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Objective:To discuss the differential effects of apolipoprotein E(APOE)gene polymorphism in the neurotoxicity-reactive astrocytes,and to provide the theoretical basis for the study of the pathogenesis of Alzheimer's disease(AD).Methods:The primary cortical astrocytes from the APOE-knockout mice(APOE-/-)were isolated and cultured in vitro,and the purity of the cells was identified by immunofluorescence staining.The human APOE3 and APOE4 recombinant over-expression plasmids were constructed and separately transfected into the primary APOE-/-astrocytes,and the APOE-/-primary cells were regarded as control.Western blotting method was used to detect the expression levels of APOE and glial fibrillary acidic protein(GFAP)proteins in the cells;enzyme-linked immunosorbent assay(ELISA)method was used to detect the APOE level in the cellular culture supernatant.The inflammatory models were prepared with the primary astrocytes transfected with APOE3 and APOE4 and co-stimulated with interleukin-1α(IL-1α),tumor necrosis factor(TNF),and complement C1q.The cells were divided into APOE3+PBS group,APOE4+PBS group,APOE3+IL-1α+TNF+ C1q group,and APOE4+IL-1α+TNF+C1q group.Cell immunofluorescence staining method was used to observe the morphology of the cells in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of glypican 4(Gpc4),glypican 6(Gpc6),thrombospondin 1(Thbs1),thrombospondin 2(Thbs2),SPARC-like protein 1(Sparcl1)and glial cell line derived neurotrophic factor(GDNF),C3,and S100 calcium binding protein B(S100B)mRNA in the cells in various groups;microsphere phagocytosis assay was used to detect the phagocytic capacities of the cells in various groups;Western blotting was used to detect the protein expression levels of B-cell lymphoma 2(Bcl-2),and cysteinyl aspartate specific protease-3(Caspase-3)proteins in the cells in various groups.Results:Compared with APOE-/-group,the expression levels of APOE and GFAP proteins in the cells and the APOE level in the cellular culture supernatant in transfected APOE3 and transfected APOE4 groups were increased(P<0.01).The fluorescence microscope observation results showed that compared with APOE3+PBS and APOE4+PBS groups,the astrocytic processes in APOE3+IL-1α +TNF+Cq1 group and APOE4+IL-1α+TNF+Cq1 group became shorter and the cell bodies became larger;compared with APOE3+IL-1α +TNF+Cq1 group,the astrocytic processes in APOE4+IL-1α +TNF+Cq1 group were even shorter.Compared with APOE3+PBS and APOE4+PBS groups,the expression levels of Gpc4,Gpc6,Thbs1,Thbs2,and Sparcl1 mRNA in the cells in APOE3+IL-1α +TNF+Cq1 group and APOE4+IL-1α +TNF+Cq1 group were significantly decreased(P<0.01);compared with APOE3+IL-1α +TNF+Cq1 group,the expression levels of Gpc4,Gpc6,Thbs1,Thbs2,and Sparcl1 mRNA in the cells in APOE4+IL-1α +TNF+Cq1 group were significantly decreased(P<0.05 or P<0.01).Compared with APOE3+PBS and APOE4+PBS groups,the expression levels of GDNF mRNA in the cells in APOE3+IL-1α+TNF+Cq1 group and APOE4+ IL-1α +TNF+Cq1 group were decreased(P<0.01),and the expression levels of C3 and S100B mRNA were increased(P<0.01);compared with APOE3+IL-1α +TNF+Cq1 group,the expression level of GDNF mRNA in the cells in APOE4+IL-1α+TNF+Cq1 group was decreased(P<0.05),and the expression levels of C3 and S100B mRNA were increased(P<0.05).Compared with APOE3+ PBS group and APOE4+PBS group,the numbers of hagocytosis of microspheres in the cells in APOE3+ IL-1α +TNF+Cq1 group and APOE4+IL-1α +TNF+Cq1 group were significantly decreased;compared with APOE3+IL-1α+TNF+Cq1 group,the number of hagocytosis of microspheres in the cells in APOE4+IL-1α+TNF+Cq1 group was significantly decreased.Compared with APOE3+PBS group and APOE4+PBS group,the expression levels of Bcl-2 protein in the cells in APOE3+IL-1α+TNF+ Cq1 group and APOE4+IL-1α +TNF+Cq1 group were decreased(P<0.05 or P<0.01)and the expression levels of Caspase-3 protein were significantly increased(P<0.01);compared with APOE3+ IL-1α+TNF+Cq1 group,the expression level of Bcl-2 protein in the cells in APOE4+IL-1α+TNF+ Cq1 group was decreased(P<0.01),and the expression level of Caspase-3 protein was increased(P<0.05).Conclusion:The APOE4 genotype has a stronger ability to induce the inflammatory factors compared with APOE3;it can lead to a neurotoxicity-reactive astrocyte phenotype,increase the neurotoxicity,affect the astrocyte apoptosis,and aggravate the neuron damage.
