Novel variation of AUH gene in a child with 3-methylpenteneduric aciduria type Ⅰ / 中华实用儿科临床杂志

Objective:To investigate the pathogenic variations of a case of 3-methylpenteneduric aciduria (MGA) type Ⅰ.Methods:Retrospective analysis gene variations of the case with MGA type Ⅰ and family members in June 2017 at Yancheng Maternal and Child Health Hospital were detected using high-throughput sequencing combined with Sanger sequencing.The pathogenicity of the novel variations was predicted using the bioinformatic method.The impact of the novel splicing variation was examined through laboratory experiments.Results:Tandem mass spectrometry and gas chromatography-mass spectrometry results diagnosed the case as MGA type Ⅰ.The compound hete-rozygous variations c. 373C>T (p.R125W) and c. 942+ 3A>G of the AUH gene were detected in the patient, which were inherited from the mother and the father, respectively.Bioinformatics analysis indicated that the c. 373C>T(p.R125W) of the AUH gene was pathogenic (3 softwares) and the R125 residue was highly conserved.Reverse transcription-PCR and Sanger sequencing analysis showed that the variation c. 942+ 3A>G caused the deletion of AUH gene exon 9, which was failed to be predicted in the 4 types of software.The patient was treated with Levocarnitine and leucine-free milk powder from 45 days after birth.The physical and mental development was normal. Conclusions:Splicing analysis of blood RNA should be considered for variants of uncertain significance in genetic diseases when the clinical diagnosis is clear.This study enriches the variation spectrum of the AUH gene.

Identification of two novel variants of the PCCB gene in a pedigree affected with propionic acidemia / 中华医学遗传学杂志

OBJECTIVE@#To detect pathogenic variants in a pedigree affected with propionic acidemia (PA).@*METHODS@#The proband was subjected to high-throughput next-generation sequencing. Suspected variants were validated by Sanger sequencing of his family members. mRNA was extracted from peripheral blood lymphocytes from the proband's father in order to verify the impact of the splicing variant by RT-PCR combined with Sanger sequencing. The pathogenicity of the missense variant was predicted by using PolyPhen-2, Mutation Taster, SIFT, COBALT and HOPE software.@*RESULTS@#The proband was found to harbor compound heterozygous variants of the PCCB gene, namely c.184-2A>G and c.733G>A (p.G245S), which were respectively inherited from his father and mother. RT-PCR combined with Sanger sequencing confirmed skipping of exon 2 during transcription. Bioinformatic analysis indicated the c.733G>A (p.G245S) variant to be damaging.@*CONCLUSION@#The two variants of the PCCB gene probably underlay the disease in this patient. Above findings have enriched the spectrum of PCCB gene variants.

Analysis of ACAT1 gene variants in a patient with β-ketothiolase deficiency / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic etiology of a child suspected for β-ketothiolase deficiency by neonatal screening.@*METHODS@#All coding exons and flanking sequences of the ACAT1 gene were subjected to targeted capture and high-throughput sequencing. Suspected variants were verified by Sanger sequencing and bioinformatic analysis.@*RESULTS@#The child was found to harbor compound heterozygous variants of the ACAT1 gene, namely c.121-3C>G and c.275G>A (p. Gly92Asp). The c.121-3C>G variant was also detected in his father and two sisters, while the c.275G>A (p. Gly92Asp) was a de novo variant. A c.334+ 172C>G (rs12226047) polymorphism was also detected in his mother and two sisters. Sanger sequencing has verified that the c.275G>A (p. Gly92Asp) and c.334+172C>G (rs12226047) variants are located on the same chromosome. Bioinformatics analysis suggested both c.121-3C>G and c.275G>A (p.G92D) variants to be damaging. Based on the American College of Medical Genetics and Genomics standards and guidelines, the c.275G>A variant of the ACAT1 gene was predicted to be pathogenic (PS2+ PM2+ PM3+ PP3+PP4), the c.121-3C>G variant to be likely pathogenic (PM2+ PM3+ PP3+PP4).@*CONCLUSION@#The c.121-3C>G and c.275G>A variants of the ACAT1 gene probably underlay the pathogenesis of the child. Above finding has enriched the variant spectrum of the ACAT1 gene.

