[Summary] A single-incision video-assisted thoracoscopic sublobular limited resection was performed for a 2-cm pulmonary nodule at the right lower lobe of the lung in August 2014.The post-operative pathological diagnosis was inflammatory myofibroblastic tumor of the lung.No recurrence and metastasis was found after one year’s follow-up.

Objectives Inborn errors of metabolism (IEM) has a diverse spectrum and different incidence in different countries, the early diagnosis at presymptomatic stage is imperative to benefic patient from sequelae. Phenylke-tonuria (PKU) / hyperphenylalaninemia (HPA) is the most common metabolism disorder in Shanghai as well as in other regions. The study is to further clarify the incidence of inborn errors of metabolism among newborn in Shanghai. Methods The dried blood spot specimens were collected from near 90 local maternity and children's hospitals or general hospitals in Shanghai. PKU/HPA screening was carried out by fluorometric method. Neonatal screening using tandem mass spectrometry was performed in one of the study centers, Xinhua neonatal screening center. Results A total of 815 160 cases were screened from 2001 - 2007 in Shanghai, the incidence of PKU/HPA was 1 : 12 351. The tetrahydrobiopterin deficiency was 12.9% among hyperphenylalaninemia patients. According to the 116 000 neonatal samples data detected by tandem mass spectrometry, 20 cases were confirmed diagnosis, including 6 kinds diseases, it was PKU/HPA, maple syrup urine disease, methylmalonicacidemia, propionic acidemia, 3-methylcrotonyl-CoA carboxylase defection, and short chain aeyl-CoA dehydrogenase deficiency. Conclusions The pilot study shown that inborn errors of metabolism neonatal screen-ing using tandem mass was 1 : 5 800 in Shanghai, PKU/HPA was the most common disease. It is expected that the expansion of newborn screening using tandem mass spectrometry could be further considered and further improving inborn errors of metabolism preventive services in Shanghai.

Adrenal Hyperplasia, Congenital ; complications ; metabolism ; Body Height ; Growth Disorders ; etiology ; Humans ; Risk Factors ; Steroid 21-Hydroxylase ; metabolism

OBJECTIVETo investigate the clinical and laboratory features of very long chain acyl-CoA dehydrogenase deficiency ( VLCADD ) and the correlations between its genotype and phenotype.

METHODEleven patients diagnosed as VLCADD of Shanghai Jiaotong University School of Medicine seen from September 2006 to May 2014 were included. There were 9 boys and 2 girls, whose age was 2 d-17 years. Analysis was performed on clinical features, routine laboratory examination, and tandem mass spectrometry (MS-MS) , gas chromatography mass spectrometry (GC-MS) and genetic analysis were conducted.

RESULTAll cases had elevated levels of blood tetradecanoylcarnitine (C14:1) recognized as the characteristic biomarker for VLCADD. The eleven patients were classified into three groups: six cases in neonatal onset group, three in infancy onset group form patients and two in late onset group. Neonatal onset patients were characterized by hypoactivity, hypoglycemia shortly after birth. Infancy onset patients presented hepatomegaly and hypoglycemia in infancy. The two adolescent patients showed initial manifestations of exercise intolerance or rhabdomyolysis. Six of the eleven patients died at the age of 2-8 months, including four neonatal onset and two infant onset patients, with one or two null mutations. The other two neonatal onset patients were diagnosed since early birth through neonatal screening and their clinical manifestation are almost normal after treatments. Among 11 patients, seventeen different mutations in the ACADVL gene were identified, with a total mutation detection rate of 95.45% (21/22 alleles), including eleven reported mutations ( p. S22X, p. G43D, p. R511Q, p. W427X, p. A213T, p. C215R, p. G222R, p. R450H, p. R456H, c. 296-297delCA, c. 1605 + 1G > T) and six novel mutations (p. S72F, p. Q100X, p. M437T, p. D466Y, c. 1315delG insAC, IVS7 + 4 A > G). The p. R450H was the most frequent mutation identified in three alleles (13.63%, 3/22 alleles), followed by p. S22X and p. D466Y mutations which were detected in two alleles (9.09%, 2/22 alleles).

CONCLUSIONThe ACADVL gene mutations were heterozygous in our patients. The mortality of neonatal onset form and infant onset form is much higher than the late onset form patients, suggesting a certain correlation between the genotype and phenotype was found. The earlier diagnosis and treatment of VLCADD are of vital importance for the improvement of the prognosis of the patients.