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Objective·To use single-cell RNA sequencing(scRNA-Seq)technology to interpret the cellular communication landscape of coronary atherosclerosis(CA),and to explore the dominant cell subsets and their key genes.Methods·The GSE131778 data set was downloaded and preprocessed,and quality controlling,dimension reduction clustering and annotation were carried out.Then cell communication analysis was conducted by using CellChat package to identify dominant cell subsets.The FindAllMarker function was used to screen differentially expressed genes(DEGs)between the dominant cell subpopulation and other cell subpopulations,and its protein-protein interaction(PPI)network was constructed.The DEGs ranked in the top five of the Degree algorithm were taken as key genes.Then,the key genes were matched and mined with the cell communication network analyzed by CellChat to obtain the ligand-receptor pairs(L-R)and the signal pathways mediated by the key genes,and the results were visualized.At the same time,the atherosclerosis mouse model was constructed and RT-PCR was used to detect the expression of key genes in carotid atherosclerosis lesions.Results·A total of 11 cell subsets were identified in CA lesions,including smooth muscle cells,endothelial cells,macrophages,monocytes,etc.Cell communication results showed that CellChat detected 70 significant L-R and 26 related signal pathways in 11 cell subsets.Smooth muscle cell was the dominant cell subgroup with the most significant interaction frequency and intensity with other cell subgroups in the active state of communication.The results of DEGs screening showed that there were 206 DEGs between smooth muscle cell subsets and other cell subsets,among which ITGB2,PTPRC,CCL2,DCN and IGF1 were identified as key genes.The results of cell communication mediated by key genes showed that CCL2 and ACKR1 formed L-R and participated in the communication network between smooth muscle cells and endothelial cells through mediating CCL signaling pathway.ITGB2 formed receptor complexes with ITGAM and ITGAX respectively,and then formed L-R with C3 to mediate the complement signal pathway,participating in the communication network among smooth muscle cells,macrophages and monocytes.The validation results of hub genes in animal experiments were consistent with the results of bioinformatics analysis.Conclusion·Smooth muscle cells are the dominant cells in the pathological process of CA,and have extensive communication networks with other cells.They can construct cellular communication networks with endothelial cells,macrophages and monocytes through CCL and complement signaling pathways mediated by CCL2-ACKR1,C3-(ITGAM+ITGB2)and C3-(ITGAX+ITGB2).
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Objective To analyze the short-term clinical efficacy and influencing factors of ustekinumab monoclonal antibody(UST)in the treatment of Crohn′s disease(CD).Methods Retrospective cohort study was used to collect the clinical data of CD patients treated with UST in the 10th People′s Hospital affiliated to Tongji University from December 2020 to October 2022.The main analysis is the short-term clinical efficacy and influencing factors of UST treatment for CD at weeks 8 and 16,And analyze the endoscopic response rate of some patients.Results A total of 91 CD patients who first used UST were included.The 8-week clinical response rate of UST treat-ment for CD was 61.5%,and the clinical response rate was 45%;The clinical response rate at 16 weeks was 71.4%,and the clinical response rate was 54.9%.56 cases underwent endoscopic re-examination in our hospital,and the endoscopic response rate at 16 weeks was 41.1%.Univariate analysis showed that fistula(including anal fistula,personal history of anal fistula,and intestinal skin fistula)is associated with clinical remission in Crohn′s disease patients at 8/16 weeks.Further multivariate COX regression analysis showed that the presence of a history of anal fistula surgery was an independent protective factor affecting clinical remission in CD patients treated with UST at 8 weeks(HR = 0.04,95%CI:0.00～0.38;P = 0.005)and 16 weeks(HR = 0.04,95%CI:0.01～0.34;P = 0.003)compared to those without fistula;Narrow lesions are an independent risk factor for 16 week clinical remission in CD patients compared to non-narrow and non-penetrating lesions(HR = 1.75,95%CI:1.08～2.84;P = 0.023).No patients were found to have stopped medication due to serious adverse reactions.Conclusions UST can improve the clinical remission and response of CD patients at 8/16 weeks,and has good short-term clinical efficacy.CD patients with a personal history of anal fistula are recommended to use UST monoclonal antibodies,while patients with stenotic lesions should be cautious in using UST monoclonal antibodies.Whether the patient has undergone surgical treatment in the past,as well as whether UST has been used on the first or non-first line,has no significant impact on clinical remission.
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The nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome is an inflammatory protein complex, and can participate into the inflammatory response. Upon activation, these inflammasomes can lead to Caspase-1 activation, thereby inducing a cascade of inflammatory factor activation and further cell pyroptosis. Excessive activation of inflammasomes will induce the overexpression of inflammatory factors, persistently triggering immune dysregulation and inflammatory chain reactions, even causing severe damage. The recent studies have confirmed a close association between retinal diseases, such as diabetic retinopathy(DR), retinal ischemia-reperfusion injury(RIRI), and proliferative vitreoretinopathy(PVR)with immune dysregulation and inflammatory responses, which is serving as crucial factors in the progression of retinal diseases. This article reviews the NLRP3 inflammasome signaling pathway and its role in the occurrence and development of retinal diseases, in order to provide new ideas for the pathogenesis and prevention of retinal diseases.
