1.Study on the role of coatomer protein Ⅰ in apolipoprotein A-1-mediated cholesterol efflux from foam cells
Weitao HONG ; Feilong ZHANG ; Jie HUANG ; Yumin LIANG ; Gengji CHEN ; Xiaojia CHEN ; Weilie MA ; Hang DING ; Zhizhen ZHANG
Chongqing Medicine 2025;54(3):580-587
Objective To establish a human monocytic THP-1 macrophage-derived foam cell model and analyze the role of the α subunit of coatomer protein Ⅰ(α-COP)in apolipoprotein A-1(apoA-1)-mediated cholesterol efflux from foam cells.Methods THP-1 cells were induced to adhere using PMA(PMA group),followed by treatment with acetylated low-density lipoprotein(Ac-LDL)to generate macrophage-derived foam cells(Ac-LDL group).Subsequent incubation with apoA-1 formed the apoA-1 group.Cholesterol efflux rates mediated by apoA-1 and intracellular lipid accumulation were quantified through liquid scintillation counting and oil red O staining.The expression patterns of α-COP were systematically analyzed using quantitative real-time PCR(qPCR),Western blotting,and laser scanning confocal microscopy.THP-1 macrophage-derived foam cells mediated by apoA-1 were transduced with scrambled shRNA(Scr group)or α-COP-specific shRNA lentivirus(α-COP shRNA group)to assess the effects of α-COP knockdown on cholesterol efflux efficiency and total cellular cholesterol content.Subcellular localization of adipopoilin(ADFP)and cholesterol probe Fil-ipin Ⅲ staining patterns were visualized via laser scanning confocal microscopy.Bioinformatics analysis of α-COP expression profiles in carotid atherosclerotic plaques was performed using datasets from the GEO data-base.Results Compared with the Ac-LDL group,the cholesterol efflux rate in the apoA-1 group was signifi-cantly increased[(9.77±0.79)%vs.(2.74±0.37)%,P<0.001].Oil red O staining demonstrated reduced lipid accumulation in foam cells of the apoA-1 group compared with the Ac-LDL group.The relative mRNA expression level of α-COP in the apoA-1 group was significantly higher than that in the Ac-LDL group(P<0.001),with corresponding elevation in protein expression(P<0.001).Fluorescence intensity analysis re-vealed increased mean fluorescence intensity of α-COP in the apoA-1 group compared with the Ac-LDL group.Following α-COP knockdown by shRNA,the apoA-1-mediated cholesterol efflux rate was significantly de-creased(P<0.05)and total intracellular cholesterol levels were increased(P<0.001)in the α-COP shRNA group compared with the Scr group.The α-COP shRNA group exhibited enhanced red fluorescence intensity of ADFP(P<0.05)and increased blue fluorescence intensity of Filipin Ⅲ(P<0.01)compared with the Scr group.Database analysis indicated that α-COP mRNA expression was lower in carotid atherosclerotic plaques than in normal arterial tissues,with reduced expression in advanced-stage plaques compared with early-stage plaques(P<0.05).Conclusion α-COP participates in apoA-1-mediated cholesterol efflux from foam cells,and interference with α-COP expression results in reduced cholesterol efflux and increased intracellular lipid accumulation.
