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

Objective:To investigate the sleep status of preschool children in Zhongshan City and the OSA-18 questionnaire scores among healthy preschool children. Methods:A cluster sampling method was adopted to select 2 222 students from 5 non-boarding kindergartens in Zhongshan from October 2022 to October 2023 as study subjects. Data were collected via an online questionnaire platform and analyzed using SPSS 27.0 statistical software. Results:A total of 1 317 valid questionnaires were collected, with an average sleep duration of（8.74±0.75） hours among children. The results of stepwise regression analysis showed that child age, prematurity, sleep latency, daily outdoor activity time, and activities within 30 minutes before bedtime were factors influencing children's nocturnal sleep duration（P<0.05）. The overall incidence of sleep disorder-related symptoms was 56.80%, including restless sleep（24.14%）, difficulty falling asleep（24.07%）, snoring with mouth breathing（14.20%）, bruxism（11.16%）, enuresis（10.55%）, sleep talking（6.08%）, limb twitching（2.74%）, and sleepwalking（0.53%）. Various factors influenced these symptoms, and binary logistic regression analysis identified allergic diseases and a smaller number of siblings at home as the most prevalent risk factors（P<0.05）. The overall OSA-18 score was 34.11±12.14, and binary logistic regression analysis identified males, allergic diseases, and tonsillar hypertrophy as risk factors for high OSA-18 scores（≥60）（P<0.05）. Conclusion:In Zhongshan City, the nocturnal sleep duration of preschool children decreases with age, with independent risk factors for this prematurity, longer sleep latency, shorter daily outdoor activity time, and the use of electronic devices within 30minute before bedtime. Allergic diseases and a smaller number of siblings at home increase the risk of sleep-disordered breathing symptoms. Males, allergic diseases, and tonsillar hypertrophy have higher OSA-18 scores.
Humans ; Child, Preschool ; Surveys and Questionnaires ; Male ; Female ; Sleep ; China/epidemiology* ; Sleep Apnea, Obstructive/epidemiology*

Nonhuman primates(NHPs)are susceptible hosts of tuberculosis(TB).After infection,TB not only spreads among monkey populations but can also spread to humans.An effective vaccine to protect NHPs from TB has not been developed.Although prevention and control protocols have matured and reduced the incidence of TB among NHPs in captivity,outbreaks continue to occur.This article summarizes the worldwide epidemiological situation of TB in monkeys in captivity and in the wild,analyzes the advantages and disadvantages of commonly used detection method,and summarizes the most common practices of TB prevention and control in NHPs.Our findings indicate that TB poses a great threat to NHPs,underscoring the importance of raising awareness of TB among NHP breeding workers and managers.Additionally,our result provide a basis for improving current management procedures and offer valuable insights for TB diagnosis,prevention,and control in NHPs in China.

Objective To investigate the effects of IGF-Ⅱ on adriamycin-induced apoptosis of hepatocellular carcinoma cell line(HepG2).Methods Cultured HepG2 cells were divided into:①group A:control group;②group B:ADM group(200 ng/ml);③group C:2 ng/ml IGF-Ⅱ+200 ns/ml ADM group;④group D:20 ng/ml IGF-Ⅱ+200 ng/ml ADM group;⑤group E:200 ng/ml IGF-Ⅱ+200 ng/ml ADM group. After HepG2 were treated for 48 h,MTT colorimetry was performed to determine the cell viability,Flow cytometry was used to detect the cell apoptosis rate,and Weastern-blot was performed to evaluate the expression of survivin. Results The cell viability of group A、B、C、D、E was respectively 0.568±0.025、0.201±0.020、0.232±0.027、0.268±0.013 and 0.304±0.019;The cell apoptosis rate of group A、B、C、D;E was respectively 6.9％±1.3％、35.4％±2.1％、31.2％±2.2％、26.4％±1.7％and 21.7％±1.9％;Survivinβ-actin of group A、B、C、D、E was respectively 0.527±0.039、0.147±0.081、0.311±0.069、0.421±0.033 and 0.469±0.031.The cell viability in IGF-Ⅱ+ADM group was better than ADM group(P＜0.01),the cell apoptosis rate in IGF-Ⅱ+ADM group was significantly lower than ADM group(P＜0.01),the expression level of survivin in IGF-Ⅱ +ADM group was significantly higher than ADM group(P＜0.01). Conclusions IGF-Ⅱ inhibits ADM-induced apoptosis of HepG2 cells,probably by up-regulating cell's survivin expression.