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OBJECTIVE To analyze the distribution characteristics of warfarin drug-gene polymorphism in Han children from Beijing area. METHODS Data of nine warfarin drug-gene loci about VKORC1 rs9923231， CYP2C9 rs1799853*2 and rs1057910*3， CYP4F2 rs2108622， APOE rs429358 and rs7412， ABCB1 rs1045642， EPHX1 rs1051740 and rs2234922 were collected from dept. of cardiovascular medicine， Children’s Hospital Affiliated to Capital Institute of Pediatrics from March 2019 to March 2023， and the population data reported in domestic and foreign literature were compared. RESULTS In Beijing area， the frequency of APOE rs429358 mutant genotype was higher in males （19.8%） than in females （13.5%）（P＜0.05）. VKORC1 rs9923231 was dominated by homozygous mutant genotype （83.3%）， which was consistent with children in Japan （82.2%）， and higher than that of predominantly Caucasian children in the UK， Sweden， the United States， and Germany （10.4%-18.3%）（P＜ 0.05）； CYP2C9 was dominated by *1/*1 type （91.9%）， which was consistent with children in Japan （94.6%）， and higher than that of predominantly Caucasian children in the UK， Sweden， the United States， and Germany （66.1%-73.4%）（P＜0.05）. The frequency of EPHX1 rs1051740 mutant genotype was higher in adults （78.5%） than in children （63.5%）（P＜0.05）. CONCLUSIONS More mutations of VKORC1 rs9923231 and ABCB1 rs1045642 are found in Han children from Beijing area. The distribution of warfarin drug-gene polymorphisms in Han children from Beijing area is different among different genders， as well as compared with other countries， and Chinese Han adults. Therefore， caution should be exercised when using the reported data.
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Background Benzo[a]pyrene (BaP) has neurotoxicity, which can induce the loss of hippocampal neurons in humans and animals and lead to spatial learning and memory dysfunction, but its mechanism is still unclear. Objective To observe the ferroptosis of mouse hippocampal neuron HT22 cells induced by 7,8-dihydroxy-9,10-epoxybenzo[a]pyrene (BPDE), an active metabolite of BaP, and to explore its potential mechanism, so as to provide a basis for the study of BaP neurotoxicity mechanism. Method Mouse hippocampal neuron HT22 cells were selected and divided into four groups: solvent control group and low, medium, and high concentration BPDE exposure groups (0.25, 0.50, and 0.75 μmol·L−1). Cell survival was detected by CCK8 method. Cell morphology and ultrastructure were observed under light and electron microscopes. The levels of reactive oxygen species (ROS) and Fe2+ were detected by fluorescence probe method. Iron, 4-hydroxynonenoic acid (4-HNE), malondialdehyde (MDA), glutathione (GSH), and glutathione peroxidase (GSH-PX) levels were detected with commercial kits. The expression levels of acyl-CoA synthase long chain family member 4 (ACSL4), cyclooxygenase 2 (COX2), solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) were detected by Western blotting. After interventions with ferroptosis inhibitors 20 μmol·L−1 deferoxamine (DFO) and 10 μmol·L−1 ethyl 3-amino-4-cyclohexylaminobenzoate (Fer-1), the cell survival rate of each BPDE exposure group and the changes of the ferroptosis characteristic indicators and protein expression levels were observed. Results With the increase of BPDE concentration, the survival rate of HT22 cells decreased gradually, and the survival rate of each BPDE group was significantly lower than that of the solvent control group (P<0.01). Under light microscope, the number of cells in the high concentration BPDE group was significantly reduced, and atrophic cells and reduced synapses were recorded. Under electron microscope, the HT22 cells in the high concentration BPDE group showed mitochondrial shrinkage, decreased crista, and increased mitochondrial membrane density. Compared with the solvent control group, the levels of intracellular lipid ROS, Fe2+, 4-HNE, and MDA significantly increased in the high concentration group (P<0.01), the GSH and GSH-PX levels were significantly decreased (P<0.01), the protein expression levels of ASCL4 and COX2 were significantly increased (P<0.01), and the protein expression levels of SCL7A11 and GPX4 were significantly decreased (P<0.01). The ferroptosis inhibitors DFO and Fer-1 significantly reversed the cell survival rate (P<0.01), the ferroptosis characteristic indicators (ROS, Fe2+, 4-HNE, MDA, GSH, and GSH-PX levels) (P<0.01), and the expression levels of ferroptosis-related proteins (ACSL4, COX2, SLC7A11, and GPX4) (P<0.01) in the high concentration BPDE group. Conclusion BPDE can induce ferroptosis in mouse hippocampal neuron HT22 cells, which may be related to the inhibition of SLC7A11/GSH/GPX4 axis and the induction of iron metabolism disorder.
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ObjectiveTo analyze the causes of death and years of life lost among patients with severe mental disorders in Jining city， in order to provide references for improving the management level of the patients. MethodsA total of 3 638 patients with severe mental disorders who were recorded in the National Information System for Severe Mental Disorders in Jining and died between January 1， 2014 and December 31， 2020 were included in the study. The general information and the status of mortality were extracted via checking management files. Thereafter， the causes of death of patients with different characteristics were discussed， and the years of life lost due to severe mental disorders was analyzed through calculating potential years of life lost （PYLL）， average years of life lost （AYLL） and potential years of life lost rate （PYLLR）. ResultsThe majority of patients who died from severe mental disorders were those with schizophrenia， accounting for 77.68% （2 826/3 638）. The most common cause of death among patients with severe mental disorders was physical illness with 1 869 cases （51.37%）. Among the selected subjects， patients with mental retardation and comorbid mental disorders had the youngest age of disease onset ［（22.49±20.14） years］， the youngest age at death ［（51.72±18.32） years］ and the longest duration of disease ［（29.26±19.35） years］. The PYLL， AYLL and PYLLR of patients with severe mental disorders in Jining were 68 941.06 person-years， 18.95 years and 382.36‰， respectively. The mental retardation and comorbid mental disorders had the highest AYLL at 27.21 years， and epilepsy-induced mental disorder had the highest PYLLR at 892.73‰. ConclusionComorbid physical illness is the main causes of death in patients with severe mental disorders in Jining city， and epilepsy-induced mental disorder have occupied the first place in terms of the years of life lost.
