OBJECTIVESince glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common hereditary hemolytic erythrocyte enzyme deficiency, most cases have single nucleotide mutations in the coding region, and current test methods for gene mutation have some missed detections, this study aimed to investigate the feasibility of RT-PCR sequencing in the detection of gene mutation in G6PD deficiency.

METHODSAccording to the G6PD/6GPD ratio, 195 children with anemia of unknown cause or who underwent physical examination between August 2013 and July 2014 were classified into G6PD-deficiency group with 130 children (G6PD/6GPD ratio <1.00) and control group with 65 children (G6PD/6GPD ratio≥1.00). The primer design and PCR amplification conditions were optimized, and RT-PCR sequencing was used to analyze the complete coding sequence and verify the genomic DNA sequence in the two groups.

RESULTSIn the G6PD-deficiency group, the detection rate of gene mutation was 100% and 13 missense mutations were detected, including one new mutation. In the control group, no missense mutation was detected in 28 boys; 13 heterozygous missense mutations, 1 homozygous same-sense mutation (C1191T) which had not been reported in China and abroad, and 14 single nucleotide polymorphisms of C1311T were detected in 37 girls. The control group showed a high rate of missed detection of G6PD deficiency (carriers) in the specimens from girls (35%, 13/37).

CONCLUSIONSRT-PCR sequencing has a high detection rate of G6PD gene mutation and a certain value in clinical diagnosis of G6PD deficiency.

Adolescent ; Child ; Child, Preschool ; Female ; Glucosephosphate Dehydrogenase ; genetics ; Glucosephosphate Dehydrogenase Deficiency ; diagnosis ; genetics ; Humans ; Infant ; Male ; Mutation ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA

Objective Establish an overall performance evaluation index system for innovative medical devices transforming project,a major science and technology project in Zhejiang province.Methods The weight of indexes was calculated using Delphi method,analytic hierarchy process and weight of percentile method.Results A three-level performance evaluation index system was established.First level included 3 indexes,as condition index,process index and result index,with weight of 0.3401、0.4042 and 0.2557 respectively.Meanwhile,second level and third level included 13 and 38 indexes.Conclusions The index system provide a performance evaluation tool for the innovative medical devices transforming project.

Objective To explore effect of endogeneous gangliosides(Gls) and integrin ?2?1 on protein phosphotyrosine expression of pp125 focal adhesion kinase (pp125FAK) after adhesion of SK-N-SH neuroblastoma cells to collagen(Col).Methods SK-N-SH cell line with high expression of integrin ?2?1 was cultured in presence of D-threo-1-phenyl-2-decanolamino-3-morphinoline-1-propanol(D-PDMP).Effect of endogeneous Gls,anti-?2 and anti-?1 monoclonal antibody on protein phosphotyrosine expression of pp125FAK during adhesion of SK-N-SH cells to Col were determined by immunoprecipitate and Western blotting.Results After 6 days,endogenous Gls in cells were almost depleted.Gls-depletion,anti?2 and anti-?1 monoclonal antibody were able to decrease pp125FAK expression of SK-N-SH cells adherent to Col respectively.GD2,the major component of neuroblastoma cell Gls could reco-ver pp125FAK expression to a certain degree.Conclusions Endogenous tumor Gls regulate protein phosphotyrosine expression of pp125FAK during adhesion of neuroblastoma cells to Col.It is suggested that tumor Gls may increase signal transduction of tumor cell integrin ?2?1 by increasing tyrosine phosphorylation of pp125FAK.

Objective To investigate the clinical states of enuresis children companied with spina bifida occulta(SBO)and study the efficient way of treatment.Methods The children with SBO were check out by X ray from a total of 121 children with bedwetting.Their parents were asked to complete the enuresis questionnaires.Urine routine test and B-ultrasound examination about kidney,bladder and ureter were also asked to be done.The clinic data of the 49 children were attained and analyzed.They were randomly divided into 2 groups,and given the controlled treatment.Group A[used 1-deamino-8-D-arginine vas-opressin(DDAVP)only] and group B(used DDAVP plus oxybutynin plus bladder training)treated for 12 weeks.Results There were totally 49 bedwetting children companied with SBO,and most of them(44 cases,89.8%)were severe type(bedwetting times≥7 times/week).Some of them coexisted with frequency,urgency,gentle urgency incontinence and microscopic hematuria(22 cases).Thirty cases were found the functional bladder capacity(FBC)decrease by B-ultrasound.The cure rates were 58.3%(group A)and 88.0%(group B)respectively.The relapse rates were 36.8%(group A)and 12.5%(group B)respectively after stopping treatment for 3 months.Conclusions SBO accounts for considerably higher rate in enuretic children.It might cause the disability of bladder function.The treatment plan with DDAVP plus oxybutynin plus bladder training can not only increase the cure rate but also lower the relapse rate.