Analysis of clinical manifestation and genetic mutations in two patients with Cornelia de Lange syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect potential mutations in two neonates suspected for Cornelia de Lange syndrome (CdLS).</p><p><b>METHODS</b>Peripheral blood samples from the neonates and their parents were collected and analyzed for CdLS-related genes using targeted sequence capture and next-generation sequencing. Suspected mutations were confirmed by direct Sanger sequencing.</p><p><b>RESULTS</b>The neonates were found to respectively carry mutations c.7219C to T and p.D2339Lfs*4 of the NIPBL gene, among which the p.D2339Lfs*4 mutation has not been reported previously. No pathogenic mutation was found in other CdLS-related genes including NIPBL, SMC1A, SMC3, RAD21 and HDAC8.</p><p><b>CONCLUSION</b>The c.7219C to T and p.D2339Lfs*4 mutations of the NIPBL gene probably account for the disease in both patients.</p>

Analysis of PHEX gene mutations in three pedigrees affected with hypophosphatemic rickets / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the molecular basis for three pedigrees affected with hypophosphatemia vitamin D resistant rickets (X-linked hypophosphatemia, XLH).</p><p><b>METHODS</b>Peripheral blood samples from the three pedigrees were collected. Following DNA extraction, the 11 exons and flanking regions of the PHEX gene were subjected to PCR amplification and direct sequencing. Pathogenicity of identified mutations was evaluated through genotype-phenotype correlation.</p><p><b>RESULTS</b>For pedigrees 1 and 2, pathogenic mutations were respectively identified in exon 8 (c.871C>T, p.R291X) and exon 15 (c.1601C>T, p.P534L) of the PHEX gene. For pedigree 3, a novel mutation (c.1234delA, p.S412Vfs*12) was found in exon 11 of the PHEX gene, which caused shift the reading frame and premature termination of protein translation.</p><p><b>CONCLUSION</b>The three mutations probably account for the XLH in the affected pedigrees. The discovery of novel mutations has enriched the spectrum of PHEX gene mutations.</p>

Detection of a patient with ring chromosome 15 by low-coverage massively parallel copy number variation sequencing / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the genetic cause for a child with developmental delay.</p><p><b>METHODS</b>The karotypes of the child and her parents were analyzed with G-banding analysis. Their genome DNA was analyzed with low-coverage massively parallel copy number variation sequencing (CNV-seq) and verified by single nucleotide polymorphism array (SNP-array).</p><p><b>RESULTS</b>The karyotype of the child was ascertained as 46,XX,r(15)(p13q26.3), while both parents showed a normal karyotype. CNV-seq and SNP-array have identified a de novo 15q26.2-q26.3 deletion in the child with a size of approximately 3.60 Mb.</p><p><b>CONCLUSION</b>The abnormal phenotype of the patient carrying the ring chromosome 15 may be attributed to the presence of the 15q26.2-q26.3 microdeletion. The deletion and haploinsufficiency of the IGF1R gene probably underlie the main clinical features of the patient.</p>

The analysis of common mutation in deafness-associated gene in 111 neonates who failed to pass newborn hearing screening / 临床儿科杂志

Objective To explore the genetic mutation in neonates who failed to pass hearing screening.Methods A total of 111 cases of neonates who failed to pass hearing screening and were conifrmed sensorineural deafness by auditory brainstem evoked potential (ABR) were randomly selected. The heel blood was collected and DNA was extracted.GJB2, SLC26A4, and 11 mutation hotspots in mitochondria gene12SrRNA were tested. The relationship between degree of hearing loss and gene mutation was analyzed.Results In 111 neonates, mutation in deafness gene were found in 24 cases (21.6%) . Among them 14 cases (12.6%) hadGJB2 gene mutation including 5 cases of 235delC single heterozygous mutation, 5 cases of 235delC, and 1 case each of 299_300delAT compound heterozygous mutation, 235delC homozygous mutation, 299_300delAT single heterozygous mutation, 176_191del16 and 235delC compound heterozygous mutation, and 299_300delAT and 508_511dupAACG compound heterozygous mutation respectively. Ten cases (9.0%) hadSLC26A4 gene mutation including 2 cases of IVS7-2A>G single heterozygous mutation, 3 cases of 1226G>A single heterozygous mutation, 2 cases of 2168A>G single heterozygous mutation, and 3 cases of IVS7-2A>G and 2168A>G compound heterozygous mutation. Mitochondrial gene mutations were not detected. Conclusions Deafness gene mutation is detected in more than 1/5 neonates who failed to pass newborn hearing screening. GJB2 gene mutation is the most commons. The implementation of hotspots deafness gene detection can improve the diagnostic rate of deafness.

