Objective To investigate the role of the O-6-methylguanine-DNA methyltransferase (MGMT) gene-3′ untranslated region (UTR) microRNA-associated single nucleotide polymorphism (miRSNP) (rs7896488 G>A) in affecting miR-4297-targeted modulation of MGMT in senescence of lung cells with polymorphic genotypes induced by fractionated low dose ionizing radiation (LDIR). Methods i) MiRSNPs were predicted and screened using bioinformatics, and DNA from two types of lung cells, A549 cells and human bronchial epithelioid cells (HBE cells), was extracted for target gene sequencing. After co-transfection of pGL3c-MGMT-3′UTR-rs7896488 G>A reporter gene recombinant plasmid, pRL-TK Vector with micrON mimic NC #22 or micrON hsa-miR-4297 mimic (set up as the mimic NC group and the miR-4297 mimic group) in these two types of lung cells, dual luciferase reporter gene assay was performed. The relative expression of MGMT mRNA was detected by real-time fluorescence quantitative polymerase chain reaction, and the relative expression of MGMT protein was detected by Western blotting. ii) These two types of lung cells were randomly divided into the control group and irradiation group, which received either 0 or 100 mGy X-rays irradiation seven times. After irradiation, the cells were transfected with either micrON mimic NC #22 or micrON hsa-miR-4297 mimic, resulting in mimic NC + control group, miR-4297 mimic + control group, mimic NC + irradiation group, and miR-4297 mimic + irradiation group. Cells were collected for senescence-associated-β-galactosidase (SA-β-Gal) staining, and the relative expression of matrix metalloproteinase-9 (MMP-9) and chemokine (C-X-C motif) ligand-1 (CXCL-1) proteins was detected via Western blotting. Results i) The rs7896488 G>A was the miRSNP located in the conserved binding region targeted by miR-4297 in the MGMT gene 3′UTR. A549 cells were the rs7896488 GG wild-type homozygous genotype, while HBE cells were the rs7896488 GA heterozygous mutant genotype. In the miR-4297 mimic group, A549 and HBE cells carrying the rs7896488 G allele showed significantly lower dual-luciferase activity compared with that in the mimic NC group (both P<0.01). However, there was no significant difference in dual-luciferase activity between the two groups in both A549 and HBE cells carrying the rs7896488 A allele (both P>0.05). The relative expression levels of MGMT mRNA and MGMT protein of A549 cells in the miR-4297 mimic group were lower than those in the mimic NC group (both P<0.05). However, there was no significant difference in MGMT mRNA and MGMT protein of HBE cells between these two groups (both P>0.05). ii) The relative activity of SA-β-Gal and the relative expression of MMP-9 and CXCL-1 proteins of A549 cells in the miR-4297 mimic+irradiation group were higher than those in the mimic NC + control group, the miR-4297 mimic + control group, and the mimic NC + irradiation group (all P<0.05). The relative activity of SA-β-Gal and the relative expression of MMP-9 and CXCL-1 proteins of HBE cells in the miR-4297 mimic + irradiation group were higher than those in the mimic NC + control group and the miR-4297 mimic + control group (all P<0.05), while there was no significant difference compared with those in the mimic NC + irradiation group (all P>0.05). Conclusion MGMT-3′UTR-miRSNP rs7896488 G>A plays a role in LDIR-induced senescence of lung cells with different polymorphic genotypes by affecting miR-4297-targeted regulation of MGMT.