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Objective:To establish a method for automatic determination of iodine level in salt by arsenic-cerium catalytic spectrophotometry using an iodine element detector (hereinafter referred to as this method), and to provide reference for in-depth study of salt iodine detection technology.Methods:This method was used to determine the iodine level in salt, and the linear range, detection limit, precision, and accuracy (determination of salt iodine standard substance GBW10006y and GBW10007y, and addition recovery experiment) of this method were determined. The iodine level of 35 salt samples was determined by this method and redox titration method recommended by the national standard, and the results were compared.Results:This method had a good linear relationship within the range of 50 - 600 μg/L standard curve, the absolute value of the correlation coefficients was > 0.999 0, and the detection limit was 5.0 mg/kg. The relative standard deviation of iodine concentration in salt samples with low, medium and high iodine concentrations were all < 6.0%. The determination results of salt iodine standard substance GBW10006y and GBW10007y were within the given value ranges; three iodine concentrations (6.0, 10.0 and 30.0 mg/kg) were added to the salt samples, with an average recovery rate of 96.7% to 105.0%, and a total average recovery rate of 100.9%. The method comparison experiment showed that there was no statistically significant difference between the salt iodine determination results of this method and the redox titration method ( t = - 1.54, P = 0.132). Conclusion:This method has the advantages of high accuracy, good precision and wide linear range in determining salt iodine, and is suitable for the detection of large quantities of samples in salt iodine monitoring.
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Systemic juvenile idiopathic arthritis is one of the common rheumatic and immune diseases in children. It has a sudden onset, obvious systemic symptoms, and lung involvement. However, systemic juvenile idiopathic arthritis with an early manifestation of pulmonary ground-glass opacities combined with macrophage activation syndrome is rare. The clinical data of a child with systemic juvenile idiopathic arthritis with pulmonary ground-glass shadow and macrophage activation syndrome who was admitted to Hubei Maternal and Child Health Care Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology in December 2021 were analyzed retrospectively in order to improve the understanding of rheumatic diseases and pulmonary lesions. The child was admitted to the hospital for 10 days due to rash and fever. Thoracic CT showed scattered ground glass like shadows in both lungs due to the prevention and control screening of COVID-19 pneumonia epidemic situation. After admission, the child was still repeatedly flaccid with high fever, accompanied by dysfunction of both lower limbs. The knee joint MRI found that there was synovitis in the knee joint, and various laboratory indicators suggested macrophage activation syndrome. After that, systemic juvenile idiopathic arthritis was diagnosed. After being treated with methylprednisolone, cyclosporine and topzumab, the clinical remission and the ground-glass shadow of the lung basically disappeared. Through the analysis of this case, it is suggested that clinicians should not ignore other diseases that cause ground glass shadow in the lung during the current epidemic of COVID-19.
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Objective To analyze the value of the size-specific dose estimation(SSDE)and volume CT dose index(CTDIvol)on dose estimations of children's head CT scans and the correlations with the body size.Methods A retrospective study was conducted on plain children's head CT scans of 1 409 patients,they were divided into six groups according to age.The age,CTDI,the head perimeter(HP)of the initial plane of the images,the anterior-posterior length(AP),lateral length(LAT),region of interest area(ARoI)and CT value(CTROI)of the intermediate plane of the images were measured retrospectively.Effect diameter(De),water equivalent diameter(Dw)and SSDE based on Dw were calculated respectively.The differences between CTDI and SSDE in measuring radiation dose and the correlations between CTDIvol,SSDE and age,HP and De were analyzed.Results There were significant differences between CTDIvol and SSDE in all groups(P＜0.001).Compared with SSDE,CTDIvol overestimated the radiation dose by-10.17％,0.69％,7.96％,12.84％,17.99％,22.36％respectively.The bigger the age,the higher CTDIvol overestimated its radiation.CTDIvol and SSDE had strong correlations with age,HP and De,the correlation coefficients CTDIvol were stronger than SSDE.Conclusion SSDE takes body size into account in the calculation,so it can reflect the radiation dose of children's head CT more accurately,CTDIvol overestimates the radiation doses except in children younger than 6 months.CTDIvol is closely collected with age,HP and De and underestimated the radiation dose,so it is more beneficial to reduce the radiation.
