AIM: To explore the effect of dihydroquercetin on visual function in mice with form deprivation myopia based on the NOD-like receptor thermoprotein domain-related protein 3(NLRP3)inflammasome pathway.METHODS: The C57BL/6 mice were randomly divided into control group and form deprivation myopia model group, and the form deprivation myopia model group was constructed by covering the right eye with a translucent eye patch. After successful modeling, the mice in the model group of form deprivation myopia were randomly divided into model group, low-, medium- and high-dose dihydroquercetin groups, and high-dose dihydroquercetin + NLRP3 agonist group. The diopter and axial length of mice in each group were detected. The kit was used to detect the levels of superoxide dismutase(SOD)and malondialdehyde(MDA)in retinal tissue. RT-qPCR was used to detect the mRNA expressions of NLRP3, apoptosis-associated spot-like protein(ASC), Caspase-1, IL-1β and IL-18 in retinal tissues. Western blot was used to detect the expression of NLRP3, ASC, cleaved Caspase-1, IL-1β and IL-18 proteins in retinal tissues. TUNEL staining was used to detect apoptosis in retinal tissue.RESULTS: Compared with the control group, the diopter of the mice in the model group decreased, and axial length increased, and the SOD decreased whereas MDA, NLRP3, ASC, Caspase-1, IL-1β, IL-18 increased, and the rate of apoptosis in retinal tissue increased(all P<0.05). Compared with the model group, the diopter of mice in the low-, medium- and high-dose dihydroquercetin groups increased, axial length shortened, the SOD increased, whereas MDA, NLRP3, ASC, Caspase-1, IL-1β, IL-18 decreased, and the rate of apoptosis in retinal tissue decreased(all P<0.05). Compared with the high-dose dihydroquercetin group, the high-dose dihydroquercetin+NLRP3 agonist group had reduced diopter, increased axial length, decreased SOD levels, elevated MDA, NLRP3, ASC, Caspase-1, IL-1β, and IL-18 levels, as well as increased apoptosis rate in retinal tissue(all P<0.05).CONCLUSION: Dihydroquercetin can improve visual function in mice with form deprivation myopia by inhibiting pyroptosis and oxidative stress responses, which may be related to the suppression of NLRP3 inflammasome. NLRP3 agonists can partially mitigate the effects of high-dose dihydroquercetin on form deprivation myopia in mice.

Bacterial infection is a major threat to global public health, and can cause serious diseases such as bacterial skin infection and foodborne diseases. It is essential to develop a new method to rapidly diagnose clinical multiple bacterial infections and monitor food microbial contamination in production sites in real-time. In this work, we developed a 4-mercaptophenylboronic acid gold nanoparticles (4-MPBA-AuNPs)-functionalized hydrogel microneedle (MPBA-H-MN) for bacteria detection in skin interstitial fluid. MPBA-H-MN could conveniently capture and enrich a variety of bacteria within 5 min. Surface enhanced Raman spectroscopy (SERS) detection was then performed and combined with machine learning technology to distinguish and identify a variety of bacteria. Overall, the capture efficiency of this method exceeded 50%. In the concentration range of 1 × 10⁷ to 1 × 10¹⁰ colony-forming units/mL (CFU/mL), the corresponding SERS intensity showed a certain linear relationship with the bacterial concentration. Using random forest (RF)-based machine learning, bacteria were effectively distinguished with an accuracy of 97.87%. In addition, the harmless disposal of used MNs by photothermal ablation was convenient, environmentally friendly, and inexpensive. This technique provided a potential method for rapid and real-time diagnosis of multiple clinical bacterial infections and for monitoring microbial contamination of food in production sites.

