Objective To prepare monoclonal antibodies against bovine CD4 and identify their basic biological characteristics. Methods Recombinant bovine CD4 (rHis-BoCD4 and rGST-BoCD4) was successfully expressed and purified by constructing a prokaryotic plasmid of bovine CD4 gene. The bovine CD4 monoclonal antibody was produced using hybridoma technology. The subtype and potency of the monoclonal antibody were identified and analyzed by ELISA, while specificity was analyzed through indirect immunofluorescence assay (IFA) and Western-blot. Results Four hybridoma cell lines, namely, 1H4, 6A10, 3F9 and 4G10, stably secreting monoclonal antibodies against BoCD4 were successfully obtained. The subclasses of the monoclonal antibodies subclass 6A10 was IgG2b and the rest of the monoclonal antibodies were of IgM type. Western-blot results showed that the four anti-bovine CD4 mAb strains were able to specifically bind to the bovine CD4 protein expressed in vitro. Indirect immunofluorescence assay showed that four monoclonal antibodies were able to specifically recognize the natural bovine CD4 protein. Flow cytometry assay showed that 3F9 was best to recognize bovine natural CD4 molecules. Conclusion Four monoclonal antibody strains with high specificity to natural bovine CD4 protein were successfully prepared, which lays the foundation for the subsequent studies on the function of bovine CD4 and diagnosis and treatment of bovine T-lymphocyte diseases.
Animals ; Antibodies, Monoclonal/isolation & purification* ; Cattle ; CD4 Antigens/genetics* ; Hybridomas/immunology* ; Antibody Specificity/immunology* ; Mice ; Mice, Inbred BALB C ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technique, Indirect

To investigate the efficacy and safety of minimal y invasive endovascular techniques treatment of hydronephrosis in pregnancy complicated by infection.
65 cases of pregnancy hydronephrosis coinfected patients with minimal y invasive endovascular treatment. Patients were fol owed up for 3-12 months, compared with 48 patients with the same period of conservative treatment .
Ureteral perforation and other surgery-related complications occurred in 65 patients in the observation group has not occurred. The patient's duration of hospitalization、the recurrence rate of postoperative infection, antibiotic duration of treatment, fever days were significantly lower than the control group(P<0.05).
Compared with conservative treatment, minimal y invasive endovascular technical treatment of gestational hydronephrosis co-infection with faster recovery, shorter hospital stay, can significantly reduce the number of days of fever patients and reduce the number of days of antibiotic use, can be used as conventional means of treatment of gestational hydronephrosis infection combined conventional.

OBJECTIVETo investigate whether the decrease in expression of interleukin-2 (IL-2) after trauma is associated with changes in DNA binding activity of nuclear factor of activated T cells (NFAT) and activator protein-1 (AP-1).

METHODSMice with closed impact injury with fracture in both hind limbs were adopted as the trauma model. Spleen lymphocytes were isolated from traumatized mice and stimulated with Con-A. Culture supernatants were assayed for IL-2 activity, and total RNA was extracted from spleen lymphocytes and assayed for IL-2 mRNA. DNA binding activity of NFAT and AP-1 were measured by electrophoretic mobility shift assay (EMSA). The expression of c-Fos, c-Jun and JunB proteins was determined by the Western blot analysis.

RESULTSDNA binding activity of NFAT and AP-1 gradually decreased to a minimum of 41% and 49%, respectively, of the control on the 4th day after injury, which was closely followed by the decline in IL-2 activity and IL-2 mRNA. A decrease in the expression of c-Fos on the 1st and 4th day after trauma had no significant effect on c-Jun expression; the increase in expression of JunB was only on the 1st day after injury.

CONCLUSIONDecreased IL-2 expression is, at least in part, due to a decline in the activation of NFAT and AP-1 in traumatized mice. The decline in DNA binding activity of NFAT and AP-1 is partly due to a trauma-induced block in the expression of c-Fos.

Animals ; Cell Nucleus ; chemistry ; DNA ; metabolism ; DNA-Binding Proteins ; metabolism ; Electrophoretic Mobility Shift Assay ; Female ; Interleukin-2 ; analysis ; genetics ; Male ; Mice ; NFATC Transcription Factors ; Nuclear Proteins ; Proto-Oncogene Proteins c-fos ; analysis ; Proto-Oncogene Proteins c-jun ; analysis ; RNA, Messenger ; analysis ; Transcription Factor AP-1 ; metabolism ; Transcription Factors ; metabolism