OBJECTIVETo evaluate the methods for collection of peripheral blood stem cells (PBSC) in children.

METHODSPeripheral blood stem cells were collected from 20 child patients and 11 donors. The patients treated with chemotherapy, received G-CSF or GM-CSF and the donors received G-CSF for mobilization. When the peripheral blood (PB) leukocyte count reached to 5 X10(9)/L,the hematopoietic stem cells were collected with CS-3000 Plus, COM TEC or COBE Spectra blood cell separators from patients and donors. For children whose weight <20 kg,HCT <24% or TBV <1 100-1 650 ml,blood cell separators were pre-injected with the same type RBCs irradiated by 25 Gy of gamma-ray and with low flow rate (10-30 ml/min). The number of CD34(+) cell was detected by flow cytometry. The relationship of number of CD34(+) cell with mononuclear cell (MNC) and processed blood volume was analyzed.

RESULTSSuccessful collection of the PBSCs with the CS- 3000 Plus (n=10), the COM TEC (n=3) and the COBE Spectra (n=18) was achieved in all the 31 cases with 1-5 aphereses used. Number of CD34(+) cells was (7.9 ±2.9) X 10(6)/kg and that of MNCs was (7.4 ±3.1) X 10(8)/kg. The total CD34(+) cell count was correlated with MNCs before aphaeresis and processed blood volume.

CONCLUSIONFor collection of high quality PBSCs, the appropriate methods should be chosen according to the body weight, TBV, mobilization of child patients/donors.

Adolescent ; Child ; Child, Preschool ; Cytapheresis ; methods ; Female ; Hematopoietic Stem Cell Mobilization ; Hematopoietic Stem Cells ; Humans ; Infant ; Male

Functional improvement to one component of the cellulase, endo-beta-1, 4-glucanase, has been a focus of the recent research in this area. We report here the saturation mutagenesis of the active site of an endoglucanase (CfEG) from termite Coptotermes formosanus. First, three dimensional structure of CfEG was built via homology modeling by using a close-related (79% homology in sequence) endo-beta-1,4-glucanase (NtEG PDB id = 1ks8) from higher termite Nasutitermes takasagoensis as a template. Second, we identified three corresponding amino acid positions at the active site of CfEG by structural superposition onto NtEG. These three putative amino acids for the active site of CfEG, i.e., Asp53, Asp56 and Glu411, were subjected to saturation mutagenesis using degenerate primers. Among the mutants, Asp53Glu and Asp56Cys showed somewhow higher activities than the wildtype, with the latter having more than 3-fold decrease in Km. Double mutation Asp53Leu/Asp56IIe showed nearly 2-fold increase in specific activity and in the same time 2-fold decrease in Km. Saturation mutagenesis to the position Glu411 produced no active mutant, even changing Glu411 explicitly into its similar amino acids such as Glu411Asp and Glu411Gln could not result in any active mutant. These imply that position Glu411 could be extremely important and therefore indispensable for CfEG functionality.
Amino Acids ; genetics ; Animals ; Cellulase ; chemistry ; genetics ; metabolism ; Isoptera ; enzymology ; Mutagenesis, Site-Directed ; Mutation

0bjective To investigate the relationships between the peripheral lung cancer and pulmonary vessels or bronchi by 16-row muhislice computed tomography(MSCT)and analyze the related factors.Methods Fifty-four patients with peripheral lung cancer confirmed pathologically underwent contrast-erdaanced MSC TI Multiplanar reformation(MPR)and maximum intensity projection(MIP)in all patients were used to demonstrate the relationships between the peripheral lung cancer and pulmonary vessels,bronchi.The relationships were categorized five types:Type 1,erupted at the edge of nodule. Type 2,erupted at the center of nodule.Type 3,penetrated through the nodule.Type 4,contacting the nodule but stretched or encased.Type 5,contacting the nodule but smoothly compressed.The pathology type,stage,size,density and location of the peripheral lung cancer were recorded and the relationships with five types were evaluated by using Chi-square test and correlation analysis.Results (1)Tumor-bronchi relationship:type 1(33,61.1%)was more often seen in≥2.0 cm and solid lesions with stage Ⅱ-Ⅳ.while Type 2(14,25.9%)was often seen in<2.0 am and part-solid or non-solid lesions with stage Ⅰ.(2)Tumor-PA relationship:Type 1 was more often seen in≥2.0 am and solid lesions with stage Ⅱ-Ⅳ.while Type 2 was often seen in part-solid or non-solid lesions with stage Ⅰ.(3)Tumor-PV relationship:type 4 was the most common type(29,53.7%).Type 2(13,24.1%)was more often seen in part-solid or non-solid lesions.(4)Tumor-bronchi relationship and tumor-PA relationship had a positive correlation(r=0.5265,P<0.01).Conclusions MSCT can demonstrate the relations between the peripheral lung cancer and bronchi.PA and PV.It is useful for the differential diagnosis and prognosis evaluation of the lung csncer.