OBJECTIVE T o compare the biological characteristics,drug resistance and pathogenicity between two strains of mucoid Pseudomonas aeruginosa and the standard strain PAO1.METHODS The strains were identified,and biofilms were detected by 96-well plates method.The bacterial drug resistance was detected by fully automatic drug susceptibility analysis system,the expression levels of RNA of virulence factors were detected by real-time fluorescent quantitative polymerase chain reaction(RT-PCR);the models of rats with pneumonia infection were established through liquid aerosol lung delivery method,the survival status of the rats was observed,and the lev-els of cytokines in bronchoalveolar lavage fluid(BALF)were detected.RESULTS NY4593,NY4605 and PAO1 strains were successfully isolated and identified.NY4593 and NY4605 showed high-yield biofilms,while PAO1 showed low-yield biofilms.The drug resistance rates of NY4593 and NY4605 were remarkably higher than those of the PAO1.The expression levels of exoT and exoY gene RNA of the NY4593 and NY4605 strains were higher than those of the PAO1 strains(P＜0.05);the expression level of exoS gene RNA of the NY4605 was lower than that of the PAO1(P＜0.05).Under the same infection dose,the PAO1 showed more powerful pathogenicity,and the secretion volumes of inflammatory factors interleukin-6(IL-6),interleuki-1β(IL-1β)and interleukin-17A(IL-17A)of the PAO1 were(2858.00±150.30)pg/ml,(7821.00±761.20)pg/ml and(1079.00±225.40)pg/ml respectively,remarkably higher than those of the NY4593 and NY4605(P＜0.05).CONCLUSION The clini-cal mucoid NY4593 and NY4605 remarkably differ from PAO1 in biology and pathogenicity.The study may facilitate deep understanding of the mechanisms of PA infection and provide guidance for clinical treatment,prevention and control.

Objective To explore the biological functions of pyroptosis-related genes in pneumonic plague using bioinformatics methods,and to evaluate their potential applicability as diagnostic markers.Methods The pneumonic plague-related dataset GSE220123 was retrieved from the Gene Expression Omnibus(GEO)database and screened for differentially expressed pyroptosis-related genes(DE-PRGs).The functions of DE-PRGs were studied via Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses,and immune infiltration analysis.The hub genes were identified via protein-protein interaction(PPI)network analysis,and further screened for key genes with sustained high expression characteristics based on differential expression analysis.The relative expression levels of the key genes were verified using the reverse transcription real-time quantitative PCR(qPCR)method.Results A total of 17 DE-PRGs were screened,and PPI network analysis revealed 7 Hub genes.Among them,Casp4 continued to be up-regulated during the course of pneumonic plague.The results of reverse transcription qPCR were consistent with the those of bioinformatic analyses.Conclusion DE-PRGs play a crucial role in the immune response of pneumonic plague,especially Casp4,which has significant applications as a diagnostic biomarker and potential therapeutic target for pneumonic plague.

Objective : To identify autophagy-related genes involved in pulmonary infection caused by the highly drug-resistant and virulent methicillin-resistant Staphylococcus aureus strain USA300 ( USA300) ，and to explore the underlying molecular mechanisms ， thereby providing potential targets for immunotherapy． Methods: The GSE220943 dataset of a USA300-induced pulmonary infection mouse model was obtained from the GEO database． Differentially expressed genes ( DEGs ) were identified using the DESeq2 package． Autophagy-related genes ( ARGs) were retrieved from the MSigDB and Autophagy databases．Weighted gene co-expression network analysis ( WGCNA) was performed to construct gene co-expression modules．Genes overlapping among DEGs，ARGs，and WGCNA modules were identified as autophagy-related DEGs．Gene Ontology ( GO) enrichment analysis was con- ducted using the clusterProfiler R package，while Kyoto Encyclopedia of Genes and Genomes ( KEGG) pathway en- richment analysis was performed via the Metascape platform．Immune cell infiltration was analyzed using the Immu- CellAI-mouse website．A protein - protein interaction ( PPI) network was constructed using the STRING database， and hub genes were identified through topological analysis in Cytoscape． Receiver operating characteristic curve ( ROC) curves were plotted via the website https: / /www．bioinformatics．com．cn． Finally，key gene expression was validated in mouse lung tissues by real-time quantitative reverse transcription PCR ( RT-qPCR) ． Results: A total of 6 135，4 075，3 680，and 2 342 differentially expressed genes ( DEGs) were identified at 12，24，48，and 96 hours post-infection，respectively．By integrating DEGs，autophagy-related genes ( ARGs) ，and WGCNA mod- ules，19 autophagy-related DEGs were identified． GO and KEGG enrichment analyses indicated that these genes were mainly involved in CD4 + T cell activation and regulation，innate immune responses，and autophagosome mem- brane formation．Immune infiltration analysis revealed that innate immune cells such as neutrophils and dendritic cells predominated during the early phase of infection，while γδ T cells and M2 macrophages became more promi- nent in the later stages．PPI network analysis identified 12 hub autophagy-related genes，among which three upreg- ulated key genes ( Eif2ak2，Ikbke，and Nfkbiz) were further confirmed．The area under the ROC curve for all three genes was 1. 000．RT-qPCR validation demonstrated significantly elevated expression of these three genes in lung tissues at 24 hours post-infection ( all P＜0. 05) ． Conclusion Eif2ak2，Ikbke，and Nfkbiz may be involved in the pulmonary infection caused by USA300 by promoting autophagy and hold promise as potential targets for immuno- therapy．

