Enterovirus A, Human ; pathogenicity ; Enterovirus Infections ; complications ; Humans ; Pulmonary Edema ; etiology ; virology

Using bioactive compounds 7-hydroxy flavone, salicylaldehyde, cinnamic acid and 4-amino-5- mercapto-1,2,4-triazoles as starting materials, three new types of flavone derivatives containing 1,2,4-triazole structure were synthesized via different step reactions. These new compounds were characterized by 1IHNMR, ESI-MS, IR and elemental analysis. Their scavenging effects on the superoxide radical (O2·-), hydroxyl radical (·OH), DPPH · radical and their total reduction activities were tested. The results showed that all of the compounds possessed some antioxidative activity at the concentration of 0.5 mg · mL(-1), but the scavenging ability of the target compounds was lower than that of the standard compound Vc.
Cinnamates ; chemistry ; Flavones ; chemical synthesis ; chemistry ; Flavonoids ; chemistry ; Free Radical Scavengers ; chemical synthesis ; chemistry ; Triazoles ; chemistry

Objective To construct the eukaryotic expression plasmid containing glyceraldehydes-3-phosphate dehydrogenase (GAPDH) and cysteine protease inhibitor ( CPI ) gene from periodic Brugia malayi (Bm) and to lay foundation for studying multivalent vaccines. Methods Total RNA was extracted from periodic Bin. The BmGAPDH and BmCPI genes were amplified by RT-PCR. The PCR product was cloned and then subeloned into eukaryotic recombinant plasmid vector pcDNA3.1 (+). pcDNA3.1 (+)/BmGAPDH/BmCPI was constructed. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification, and were transformed into HeLa cell subsequently. The transient expression of BmGAPDH and BmCPI were examined by RT-PCR. The expressed protein was identified by sodium dodeeylsulphate-polyacrylamide gel electrophoresis(SDS-PAGE). Results Two specific bands of around 877 bp of BmGAPDH and 621 bp of BmCPI were amplified, consistent with the expected value. The same bands were obtained by double restriction enzyme digestion of recombinant plasmids or PCR using recombinant plasmid as template. BmGAPDH and BmCPI mRNA were highly expressed in transfeeted HeLa cell. The relative molecular mass (Mr) of the recombinant protein was about 54 × 103. Conclusion The recombinant eukaryotic expression plasmid pcDNA3.1 (+)/BmGAPDH/BmCPI has been constructed successfully and the protein is expressed correctly in mammalian cell.

OBJECTIVEAccurate and reliable assessment of renal function is important in the management of children with chronic kidney disease (CKD). Glomerular filtration rate (GFR) is the best index of assessing kidney function. For assessment of GFR, both gold standard tests and prediction equations have been used. The well-known 24-hour endogenous creatinine clearance (Ccr), the Schwartz formula and the Filler formula are increasingly used in daily clinical practice. However, there are few studies on the applicability of these prediction equations for estimating GFR in Chinese children with CKD. The aim of this study was to compare these prediction equations estimating GFR with an isotope clearance method [isotope glomerular filtration rate (rGFR)] in such patients.

METHODChildren aged 1-16 years who underwent isotope (99m)Tc-diethylenetriaminepentaacetic acid ((99m)Tc-DTPA) GFR testing (Gates' method) between the year of 2002 and 2005 were studied retrospectively. GFR was estimated using: (1) 24-hour Ccr, which was calculated using the standard formula: [urine creatinine (milligrammes per millilitre) × 24-hour urine volume/serum creatinine (milligrammes per millilitre) × 1440] × [1.73 (m(2))/body surface area (m(2))]; (2) the Schwartz formula, which is: eGFR (ml/min per 1.73 m(2)) = k × height (centimetres)/serum creatinine (micromoles per litre), where k is 62 in males at 13 years of age and older, 40 in infants, and 49 in all other children; and (3) the Filler formula, which is: logGFR = 1.962 + [1.123 × log(1/Cys C)], where cystatin C is measured in milligrammes per litre. Serum and urinary creatinine levels were detected by alkaline kinetic method. Serum cystatin C was analysed by particle-enhanced immunoturbidimetric assay. Bias and precision were evaluated.

RESULTThirty subjects (18 males and 12 females; mean age 9.4 years) fulfilling both inclusion criteria and exclusion criteria were included in this study. The mean (SD) rGFR was 81.57 (36.92) ml/min per 1.73 m(2); 18 subjects were in CKD stage I, 8 in CKD stage II, 8 in CKD stage III, and 1 in CKD stage IV. Only the mean 24 h Ccr-eGFR was slightly higher than rGFR (0.4 ml/min per 1.73 m(2) higher). Within 95% limits of agreement, the maximum absolute value of bias was about 50 ml/min per 1.73 m(2). Accuracy (estimated GFR values within ± 30% of rGFR) for all formulae was poor, ranging from 23.3% to 43.3%. All formulae overestimate or underestimate rGFR in different CKD stages.

