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.

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.

Objective To investigate the factors of families related to the outcome of rehabilitation for cerebral palsy children. Methods Families of 153 cerebral palsy children were investigated with questionnaires of family condition, knowledge of cerebral palsy, compliance,and the Eysenck Personality Questionnaire, Marital Quality Questionnaire, Social Support Scale, Symptom Check List, Happiness Index.The children were assessed with Comprehensive Assessment for Disable Children before and after rehabilitation. Results There were significant differences of compliance, knowledge of cerebral palsy, being of other healthy children, personality of extraversion/introversion and lying,and mental symptom between the families of the cerebral palsy children improved or not after rehabilitation (P<0.05). Multivariate Logistic regression analysis showed that, being of other healthy children, personality of extraversion/introversion and mental symptom were the independent factors. Conclusion It may relate to the outcome of rehabilitation for cerebral palsy children of family condition, personality and mental health of the families, which need further intervention.

Objective:To investigate the clinicopathological features and prognosis of IgA nephropathy (IgAN) patients with gross hematuria.Methods:The clinical and pathological data of 490 primary IgAN patients admitted in the Affiliated Hospital of Qingdao University from January 2010 to June 2019 were analyzed. Patients were divided into gross hematuria group and non-gross hematuria group. The clinical and pathological features and prognosis were compared between the two groups. All patients were diagnosed by kidney biopsy, and followed up until June 30, 2020. Kaplan-Meier survival curve was used to examine the renal survival,and Cox regression model was used to analyze the risk factors affecting renal survival in two groups.Results:Among 490 patients there were 111 patients (22.7%) in the gross hematuria group and 379 patients (77.3%) in the non-gross hematuria group. Age, hypertension, 24-h urine protein, serum creatinine, blood uric acid, blood triglycerides, total blood cholesterol level, mesangial cell hyperplasia (M1), the proportion of renal tubular atrophy or renal interstitial fibrosis (T1/2) in gross hematuria group were lower than those in non-gross hematuria group ( P<0.05), while the estimated glomerular filtration rate (eGFR) in gross hematuria group was higher than those in non-gross hematuria group ( P<0.05). Four hundred and twenty six patients (86.9%) were followed up for at least 6 months, including 93 patients in gross hematuria group and 333 patients in non-gross hematuria group. There was no statistically significant difference in treatment method between the two groups ( P>0.05). The incidence of end-point events in non-gross hematuria group was higher than that in gross hematuria group [18.6%(62/333) vs. 6.5%(6/93), χ2=8.023, P<0.05]. Kaplan-Meier survival analysis showed that the cumulative renal survival rate of the gross hematuria group was higher than that of the non gross hematuria group (χ2=11.44, P<0.001). Multivariate Cox regression analysis showed that urine protein>1 g/24 h, eGFR<60 ml·min -1·(1.73 m 2) -1, hypertension, hyperuricemia and the elevated serum IgA/C3 were risk factors for renal survival [ HR(95% CI)=3.457(1.137-10.514),2.736(1.073-6.977),2.720(1.144-6.465),2.789(1.102-7.060),1.070(1.009-1.135), all P<0.05]. Conclusions:IgAN patients with gross hematuria has less severe kidney damage and higher cumulative renal survival rate than non-gross hematuria patients. Urinary protein>1.0 g/d, hypertension, hyperuricemia and the elevated serum IgA/C3 are risk factors for adverse end-point events, to which timely attention and corresponding treatment should be given.

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 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 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 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.