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Objective To explore the mediation effect of emotional intelligence in social support and subjective happiness,and to provide practical guidance for scientific management of nursing talents in hemodialysis center.Methods Using a cross-sectional study,by a proportional stratified sampling method,from October 2022 to January 2023,800 hemodialysis nurses in Guangzhou area were selected as the respondents,using the general data adjustment table,general well-being schedule(GWB),social support rating scale(SSRS),and wong law emotional intelligence scale(WLEIS-C).The pearson correlation was used to analyze the correlation between emotional intelligence,social support and subjective happiness of hemodialysis nurses in Guangzhou;the process macro program was used to explore the mediation effect of emotional intelligence in social support and subjective happiness in Guangzhou.Results 707 valid questionnaires were collected,and the effective recovery rate was 88.38%.The total score of subjective well-being of hemodialysis nurses in Guangzhou was(75.67±8.17),the total score of emotional intelligence(82.29±16.20),and the total score of social support(38.76±8.40).The total score of social support was positively associated with the total score of subjective well-being(r=0.517,P<0.01)and the total score of emotional intelligence(r=0.633,P<0.01),the total score of emotional intelligence was positively related to the total score of subjective well-being(r=0.634,P<0.01).Social support had a direct effect on subjective well-being(β=0.165,95%CI:0.103-0.261),and indirectly affected it through the partial mediation effect of emotional intelligence(β=0.095,95%CI:0.069-0.142),and the indirect mediation effect accounted for 36.54%of the total effect.Conclusion Guangzhou area hemodialysis nurses subjective well-being is in upper level,and emotional intelligence in hemodialysis nurses social support and subjective happiness plays intermediary effect,managers should focus on hemodialysis nurses emotional intelligence,take various measures to improve their emotional intelligence level,enhance social support,so as to improve hemodialysis nurses subjective well-being.
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Objective To investigate the effect of long-chain non-coding RNA(lncRNA)AP000892.6 on the migration,inva-sion and epithelial-mesenchymal transition of bladder cancer cells and the related molecular mechanism.Methods The expression of AP000892.6 in bladder cancer and adjacent tissues was analyzed using GEPIA dataset.The expression levels of AP000892.6 in bladder cancer cell lines 647V,T24,UMUC3,5637,RT4 and normal bladder epithelial cells SV-HUC-1 were detected by real-time quanti-tative polymerase chain reaction(RT-qPCR).The AP000892.6 overexpression plasmid or negative control plasmid was transfected into T24 cells,and the experiment was divided into AP000892.6 group and NC group.The plasmid transfection efficiency was verified by RT-qPCR.The migration and invasion abilities of transfected T24 cells were detected by cell scratch method and Transwell assay,re-spectively.The downstream target gene of AP000892.6 was predicted and validated using bioinformatics method and dual-luciferase re-porter gene experiment.The expression of AP000892.6downstream target gene was detected by RT-qPCR.Western blot was used to de-tect the expression of epithelial phenotype proteins E-cadherin,ZO-1 and mesenchymal phenotype proteins Vimentin,N-cadherin and Twist.Results Compared with adjacent tissues,the expression of AP000892.6 was decreased in bladder cancer tissues(P＜0.01).Compared with SV-HUC-1 cells,the expression of AP000892.6 was decreased in bladder cancer cell lines(P＜0.05),and the rela-tive expression level of AP000892.6 in T24 cells was the lowest(P＜0.01).Compared with the NC group,overexpression of AP000892.6 inhibited the migration(P＜0.01)and invasive ability(P＜0.01)of bladder cancer T24 cells.Dual-luciferase reporter gene analysis proved that miR-616-5p was the target gene of AP000892.6(P＜0.01).Compared with the NC group,AP000892.6 negatively regu-lated the expression of miR-616-5p(P＜0.01).Compared with the NC group,the expression of epithelial phenotype proteins E-cad-herin and ZO-1 were up-regulated in T24 cells of AP000892.6group,and the expression of mesenchymal phenotype proteins Vimentin,N-cadherin and Twist were down-regulated.Conclusion AP000892.6 inhibits the migration,invasion and epithelial-mesenchymal transition process of bladder cancer T24 cells by targeting miR-616-5p.
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Objective@#To explore the association between dietary inflammatory index (DII) and metabolic syndrome (MetS) and its components among children aged 6-14 years in Beijing, so as to provide a reference for preventing MetS.@*Methods@#A cross sectional study was carried out in 2 086 records of 1 832 children from the 2017 and 2019 Nutrition and Health Surveillance in Primary and Secondary school students of Beijing (NHSPSB). Three day consecutive 24 hour dietary recalls combined with weighing household cooking oils and condiments were used to collect dietary intake and calculate DII. MetS was diagnosed according to "Definition and Suggestion on the Metabolic Syndrome of Chinese Children and Adolescent". The Generalized estimating equations (GEEs) models were used to analyze the association between DII and the presence of MetS and its components (abdominal obesity, high triglyceride, low high density lipoprotein cholesterol, hypertension, and hyperglycemia).@*Results@#The mean DII score was (1.64±1.07) for the included children. No significant association was found between DII scores and the likelihood of MetS (per 1 point increment: OR =1.16, 95% CI =0.92-1.48, P >0.05). In terms of the components of MetS, DII scores were positively associated with the odds of high triglyceride (per 1 point increment: OR =1.17, 95% CI =1.01-1.36, P <0.05). There was no statistically significant difference in the association among different age groups ( P >0.05). No significant associations were observed between DII and other MetS components( P >0.05).@*Conclusion@#DII scores may not be correlated with the risk of MetS, but proinflammatory diet might increase the risk of high triglyceride. DII score in childhood should be emphasized to identify and prevent MetS as soon as possible.
