Adolescent ; Child ; Child, Preschool ; Critical Care ; Fatty Acid-Binding Proteins ; blood ; metabolism ; urine ; Female ; Gastrointestinal Diseases ; blood ; urine ; Humans ; Male ; Metabolic Diseases ; blood ; urine

Objective To observe the changes of serum ferritin (SF)in neonates with mechanical ventilation and its clinical significance in the ventilation-induced lung injury (VILI). Methods The study was carried out in 36 neonates with mechanical ventilation and 31 neonates without mechanical ventilation in neonate intensive care unit(NICU).SF level in venous blood was measured on 1,24,48,72 hours after mechanical ventilation and 24 hours after mechanical ventilation removal by radioimmunoassay (RIA ).SF level of non-mechanical ventilation group were determined at the same time. Results SF levels in mechanical ventilation groups were significantly higher than those of control group (P

Objective To explore the clinical significance of monocyte chemoattractant protein-1(MCP-1)in children with Henoch-Schonlein purpura nephritis(HSPN).At the same time compare the association between serum and urine MCP-1,to investigate the impact of the both on them in children with HSPN.Methods Serum and urine MCP-1 were measured by enzyme linked immunosorbent assay in 50 children with Henoch-Schonlein purpura(HSP)(25 cases of them patients with renal injures),and 25 healthy children,the changes of serum and urine MCP-1 were compared;at the same time serum urea nitrogen,creatinine,urinary albumin,urine N-acetyl-D-glucosaminidase(NAG),urine ?2-MG,24 hours urinary levels of protein were investigated in children with HSPN by analyzing the correlation between these indicators and serum and urine MCP-1;urine MCP-1 in HSPN group were measured in recovery period,and were compared with urine MCP-1 in HSP group and HSPN group in acute period.Results 1.The expressions of urine MCP-1 was significantly higher in HSPN group than those in HSP group and healthy controls(P0.05).2.Urine MCP-1 levels were associated with proteinuria in children with HSPN,but serum MCP-1 levels had nothing to do with HSPN.3.There was a close correlation between urine MCP-1 expression and urinary albumin,urine NAG,urine ?2-MG and 24 hours urinary levels of protein,but the expression of urine MCP-1 levels were not correlated with the serum urea nitrogen and creatinine.4.There was statistical significance in urine MCP-1 in acute and recovery periods with HSPN group(P

Biomarkers ; blood ; Child ; Child, Preschool ; Critical Illness ; Early Diagnosis ; Enzyme-Linked Immunosorbent Assay ; Female ; Gastrointestinal Diseases ; blood ; diagnosis ; etiology ; Humans ; Infant ; Male ; Peptides ; blood ; Severity of Illness Index ; Trefoil Factor-2

A promoter-trap vector pGBT14 for selecting promoters of fungus gene was constructed with E. coli-yeast shuttling plasmid pGBT9. Using this vector, a0. 5-2. 0kb chromosomal DNA library of Cepholosporium acremonium was constructed, and twenty four DNA fragments with promoter function in Saccharomyces oerevisiae Y153 were selected from this DNA library. And the promoter function of these DNA fragments was analyzed.

TDP-4-ketohexulose reductase, encoded by dnmV, is important in daunorubicin biosynthesis. To obtain a daunorubicin block mutant, double cross-over plasmid pYG817 was constructed by inserting apramycin resistant gene and amplified dnmV together with upstream dnmU into vector pUC18. dnmV was successfully disrupted after transformation of daunorubicin-producing strain SIPI-1482 by pYG817. Daunorubicin was absent from metabolites of the resulting transformant, and its biosynthesis can be reconstituted by introducing dnmV expression plasmid into the disruptant, although the yield is lower than wild-type SIPI-1482, according to HPLC analysis. This mutant can be a good candidate for production of anthracycline such as epi-daunorubicin by introducing exogenous gene into the host.

To improve the growth enhancement activity of Vitreoscilla hemoglobin(VHb), Vitreoscilla hemoglobin gene(vgb) was mutated by error-prone PCR and then reconstituted by DNA shuffling. The shuffling library was constructed by inserting the shuffled genes into the downstream of vgb natural promoter and transforming them into E.coli DH5?. Mutated active VHb proteins were first screened in test tubes according to host cell pellets color and then in shake flasks according to host pellets wet weight .One active mutant protein, VHb′042506, was obtained after second screening. It could increased the host wet weight by 31.25% and 58.75% than that of the control which bearing natural VHb under microaerobic and extremely microaerobic conditions, respectively. Sequencing and alignment results showed that 11 nucleotides were mutated, thus resulted in 4 amino acids changes occurred in this mutant protein. CO difference spectrum test also indicated that it had higher specific absorption.

A novel gene,located between dnrX and drrB in the genome of daunorubicin-producing strain Streptomyces coeruleorubidus SIPI-1482,was cloned and named as dauW.The full sequence of dauW was submitted to GenBank(Accession No.EF523565).Blast result indicated that it showed high homology with dnrW in GenBank.The exact function of dauW is as yet unknown despite the possibility that it might belong to a family of FAD-dependent oxidoreductases on the basis of conserved domain analysis.dauW was cloned into expression plasmids pET-28a(+)and pET-32a(+),respectively,and was successfully expressed in E.coli DE3 after induction with IPTG.The preliminary results of the expression of dauW suggested that it might be involved in the self resistance in Streptomyces coeruleorubidus due to the increased resistance to daunorubicin in the E.coli host.

TTI gene coding for Tsetse thrombin inhibitor was modified with E.coli bias codon and expressed in Escherichia coli with high efficiency.Recombinant protein was purified to more than 98% purity.Assay for enzyme activity determination was set up.The result showed that the fusion protein exhibited inhibiting activity for thrombin.Inhibitory rate of purified TTI was 73% when concentration of thrombin and substrate was 10U/ml and 250?mol/L respectively.Inhibition pattern was determined as competitive with Ki at 35?mol/L.

?-lactamase inhibitor research is popular for its potential on ?-lactam antibiotics resistant strain.A ?-lactamase binding peptide SIPIS04-01 was obtained by the yeast two-hybid system.In vitro assay showed that it can inhibit the ?-lactamase activity.In order to improve the expression level of the recombinant peptide,a two-copy expression plasmid pYG563 was constructed by random orientation tandem repeat method after codon modification,the two-copy plasmid was successfully expressed and the product was increased by 48.4% than that of one-copy plasmid.Purified peptide showed inhibitory activity against TEM-1 ?-lactamase in vitro and the inhhibitory constant Ki was measured.