Molecular and prenatal diagnosis of a family with Fanconi anemia by next generation sequencing / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To provide prenatal diagnosis for a pregnant woman who had given birth to a child with Fanconi anemia with combined next-generation sequencing (NGS) and Sanger sequencing.</p><p><b>METHODS</b>For the affected child, potential mutations of the FANCA gene were analyzed with NGS. Suspected mutation was verified with Sanger sequencing. For prenatal diagnosis, genomic DNA was extracted from cultured fetal amniotic fluid cells and subjected to analysis of the same mutations.</p><p><b>RESULTS</b>A low-frequency frameshifting mutation c.989_995del7 (p.H330LfsX2, inherited from his father) and a truncating mutation c.3971C>T (p.P1324L, inherited from his mother) have been identified in the affected child and considered to be pathogenic. The two mutations were subsequently verified by Sanger sequencing. Upon prenatal diagnosis, the fetus was found to carry two mutations.</p><p><b>CONCLUSION</b>The combined next-generation sequencing and Sanger sequencing can reduce the time for diagnosis and identify subtypes of Fanconi anemia and the mutational sites, which has enabled reliable prenatal diagnosis of this disease.</p>

Prenatal diagnosis of methylmalonic acidemia by amniotic fluid metabolites analysis using mass spectrometry / 中华围产医学杂志

Objective To investigate the effect of tandem mass spectrometry and gas chromatography-mass spectrometry to make prenatal diagnosis of methylmalonic acidemia (MMA) by detecting organic acid and acylcarnitine in amniotic fluid.Methods From October 11,2007 to December 20,2014,131 pregnant women with MMA proband received prenatal diagnosis of MMA in Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine (case group).Another 120 cases of pregnant women for conventional prenatal diagnosis at the same period were as control group.The pregnant women of two groups had the amniocentesis at 16 to 20 weeks of gestation.The levels of propionylcarnitine(C3) and acetylcarnitine(C2)in amniotic fluid were detected by tandem mass spectrometry.The methylmalonic acid and methylcitrate acid were detected by gas chromatography-mass spectrometry.MMA gene of cells in amniotic fluid of eighty fetuses with proband clearly diagnosed were detected by gene testing.Data were analyzed by Wilcoxon test.Results In case group,29 fetuses were found positive for higher level of C3,C3/C2,methylmalonic acid and methylcitrate acid compared with normal reference value,and the detected rate of fetal MMA was 22.1％(29/131).The levels of C3 and C3 / C2 in amniotic fluid of these 29 cases were higher than those in control group[8.13(2.42-16.70) vs 1.04(0.52-3.40) μmol/L,Z =-8.313; 0.77(0.30-1.79) vs 0.10(0.05-0.22),Z=-8.374; P ＜ 0.05 respectively].The levels of methylmalonic acid and methylcitrate acid were also higher[9.13(1.68-61.78) vs 0.00(0.00-1.31) mmol/mol Crea,Z=-11.348; 0.58(0.00-1.90) vs 0.05(0.00-0.52) mmol/mol Crea,Z=-6.632,P ＜ 0.05 respectively].For the other 102 cases in case group,the levels of C3,C3/C2,methylmalonic acid and methylcitrate acid were not higher than normal reference value,and were similar to those in control group (P ＞ 0.05); while they were lower than those of positive MMA fetuses (all P ＜ 0.05).Among 29 positive fetuses,16 fetuses were detected MMA gene,five were diagnosed as MUT forms of MMA and 11 were MMACHC forms of MMA.In 102 MMA negative fetuses,64 fetuses were detected MMA gene,44 were found one mutant site and 20 were found no gene mutation.The coincidence rate between gene detecting and mass spectrometry was 100％(80/80).Conclusions Mass spectrometry could be used to measure the C3,methylmalonic acid and methylcitrate acid levels in amniotic fluid of pregnant women with MMA proband to make prenatal diagnosis.