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ObjectiveTo investigate the inhibitory effects and mechanism of the compound Phyllanthus urinaria Ⅱ （CPU Ⅱ）on the growth of transplanted hepatocellular carcinoma Hep3B2.1-7 （Short for Hep3R） cells in nude mice. MethodAfter the establishment of a xenograft model of hepatocellular carcinoma Hep3B cells in mice， the model mice were randomly divided into a model group， a high-dose CPU Ⅱ group （57.5 g·kg-1）， a low-dose CPU Ⅱ group （28.75 g·kg-1）， and a 5-fluorouracil （5-FU） group （0.025 g·kg-1）， with eight mice in each group. The mice in the high- and low-dose CPU Ⅱ groups were treated with drugs by gavage， once per day， and those in the model group were treated with the same volume of normal saline. The mice in the 5-FU group were treated by 5-FU by intraperitoneal injection， once every other day. After 28 days of administration， mice were sacrificed， and transplanted tumors were collected. Immunohistochemistry （IHC） was used to detect the expression of proliferating cell nuclear antigen （PCNA） of tumor tissues. Terminal-deoxynucleotidyl transferase-mediated nick end labeling （TUNEL） was used to detect cell apoptosis of tumor tissues. The mRNA expression of miR-122 and insulin-like growth factor 1 receptor （IGF-1R） in tumor tissues was detected by Real-time quantitative PCR （Real-time PCR）. The protein expression of CCAAT/enhancer-binding protein α （C/EBPα）， hepatocyte nuclear factor-4α （HNF-4α）， and IGF-1R in tumor tissues was detected by Western blot. ResultThe tumor suppression rates of the high- and low-dose CPU Ⅱ groups and the 5-FU group were 74.90%， 63.62%， and 64.15%， respectively. Compared with the model group， the CPU Ⅱ groups and the 5-FU group showed reduced weight （P<0.01） and volume of tumors （P<0.01）， decreased PCNA positive cells， shallow staining， increased apoptosis cells of transplanted tumor tissues （P<0.05， P<0.01）， increased expression of mRNA expression of miR-122 （P<0.01）， down-regulated mRNA expression of IGF-1R （P<0.01）， and up-regulated protein expression of C/EBPα and HNF-4α in nude mouse transplanted tumor tissues （P<0.01）. The expression of IGF-1R protein in the high-dose CPU Ⅱ group was down-regulated （P<0.05）. Compared with the low-dose CPU Ⅱ group， the high-dose CPU Ⅱ group showed increased apoptotic cells （P<0.01）， up-regulated mRNA expression of miR-122 （P<0.01）， and increased expression of C/EBPα and HNF-4α proteins （P<0.01）. ConclusionCPU Ⅱ has an obvious inhibitory effect on the growth of transplanted hepatocellular carcinoma Hep3B cells in nude mice. The mechanism of action is related to enhancing the expression of transcription factors HNF-4α and C/EBPα， thereby promoting the expression of miR-122 and inhibiting the expression of its target gene IGF-1R.
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【Objective】 To analyze the current situation of direct exemption of clinical blood fess for voluntary blood donors and their family members in Henan Province, in order to improve and fully implement the policy. 【Methods】 According to the policy on blood fees exemption issued by China and Henan Province in 2019,the data of hospitals in 18 prefecture-level cities in Henan from 2020 to 2021 were continuously collected from the system of clinical blood fees exemption,including the way of exemption,the number of people (times) of exemption,exemption amount, the proportion of blood fees exemption and the total exemption rate. The experience gained in the past two years after the implementation of the policy was summarized,and the existing problems and causes were analyzed. 【Results】 The rates of direct exemption of blood fees in Henan Province in 2020 and 2021 were 34.53% (8 709/25 221) and 71.68%(23 587/32 906) (P<0.05) ,respectively. In 2021, the direct exemption rate of blood fees in 18 cities was 6.20% (83/1 370) to 88.50% (1 332/1 505) [ (47.35±41.15)%],and increased month by month from 43.19% (1 183/2 507) in January to 83.15% (2 097/2 522) in August, then remained stable at a similar level to August from September to December, with 83.43% (2 744/3 289) in December as the highest for the year. 【Conclusion】 The implementation of the policy of blood fees exemption showed significant effectiveness, which has effectively promoted the development of voluntary blood donation in Henan. However, there is still room for improving the policy in some cities, which is expected to further increase the direct exemption rate of the city and the whole province.
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Objective:To explore the correlation between differential immune-related genes(DIRGs)and immune cells in the progression of atherosclerosis(AS)and general rule of Chinese medicine for intervening DIRGs.Methods:Firstly,GSE28829 data set was obtained from GEO database,and immune-related genes were downloaded from ImmPort database and MSigDB database.Secondly,differentially expressed genes between early AS plaques(EAP)and advanced AS plaques(AAP)of GSE28829 were screened by limma package,and their intersections with immune-related genes were known as DIRGs.clusterProfiler package was used to enrich the DIRGs.Protein interaction network of DIRGs was constructed and Hub genes were screened.Then,the infiltration patterns of 22 kinds of immune cells were analyzed by CIBERSORT to screen differential immune cells,and the correlation between them and Hub genes were analyzed by Pearson method.Finally,Coremine Medical database was used to predict Chinese herbs for in-tervening DIRGs,the property and flavor of Chinese herbs were collected.Results:Total 63 DIRGs were obtained,and 10 Hub genes(CD86,TLR2,TYROBP,CCR1,ITGB2,CCL2,CCL4,CSF1R,CXCR4,CTSS)were screened out.Enrichment analysis results showed that the molecular functions,biological processes and signaling pathways of DIRGs were closely related to immune regulation.Analysis of immune cell infiltration showed that proportions of regulatory T cells,activated dendritic cells and resting mast cells in EAP were increased.Proportions of memory B cells,γδ T cells,M0 macrophages and M2 macrophages were increased in AAP.Correla-tion analysis showed that CD86 was positively correlated with M2 macrophages in EAP.In AAP,CD86,CTSS,CXCR4,CSF1R,ITGB2 and TYROBP were positively correlated with M0 macrophages,while CCL4 and CCL2 were negatively correlated with resting mast cells.Results of frequency showed that Chinese herbs for intervening DIRGs mainly distributed to liver and lung meridians,and their property and taste were cold and bitter.Conclusion:This study found that in the progression of AS,10 DIRGs were the most important,and 7 kinds of immune cells were dysregulated,among which CD86,CTSS,CXCR4,CSF1R,ITGB2,TYROBP,CCL4 and CCL2 were correlated with resting mast cells,M0 and M1 macrophages.At the same time,immune mechanism of AS progression was closely related to liver meridian,lung meridian,bitter,sweet,cold and warm.The results can provide reference and ideas for Chinese herbs clinical prescription to treat AS and further exploratory the immunological mechanism of AS progression.