Acyl-CoA Dehydrogenase, Long-Chain ; deficiency ; genetics ; Adolescent ; Age of Onset ; Alleles ; Asian Continental Ancestry Group ; Child ; Child, Preschool ; China ; Female ; Genetic Testing ; Genotype ; Heterozygote ; Humans ; Infant ; Infant, Newborn ; Lipid Metabolism, Inborn Errors ; complications ; genetics ; Male ; Mitochondrial Diseases ; complications ; genetics ; Muscular Diseases ; complications ; genetics ; Mutation ; Neonatal Screening ; Phenotype ; Prognosis ; Rhabdomyolysis ; etiology ; Spectrum Analysis ; Tandem Mass Spectrometry

Objective To evaluate the predictive value of modified early warning score (MEWS) and SMART-COP score on mechanical ventilation in patients with severe influenza A H1N1. Methods Fifty cases diagnosed with severe influenza A H1N1 were retrospectively analyzed. The MEWS and SMART-COP score were calculated. The area under the receiver operating characteristic (ROC) curve (AUC) was evaluated using ROC curve. MEWS, SMART-COP score and AUC were analyzed by Z test. Results The AUCs of MEWS and SMART-COP score for predicting mechanical ventilation were 0. 923 and 0. 889, respectively, which were not significantly different (Z=0. 548, P =0. 584).Conclusion Both of MEWS and SMART-COP score are predictive factors of mechanical ventilation in the patients with severe influenza A H1N1.

Objective To analyze gene mutations of a Niemann-Pick disease type C (NPC) proband,and carry out prenatal diagnosis for the family.Methods The coding regions of NPC1 gene in the proband (late-infantile form) and white blood cell (WBC) in peripheral blood of its parents were amplified by polymerase chain reaction and direct DNA sequencing in both directions was performed.The sequencing results were compared with human NPC1 gene sequence (NM_000271) in GenBank,and sequences of mutated exons were determined.Direct sequencing was used on 50 normal Chinese individuals' DNA samples (control) to exclude mutation's single nucleotide polymorphism (SNP).An inter-species alignment of homologous NPC1 proteins was performed using ClustalX 1.81 software.During the second pregnancy of the proband's mother,the amniotic fluid was obtained at 18 weeks of gestation and the amniocytes were cultured for gene mutation analysis.Neonate's DNA of WBC in peripheral blood was also extracted for NPC1 gene analysis.Results Mutation analysis of NPC1 gene revealed two novel heterozygous mutations (c.2284-2287 delCTCT and p.V959G) in the proband,which originated from her father and mother,respectively.These two mutations were absent in the control,suggesting that these mutations were not SNP.While comparing with the amino acid in NPC1 protein of human,mouse,rat,rabbit,cat and pig,it revealed that p.V959 belonged to a conservative amino acid region and the missense mutation of p.V959G may perturb the function of NPC protein.Neither mutation was found in DNA from amniotic fluid or from the cultivated amniocytes in the second pregnancy,suggesting a normal fetus.c.2284-2287 delCTCT and p.V959G mutation were not found in NPC1 gene analysis of WBC in peripheral blood of the neonate,which was consistent with the prenatal diagnosis.Conclusions PCR-direct sequencing could be used as genetic diagnosis for NPC proband and prenatal diagnosis for its family.The mutation p.V959G may be correlated to late infantile form of NPC.

Objective Sphingolipidoses are a group of rare genetic disorders caused by catabolism defect of sphingolipids by lysosomal hydrolases with diverse presentations,and represent an important health problem to almost all ethnic populations. To date,there is no epidemiologic study on the prevalence of sphinglipidoses,individually,or as a group,in China. We set up a series of enzymatic assays that could make definite diagnoses with the aim to collect data for an epidemiologic investigation of sphingolipidoses and also pave the way to prenatal diagnosis to decrease the rate of inborn error of metabolism. Methods Patients with suspected sphingolipidosis were recruited from pediatric endocrinology and inherited metabolism outpatient clinics of Xinhua Hospital. Leukocytes were isolated with dextran from peripheral bloods. Activities of leukocyte acid β-glucosidase,acid sphingomyelinase,arylsulphatase A,galacto-cerebrosidase,beta-galactosidase were measured using their specific artificial fluorescent substrates,while arylsulfatase A was determined by a colorimetric assay with dipotassium 2-hydroxy-5-nitrophenyl sulfate as the substrate. Results In one year,we identified 17 patients with 5 different kinds of sphingolipidoses,including 3 patients with Gaucher disease,9 patients with Niemann-Pick type A/B,2 patients with metachromatic leukodystrophy,2 patients with Krabbe disease,and 1 patient with GM1 gangliosidosis. We made brief descriptions of disease characters of each different kind disease and compared our results with findings of other ethnic groups. Conclusions Sphinglipidoses was markedly under-diagnosed in China and general pediatricians should be alerted to sphinglipidoses.

