Objective To explore influence of long-term inhaled glucocoticoids(IGS) on soluble intercellular adhesion molecule-1(sICAM-1)) in children with bronchial asthma.Methods Enzyme-linked immunosorbent assay(ELISA) method was used to detect the serum sICAM-1 level in 36 healthy children and 29 children with bronchial asthma(untreated and post-treated for 3,6 and 12 months).Results 1.Serum sICAM-1 level was significantly elevated in children with asthma and significantly higher than that in normal control group(P

Objective:To compare the two-dimensional electrophoresis(2-DE) maps of rat spinal cord protein extracted by two different solution systems.Methods: Adult rat spinal cord protein was precipitated with 10% trichloracetic acid in acetone and resuspended in 8 mol/L urea plus 4%CHAPS (A solution) or, 5 mol/L urea, 2 mol/L thiourea, 2%CHAPS plus 2%SB3-10 (B solution). One hundred and fifty micrograms of protein was loaded on 18 cm IPG strip holder and run isoelectric focusing electrophoresis as the first dimension, then horizontal SDS-PAGE as the second dimension. Protein spots were visualized by silver stain.Results:There were 1 059 and 1 023 protein spots in each map, of which 790 spots were matched in two maps. There were 269 and 233 spots exclusively extracted by A and B solutions, respectively. Taken together, 1292 different spots were totally obtained by A and B solutions.Conclusion: Integrating protein spots extracted by different solution systems is beneficial for achieving intact 2-DE map of tissues.

Objective To study the uncertainty and traceability of HBV DNA assays and discuss the comparability of results among different detection systems. Methods Different detecting systems were used to detect HBV DNA using the national standard substance as "quality control substance". The uncertainty of the results was evaluated referring "Guidelines for estimating and reporting measurement uncerTAinty of chemical test results" of NATA The results were traced back to the national standard substance. According to the CLSI document EP9-A2, the results were analyzed and subjected to bias estimation with the t(0.05sv) √u2b1+ u2b2 as the criterion clinically accepted to investigate the comparability of different detecting systems. Results The means (-y) measured by 3 HBV DNA assay systems were 6.15,5.88,and 6.31 lg(kIU/L) respectively. Except system A,both the biases of system B and C had statistical significance (all P ＜ 0. 05) and expanded uncertainty of three detection systems was varied, but the difference was within the maximum acceptable range (± 0. 5) of the external quality assessment by National Center for Clinical Laboratory. Being traceable to national standard substance, the results of HBV DNA of the three detecting systems were (5.45 ± 1.23), (5.55 ± 1.32) and (5.42 ± 1.25) lg(kIU/L), respectively.There was significant difference among three systems (F = 5.63, P ＜ 0. 05). Comparing system A and B,there was significant difference in statistic (q = 5. 12, P ＜ 0. 05) and the difference between system B and C also had statistically significant (q = 6. 85, P ＜ 0. 05), but the results between system A and C had no statistical difference (q = 1.85,P ＞ 0. 05). Among these three systems, the difference of any two detection systems had no statistical significance (all P ＞ 0. 05). It showed that system bias was acceptable in clinical application and the results between different systems were comparable. Conclusions It is necessary to estimate the uncertainty and traceability when comparing the HBV DNA assay among the different labs. It also needs to estimate the bias of different systems and evaluate the clinical acceptability to ensure the accuracy and comparability of the results.

Objective To establish a flow cytometric measurement of detecting minimal residual disease(MRD) according to the leukemia-associated immunophenotypes in children with acute lymphoblastic leukemia(ALL) and to explore the significance of MRD detection in ALL children for a individualized treatment. Methods A variety of four-color fluorescent antibody combinations were used to investigate the children's normal bone marrow. The normal bone marrow pattern at two-parameter plots was established to identify the residual tumor cells, seventy-five bone marrow samples from newly diagnosed ALL children were analyzed with four-color cytometry to determined the optimal combinations which can clearly distinguish the tumor cells from normal cells. The bone marrow samples were monitored with the combination panel in 60 patients at the end of induction therapy and follow-up treatment. Cytomorphology test, PCR amplification of 29 fusion genes as well as IgG and TCR gene rearrangements were performed simultaneously. Results Sixty-nine cases (92.0%) could be identified for effective antibody combinations to monitor MRD by four-color cytometry. Fusion genes or IgG and T cell receptor (TCR) gene rearrangements can be detected in 21 cases (28.0%) to monitor MRD by PCR. No MRD can be detected in 25 bone marrow samples at the end of induction therapy and follow-up treatment. Four-color cytometry could detect as low as 0.021%-4.130% residual leukemia cells. Conclusion MRD can be monitored by flow cytometry which is faster than PCR, and the sensitivity is superior to morphology method.