Objective To investigate the effects of fractionated low-dose ionizing radiation (LDIR) on the ferroptosis in human bronchial epithelial (HBE) cells as well as the associated differentially expressed genes (DEGs), biological processes, and signaling pathways. Methods HBE cells were exposed to different single doses of X-ray irradiation (0, 25, 50, 75, and 100 mGy) for 24, 48, and 72 h, respectively. The change in cell viability was detected by MTT assay. Cells were irradiated with 0, 25, 50, and 100 mGy X-rays 5 times, with 48 h between each irradiation and a dose rate of 50 mGy/min. Cells were harvested 24 h after irradiation for the measurement of the expression of ferroptosis-related genes SLC7A11 and GPX4 at the mRNA and protein levels, cellular iron content, and the expression of FTH1 and FTL mRNAs. High-throughput sequencing was used to screen for the DEGs in each dose group, followed by Gene Ontology-Biological Process (GO-BP) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and Gene Set Enrichment Analysis (GSEA). Results Compared with the control group, single-dose LDIR significantly increased cell proliferation at 75 mGy after 24 h (P < 0.05), at 50, 75, and 100 mGy after 48 h (P < 0.05), and at 75 and 100 mGy after 72 h (P < 0.05). Compared with the control group, at the end of the fifth fractionated LDIR, SLC7A11 and GPX4 mRNAs decreased at all doses (P < 0.05), SLC7A11 protein decreased at all doses, GPX4 protein decreased at 25 and 100 mGy, iron content increased at all doses, and FTH1 and FTL mRNAs decreased at all doses (P< 0.05). Sequencing analysis identified 248, 30, and 291 DEGs and 10, 2, and 9 ferroptosis-associated genes at the three doses compared to the control. Gene Ontology-Biological Process analysis showed that DEGs were mainly enriched in biological processes such as response to lipids, cell death, and response to unfolded proteins. Kyoto Encyclopedia of Genes and Genomes analysis showed that DEGs were mainly enriched in the JAK-STAT signaling pathway, lipids and atherosclerosis, ferroptosis, protein processing in the endoplasmic reticulum, and FoxO signaling pathway. Gene set enrichment analysis showed that DEGs were mainly enriched in ferroptosis, fatty acid degradation, and glutathione metabolism. Conclusion Fractionated low-dose radiation induced ferroptosis in HBE cells, and DEGs were predominantly enriched in biological processes and signaling pathways related to inflammation, ferroptosis, and endoplasmic reticulum stress.

Objective:The epidemiological characteristics of tuberculosis deaths among Chinese residents from 2006 to 2021 were analyzed, and the tuberculosis mortality rate from 2022 to 2027 was predicted to provide a reference for tuberculosis prevention and control in China.Methods:The data set of tuberculosis deaths from 2006 to 2021 was published regularly by the China CDC, and the crude mortality rate (CMR) and age-standardized mortality rates (ASMR) were calculated according to the population structure of China in 2000. The distribution characteristics of age, sex, region, and time of tuberculosis deaths were analyzed, the Joinpoint regression analysis model was used to analyze the changing trend, and the grey model was applied to predict CMR and ASMR from 2022 to 2027.Results:From 2006 to 2021, the CMR and ASMR of tuberculosis showed a downward trend among males and females, urban and rural areas, and all age groups, in a word, all the Chinese residents. Except for the age group ≥85 years old, the mortality trend was insignificant. In the eastern, central, or western regions. CMR and ASMR were significantly higher in males than in females.CMR and ASMR were significantly lower in urban areas than in rural areas. In general, active tuberculosis patients present a higher mortality rate. The CMR and ASMR in the western region were higher than those in the eastern and central regions and lower in the eastern region than in the central region, but the differences were less obvious. The ASMR of the eastern cities was lower than that of the central and western regions, and the ASMR of the central cities was higher than that of the western region from 2006 to 2009 and 2012 and lower than that of the western region in other years. The ASMR in the western countryside was higher than that in the eastern and central regions and lower in the eastern part than in the central region, but the difference was not obvious. The grey model prediction results show that the CMR (/100 000) of Chinese residents from 2022 to 2027 is 1.585, 1.471, 1.360, 1.250, 1.143, and 1.038, and the ASMR (/100 000) is 0.779, 0.653, 0.531, 0.411, 0.295 and 0.181, respectively.Conclusions:The CMR and ASMR of tuberculosis will continue to decline, and extraordinary achievements have been made in tuberculosis prevention and control in Chinese residents from 2006 to 2021 and, presumably, from 2022 to 2027. However, tuberculosis screening and treatment programs in the western region, men, the elderly population, and rural areas should be further strengthened, and targeted prevention and control measures should be formulated to reduce mortality.

