Background Liver X receptors (LXRs) are members of the nuclear hormone receptor superfamily which involve in energy metabolism of intracellular cholesterol and glucose regulation.Objective To investigate the effect of Liver X receptors (LXRs) agonists on the hypertrophy induced by angiotensin Ⅱ(Ang Ⅱ) in the murine HL-1 cardiomyocytes in vitro.Methods Hypertrophy in murine HL-1 cardiomyocytes was induced by Ang Ⅱ and treated with LXRs agonist T0901317 (1 ?mol/L).Immunofluorescent staining was carried out to identify the HL-1 cells.The surface area of HL-1 cells was analyzed by using NIH Image J software.The synthetic rate of protein in HL-1 cells was detected by {}3H leucine incorporation.The mRNA level of atrial natriuretic peptide (ANP) was measured by quantitative realtime PCR.Results HL-1 cells hypertrophy induced by Ang Ⅱ were manifested by the increases in surface area,mRNA expression of ANP,and {}3H leucine incorporation(control group vs Ang Ⅱ group:cell surface:1.00?0.16 vs 2.00?0.21,ANP mRNA:1.00?0.02 vs 1.58?0.27,{}3H leucine incorporation:1.00?0.03 vs 1.44?0.07,respectively,all P

Objective: Zhiyu presciption is composed of Whitmania pigra (Whitman) and Cornus officinalis Sieb. et Zucc. etc. It treats mainly for diabetic peripheral neuritis. To study the pharmacologic effects of Zhiyu prescription in this paper. Methods:The analgesic effect of Zhiyu presciption was examined through writhe method in mice, and the functions of Zhiyu prescription on extrinsic thrombosia and platelet aggregation in normal rats were observed. The model of alloxan induced diabetes in mice was established, the effect of Zhiyu prescription on blood glucose level was detected. The functions of nervus electrophysiology were examined on the model rats of diabetic peripheral neuritis induced by streptozotocin. Results: Zhiyu prescription could decrease the numbers of writhes of mice caused by acetic acid, reduce the blood glucose level in alloxan induced diabetic mice, improve the nervus conduction velocity of motor nerve.Conclusion: Zhiyu prescription is effective in clinical treatment of diabetic peripheral neuritis.

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.

Objective To explore the distribution of inflam m atory cells and positive expression of P-se-lectin glycoprotein ligand-1 (PSG L-1) in infant brainstem tissue from hand-foot-m outh disease related fatal brainstem encephalitis. Methods Tw enty brainstem sam ples from infants suffered from brainstem en-cephalitis w ere collected as the experim ental group. Ten brainstem sam ples from infants died of non-brain diseases and injuries w ere collected as the control group. The distribution of inflam m atory cells and the expression of PSG L-1 in the tw o groups w ere exam ined by im m unohistochem ical m ethod. The characteristics of the positive cells w ere observed. Results In brainstem tissue of the experim ental group, there w ere sleeve infiltrations of inflam m atory cells around the vessels and in the glial nodule. Microglia was the m ost and following was neutrophils around the vessels and in the glial nodule. There was a significant statistical difference am ong m icroglias, neutrophils and lym phocytes (P<0.05). There was no sleeve infiltration in the control group. PSG L-1 protein was expressed w idely in inflam m atory cells in the experim ental group, especially in the inflam m atory cells around the vessels and in the glial nodule. B ut PSG L-1 positive staining could be observed significantly less in the control group com paring with the experim ental group (P<0.05). Conclusion Microglia is the m ain type of inflam m atory cells involved in the progress of the fatal disease. Moreover, PSG L-1 could participate in the pathogenesis of hand-foot-m outh disease related fatal brainstem encephalitis.

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 investigate changes in expressions of activation antigens CD69 and HLA-DR in CD3+T lymphocytes in peripheral blood and skin lesions in patients with psoriasis vulgaris. Methods Peripheral blood samples were obtained from 20 patients with psoriasis vulgaris and 20 healthy controls, and skin specimens from the lesions of 15 out of the 20 patients and 10 healthy controls. Flow cytometry was performed to quantify the expressions of CD69 and HLA-DR in peripheral blood CD3+T cells, and an immunohistochemical study to measure the expression of HLA-DR in skin specimens. Statistical analysis was carried out by a two-sample t-test and Pearson correlation analysis with the SPSS 19.0 software. Results Compared with the healthy controls, the patients with psoriasis vulgaris showed increased expression rates of CD69 (4.70%± 1.90%vs. 1.56%± 0.95%, t=6.629, P<0.01)and HLA-DR (8.97%± 1.79% vs. 3.02% ± 1.15%, t= 6.204, P< 0.01)in peripheral blood. Pearson correlation analysis revealed that the percentage of CD3+HLA-DR+cells in peripheral blood was positively correlated with the psoriasis area and severity index (PASI)score (r=0.5626, P<0.05). The expression rate of HLA-DR was significantly higher in the dermis (64.87%± 17.31%vs. 19.80%± 5.69%, t=7.916, P<0.01), but lower in the epidermis(11.80%± 5.55%vs. 27.40%± 8.61%, t=5.479, P<0.01)in the psoriatic specimens compared with the control specimens. Immunohistochemically, HLA-DR was widely expressed in the dermis of psoriatic lesions, but mainly distributed around blood vessels in the control skin. Conclusions There is an aberrant activation of CD3+T cells in peripheral blood and inflammatory cells in skin lesions in patients with psoriasis vulgaris, and the percentage of CD3 +HLA-DR+ cells in peripheral blood is correlated with the severity of psoriasis vulagaris.

Objective To observe the effect of ultraviolet irradiation comparising 95％ ultraviolet A (UVA)and 5％ ultraviolet B(UVB)on the proliferation of human epidermal keratinocytes(HEKs),in hope to offer a basis for the construction of a photodamaged skin model induced by sunlight.Methods HEKs were isolated from foreskin tissue and cultured in vitro.After several passages,the HEKs were irradiated with different doses(0,2.5,5,10,20,30,40,60 J/cm2)of UV comprising 95％ UVA and 5％ UVB.Methyl thiazolyl tetrazolium(MTT)assay was used to evaluate cell viability after 24 hours of additional culture.SPSS 17.0 software was used to calculate the median lethal dose(LD50)of ultraviolet radiation in HEKs.Results The proliferation of HEKs was inhibited by 0,1.03％,6.60％,17.28％,31.28％,49.59％,59.67％ and 70.99％ respectively after irradiation with UV of 0,2.5,5,10,20,30,40 and 60 J/cm2.A significant inhibition of cell proliferation was observed in HEKs irradiated with UV at a dose of no lower than 10 J/cm2 compared with unirradiated HEKs(F =62.11,P ＜ 0.05).The LD50 of UV in HEKs was 31.31 J/cm2.Conclusions Aas the dose of UV irradiation increases,the proliferative activity of HEKs decreases,with the LD50 of UV being 31.31 J/cm2.

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