Bacterial infection is a major threat to global public health,and can cause serious diseases such as bacterial skin infection and foodborne diseases.It is essential to develop a new method to rapidly di-agnose clinical multiple bacterial infections and monitor food microbial contamination in production sites in real-time.In this work,we developed a 4-mercaptophenylboronic acid gold nanoparticles(4-MPBA-AuNPs)-functionalized hydrogel microneedle(MPBA-H-MN)for bacteria detection in skin inter-stitial fluid.MPBA-H-MN could conveniently capture and enrich a variety of bacteria within 5 min.Surface enhanced Raman spectroscopy(SERS)detection was then performed and combined with ma-chine learning technology to distinguish and identify a variety of bacteria.Overall,the capture efficiency of this method exceeded 50％.In the concentration range of 1 × 10 7 to 1 × 10 10 colony-forming units/mL(CFU/mL),the corresponding SERS intensity showed a certain linear relationship with the bacterial concentration.Using random forest(RF)-based machine learning,bacteria were effectively distinguished with an accuracy of 97.87％.In addition,the harmless disposal of used MNs by photothermal ablation was convenient,environmentally friendly,and inexpensive.This technique provided a potential method for rapid and real-time diagnosis of multiple clinical bacterial infections and for monitoring microbial contamination of food in production sites.