The gene (eg1) encoding for novel endoglucanase 1 was cloned previously from Chinese straw mushroom Volvariella volvacea. EG1 has high thermal stability and optimal pH at neutral and shows great potential in textile and paper industry applications. To improve the expression level of EG1 in Pichia pastoris, the increasing copy number of clone, and its high cell density fermentation in 3.2L fermenter for its high-level expression were investigated in this work. By electro-transformation of pPICZalphaB-egl into GS115EG11 integrated with single copy of eg1 gene, A resistant transformant with 3.8 times higher level expression than GS115EG11 was screened from YPDSZ plate containing 2000 microg/mL of Zeocin. The effect of initial cell density, pH and methanol on its expression and biomass accumulation was evaluated in shaking culture. Optimal EG1 production was observed when initial cell density OD600 was 5.0. EG1 production and biomass accumulation did not seem to vary when cells were induced at different pH values. Both of EG1 and cell density were found to increase with higher methanol concentrations, reaching 62.48 IU/mL and 31.7 (OD600) respectively after 120 h induction with 2.0% (V/V) methanol compared to 30.24 IU/mL and 17.79 (OD60) with 0.25% methanol induction. EG1 expression was further increased by 6.4 times higher than shaking culture after 95.5 hours induction with methanol in fed-batch fermentation, so totally 34 times higher than that for GS115EG11 was achieved by screening of high Zeocin resistant clone and high cell density fermentation. The production of EG1 with 543.36IU/mL CMC activity and 8.80mg/mL protein expression was obtained in Pichia pastoris.
Cellulase ; biosynthesis ; genetics ; Cloning, Molecular ; Culture Media ; Electroporation ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Fungal Proteins ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Volvariella ; enzymology ; genetics

OBJECTIVETo investigate the viral pathogens of acute respiratory infection (ARI) in hospitalized children from Suzhou of China.

METHODSThe nasopharyngeal aspirate samples were obtained from 1,668 hospitalized children with ARI. Common respiratory viruses, including respiratory syncytial virus (RSV), influenza virus A and B, parainfluenza viruses 1, 2 and 3 and adenovirus, were detected using direct immunofluorescence. Human metapneumovirus (hMPV) gene fragments were detected by RT-PCR.

RESULTSViral agents were identified in 597 cases (35.8%). RSV was the most frequent (17.6%). RSV infection is more common in children less than 1 year old. A highest detection rate of RSV was found during winter and spring. hMPV was detected in 10.6% of the cases, with a peak detection rate between March and May. Single viral infection was found in 561 cases (33.6%) and mixed viral infection in 36 cases (including 27 cases at age of less than 1 year). RSV and hMPV co-infection was common (n=22).

CONCLUSIONSRSV is common pathogen of ARI in children from Suzhou. RSV viral activity peaks during winter and spring. The children at age of less than 1 year are susceptible to RSV. hMPV is also an important pathogen of ARI, with a peak detection rate between March and May. Mixed viral infection is common in children less than 1 year old.

Acute Disease ; Age Factors ; Child ; Child, Hospitalized ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Male ; Metapneumovirus ; isolation & purification ; Nasopharynx ; virology ; Respiratory Syncytial Virus, Human ; isolation & purification ; Respiratory Tract Infections ; epidemiology ; virology ; Seasons

OBJECTIVETo evaluate the clinical and laboratory features of pediatric inv(16) acute myeloid leukemia (AML) retrospectively.

METHODDual color fluorescence in situ hybridization (D-FISH) using a LSI CBFβ inv(16) break apart probe labeled by Spectrum red and Spectrum green was performed in 15 acute myeloid leukemia cases, including 13 cases with or without abnormal eosinophils but with positive core binding factor β (CBFβ)-MYH11 fusion transcript detected by RT-PCR, and 2 cases with trisomy 8 (+8). The results were compared with the morphology, immunophenotype, karyotype and RT-PCR.