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Objective:To investigate the clinical and genetic characteristics of genetic and metabolic infantile cholestatic hepatopathy (ICH), and to provide evidence for its diagnosis and treatment.Methods:Clinical data and follow-up outcomes of hospitalized children diagnosed with ICH in the Department of Gastroenterology, Children′s Hospital, Capital Institute of Pediatrics from January 2014 to December 2019 were retrospectively analyzed.Among the 80 children, 27 were female and 53 were male, with a mean age of onset of (39±18) days old.Children with confirmed etiology by high-throughput sequencing analysis were included in the genetic metabolic group (44 cases), and those with idiopathic neonatal cholestasis(INC) of unknown etiology after the systematic examination were included in the INC group (36 cases). The t-test or independent sample rank sum test was used to compare the laboratory test results and biochemical indexes.The infection rate of cytomegalovirus was compared by the Chi- square test. Results:(1) A total of 80 cases were included, and 44 cases (55.0%)were confirmed as INC by high-throughput sequencing.Among those with a positive molecular diagnosis, there were 23 cases of citrin deficiency (CD), 10 cases of Alagille syndrome (ALGS), 6 cases of progressive familial intrahepatic cholestasis (PFIC), 2 cases of congenital bile acid synthesis defect, 2 cases of Nieman Pick disease, and 1 case of cystic fibrosis.(2) Serum total bile acid (TBA) and activated partial prothrombin time (APTT) levels in the genetic metabolic group were significantly higher than those in the INC group (all P<0.05). TBA and APTT levels in genetic metabolites were 180.6 (115.5, 271.6) μmol/L and 40.6 (37.1, 45.2) s, respectively, which were 123.3 (98.8, 163.4) μmol/L and 34.8 (31.7, 40.1) s in INC group, respectively.There was no significant difference in the cytomegalovirus infection rate between the 2 groups ( P>0.05). (3)The pathological examination of liver tissue in the genetic metabolic group was worse than that in the INC group, with spot-like and fusion focal-like necrosis, and 5 cases (4 cases of ALGS and 1 case of CD) showed a reduced number of bile ducts in the portal area and lumen stenosis. Conclusions:CD, ALGS and PFIC are the common causes of genetic and metabolic ICH.Fundamental cause of cholestasis should be actively examined in children with cytomegalovirus infection.High-throughput sequencing is of great significance in the accurate diagnosis of ICH.
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Objective:To investigate the effect of 1 064-nm Q-switched Nd:YAG laser at different energy settings on cell viability, protease activity and structures of Malassezia furfur. Methods:Cultured standard strains of Malassezia furfur were divided into several groups to be irradiated with 1 064-nm Q-switched Nd:YAG laser at different energies of 0 (control group) , 500, 600, 700, 800 and 900 mJ, respectively. Then, fungal suspensions in the above groups were inoculated onto the Leeming & Notman medium separately. After 7-day culture, the diameter and number of colonies were measured to evaluate the fungal cell viability, the protease activity was measured by using the whole-milk plate medium, and the ultrastructure of Malassezia furfur in each group was observed by transmission electron microscopy. One-way analysis of variance was used for comparisons among multiple groups, least significant difference- t test for multiple comparisons, and Pearson correlation analysis for analyzing correlations of laser energy with colony diameter, number and protease activity. Results:The colony diameter and number both significantly differed among the control group, 500-, 600-, 700-, 800- and 900-mJ groups (colony diameter: 4.05 ± 0.69, 3.76 ± 0.51, 3.28 ± 0.41, 3.09 ± 0.72, 2.54 ± 0.64 and 2.43 ± 0.41 mm, respectively; colony number: 4 787 ± 597, 4 287 ± 761, 1 879 ± 275, 1 082 ± 248 and 209 ± 42, 72 ± 31 colony-forming units, respectively; F = 14.83, 231.85, respectively, both P < 0.05) , and were significantly decreased in the 600-, 700-, 800- and 900-mJ groups compared with the control group (all P < 0.05) . The laser energy was negatively correlated with the colony diameter and number ( r = -0.67, -0.91, respectively, both P < 0.05) . The protease activity significantly differed among the control group, 500-, 700- and 900-mJ groups ( F = 346.60, P < 0.05) , and was significantly lower in the 700- and 900-mJ groups than in the control group (both P < 0.05) . There was a negative correlation between the laser energy and protease activity ( r = -0.94, P < 0.05) . Transmission electron microscopy showed intact fungal structures in the control group, relatively intact fungal structures in the 500-mJ group, and obviously damaged fungal structures in the 600- to 900-mJ groups, and the greater the laser energy, the more severely the fungal structures were damaged. Conclusion:The 1 064-nm Q-switched Nd:YAG laser could affect the cell viability of and protease activity in Malassezia furfur, and damage its structures.