2.Clinical features of pertussis in 248 hospitalized children and risk factors of severe pertussis
Jiyan ZHANG ; Yufei ZHUO ; Yanping CHEN ; Fang FAN ; Chengjuan WANG ; Wu ZHOU ; Gengji XIAO ; Fangzhao CHEN
Chinese Journal of Applied Clinical Pediatrics 2023;38(4):275-280
Objective:To investigate the clinical features of pertussis in children and analyze the risk factors of severe pertussis.Methods:The clinical data of 248 children with pertussis hospitalized in Hunan Children′s Hospital from March 2018 to March 2022 were analyzed retrospectively.According to the age at admission, the patients were divided into two groups: ≤3 months and > 3 months.According to the patient′s condition, they were classified into ordinary group and severe group.According to the pathogens detected, the children were divided into single infection group and mixed infection group.The independent sample t-test, chi- square test were used to analyze the clinical indexes of the infants in above groups. Results:(1)Of 248 hospitalized children with pertussis, 204 cases (82.2%) were less than 1 year old, 92 cases (37.0%) had contact with a coughing family member before, and 169 cases (68.1%) were unvaccinated.Among 248 children, 193 cases (77.8%) had an elevated white blood cell count, and 145 cases (58.4%) had mixed infections.The most common pathogen was respiratory syncytial virus [29/248(11.6%)]. About 173 cases (69.7%) had concurrent pneumonia, and 35 cases (14.1%) had pulmonary consolidation.(2)Compared with the group > 3 months of age, more patients in the group ≤3 months of age had contact with a coughing family member before, and suffered from cyanosis, dyspnea, respiratory failure, heart failure and pertussis encephalopathy ( χ2=4.612, 20.810, 7.882, 16.617, 13.740, 7.846, all P<0.05). The proportions of patients in the group ≤3 months of age required intensive care unit(ICU) hospitalization and mechanical ventilation were higher than those in the group > 3 months of age ( χ2=14.810, 21.436, all P<0.05). The mortality of the group ≤3 months of age was higher than that of the group >3 months of age ( χ2=12.016, P<0.05). Children ≤3 months of age had a higher WBC level [(27.83±27.70)×10 9/L vs.(23.34±15.28)×10 9/L, t=22.244, P<0.001], longer duration of spasmodic cough [(16.56±9.33) d vs.(15.06±6.16) d, t=10.145, P=0.002] and longer hospitalization time [(11.47±10.48) d vs.(9.48±4.80) d, t=20.050, P<0.001] than those >3 months of age.(3)Compared with the ordinary group, a higher proportion of children in the severe pertussis group were under 3 months old, and had not been vaccinated against pertussis vaccine ( χ2=14.803, 4.475, all P<0.05). The ratio of patients with dyspnea, an lymphocyte count/neutral cell(LC/NC) ratio <1, mixed infections, lung consolidation and pleural effusion in the severe pertussis group was higher than that in the ordinary group ( χ2=116.940, 43.625, 13.253, 106.370, 11.874, all P<0.05). The patients in the severe pertussis group had a higher WBC [(61.66±29.63)×10 9/L vs.(18.83±10.00)×10 9/L, t=112.580, P<0.001] and a lower LC (0.494±0.186 vs.0.676±0.132, t=13.752, P<0.001) than those in the ordinary group.(4)Compared with the single infection group, the proportions of children with fever, dyspnea, fine moist lung rales, an LC/NC ratio <1, and lung consolidation were higher in the mixed infection group ( χ2=8.909, 6.804, 7.563, 8.420, 12.458, all P<0.05). More children in the mixed infection group required ICU hospitalization and mechanical ventilation than those in the single infection group ( χ2=11.677, 7.397, all P<0.05). The mixed infection group had higher respiratory failure and death rates than the single infection group ( χ2=7.980, 4.267, all P<0.05). Compared with the single infection group, the mixed infection group had a higher WBC level [(27.73±24.13)×10 9/L vs.(21.25±14.65)×10 9/L, t=13.318, P<0.001], longer hospitalization time [(11.593±9.010) d vs.(8.339±4.047) d, t=17.283, P<0.001], and a smaller LC ratio (0.626±0.165 vs.0.684±0.132, t=7.997, P=0.005). (5) Logistic regression analysis showed that age ≤3 months, peak WBC and dyspnea were risk factors of severe pertussis. Conclusions:Hospitalized pertussis children are prone to pneumonia and pulmonary consolidation.Patients aged ≤3 months with a large WBC and dyspnea easily develop into severe pertussis.Monitoring blood routine is helpful for judging the severity of the disease.Mixed infections increase the incidence of complications and can impair the treatment effect.

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