OBJECTIVETo evaluate the association of sperm mobility parameters assessed by computer-assisted sperm analysis (CASA) with the rates of normal fertilization, oocyte cleavage and excellent embryos in in vitro fertilization (IVF) cycles.

METHODSA total of 288 infertile women undergoing IVF cycles patients were divided into two groups according to the normal fertilization rate (≥50% and <50%), cleavage rate (≥90% and <90%), or excellent embryo rates (≥50% and <50%). The means of the sperm motility parameters analyzed by CASA twice before oocyte retrieval were recorded and analyzed using t-test in relation to the rates of normal fertilization, cleavage and excellent embryos in IVF cycles.

RESULTSThe mean curvilinear velocity (VCL), average path velocity (VAP), and amplitude of lateral head displacement (ALH) were significantly higher in women with a normal fertilization rate of ≥50% than in those with a normal fertilization rate of <50% (P<0.05). Women with an oocyte cleavage rate of ≥90% had significantly higher VCL and VAP than those with a cleavage rate of <90% (P<0.05). The VCL, straight line velocity (VSL), VAP, linearity, straightness, wobble coefficient, ALH, or beat-cross frequency showed no significant differences between women with excellent embryo rates of ≥50% and <50% (P>0.05).

CONCLUSIONThe sperm motility parameters assessed using CASA are associated with normal fertilization and oocyte cleavage rates but not with excellent embryo rate in IVF cycles.

Adult ; Female ; Fertilization in Vitro ; methods ; Humans ; Infertility, Female ; therapy ; Male ; Middle Aged ; Semen Analysis ; Sperm Motility ; Therapy, Computer-Assisted

OBJECTIVETo construct full-length human bladder cancer-specific antibody libraries for efficient display of full-length antibodies on the surface of mammalian cells.

METHODSThe total RNA was isolated from peripheral blood mononuclear cells from patients with bladder cancer. The repertoires of IgG1 heavy chain variable region (VH) and Kappa light chain were amplified by RT-PCR using specific primers. The antibody genes were inserted into the vector pDGB-HC-TM to construct the bladder-cancer-specific antibody libraries of heavy chains and light chains. Ten clones from each library were randomly picked for gene sequencing and transient transfection into FCHO cells to analyze antibody display on mammalian cell surface by flow cytometry after staining with corresponding fluorescent labeled antibodies.

RESULTSThe libraries of bladder-cancer-specific antibody heavy chain (IgG1) and light chain (LCk) were successfully constructed. Seven out of the 10 clones randomly selected from the heavy chain library and 9 out of the 10 clones from the light chain library showed correct open reading frame, coding for 7 unique VH and 9 unique LCk. The combinatory library size reached 3.32×10(11).

CONCLUSIONWe have successfully constructed a full-length human bladder-cancer-specific antibody library with a combinatory diversity of 3.32×10(11) based on mammalian display technology, which can be used for screening monoclonal antibodies against bladder-cancer-associated antigens.

Amino Acid Sequence ; Animals ; Antibodies ; genetics ; Cell Surface Display Techniques ; Gene Library ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin kappa-Chains ; genetics ; Peptide Library ; Urinary Bladder Neoplasms ; genetics ; immunology