Clinical Observation of Salvia miltiorrhiza for Severe Craniocerebral Injury Patients after Operation / 中国药房

OBJECTIVE:To observe the clinical efficacy and sequela of Salvia miltiorrhiza treatment for severe craniocere-bral injury patients after operation. METHODS:60 severe craniocerebral injury patients were randomly divided into control group and treatment group with 30 cases in each group. Control group was given routine and symptomatic treatment after clearance of hematoma by craniotomy or decompressive craniectomy according to the patient’s condition;treatment group was additionally giv-en intravenous injection of Danshen injection 20 ml/d for 7-10 days 3-5 days after operation without hemorrhagic tendency,on the basis of control group,and then given Compound danshen dropping pills instead (10 pills/time,3 times/d) for one month. GCS score,GOS score,intracranial pressure and brain CT,the levels of ET and CRP were compared between 2 groups after op-eration. The occurrence of sequelae were evaluated 3 months after operation. RESULTS:After the operation,S. miltiorrhiza treat-ment could increase GCS score and GOS score,reduced intracranial pressure and brain edema of craniocerebral injury patients, and the improvement of clinical symptoms in treatment group was significantly faster than in control group,with statistical signifi-cance(P<0.05 or P<0.01). The postoperative S. miltiorrhiza treatment could decrease the levels of ET and CRP in patients with craniocerebral injury in the time-dependent manner,the decreased of treatment group was significantly faster than that of control group,with statistical significance (P<0.05 or P<0.01). The effective of postoperative sequelae improvement was 93.3% in treatment group,which was significantly higher than that of control group(76.7%),with statistical significance(P<0.05). CON-CLUSIONS:Postoperative S. miltiorrhiza treatment can improve the clinical symptoms of severe craniocerebral injury patients,re-duce the incidence of complication and improve the prognosis. The effect may be related to the decrease of plasma ET and CRP levels.

The biomechanical analysis of the etiologies of spontaneous pneumothorax in flat-chest teenagers / 中华胸心血管外科杂志

Objective To investigate the etiologies of spontaneous pneumothorax in flat-chest teenagers. Methods 139 teenagers with flat chest and spontaneous pneumothorax were designed as the study group, while 152 teenagers without spontaneous pneumothorax during the same period were taken as control. The ratio of thoracic transverse diameters to anterior-posterior diameters, and the ratio of tracheal length to transverse diameters were measured respectively by CT scan. The resected pulmonary tissues in 47 cases with spontaneous pneumonthorax and in 47 cases without spontaneous pneumonthorax underwent various examinations, including tension, compression, and bending tests. Student t-test was employed to perform the statistical analysis. Results The ratios of thoracic transverse diameters to anterior-posterior diameters and the ratios of tracheal length to transverse diameters in the study group were remarkably higher than those were in the control group, with significant statistical difference (P

Acute retroperitoneal colon perforation: a report of 20 cases / 中国普通外科杂志

Objective To explore the clinical features of acute retroperitoneal colon perforation. Methods A retrospective analysis and summary were made on the clinical data of 20 cases of various retroperitoneal colon perforation. Results All of the 20 cases had been misdiagnosed preoperatively. Eighteen cases received surgical treatment, of which 5 cases underwent one-stage tumor resection, 5 cases had two-stage tumor resection and anastomosis;of 8 cases with traumatic perforation,7 were treated by simple suture, 1 case underwent two-stage colonic anastomosis; 2 cases died,all the 2 cases were treated conservatively.Conclusions There are many differences between retroperitoneal colon perforation and intra-abdominal perforation in clinical symptoms, signs and supplementary examinations. Diagnosis of retroperitoneal colon perforation is difficult and the rate of misdiagnosis is high, so this condition must be highly regarded.Surgery is the best option to treat this disease.

The clinical study on thymoma associated with pure red cell aplasia: report of five cases / 中华胸心血管外科杂志

Objective To present the clinic characteristics of thymoma associated with pure red cell aplasia. Methods Between 1979 and 2004, 5 patients with thymoma associated with pure red cell aplasia were surgically treated. Results The age of the patients ranged from 43 to 68 years, mean age 54.6 years. The incidence of thymoma associated with pure red cell aplasia was 2.7% (5/185) in the contemporary series. All five patients underwent radical operation. The early results of thymomectomy were excellent, 4 cases are long-term survivals, and recurrence of PRCA was found in one patient 2 months after operation and died 31 months later. Conclusion Thymoma associated with PRCA is a rare autoimmune disease. Surgery for thymoma is still the top priority in the selection of treatment. Incretion and immunosuppressor may be considered for postoperative persistent PRCA cases.