RESULTMorphologically, 12 cases were diagnosed as M(4)EO, 2 as M(4), and 1 as M(2a). Immunophenotypically, all 13 AML cases with inv(16) showed positive expression of CD(13) and CD(33), but without the expression of any lymphoid lineage antigens. Karyotyping analysis with G-banding detected inv(16) in 10 AML cases, including 9 M(4)EO cases and 1 M(2a), but only 5 positive cases were detected using R-banding technique. Among them, 2 cases had simultaneous +8 and trisomy22 (+22), one had +22 only in addition to inv(16). D-FISH revealed a CBFβ-MYH11 rearrangement in 13 cases of AML with positive RT-PCR results, and the mean positive rate of cell detection was 55.15% (range 37.0% - 86.0%). The complete remission rate (CR) and median survival period in this series of inv(16) AML were 81.5%and 11 months, respectively, of whom, 8 cases were still in CR. Relapse and karyotypic evolution were seen in case 5 with +8, +22 in addition to inv(16).

CONCLUSIONAML with inv(16) is a special subtype. Most cases belong to M(4)EO. Its prognosis is good in general, but it seems to be an unfavorable feature for AML with inv(16) and +8, +22 simultaneously, especially with karyotypic evolution. For detection of inv(16), G-banding technique is evidently superior to R-banding technique. D-FISH combined with RT-PCR are more sensitive and reliable than chromosome banding analysis.

Adolescent ; Child ; Child, Preschool ; Chromosome Deletion ; Chromosome Inversion ; Chromosomes, Human, Pair 16 ; genetics ; Eosinophilia ; pathology ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Infant ; Karyotyping ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; Male ; Prognosis ; Retrospective Studies ; Reverse Transcriptase Polymerase Chain Reaction

Along with the population aging in China, patients with lumbar spinal stenosis(LSS) caused by recessive change incessantly increase. At present, there is no adequate evidence to recommend any specific nonoperative treatment for LSS, and surgery is still an effective method. The cilincal symptoms of the patients without conservative treatment got improvement after surgery, which is the strongest evidence base. Spinal instability after simple decompression promotes the development of fusion technique, and the accelerated adjacent segment degeneration and no relief in symptoms after fusion lead to dynamic fixation technology emerge as the times require. Patients with spinal canal decompression whether need bone fusion or not is still controversial. For the past few years, the operation of simple decompression for LSS obviously decreased, whereas the decompression plus fusion surgery showed sustainable growth. Decompression complicated with fusion was more and more adopted in LSS, in order to reduce the hidden risk of spinal instability and deformity. Although decompressive operation has determinate effect, now it is still unclear if the therapeutic effect of decompression complicated with fusion is better than simple decompression. This article reviews the current studies to explore whether decompression plus bone fusion is applicable for LSS. To further explore the best choice of surgical treatment for LSS, we focused on evidence-based therapeutic options.

OBJECTIVETo investigate the effect of cell surface sialic acid and its linkage on the cell-cell and cell-matrix adhesion of mammary carcinoma cells MD-MB-435.

METHODSMD-MB-435 cells were sense-transfected with ST6Gal I cDNA or antisense-transfected with part of the ST6Gal I sequence inserted in pcDNA 3.1 vector, with mock transfection with pcDNA3.1 vector as the control. The cell surface alpha2, 6-linked sialylation was determined by fluorescence-activated cell sorting (FACS) using lectin SNA (Sambucus nigra agglutinin specific to alpha2, 6-linked sialic acid on N-linked glycoprotein). A significantly increased alpha2, 6-sialylation subclone in sense-transfectants and a decreased alpha2, 6-sialylation subclone in antisense-transfectants were selected for further examination of cell-cell and cell-matrix (collagen IV) adhesion. The transfectants were also treated with sialidase to compare the capacity of cell adhesion affected by cell surface sialylation.

RESULTSSense-transfection subclone showed a reduced cell-cell aggregation but enhanced cell-matrix adhesion. In contrast, the antisense-transfection subclone exhibited increased cell-cell aggregation and decreased cell-matrix adhesion. After treatment with sialidase, the cell-matrix adhesion of all the transfectants and the parental MDA-MB-435 cells were significantly reduced to the level of 31%-57% of untreated cells.

CONCLUSIONCell surface sialic acid and alpha2, 6-linked sialylation play an important role in cell-cell and cell-matrix adhesion of mammary carcinoma cell MDA-MB-435.

Antigens, CD ; genetics ; metabolism ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Adhesion ; Cell Line, Tumor ; Cell Membrane ; metabolism ; Cell-Matrix Junctions ; metabolism ; Collagen Type IV ; metabolism ; Extracellular Matrix ; metabolism ; Humans ; N-Acetylneuraminic Acid ; metabolism ; Sialyltransferases ; genetics ; metabolism ; Transfection