Objective To investigate the dynamic expressions and clinical significance of CD141, CD31 and CD95 on circulating endothelial cells (CEC) in febrile and polyuria phases of patients with hemorrhagic fever with renal syndrome (HFRS). Methods Expressions of CD141, CD31 and CD95 in the peripheral blood of patients with HFRS in febrile and polyuria phases were detected by flow cytometry. Comparisons among groups were done by one-factor analysis of variance. Results The percentages of CD141+ CD31+ cells in the peripheral blood cells from patients with HFRS in febrile and polyuria phases were 9.47% ±1.98 % and 8. 26% ±1.55 %, respectively, which were both higher than that (7.05%±1.45%) in healthy controls (F=8. 42; P=0. 000 and P=0. 029, respectively), and that in febrile phase was higher than that in polyuria phase (P = 0. 048). The mean fluorescent intensity (MFI) of CD95 on CEC of HFRS patients in febrile and polyuria phases were both significantly higher than that in healthy controls (F=19. 93; P=0. 000 and P=0. 000 respectively), and that in febrile phase was higher than that in polyuria phase (P=0. 049). In the febrile phase of HFRS,the MFI of CD95+ on CEC in patients with all clinical types were all higher than that in healthy controls (F= 17. 36; all P=0. 000), and that in severe (critical) type was the highest and higher than those in mild type and moderate type (P=0. 002 and P=0. 009, respectively). Conclusion The proportion of CEC and expression of CD95 on CEC are possibly related with the phase and severity of HFRS.

Objective To study the expression of phosphorylated serine 910 of focal adhesion kinase(FAKps910)in circulation endothelial cells(CEC)from patients with hemorrhagic fever with renal syndrome(HFRS).Methods Fifty HFRS patients were involved in the study and divided into mild(n=17),moderate(n=20)and severe(n=13)groups according to patients' conditions. Twenty healthy volunteers were enrolled as control group. CEC were isolated by Percoll density gradient centrifugation method. The expression of FAKps910 in specific APC-CD31+ CEC was measured by flow cytometry. The data were analyzed by one-way ANOVA. Results The positive rates of FAKps910 in CEC were 59.87%±9.58% in early febrile stage patients and 21.14%±2.53% in the healthy controls, while it was 11.64%±2.17%in the later febrile or hypotension stage patients(F=262.31,P＜0.01).It gradually restored to normal afterwards. The positive rates of FAKps910 in CEC were 17.45%±2.64%,13.84%±2.54% and 7.47%±2.57%,respectively in mild, morderate and severe groups in the later febrile or hypotension stage, which were all significantly different from that in control group(F=52.642,P＜0.01).Conclusions FAK in CEC iS related with the clinical severity of HFRS patients. FAK may be involved in intergrin β3-mediated endothelial injury during Hantavirus infection.

Objective To investigate the feasibility of buccal mucosa swab method to isolate genomic DNA for au-tism spectrum disorders (ASD)-related genetic screening. Methods Buccal mucosa swabs and blood were collected from 41 children with ASD. Genomic DNA was extracted from either blood by using a commercial genomic DNA kit or buccal mucosa swab by using phenol-chloroform-isoamyl alcohol method. The concentration, total quality and purity of genomic DNA were compared between these two methods. Genotyping of the ASD-related methylenetetra-hydrofolate reductase (MTHFR) gene C677T locus was analyzed using PCR-restriction enzymatic digestion and sanger sequencing was per-formed for validation. Results The total quality [(5.87±2.58)μg vs. (2.00±0.92)μg] and concentration [(143.25±72.78) mg/L vs. (66.68±24.43) mg/L] of genomic DNA extracted from buccal mucosa swab were higher than that form blood (P<0.05), while the purity was not significantly different between these two methods (P>0.05). Genotyping analysis of MTHFR was also consistent between these two methods. Conclusion Buccal mucosa swab is a simple, non-invasive and reliable meth-od to obtain genomic DNA, which can partially replace blood for analysis of ASD-related gene polymorphisms.

OBJECTIVE: To explore the clinical and genetic features of a child with primary ciliary dyskinesia. METHODS: Genomic DNA of the child and her parents was extracted and subjected to targeted gene capture and next generation sequencing. Suspected mutation was verified by Sanger sequencing, with its nature and impact predicted by Bioinformatic analysis. RESULTS: Clinical manifestations of the child mainly included severe pneumonia, bronchiectasia, nasosinusitis and pneumothorax. DNA sequencing showed that she has carried compound heterozygous mutations of the CCNO gene, namely c.848T>C (p.L283P) and c.262_263 insGGCCCGGCCC (p.Q88Rfs*51), which were respectively inherited from her mother and father. CONCLUSION The child was diagnosed with primary ciliary dyskinesia caused by the compound heterozygous mutations of the CCNO gene.
Base Sequence ; Female ; High-Throughput Nucleotide Sequencing ; Humans ; Kartagener Syndrome ; genetics ; Male ; Mutation ; Sequence Analysis, DNA

OBJECTIVE: To explore the genetic cause for a child with growth retardation by next generation sequencing (NGS). METHODS: Clinical data of the patient was collected. Peripheral venous blood samples were taken from the neonate and his parents. Targeted capturing and NGS were carried out to detect mutations of genes associated with inborn errors of metabolism. Suspected mutations were validated by Sanger sequencing. RESULTS: The 15-month-old female patient was admitted to hospital for growth retardation for 4 months. Hypomyelination was found upon cranium MRI. Genetic testing revealed two novel insertional mutations in the GLB1 gene in the patient, namely c.2006-2007insT and c.475-476 insGGTCC. CONCLUSION The c.2006-2007insT and c.475-476 insGGTCC mutations of the GLB1 gene probably underlie the GM1 gangliosidosis resulting in the growth retardation in the child.
Female ; Gangliosidosis, GM1 ; genetics ; Humans ; Infant ; Infant, Newborn ; Mutation ; Pedigree ; beta-Galactosidase ; genetics