CONCLUSIONIn Chinese children with CKD, there was a significant difference between measured GFR and estimated GFR using 24h Ccr, Schwartz formula and Filler formula. More suitable GFR predictive equations to assess glomerular function of such patients should be developed.

Adolescent ; Child ; Child, Preschool ; Female ; Glomerular Filtration Rate ; Humans ; Infant ; Kidney Failure, Chronic ; physiopathology ; Kidney Function Tests ; Male ; Renal Insufficiency, Chronic ; physiopathology

Objective To learn the quality of life (QOL) and mental health status of the physically disabled people in Zhejiang Province,in order to provide evidence for improving the quality of life and mental health of the physically disabled people.Methods Using questionnaires and interviews to make household surveys of 950 physically disabled people.The questionnaire included the brief quality of life scale (SF-36) and the general health questionnaire (GHQ-28) in the anxiety and depression scale,physical and mental health status.The basic information and other living conditions of physically disabled people were collected through interviews.Results A total of 907 valid questionnaires were recovered,and the effective response rate was 95.47％.Scores of QOL of 907 physically disabled people was 60.89,and anxiety and depression scores were more than 4 points and there was a high score on anxious and depression.The scores of quality of life,anxiety and depression were not statistically significant between males and females(P ＞0.05).There were significant differences in all dimensions of SF-36 and anxiety and depression in different levels of physically disabled people(P ＜ 0.01).There was a significant negative correlation between the quality of life and anxiety and depression in physically disabled people(rs =-0.626、-0.639,P ＜ 0.01).The multiple linear regression showed that physiological function,role physical,physical pain and general health were the influencing factors of anxiety of the physically disabled people,and role physical,physical pain and general health were the influencing factors of depression of the physically disabled people.Condusion The quality of life of physically disabled people were comparably poor,and presented anxiety,depression and other psychological problems.Physical function status directly affects the quality of life and mental health status.

Objective To clone and sequence the cysteine protease inhibitor gene of periodic Brugia malayi(BmCPI) and predict B-cell epitopes in amino acide sequence of BmCPI in order to provide basis for further study the expression of BmCPI and its function. Methods Total RNA was extracted from periodic Brugia malayi.A couple of specific primers were designed on the basis of known sequences of cysteine protease inhibitor gene from BmCPI. The desired gene was amplified by PCR technique from cDNA. The PCR products were purified and cloned into plasmid pGEM-T by T-A cloning method, transformed into Escherichia coli(E, coli) strain DH5α. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification. Five parameters and methods were used to predict B-cell epitopes in amino acide sequence of BmCPI. Results For RT-PCR, a specific band of around 621 bp was amplified. The same band was obtained by double restriction of recombinant plasmids or PCR using recombinant plasmid as template. The result of DNA sequencing showed that BmCPI shares 99% nucleotide sequence identity with that of published sequence. It showed that B-cell epitopes were probably at or adjacent to 23 - 32, 50 - 79 and 117 - 126 in its amino acide sequence. Conclusions pGEM-BmCPI is successfully constructed and sequenced, anticipated objective is reached and conditions is provided for further study of BmCPI expression and its function.

Increasing evidence has revealed that maternal cytomegalovirus (CMV) infection may be associated with neurodevelopmental disorders in offspring.Potential relevance between the placental inflammation and CMV-related autism has been reported by clinical observation.Meanwhile,abnormal expression of Toll-like receptor 2 (TLR2) and TLR4 in placenta of patients with chorioamnionitis was observed in multiple studies.IL-6 and IL-10 are two important maternal inflammatory mediators involved in neurodevelopmental disorders.To investigate whether murine CMV (MCMV) infection causes alterations in placental IL-6/10 and TLR2/4 levels,we analyzed the dynamic changes in gene expression of TLR2/4 and IL-6/10 in placentas following acute MCMV infection.Mouse model of acute MCMV infection during pregnancy was created,and pre-pregnant MCMV infected,lipopolysaccharide (LPS)-treated and uninfected mice were used as controls.At E13.5,E14.5 and E18.5,placentas and fetal brains were harvested and mRNA expression levels of placental TLR2/4 and IL-6/10 were analyzed.The results showed that after acute MCMV infection,the expression levels of placental TLR2/4 and IL-6 were elevated at E13.5,accompanied by obvious placental inflammation and reduction of placenta and fetal brain weights.However,LPS 50 μg/kg could decrease the IL-6 expression at E13.5 and E14.5.This suggests that acute MCMV infection during pregnancy could up-regulate the gene expression of TLR2/4 in placental trophoblasts and activate them to produce more proinflammatory cytokine IL-6.High dose of LPS stimulation (50 tg/kg) during pregnancy can lead to down-regulation of IL-6 levels in the late stage.Imbalance ofIL-6 expression in placenta might be associated with the neurodevelopmental disorders in progeny.