Objective To explore the therapeutic mechanism of compound Yuye Decoction against diabetic cardiomyopathy(DCM).Methods Drugbank,Gene Cards,OMIM and PharmGKb databases were used to obtain DCM-related targets,and the core targets were identified and functionally annotated by protein-protein interaction network analysis followed by GO and KEGG enrichment analysis.The"Traditional Chinese Medicine-Key Component-Key Target-Key Pathway"network was constructed using Cytoscape 3.9.1,and molecular docking was carried out for the key components and the core targets.In the animal experiment,Wistar rat models of DCM were treated with normal saline or Yuye Decoction by gavage at low(0.29 g/kg)and high(1.15 g/kg)doses for 8 weeks,and the changes in cardiac electrophysiology and histopathology were evaluated.The changes in serum levels of LDH,CK,and CK-MB were examined,and myocardial expressions of PI3K,P-PI3K,Akt,P-AKT,BAX,IL-6,and TNF-α were detected using Western blotting.Results We identified 61 active compounds in Yuye Decoction with 1057 targets,3682 DCM-related disease targets,and 551 common targets between them.Enrichment of the core targets suggested that apoptosis,inflammation and the PI3K/Akt pathways were the key signaling pathways for DCM treatment.Molecular docking studies showed that the active components in Yuye Decoction including gold amidohydroxyethyl ester and kaempferol had strong binding activities with AKT1 and PIK3R1.In DCM rat models,treatment with Yuye Decoction significantly alleviated myocardial pathologies,reduced serum levels of LDH,CK and CK-MB,lowered myocardial expressions of BAX,IL-6 and TNF-α,and increased the expressions of P-PI3K and P-AKT.Conclusion The therapeutic effect of compound Yuye Decoction against DCM is mediated by its multiple active components that act on multiple targets and pathways to inhibit cardiomyocyte apoptosis and inflammatory response by regulating the PI3K/Akt signaling pathway.

Objective To explore the therapeutic mechanism of compound Yuye Decoction against diabetic cardiomyopathy(DCM).Methods Drugbank,Gene Cards,OMIM and PharmGKb databases were used to obtain DCM-related targets,and the core targets were identified and functionally annotated by protein-protein interaction network analysis followed by GO and KEGG enrichment analysis.The"Traditional Chinese Medicine-Key Component-Key Target-Key Pathway"network was constructed using Cytoscape 3.9.1,and molecular docking was carried out for the key components and the core targets.In the animal experiment,Wistar rat models of DCM were treated with normal saline or Yuye Decoction by gavage at low(0.29 g/kg)and high(1.15 g/kg)doses for 8 weeks,and the changes in cardiac electrophysiology and histopathology were evaluated.The changes in serum levels of LDH,CK,and CK-MB were examined,and myocardial expressions of PI3K,P-PI3K,Akt,P-AKT,BAX,IL-6,and TNF-α were detected using Western blotting.Results We identified 61 active compounds in Yuye Decoction with 1057 targets,3682 DCM-related disease targets,and 551 common targets between them.Enrichment of the core targets suggested that apoptosis,inflammation and the PI3K/Akt pathways were the key signaling pathways for DCM treatment.Molecular docking studies showed that the active components in Yuye Decoction including gold amidohydroxyethyl ester and kaempferol had strong binding activities with AKT1 and PIK3R1.In DCM rat models,treatment with Yuye Decoction significantly alleviated myocardial pathologies,reduced serum levels of LDH,CK and CK-MB,lowered myocardial expressions of BAX,IL-6 and TNF-α,and increased the expressions of P-PI3K and P-AKT.Conclusion The therapeutic effect of compound Yuye Decoction against DCM is mediated by its multiple active components that act on multiple targets and pathways to inhibit cardiomyocyte apoptosis and inflammatory response by regulating the PI3K/Akt signaling pathway.