Objective:To investigate the influencing factors of adverse reactions of reconstructive flaps after postoperative precision radiation therapy for head and neck tumors.Methods:Medical records of 46 patients who underwent radiotherapy after flap reconstruction for head and neck tumors in the Affiliated Hospital of Inner Mongolia Medical University between January 2016 and October 2022 were retrospectively analyzed. The overall survival (OS), local regional control (LRC) and dosimetric parameters after radiotherapy were analyzed. The adverse reactions mainly including radiation dermatitis and flap necrosis at 3 and 6 months after radiotherapy, flap atrophy, flap fibrosis, dysphagia and chewing dysfunction at 12 and 24 months after radiotherapy were recorded. Adverse reactions were graded using the common terminology criteria for adverse events (CTCAE) version 5.0. Quantitative data were analyzed by one-way analysis of variance or rank-sum test. Qualitative data were analyzed by chi-square test or Fisher's exact test. Multivariate analysis of influencing factors of adverse reactions was performed using binary logistic stepwise regression.Results:All 46 patients were aged 57.6 years on average. The median follow-up time was 65 months (12-100 months). After the follow-up, 22 patients died, 6 recurred, and 7 had distant metastases. The 5-year OS rate was 48% and 5-year LRC rate was 69%. The incidence of radiation dermatitis was decreased over time after the end of radiotherapy: 80% (37/46) and 65% (30/46) at 3 months and 6 months after radiotherapy, respectively. The incidence of grade 3 radiation dermatitis was 11% (5/46) and 0 at 3 and 6 months after radiotherapy, respectively. The incidence of grade 2 flap necrosis was 4% (2/46) at 3 months after radiotherapy. At 12 months after radiotherapy, the incidence of flap atrophy and fibrosis was 83% (38/46) and 67% (31/46) , and the incidence of grade 3 flap atrophy and fibrosis was equally 4% (2/46) , respectively. All patients had dysphagia and chewing dysfunction at 12 months after radiotherapy. At 24 months after radiotherapy, 83% (38/46) and 61% (28/46) of patients still had dysphagia and chewing dysfunction. Multivariate regression analysis showed that irradiated flap volume, total radiotherapy dose, pharyngeal constrictor D mean and pharyngeal constrictor V 60 Gy, the interval between surgery and radiotherapy, gender and age were the independent influencing factors of adverse reactions of reconstructive flaps after precision radiation therapy for head and neck tumors. Conclusions:For patients undergoing flap reconstruction for head and neck tumors, under the premise of ensuring tumor control, the dose to normal tissue and flap blood supply area should be strictly limited, the irradiated volume should be reduced, and the interval between surgery and radiotherapy should be shortened (≤6 weeks) to reduce the risk of adverse reactions. Hypo-fractionated radiotherapy plan should be formulated or the total radiotherapy dose should be appropriately adjusted for elderly patients to reduce long-term complications.

OBJECTIVE T o compare the biological characteristics,drug resistance and pathogenicity between two strains of mucoid Pseudomonas aeruginosa and the standard strain PAO1.METHODS The strains were identified,and biofilms were detected by 96-well plates method.The bacterial drug resistance was detected by fully automatic drug susceptibility analysis system,the expression levels of RNA of virulence factors were detected by real-time fluorescent quantitative polymerase chain reaction(RT-PCR);the models of rats with pneumonia infection were established through liquid aerosol lung delivery method,the survival status of the rats was observed,and the lev-els of cytokines in bronchoalveolar lavage fluid(BALF)were detected.RESULTS NY4593,NY4605 and PAO1 strains were successfully isolated and identified.NY4593 and NY4605 showed high-yield biofilms,while PAO1 showed low-yield biofilms.The drug resistance rates of NY4593 and NY4605 were remarkably higher than those of the PAO1.The expression levels of exoT and exoY gene RNA of the NY4593 and NY4605 strains were higher than those of the PAO1 strains(P＜0.05);the expression level of exoS gene RNA of the NY4605 was lower than that of the PAO1(P＜0.05).Under the same infection dose,the PAO1 showed more powerful pathogenicity,and the secretion volumes of inflammatory factors interleukin-6(IL-6),interleuki-1β(IL-1β)and interleukin-17A(IL-17A)of the PAO1 were(2858.00±150.30)pg/ml,(7821.00±761.20)pg/ml and(1079.00±225.40)pg/ml respectively,remarkably higher than those of the NY4593 and NY4605(P＜0.05).CONCLUSION The clini-cal mucoid NY4593 and NY4605 remarkably differ from PAO1 in biology and pathogenicity.The study may facilitate deep understanding of the mechanisms of PA infection and provide guidance for clinical treatment,prevention and control.