OBJECTIVE: To explore the genetic basis for a neonate featuring hyperammonemia. METHODS: The patient was examined and tested by tandem mass spectrometry and next generation sequencing (NGS). Suspected mutations were confirmed by Sanger sequencing of the proband and her parents. Potential impact of the mutation was predicted with SIFT, PolyPhen-2 and MutationTaste software. RESULTS: Plasma ammonia and alanine were significantly increased in the proband, while serum citrulline was decreased. The neonate was found to harbor compound heterozygous mutations of the CPS1 gene [c.1631C>T(p.T544M) and c.1981G>T(p.G661C)], which were respectively inherited from her father and mother. CONCLUSION The carbamoyl phosphate synthetase I deficiency of the proband can probably be attributed to the mutations of the CPS1 gene. Above finding has expanded the spectrum of CPS1 mutations in association with carbamoyl phosphate synthetase I deficiency.
Carbamoyl-Phosphate Synthase (Ammonia) ; genetics ; Carbamoyl-Phosphate Synthase I Deficiency Disease ; genetics ; Female ; High-Throughput Nucleotide Sequencing ; Humans ; Hyperammonemia ; diagnosis ; genetics ; Infant, Newborn ; Mutation

OBJECTIVE: To carry out variant analysis for a Chinese boy featuring dyshormonogenesis due to congenital hypothyroidism. METHODS: DNA of the patient and his parents was extracted and sequenced by high-throughput sequencing. The results were validated with Sanger sequencing and analyzed with Bioinformatics software. RESULTS: Sequencing result showed that the patient has carried compound variants of c.2654G>T(p.Arg885Leu) and c.943G>T(p.Gly315X) of the DUOX2 gene, which were inherited respectively from his mother and father. CONCLUSION The missense mutation c.2654G>T and nonsense mutation c.943G>T probably underlie the disease in this child.
Child ; Congenital Hypothyroidism ; diagnosis ; genetics ; Dual Oxidases ; genetics ; High-Throughput Nucleotide Sequencing ; Humans ; Male ; Mutation, Missense

Objective To discuss clinical characteristics of a family with Xia-Gibbs syndrome, test and analyze the mutation of their pathogenic gene, and to explore the clinical and genetic characteristics of Xia-Gibbs syndrome. Methods A patient with unexplained developmental retardation was clinically examined and the medical history of his family was collected. Genetic detection was performed to analyze his genetic causes. Results The proband, who was two-year and 1-month old, displayed unusual facies, hypotonia and unexplained developmental retardation. Brain MRI showed leukodystrophy. Other members of his family had no similar medical history. And the proband was found to carry the de novo mutation of c.1073dupC in AHDC1 gene. Conclusion This is the first case with Xia-Gibbs syndrome caused by AHDC1 mutation in China, which has a great significance in studying the correlation between genotype and phenotype.

OBJECTIVETo delineate the clinical and genetic characteristics of a girl featuring motor retardation, language retardation and regression, and light persisting diarrhea.

METHODSThe patient was clinically examined and tested by tandem mass spectrometry and next generation sequencing.

RESULTSThe proband could not stand and walk alone, and had light persisting diarrhea. She manifested language development retardation and regression. Laboratory tests were all normal, but the screening of metabolic disorders for urine and blood showed deficiency of short chain coenzyme A dehydrogenase due to elevated ethylmalonic acid and butyryl carnitine. By next generation sequencing, two compound heterozygous mutations of the ETHE1 gene, c.2T>A and c.488G>A, were discovered in the proband, which were respectively inherited from her father and mother. Bioinformatics analysis predicted both mutations to be pathogenic. The patient was diagnosed with ethylmalonic encephalopathy. Vitamin B1, B2, Coenzyme Q10, and L-carnitine were prescribed. The patient deteriorated and required liver transplantation at 4-year-1-month.

CONCLUSIONBased on the clinical and genetic analysis, the proband was diagnosed with ethylmalonic encephalopathy caused by ETHE1 gene mutation. Next generation sequencing has provided a powerful tool for the diagnosis of such disorders.

OBJECTIVETo analyze the clinical features and genetic basis of a female neonate with muscle weakness, abnormal brain magnetic resonance imaging and elevated blood lactate.

METHODSThe patient was subjected to clinical and laboratory examination. Next generation sequencing was carried out for the patient and her relatives.

RESULTSThe proband was diagnosed as small for gestational age, with clinical features including muscle weakness, abnormal brain magnetic resonance imaging, increased blood lactate, and acidosis. By genetic testing, a de novo PDHA1 mutation c.1133G to A (p.R378H) was identified, which was known to be pathogenic. The patient was diagnosed with pyruvate dehydrogenase complex deficiency disease (PDCDD), for which vitamin B1, coenzyme Q10, and L-carnitine were prescribed, and a ketogenic diet was recommended. Follow-up at 4-month-7-day found that her blood lactic acid was reduced to normal but her muscle tone was still low.

CONCLUSIONThe proband was diagnosed as PDCDD caused by a PDHA1 missense mutation. NGS has provided a powerful tool for the diagnosis of such diseases.