Objective To investigate the status of metabolically unhealthy status (MUS) and its influencing factors in male residents living around a uranium mine in Guangdong Province. Methods A total of 867 local male residents born and living around a uranium mine in Guangdong Province were selected as the study subjects using two-stage random sampling method. The residents were divided into 10 km- and 20 km- radius groups, according to their living distance <10 km or 10-20 km from the uranium mine. Blood pressure, fasting blood glucose, and blood lipid levels were tested among the study subjects. The influencing factors of MUS and its components were analyzed by multiple logistic regression. Results The detection rate of MUS was 42.2%. The detection rates of MUS component such as elevated diastolic blood pressure, elevated systolic blood pressure, elevated fasting blood glucose, elevated triglyceride and low high-density lipoprotein cholesterol (HDL-C) were 48.8%, 60.9%, 11.9%, 40.7% and 19.3%, respectively. The MUS detection rate in the 10 km group was lower than that in the 20 km group (38.5% vs 45.9%, P<0.05). The result of multivariate logistic regression analysis showed that subjects aged >50 years had a higher risk of MUS and elevated systolic blood pressure than those aged ≤ 50 years (all P<0.05). Drinkers had a higher risk of MUS, elevated systolic blood pressure, elevated diastolic blood pressure, elevated triglyceride, and low HDL-C than non-drinkers (all P<0.05). Those who ate fruit occasionally had a higher risk of MUS than those who ate fruit regularly (P<0.05). Overweight and obese individuals had a higher risk of MUS, elevated systolic blood pressure, elevated diastolic blood pressure, elevated triglyceride, and low HDL-C than those with normal body mass (all P<0.05). Individuals in the 20 km group had a higher risk of MUS and low HDL-C than the 10 km group (all P<0.05). Conclusion The risk factors of MUS detection among male residents living around the uranium mines are age > 50 years, drinking, occasional fruit intake, and being overweight or obese. Age > 50 years, drinking alcohol, overweight and obesity can affect the detection of MUS components in different degrees. Environmental radiation levels have not been identified as a risk factor for MUS in these study subjects.

Objective:To identify XMA09,a broadly neutralizing antibody against severe acute respiratory syndrome coronavi-rus 2(SARS-CoV-2),and to explore the potential mechanism of XMA09 to broadly neutralize variants of SARS-CoV-2,so as to pro-vide a reference for vaccine design and broadly neutralizing antibody screening against SARS-CoV-2.Methods:XMA09 protein was expressed and purified by ExpiCHO expression system and Protein A chromatography column.The key amino acid sites on receptor binding domain(RBD)recognized by XMA09 were identified by Cryo-EM,and the affinity of XMA09 to Spike protein of SARS-CoV-2 and its variants was detected by indirect ELISA and Surface Plasmon Resonance(SPR).The pseudovirus neutralization assay based on vesicular stomatitis virus(VSV)was used to detect the neutralization ability of XMA09 to wild-type and variants of SARS-CoV-2.Results:In this study,it was found that the epitopes recognized by XMA09 were conservative,who has an excellent binding ability to Spike protein of many kinds of variants,and could neutralize the variants of concern(VOCs),including the widespread BA.4/5.Conclusion:XMA09 is a broadly neutralizing antibody,which has the potential to be used as a therapeutic antibody against SARS-CoV-2,and can provide reference for broad-spectrum vaccine design and antibody drug development against SARS-CoV-2.

African swine fever(ASF)is an acute and highly pathogenic hemorrhagic disease of pigs,causing huge economic losses to pig industry.In order to quantitatively detect clinical samples of ASF and inactivated ASFV antigens,the IgG of ASF positive serum was used as capture anti-body and the HRP-labeled p72 monoclonal antibody was used as detecting antibody.The standard curve was drawn with the cell-cultured ASFV,and a sandwich ELISA detection of antigen was es-tablished.The specificity,sensitivity and stability of the method were evaluated.The effects of dif-ferent inactivation methods and adjuvant addition on antigen detection were further evaluated.The results showed that the minimum detection limits of the recombinant protein and the ASFV were 0.1 mg/L and 103.7 TCID50/mL,respectively.There was no cross-reaction with five common porcine pathogenic viruses,and the coefficient variations between batches was less than 10％.The total co-incidence rate with real-time fluorescence quantitative PCR was 92％(23/25).The sensitivity of antigen detection was significantly reduced when antigen was treated by BEI inactivation,and the detection results were severely interfered by aluminum adjuvant and nano-adjuvant.In summary,the sandwich ELISA antigen detection method established is specific,sensitive,and repeatable,with a good consistency to the qPCR method,which provides an effective clinical diagnostic meth-od for ASFV antigen.