OBJECTIVETo study the association of functional bladder capacity with the severity of bedwetting in children with nocturnal enuresis.

METHODSA questionnaire investigation was performed in 1 500 children with nocturnal enuresis and the functional bladder capacity was examined by B-ultrasound.

RESULTSThe ratio of males to females was 1.3:1. The majority of patients (87%) were in an age range of 5-10 years, followed by the 10-14 years group (12%), and the 15-18 years group (1%). Six hundred and thirty-seven patients (42.4%) showed a decreased functional bladder capacity (less than 50% of normal level). The patients were classified into four groups according to the severity of bedwetting (from severe to mild): > or =2 times per night (n=53, 3.5%), > or =7 times per week (n=969, 64.6%), 3-6 times per week (n=380, 25.3%) and 1-2 times per week (n=98, 6.5%). The incidence of the reduction in functional bladder capacity in the above four groups was 79.2%, 48.3%, 29.7% and 14.3% respectively and a significant difference was noted among the four groups.

CONCLUSIONSMost of children with nocturnal enuresis showed decreased functional bladder capacity. Functional bladder capacity is associated with the severity of bedwetting in children with nocturnal enuresis.

Child ; Child, Preschool ; Female ; Humans ; Male ; Nocturnal Enuresis ; physiopathology ; Urinary Bladder ; physiopathology

Child ; DNA Methylation ; Humans ; Leukemia ; classification ; drug therapy ; genetics ; Neoplasm, Residual ; Prognosis

OBJECTIVETo study the effect of Fuzheng Sanjie recipe in regulating tumor-associated macrophages (TAMs) in Lewis lung cancer mice.

METHODEfforts were made to establish the Lewis lung cancer mouse model, weigh tumors and calculate the anti-tumor rate. The immunohistochemical method was used to examine the infiltration degree of CD68 + in tumor tissues in each group. ELISA was used to examine the content of IFN-γ, TGF-β, IL-4, IL-13, IL-6, IL-10, IL-12, TNF-α in mice serum.

RESULTCompared with the tumor-bearing model group, all of the other groups showed higher tumor inhibition rates, i. e. 50.28% for the DDP group, 34.37% for the TCM-preventing group and 66.76% for the Chinese and western medicine group, with statistical difference (P < 0.05), but without statistical difference in the infiltration degree of CD68+. The expressions of the IFN-γ, IL-6, IL-12 in tumor-bearing groups were lower than that in the blank control group, but with higher contents of IL-4, IL-13, TGF-β. Intervened with different drugs, there were significant differences in content among some relevant cytokines (P < 0.05), as well as statistical differences among the TCM prevention group, the Chinese and western medicine group and the tumor-bearing control group (P <0. 05) , but without statistical difference in TNF-α and IL-10 content from the tumor-bearing control group (P < 0.05).

CONCLUSIONFuzheng Sanjie recipe could reverse the immune remodeling effect and control the tumor growth by down-regulating the expressions of IL-4, IL-13, TGF-α in lung cancer immune microenvironment and up-regulating the expression of IFN-γ.

Animals ; Cell Line, Tumor ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Interleukin-10 ; blood ; Interleukin-12 ; blood ; Interleukin-13 ; blood ; Lung Neoplasms ; blood ; drug therapy ; immunology ; Macrophages ; drug effects ; immunology ; Male ; Mice ; Mice, Inbred C57BL ; Transforming Growth Factor beta ; blood ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVEThis study examined the changes of regional cerebral oxygen saturation (rSO2) by noninvasive near infrared spectrophotometry in neonates with meconium aspiration syndrome (MAS).

METHODSSeventy-three full neonates with MAS were divided into three groups by respiratory symptoms: asymptomatic group (group 1, n=38), common group (group 2, n=28) and severe group (group 3, n=7). Near infrared spectrophotometry was used to measure the cerebral rSO2 on days 1, 3, 5 and 7 after birth. Thirty healthy full-term newborns served as the Control group.

RESULTSThe cerebral rSO2 of group 1 decreased significantly compared with that of the Control group between days 1 and 3 (P < 0.05). The cerebral rSO2 of group 2 or group 3 was significantly lower than that of group 1 and the Control group on days 1, 3 and 5 (P < 0.05). The MAS patients with mild hypoxic-ischemic encephalopathy (HIE) had significantly higher brain rSO2 levels than those with medium or severe HIE on days 2, 3 and 5 (P < 0.05).

CONCLUSIONSThe cerebral rSO2 decreased in neonates with MAS. The values for rSO2 correlated with the severity of HIE in MAS patients.

Brain ; metabolism ; Female ; Humans ; Hypoxia-Ischemia, Brain ; metabolism ; Infant, Newborn ; Male ; Meconium Aspiration Syndrome ; metabolism ; Oxygen ; metabolism ; Spectroscopy, Near-Infrared