second day of menstruation,continuous 21 days for one period of treatment for 3 courses.Twenty patients in surgery group,without any medicine used preoperatively.All patients were followed by 12 months and compare their surgery time,blood loss,recovery,visual analog scale (VAS),and recurrence and so on.Results The operating time were (68 ± 18) min in GnRH-a group,(80 ± 21) min in Marvelon group and (90± 24) min in surgery group.The amount of bleeding were (118 ± 15) ml in GnRh-a group,(161 ± 18) ml in Marvelon group and (193 ± 13) ml in surgery group.There was significant lower in the operating time and amount of bleeding in GnRH-a group than those in other two groups (P ＜ 0.05).The activity time and the anus exhaust time were shorter in patients in GnRh-a group than those in the other two groups significantly(P ＜ 0.05).When followed up in 12 months after treatment,visual analogue scale had dropped from 3.8 (1.9-6.8) to 1.9 (1.1-2.8) in GnRh-a group,from 2.7 (1.3-5.5) to 1.8 (1.2-3.2) in Marvelon group and from 1.9(1.0-4.9) to 1.6(1.0-3.6) in surgery group.It was showed the most remarkable decreased VAS in GnRHa group when compared with the other two groups(P ＜ 0.05).The recurrence rates were 12％ (3/25) in GnRH-a group,22％ (5/23) in Marvelon group and 25 ％ (5/25) in surgery group.It was found that the most significant lower recurrence was in GnRH-a group when compared with the other two groups (P ＜ 0.05).Conclusions It was safe and efficacy that GnRH-a combined with laparoscopic conservative surgery were used in treatment of endometriosis.It could bring shorter operation time,less intraoperative blood loss,quick postoperative recover,the lower recurrence rate.

Objective To detect the expression of heparanase (HPA) in cervical cancer cells and investigate the impact of quercetin on the expression of HPA,and the molecular mechanism that quercetin inhibits the growth of cervical cancer cells.Methods The experimental groups included cervical cancer cell lines (HeLa and Caski) exposed to different concentrations of quercetin (20,40 and 80 μmol/L) in the culture medium.The control groups included a negative control group,which was cultured with RPMI 1640 only,and a positive control group,in which cervical cancer cells were transfected with HPA small interference RNA (siRNA) to silence HPA expression.The cellular expression levels of HPA were detected with fluorescence quantitative real-time PCR and western blot analysis at 24,48 and 72 hours after treatment.Results (1) HPA was significantly expressed in both cervical cancer cell lines (HeLa and Caski),and it exists both nucleus and cytoplasm.(2)The real-time PCR shows as follows:as the quercetin concentration increased (20,40 and 80 μmol/L),the mRNA expression level of HPA decreased (P ＜0.01),in which the inhibition of HPA expression was concentration dependent.In addition,the inhibition of HPA expression was also time dependent.As time growth,the expression level of HPA mRNA (24,48 and 72 hours) in HeLa and Caski cells decreased (P ＜ 0.01).Compared with negative control group,the expression level of HPA mRNA decreased in different concentrations of quercetin (40 and 80 μmol/L) in both HeLa and Caski cells (all P ＜ 0.05) ; Compared with positive control group,the expression level of HPA mRNA expressed no obvious difference in quercetin (80 μmol/L) group (P ＞ 0.05) in HeLa cells,while it was opposite in Caski cells(P ＜0.01).(3)The result of western blot shown that,as the quercetin concentration increased(20,40 and 80 μmol/L)and time growth (24,48 and 72 hours),the expression level of HPA protein decreased (P ＜ 0.01),and the inhibition of HPA protein expression was concentration and time dependent.Compared with negative control group,the expression level of HPA protein decreased in different concentrations of quercetin (40 and 80 μmol/L) in both HeLa and Caski cells (all P ＜ 0.05) ;Conpared with positive control group,the expression level of HPA protein expressed no obvious difference in quercetin (80 μmol/L) group (all P ＞ 0.05) in both HeLa cells and Caski cells (all P＞0.05).Conclusion Quercetin could inhibits the expression of HPA in cervical carcinoma cell lines,which inhibition is concentration and time dependent.