Objective To investigate the effect of docosahexaenoic acid(DHA)on human colon cancer cell line HT-29 and underlying mechanism.Methods Human colon cancer cell line HT-29 was incubated with DMSO(control),DHA(25,50,100 μmol/L)and 100 μmol/L DHA and/or 30 μmol/L 740Y-P.Proliferation was examined by MITT;apoptosis was detected by annexin V-FITC/PI.Western blot was used for detection of protein expression of Bcl-2,Bax apoptosis-related protein and PI3K/Akt/mTOR pathway,and RT-qPCR was used for checking mRNA expression of NLRP3/Caspase-1/IL-1β pathway.Results Compared with the control group,DHA 25,50,and 100 μmol/L treatment of HT-29 cells resulted in decreased cell survival(P＜0.05),increased apoptosis(P＜0.05),decreased Bcl-2/Bax ratio(P＜0.05)and decreased phosphorylation of PI3K,Akt and mTOR in HT-29 cells(P＜0.05 or P＜0.01).Expressions of NLRP3,Caspase-1 and IL-1 β mRNA were decreased(P＜0.05).In addition,cell viability,protein phos-phorylation(p-PI3K,p-Akt,p-mTOR)and relative mRNA expression of NLRP3,Caspase 1,and IL-1β were lower in HT-29 cells which were co-incubated with DHA 100 μmol/L and 740Y-P 30 μmol/L than those in the control group(P＜0.05 or P＜0.01)and 740Y-P 30 μmol/L group(P＜0.05),while higher than that of DHA 100 μmol/L group(P＜0.05 or P＜0.01).Conclusions DHA inhibits the proliferation of human colon cancer cell line HT-29,its mecha-nism is potentially related to the inhibition of PI3K/Akt/mTOR and NLRP3/Caspase-1/IL-1 β signaling pathways.

Objective This study aims to investigate the structure and anti-angiogenic activity of homogeneous polysaccharides in Honeysuckle flowers,constructing the theoretical basis for its widespread application.Methods Homogeneous Honeysuckle polysaccharide LF-02-2 was obtained through water extraction,alcohol precipitation,anion exchange chromatography,and gel permeation chromatography.The structure of LF-02-2 was deduced through molecular weight determination,monosaccharide composition analysis,sugar residue linkage analysis,partial acid hydrolysis,and sugar aldonic acid reduction combined with NMR data.Additionally,in vitro tube formation experiments using human microvascular endothelial cells(HMEC-1)were conducted to assess its anti-angiogenic activity.Results Structural analysis of LF-02-2 revealed a homogeneous polysaccharide with a weight-average molecular weight of 74.1 kDa.The monosaccharide composition included rhamnose(Rha),galactose(Gal),galacturonic acid(GalA),and arabinose(Ara)in molar ratios of 10.43:14.94:6.66:67.97.The main chain consisted of 1,4-linked α-Galp A,1,2-linked α-Rhap,and 1,2,4-linked α-Rhap.Branches were connected to O-4 of 1,2,4-linked α-Rhap and included β-Galp with terminal connections and 1,4,6-linked β-Galp and α-Araf with terminal and 1,5-linked connections.Tube formation experiments demonstrated that LF-02-2 significantly inhibited tube formation in human microvascular endothelial cells(HMEC).Conclusion LF-02-2,an RG-I pectic polysaccharide from Honeysuckle flowers,exhibited significant anti-angiogenic activity,suggesting its potential for development as an anti-angiogenic drug.