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 genetic basis for a child featuring global developmental delay. METHODS: DNA was extracted from peripheral blood sample taken from the patient and subjected to whole exome sequencing. Suspected variants were verified by Sanger sequencing of his family members. RESULTS: A heterozygous c.239T>C (p.Ile80Thr) variant of the GNB1 gene was detected in the proband, which was a verified to be de novo in origin. CONCLUSION The heterozygous c.239T>C (p.Ile80Thr) variant of the GNB1 gene probably underlay the disease in this child.
Arthrogryposis ; Child ; Family ; GTP-Binding Protein beta Subunits ; Heterozygote ; Humans ; Intellectual Disability/genetics* ; Whole Exome Sequencing

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.

OBJECTIVETo explore the clinical and genetic features of a Chinese boy featuring X-linked mental retardation.

METHODSClinical features of the patient were analyzed. The DNA of the patient and his parents was extracted and sequenced by next generation sequencing. The results were validated and analyzed with software.

RESULTSThe child displayed X-linked mental retardation. Sequencing showed the patient has carried a c.455T>C (p.L152P) mutation of the GRIA3 gene inherited from his mother.

CONCLUSIONThe c.455T>C (p.L152P) mutation of the GRIA3 gene probably underlies the X-linked mental retardation in this child.

Child, Preschool ; High-Throughput Nucleotide Sequencing ; methods ; Humans ; Male ; Mental Retardation, X-Linked ; genetics ; Mutation ; Receptors, AMPA ; genetics

OBJECTIVETo explore the genetic basis of two neonates suspected for galactosemia.

METHODSNext generation sequencing(NGS) was used to screen the whole exome of the neonates. Suspected mutation was validated by PCR and Sanger sequencing. Potential impact of novel mutation was predicted by using PolyPhen-2, MutationTaste and SIFT software.

RESULTSBoth neonates harbored compound heterozygous mutations of the GALT gene inherited from their parents. One has inherited two novel mutations c.564G>C(p.Q188H) and c.116A>T(p.D39V) respectively from his father and mother. The other has inherited mutations c.754C>T(p.Q252X) and c.904+1G>T from her father and mother, respectively.

CONCLUSIONThe galactosemia in the two neonates may be attributed to compound heterozygous mutations of the GALT gene. This is the first domestic report of using the NGS for the diagnosis of galactosemia.

Female ; Galactosemias ; diagnosis ; Heterozygote ; High-Throughput Nucleotide Sequencing ; methods ; Humans ; Infant, Newborn ; Male ; Mutation ; UTP-Hexose-1-Phosphate Uridylyltransferase ; genetics