[Treatment of complicated hypospadias in children].

OBJECTIVE: To study the operation results of complicated hypospadias. METHODS: 29 cases of complicated hypospadias in recent 5 years were studied retrospectively. All cases were operated for the second time, methods used were urethroplasty with pedicled prepuce or transmitted penis prepuce tube, urethroplasty with penis-scrotum skin tube in-situ and urethroplasty with combination of scrotum skin tube and transmitted prepuce tube. RESULTS: Except 2 fistulae sequela occurred after urethroplasty with penis skin tube in-situ in 1 case, all the other 28 cases were a success, the success rate was 96.5%. CONCLUSIONS: The key point for success is precise appraisal before operation and appropriate choice of operation method.

Critical factors in basal cell adhesion molecule/lutheran-mediated adhesion to laminin.

Basal cell adhesion molecule (B-CAM) and Lutheran (LU) are two spliceoforms of a single immunoglobulin superfamily protein containing five Ig domains and comprise the sickle (SS) red cell receptor for laminin. We have now analyzed laminin binding to murine erythroleukemia cells transfected with various human B-CAM/LU constructs. B-CAM and LU bound equally well to laminin, indicating that the longer cytoplasmic tail of LU is not required for binding. However, binding of soluble laminin did require the presence of the membrane-proximal fifth immunoglobulin superfamily (IgSF) domain of LU, while deletion of IgSF domains 1, 2, 3, or 4 individually or together did not abrogate laminin binding. Under flow conditions, MEL cells expressing B-CAM, LU, and LU lacking domains 1, 2, 3, or 4 adhered to immobilized laminin with critical shear stresses over 10 dynes/cm2. However, MEL cells expressing LU lacking domain 5 bound to laminin poorly (critical shear stress = 2.3 dynes/cm2). Moreover, expression of only IgSF domain 5 of LU was sufficient to mediate MEL cell adhesion to immobilized laminin (critical shear stress >10 dynes/cm2). Finally, Scatchard analysis showed that SS red cells had an average of 67% more B-CAM/LU than normal red cells, and low density red cells from sickle cell disease patients expressed 40-55% more B-CAM/LU than high density SS red cells. B-CAM/LU copy number thus may also play a role in the abnormal adhesion of SS red cells to laminin.

Effect of human C-reactive protein on chemokine and chemotactic factor-induced neutrophil chemotaxis and signaling.

C-reactive protein (CRP) is a unique serum pentraxin and the prototype acute phase reactant. CRP is a ligand for specific receptors on phagocytic leukocytes, and mediates activation reactions of monocytes/macrophages, but inhibits the respiratory burst of neutrophils (PMN). Since CRP selectively accumulates at inflammatory sites in which IL-8 is also produced, we tested the effects of CRP on the responsiveness of PMN to IL-8 and the bacterial chemotactic peptide, FMLP-phenylalanine (FMLPP). Purified human CRP inhibited the chemotactic response of PMN to IL-8 and FMLPP. A mouse IgM mAb that was generated against the leukocyte CRP receptor (CRP-R) also inhibited the chemotactic response. Incubation of purified CRP with activated PMN generated CRP-derived peptides that also inhibited chemotaxis. A synthetic CRP peptide (residues 27-38) that binds to the CRP-R had weak chemotactic activity, whereas two other CRP synthetic peptides (residues 174-185 and 191-205) inhibited chemotaxis of PMNs to both IL-8 and FMLPP. CRP did not alter receptor-specific binding of IL-8, but exerted its effect at the level of signaling. CRP augmented both IL-8- and FMLPP-induced mitogen-activated protein kinase (extracellular signal-regulated kinase-2) activity. CRP at acute phase levels increased both agonist-induced and noninduced phosphatidylinositol-3 kinase activity. The results suggest a role for CRP as a regulator of leukocyte infiltration at inflammatory sites.

Basal cell adhesion molecule/lutheran protein. The receptor critical for sickle cell adhesion to laminin.

Sickle red cells bind significant amounts of soluble laminin, whereas normal red cells do not. Solid phase assays demonstrate that B-CAM/LU binds laminin on intact sickle red cells and that red cell B-CAM/LU binds immobilized laminin, whereas another putative laminin binding protein, CD44, does not. Ligand blots also identify B-CAM/LU as the only erythrocyte membrane protein(s) that binds laminin. Finally, transfection of murine erythroleukemia cells with human B-CAM cDNA induces binding of both soluble and immobilized laminin. Thus, B-CAM/LU appears to be the major laminin-binding protein of sickle red cells. Previously reported overexpression of B-CAM/LU by epithelial cancer cells suggests that this protein may also serve as a laminin receptor in malignant tumors.

Binding site on human C-reactive protein (CRP) recognized by the leukocyte CRP-receptor.

C-reactive protein (CRP), the prototypical inflammatory acute phase reactant in humans, interacts with monocytes and neutrophils via a specific receptor. To map the site on CRP recognized by the CRP receptor (CRP-R), synthetic peptides corresponding to the surface region on each of the five identical subunits were tested as competitors vs. [125I]-CRP for cell binding. A peptide of residues 27-38 (TKPLKAFTVCLH) efficiently inhibited CRP binding when compared to other nonoverlapping peptides. This peptide was termed the cell-binding peptide (CB-Pep). The F(ab')2 of an IgG Ab to the CB-Pep specifically inhibited CRP binding upon reacting with the ligand. Competitive binding studies with synthetic peptides truncated from either the NH2- or COOH-terminus of the CB-Pep revealed that the minimum length recognized by the CRP-R consisted of residues 31-36: KAFTVC. Conservative substitutions of residues within the CB-Pep indicated that the four residues AFTV were critical for CRP-R binding. The CB-Pep also inhibited induced superoxide generation by HL-60 granulocytes. The minimum length required for the inhibition was also KAFTVC; however, only Phe-33 and Leu-37 were critical residues in this assay. Anti-CB-Pep IgG Ab reacted more extensively with heat-modified CRP, suggesting that an altered conformation of CRP is preferentially recognized by the CRP-R. The results suggest that this contiguous sequence on a beta-strand on one face of each of five subunits of the CRP pentamer serves as a unique recognition motif for inflammatory leukocytes.