Objective Currently, there is no commercially available quantitative detection kit for the main Felis domestic allergen (Fel d 1) in China. To establish a rapid detection method for Fel d 1, this study aims to prepare monoclonal antibodies against Fel d 1 protein. Methods The codon preference of Escherichia coli was utilized to optimize and synthesize the Fel d 1 gene. The prokaryotic expression plasmid pET-28a-Fel d 1 was constructed and used to express and purify the recombinant Fel d 1 protein. Subsequently, the recombinant protein was immunized into BALB/c mice and monoclonal antibodies (mAbs) were prepared by the hybridoma technique. An indirect ELISA was established using the recombinant Fel d 1 as the coating antigen, and hybridoma cell lines were screened for positive clones. The specificity and antigenic epitopes of the mAbs were confirmed by Western blot analysis. Finally, the selected hybridoma cells were injected into the peritoneal cavities of BALB/c mice for large-scale monoclonal antibody production. Results The recombinant plasmid pET-28a-Fel d 1 was successfully constructed, and soluble Fel d 1 protein was obtained after optimizing the expression conditions. Western blot and antibody titer assays confirmed the successful isolation of two hybridoma cell lines, 7D11 and 5H4, which stably secreted mAbs specific to Fel d 1. Antibody characterization revealed that the 5H4 mAb was of the IgG2a subtype and could recognize the amino acid region 105-163 of Fel d 1, while the 7D11 mAb was the IgG1 subtype and could recognize the amino acid region 1-59. Conclusion The high-purity recombinant Fel d 1 protein produced in this study provides a promising alternative for clinical immunotherapy of cat allergies. Furthermore, the monoclonal antibody prepared in this experiment lays a material foundation for the in-depth study of the biological function of Fel d 1 and the development of ELISA detection.
Animals ; Antibodies, Monoclonal/biosynthesis* ; Mice, Inbred BALB C ; Cats ; Mice ; Allergens/genetics* ; Glycoproteins/genetics* ; Enzyme-Linked Immunosorbent Assay ; Hybridomas/immunology* ; Recombinant Proteins/genetics* ; Female ; Antibody Specificity

Objective To observe the effects of atorvastatin calcium on serum tumor necrosis factor alpha (TNF)-α and C-reactive protein (CRP) in patients with acute cerebral infarction (ACI), so as to approach the mechanism of atorvastatin calcium inhibiting ACI inflammatory injury. Methods Eighty-four patients with ACI were randomly divided into 2 groups: group B (42 cases, treated with antiplatelet therapy and improving cerebral circulation), group A(42 cases, treated with atorvastatin calcium 20 mg/d after the onset of ACI for 28 days on the base of group B). TNF- α and CRP were detected before treatment and in the 3rd,7th day after treatment. The European stroke scale (ESS) was evaluated on the same time. A healthy control group (group C, 16 cases) was also included in the study. Results The peak of CRP and TNF-α levels were observed in the 3rd and 7th day after treatment respectively, and the levels of group A were lower than those of group B [(13.00 ± 2.45) mg/L vs (19.21 ± 3.67) mg/L,(19.79 ± 11.01) ng/L vs (30.69 ± 18.47) ng/L, P < 0.05]. In the 7th day after treatment, the scores of ESS was higher in group A than that in group B [(79.19 ± 30.59) scores vs (63.91 ± 27.87) scores, P < 0.05]. Conclusions Atorvastatin calcium can prevent the increase of serum TNF-α and CRP, and it has anti-inflammatory effect. Atorvastatin calcium may have the role of neuroprotection besides lipid-lowering.

0.05),but the level of P- selectin in treatment group were obviously decreased compared with control group after the seventh and fourteenth day(P

Objective To investigate the effects of Polysaccharide Sulfate (PSS) on plasma von Willebrand factor (vWF) and thrombomodulin (TM) in patients with acute cerebral infarction.Methods 64 patients with acute cerebral infarction were randomly assigned to receive intravenous drip of PSS (150 mg/d for 14 days, 32 cases, PSS group) or Troxerutin (600 mg/d for 14 days, 32 cases, control group). All of the patients were tested for plasmatic levels of vWF and TM prior to and at 7 d, 14 d after treatment.Results The levels of vWF in PSS group at 7 d, 14 d after treatment were obviously decreased compared with control group (all P