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 understand the current situation and differences in physical activity among children and adolescents in different regions of China, so as to provide a scientific reference for promoting the level of overall physical activity promotion among children and adolescents in China. Methods: From June to December 2023, the student satisfaction assessment questionnaire for the national children and adolescents theme fitness activity "Run, Youth!" was conducted, which was administered via a combined online and offline questionnaire survey to explore 397 013 students participation in the "Run, Youth!" activity in 2023. Descriptive statistics, Logistic regression, and other approaches were utilized for data analysis. Results: There were notable disparities in the rates of children and adolescents meeting the physical activity standards among northeast China, northwest China, north China, east China, central China and southwest China (10.27%, 11.79%, 9.32%, 8.48%, 8.81%, 7.30%) ( χ 2=33.48, P <0.05). The results of multivariate Logistic regression analysis showed that rate of recommended level of physical activity among children in northwest China, boys, children aged 16-18, children in urban areas were relatively high ( OR =1.12, 1.68, 0.60, 1.88, P < 0.05 ). After adjusting for confounding factors, the results of binary Logistic regression analysis showed that there was an interactive effect relationship between the interaction of region, age, gender, and urban-rural distribution and physical activity among children and adolescents ( OR=0.51, P <0.01). Based on gender analysis, weekly physical activity duration of boys in each region surpassed that of girls; based on age analysis, except for the central and southwestern regions, weekly physical activity duration of children and adolescents gradually increased with age in other regions; based on the proportion of urban and rural areas, weekly physical activity duration of children and adolescents in urban areas across all regions exceeded that in rural areas. Conclusions The attainment rate of sufficient physical activity among children and adolescents in northwest China is the highest, and the differences between regions are not significant. The participation and level of physical activity of male students in each region surpass those of females and the attainment rate of physical activity of physical activity of urban children and adolescents is higher than that of those in rural areas.

Objective To evaluate the in vitro antimicrobial activity of tigecycline in combination with imipenem against multi-drug resistant and pan-drug resistant Acinetobacter baumannii isolates,so as to discuss the feasibility of drug combination and provide the basis for chnical rational use of antimicrobial agents.Methods Sixteen multi-drug resistant and pan-drug resistant Acinetobacter baumannii isolates were collected between January and April in 2015 from all kinds of infected specimens of Nanjing Drun Tower Hospital.The protocol was designed by checkerboard method,and the minimum inhibitory concentration (MIC) of antibiotics was determined by microdilution broth method,and the fractional inhibitory concentration (FIC) index was calculated according to MIC results.Results The average value of MIC (MICG),MIC50,MIC90 of tigecycline and imipenem single were 1.73,1,4 μg·mL-1and 31.00,32,64 μg·mL-1.When tigecycline was combined with imipenem,MICG,MIC50,MIC90 of tigecycline and imipenem were 0.24,0.25,0.50 μg·mL-1 and 8.16,8.00,16.00 μg·mL-1,respectively.Compared with the drug single use groups,MIC was significandy decreased in the drug combination group.In 6 strains (37.50％),synergy effect (FIC≤0.5) was observed,and in 10 strains (62.50％),additive effect (0.5 ＜ FIC ≤ 1) was found.No negative and independent effects were shown.Conclusion Both additive and synergistic action is observed when tigecyclineis combined with imipenem against multi-drug resistant and pan-drug resistant Acinetobacter baumannii isolates.No negative and independent effects are shown.This combination use against multi-drug resistant and pan-drug resistant Acinetobacter baumannii may be an effective therapy for clinical treatment.

ObjectiveTo investigate the initial symptoms, onset time and predisposing factors of acute cerebral haemorrhage in order to more effectively prevent cerebral haemorrhage. Methods 728 cases with acute cerebral haemorrhage were collected in recent 8 years. The initial symptoms,onset time and predisposing factors of all the patients were investigated. ResultsAcute cerebral haemorrhage in 582 caces (79.9%) occurred at time between 6am and 18pm and 146 cases (20%) at night. The main predisposing causes of acute cerebral haemorrhage were the playing of chess, cards and mahjong (16.1%), quarrels (12.8%), overwork (11.7%), alcoholism (7.2%) and so on. The most common initial symptom was hemianesthesia, accounting for 16.2%, others were vertigo (13.0%), headache ( 11.4%), amaurosis fugax ( 9.3%), speech ambiguity ( 8.6%) and transitory visual disturbance (7.9%).ConclusionsThe preventive measures of cerebral haemorrhage should be focused on daytime, especially in the morning. Sixteen locations where the cerebral haemorrhage is prone to happen should be paid more attention to by the patients with hypertension and the medical staff. Thirteen symptoms,such as hemianesthesia and vertigo,are the aurae of cerebral haemorrhage.