Objective Establish a method for measuring the activities of Galactocerebrosidase (GALC), α-Glucosidase(GAA), α-Galactosidase (GLA) and α-L-Iduronidase (IDUA) in dried blood spots specimen by tandem mass spectrometry ( MS/MS ).Methods A total of 2175 dried blood spot samples forinborn errors of metabolism in neonatalscreening center of Shanghai Xinhua hospital were collected in July and November, 2013.And twenty dried blood spot samples from patients withlysosomalstorage disorders( LSDs) of Shanghai Xin Hua Hospital were collected from September 2012 to January 2014.The extraction of DBS was incubated with enzyme substrates and internal standards.After liquid-liquid and solid-phase extraction, the extraction solution was dried under nitrogen and reconstituted.Then enzyme reaction products and internal standards were analyzed by MS/MS.Linearity, precision, accuracy and the limit of detection were evaluated.2175 dried blood spot samples were detected to establish the normal reference range for the activities of four enzymes according to 0.5th to 99.5th percentiles.20 specimens from patients withLSDs were detected to verify the reference range inclinical judgment.Results The intraassay and interassay precisions ranged from 1.7%to 11.8%, and the intraassay and interassay accuracies ranged from 85%to 115%.The linear coefficients for measured concentration of enzyme products/internal standards and theoretical concentration were 0.997-0.999.The limits of detection forGALC, GAA, GLA and GLA were 0.03 μmol/(L· h), 0.09 μmol/(L· h), 0.12 μmol/(L· h) and 0.16 μmol/(L· h) .The normal reference values for GALC, GAA, GLA and GLAwere 0.51-8.51μmol( L· h) ,1.99-22.22μmol/( L· h),1.68-41.59 μmol/(L· h) and 2.36-19.21 μmol/(L· h).The enzymes of 20 patients with LSDs were remarkably decreased compared to the normal range.The Krabbe, Pompe, Fabry, MPSⅠpatients can be effectively detected by this MS/MS method.Conclusions A MS/MS method for measuring GALC, GAA, GLA and IDUA enzyme activities in DBShas been established.

Objective To derive and experiment validate the probability of exclusion (PE) formulas in the cases of standard triplet parentage testing.Methods The formulas were derived voluntarily based on the PE definition:PE=∑Pi2（1-Pi）2+∑PiPj(1-Pi)3+ ∑PiPj（1-Pj）3+∑PiPj（Pi+Pj）(1-Pi-Pj)2.This formula was compared with the 5 formulas (1)-(5) reported previously,and the PE values of 19 autosomal STR loci in AGCU EX20 system were calculated.Based on 1 000 samples of single-parentage cases and 1 000 unrelated individuals,the real experiment was designed and the real experiment results of PE were calculated.Ten million pairs of simulated biological mother and son and 10 million random individuals were gained by random simulation method,and the simulated experiment was designed and the simulated values of PE were calculated.In 19 STR loci,the sum of all allele frequency (S) was calculated,and the formula values of PE were compared with the values of real and simulated experiments.Results If S=I,the calculation values of formula (1),(2),(5) and (6) were quite the same,which accord with the double verification of real and simulated experiments.If S≠ 1,there was a minor error in the calculation results of formula (1),(2),(5) and (6),while which had a large error in formula (3) and (4).Conclusion The formula (6) derived in present study and the classical formulas (1),(2) and (5) can be applied to the standard triplet parentage testing.The S value has a certain influence on PE calculation.

In order to provide effective tool for further studying of the function of HCMV genome, developing of molecular vaccine and diagnostic reagents. Extraction of HCMV mRNA from HF cell infected by HCMV AD169 strain for 96h was reverse transcripted into cDNA, then was cloned into EcoR I-digested lambda gt11. HCMV AD169 strain cDNA expressing library has been constructed after packaging. The volume and the recombination rate of the prime cDNA expressing libraries was 3.6?10 6 and 96%, 168 positive clones of HCMV were screened by immune blotting with anti-HCMV mouse convalescent sera, 34 positive clones were obtained by dot nucleic acid hybridization with DIG-labled HCMV pp65 gene probe. 2 positive clones were amplified by HCMV pp65 all length primer. The PCR product has been tested by southern-blotting.The PCR product was sequenced and was taken as homology comparison by DNASIS software,and the homology is 98%.To lay the foundation of furher cloning,expressing the pp65 gene,further studying of the function of the pp65 prodct.