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Objective:To investigate the expression of long non-coding RNA (lncRNA) CALCOCO1 in bladder cancer tissue and its effect on the proliferation and migration of bladder cancer cells by regulating miR-200a-3p.Methods:The relative expression levels of CALCOCO1 in bladder cancer tissues and adjacent tissues were analyzed by TCGA database. Human bladder cancer cells UM-UC-3 were selected, and the cells were divided into negative control group and CALCOCO1 group, and NC plasmid and CALCOCO1 plasmid were transfected into UM-UC-3 cells respectively. The expression level of CALCOCO1 in each group was detected by quantitative real-time polymerase chain reaction (qRT-PCR). The proliferation and migration ability of UM-UC-3 cells were detected by MTT assay and Transwell migration assay. Bioinformatics technology was used to predict and dual-luciferase reporter gene experiments to verify the targeting relationship between CALCOCO1 and miR-200a-3p. The expression levels of miR-200a-3p in UM-UC-3 cells in each group were detected by qRT-PCR. Western blotting was used to detect the expression of UM-UC-3 cells proliferation and migration phenotype in each group. Measurement data were expressed as mean ± standard deviation ( ± s), t-test was used for comparison between two groups, and repeated measurement analysis of variance was used for comparison at different time. Results:Compared with adjacent tissues, the relative expression level of CALCOCO1 in bladder cancer tissues was significantly lower, the difference was statistically significant( P<0.01). The relative expression of CALCOCO1 in UM-UC-3 cells in CALCOCO1 group and negative control group was 9.66±2.51 and 1.07±0.59, respectively. The relative expression level of CALCOCO1 in CALCOCO1 group was significantly higher than that in negative control group, the difference was statistically significant ( P<0.01). Compared with the negative control group, the proliferation activity of UM-UC-3 cells in the CALCOCO1 group was decreased ( P<0.05), and the migration number of UM-UC-3 cells was significantly decreased ( P<0.01). CALCOCO1 had a binding site with miR-200a-3p ( P<0.01). The relative expression of miR-200a-3p in UM-UC-3 cells in CALCOCO1 group and negative control group was 1.02 ± 0.31 and 5.79 ± 1.68, respectively, the difference was statistically significant ( P<0.01). Compared with the negative control group, the expression levels of proliferation phenotype proteins CCNB1, CCNE1 and CCND2 in UM-UC-3 cells in CALCOCO1 group decreased, and the expression levels of migration phenotype proteins FOXC2 and Fibronectin decreased. Conclusion:The expression of CALCOCO1 is down-regulated in bladder cancer tissue, promoting the expression of CALCOCO1 can inhibit the proliferation and migration of bladder cancer UM-UC-3 cells through targeted down-regulation of miR-200a-3p expression.
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Objective:To explore the clinical characteristics of neonate with arrhythmia and the potential risk factors for cardiac insufficiency.Methods:Research data were collected from the cases of neonate with arrhythmia from January 2017 to June 2021 at the Neonatology Department at Children′s Hospital Capital Institute of Pediatrics.A retrospective analysis was conducted to summarize the clinical features and analyze the risk factors of arrhythmia leading to cardiac insufficiency.Results:A total of 90 cases were enrolled, with a male to female ratio of 1.43∶1(53∶37), and the median gestational age was 39(37, 40)weeks; with 16(17.8%) premature infants, 74 full-term infants(82.2%), and the median onset age was 3(0, 11)days.In this group, 63.3% cases(57/90)were irregular rhythmic arrhythmias, of which 66.7%(38/57)were atrial premature contractions, 23.3%(21/90) were tachyarrhythmias, and 13.3%(12/90)were bradyarrhythmias.Additionally, 80.0%(72/90)of the cases had no typical clinical manifestations.Besides, 37.7%(34/90)of the patients had single infection factor, and 4.4%(4/90)of the patients had infection with perinatal hypoxia asphyxia.The ratio of the premature birth, maternal autoimmune diseases and the tachyarrhythmia were higher in heart failure group than those in normal heart function group(71.4% vs.13.3%, P<0.001; 28.6% vs.3.6%, P=0.006; 100% vs.16.9%, P<0.001). The random forest model showed that the factors of premature delivery and maternal autoimmune disease had the greatest influence on cardiac function, and the analysis of 21 tachyarrhythmias showed that the duration of arrhythmia had the greatest influence on cardiac function. Conclusion:Atrial premature contraction is the most common in neonate with arrhythmia, and with non-typical clinical manifestations.The infection and the infection combined with perinatal asphyxia are the main influencing factors of neonate with arrhythmia.Premature delivery, maternal autoimmune disease and the duration of tachyarrhythmias are the main risk factors of neonatal arrhythmias leading to cardiac dysfunction.