OBJECTIVETo investigate the genetic polymorphism of human cytomegalovirus (HCMV) glycoprotein H (gH) from infantile clinical isolates, to analyze the genotypic distribution of gH in different diseases of HCMV infection and try to find the correlations between the diseases and genotypes.

METHODFresh urine specimens were collected from the hospitalized children with different diseases whose blood HCMV-IgM and HCMV-IgG were positive. Virus was isolated from these specimens. Glycoprotein H of harvest clinical isolates was genotyped by nested-PCR combined with restriction fragment length polymorphism (RFLP), the purified PCR products were digested by restriction endonuclease HhaI. The digested products were genotyped by polyacrylamide gel electrophoresis and silver staining. Classification and results of sequencing were compared.

RESULTTotally 102 HCMV clinical isolates were obtained. Glycoprotein H gene of these clinical isolates (43 cases had infantile hepatitis syndrome, 38 cases had anicteric hepatitis, 13 pneumonia, 7 thrombocytopenic purpura, and 1 congenital CMV infection) were positive by nested-PCR, whose positive rate was 100%. The results showed that 62 strains were gH1 genotypes (60.8%), while 40 strains were gH2 (39.2%), mixed type or new genotype was not observed. In infantile hepatitis syndrome (26 clinical isolates were gH1 genotypes, 17 clinical isolates were gH2 genotypes), anicteric hepatitis (25 were gH1, 13 were gH2) and pneumonia (9 were gH1, 4 were gH2), the distribution of HCMV gH genotypes of infantile clinical isolates was consistent with the overall trend (χ(2) = 0.357, P > 0.05). However , the gH2 was more common than gH1 in the clinical isolates of patients with thrombocytopenic purpura (6 were gH2, 1 were gH2, χ(2) = 6.083, P < 0.05).

CONCLUSIONGenotype 1 was the dominant genotype of glycoprotein H in HCMV clinical isolates from our hospital infants. There was no significant difference between the distribution of gH genotypes in infantile hepatitis syndrome, anicteric hepatitis and pneumonia. However, gH2 was the dominant genotype in thrombocytopenic purpura. These findings suggested that there may be a certain relevance between gH genotype and different clinical manifestations.

Amino Acid Sequence ; Base Sequence ; Child, Preschool ; Cytomegalovirus ; classification ; genetics ; isolation & purification ; Cytomegalovirus Infections ; virology ; DNA Primers ; DNA, Viral ; genetics ; Female ; Genotype ; Hepatitis ; virology ; Humans ; Infant ; Infant, Newborn ; Male ; Pneumonia, Viral ; virology ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Urine ; virology ; Viral Envelope Proteins ; genetics

Biomedical Research ; Child ; China ; Cytomegalovirus Infections ; diagnosis ; Diarrhea ; diagnosis ; therapy ; Hepatitis ; diagnosis ; therapy ; Hepatitis B ; transmission ; Humans ; Infectious Disease Medicine ; classification ; Infectious Disease Transmission, Vertical ; prevention & control ; Pediatrics ; classification

BACKGROUNDMurine cytomegalovirus (MCMV) early protein M112-113 is involved in viral DNA replication and believed to play a crucial role in the viral pathogenesis. To investigate the biological function of M112-113 protein in the pathogenesis of the brain disorders caused by cytomegalovirus (CMV), a screening for proteins interacting with M112-113 was performed by a yeast two-hybrid system.

METHODSBait plasmid pGBKT7-M112-113 was constructed and transformed into AH109 yeast. After confirmation of the expression of MCMV M112-113 in yeast, the bait yeast was mated with a prey yeast containing mouse brain cDNA library plasmid to screen the proteins interacting with M112-113. Interactions between M112-113 and the obtained proteins were verified by yeast two-hybrid assay and chemiluminescent co-immunoprecipitaion.

RESULTSTwo proteins interacting with M112-113 were identified, including metastasis-associated 1 (MTA1) and zinc finger, CCHC domain containing 18 (ZCCHC18). M112-113 protein could interact with MTA1 or ZCCHC18 in yeast and mammalian cells.

CONCLUSIONThe interactions of M112-113 with MTA1 or ZCCHC18 may be related to the pathogenesis of MCMV-associated disease in central nervous system.

Animals ; Brain ; metabolism ; Cell Line ; Humans ; Immunoprecipitation ; Mice ; Muromegalovirus ; metabolism ; Plasmids ; Protein Binding ; Two-Hybrid System Techniques ; Viral Proteins ; metabolism