Objective:To investigate the influencing factors of adverse reactions of reconstructive flaps after postoperative precision radiation therapy for head and neck tumors.Methods:Medical records of 46 patients who underwent radiotherapy after flap reconstruction for head and neck tumors in the Affiliated Hospital of Inner Mongolia Medical University between January 2016 and October 2022 were retrospectively analyzed. The overall survival (OS), local regional control (LRC) and dosimetric parameters after radiotherapy were analyzed. The adverse reactions mainly including radiation dermatitis and flap necrosis at 3 and 6 months after radiotherapy, flap atrophy, flap fibrosis, dysphagia and chewing dysfunction at 12 and 24 months after radiotherapy were recorded. Adverse reactions were graded using the common terminology criteria for adverse events (CTCAE) version 5.0. Quantitative data were analyzed by one-way analysis of variance or rank-sum test. Qualitative data were analyzed by chi-square test or Fisher's exact test. Multivariate analysis of influencing factors of adverse reactions was performed using binary logistic stepwise regression.Results:All 46 patients were aged 57.6 years on average. The median follow-up time was 65 months (12-100 months). After the follow-up, 22 patients died, 6 recurred, and 7 had distant metastases. The 5-year OS rate was 48% and 5-year LRC rate was 69%. The incidence of radiation dermatitis was decreased over time after the end of radiotherapy: 80% (37/46) and 65% (30/46) at 3 months and 6 months after radiotherapy, respectively. The incidence of grade 3 radiation dermatitis was 11% (5/46) and 0 at 3 and 6 months after radiotherapy, respectively. The incidence of grade 2 flap necrosis was 4% (2/46) at 3 months after radiotherapy. At 12 months after radiotherapy, the incidence of flap atrophy and fibrosis was 83% (38/46) and 67% (31/46) , and the incidence of grade 3 flap atrophy and fibrosis was equally 4% (2/46) , respectively. All patients had dysphagia and chewing dysfunction at 12 months after radiotherapy. At 24 months after radiotherapy, 83% (38/46) and 61% (28/46) of patients still had dysphagia and chewing dysfunction. Multivariate regression analysis showed that irradiated flap volume, total radiotherapy dose, pharyngeal constrictor D mean and pharyngeal constrictor V 60 Gy, the interval between surgery and radiotherapy, gender and age were the independent influencing factors of adverse reactions of reconstructive flaps after precision radiation therapy for head and neck tumors. Conclusions:For patients undergoing flap reconstruction for head and neck tumors, under the premise of ensuring tumor control, the dose to normal tissue and flap blood supply area should be strictly limited, the irradiated volume should be reduced, and the interval between surgery and radiotherapy should be shortened (≤6 weeks) to reduce the risk of adverse reactions. Hypo-fractionated radiotherapy plan should be formulated or the total radiotherapy dose should be appropriately adjusted for elderly patients to reduce long-term complications.

Objective:To investigate the role and mechanism of mitochondrial DNA N6-methyladenine (6mA) in the hippocampal neurons in vascular cognitive impairment.Methods:(1) In vivo experiments: SPF male rats were randomly divided into sham-operated group and chronic cerebral hypoperfusion (CCH) group ( n=12). CCH models in the CCH group were established by ligating bilateral carotid arteries, while rats in the sham-operated group were only bilaterally dissected without ligation. Exploratory ability was detected by open field test 50 d after modeling, cognitive function was evaluated by novel object recognition test 51-53 d after modeling, and learning and memory abilities were tested by Morris water maze 54-59 d after modeling. And then, rats were sacrificed; ATP concentration and reactive oxygen species (ROS) level in the hippocampal tissues were detected, and neuron apoptosis in the hippocampal CA1 area was detected by TUNEL. (2) In vitro experiments: HT-22 cells were divided into normal control (NC) group, oxygen-glucose deprivation (OGD) group, OGD+siControl group, and OGD+siMETTL4 group. Cells in the NC group were cultured routinely, cells in the OGD group were subjected to low sugar and low oxygen for 12 h, and cells in the OGD+siControl group and OGD+siMETTL4 group were, respectively, transfected with NC-siRNA or METTL4-siRNA after being subjected to low sugar and low oxygen for 12 h. Mitochondria morphology was observed by transmission electron microscopy, ROS was detected by flow cytometry, mitochondria membrane potential was detected by JC-1 fluorescent staining, and mitochondrial complex I and III activity was detected by kit. (3) In vivo and in vitro experiments: METTL4 and DNA 6mA expressions in neuronal mitochondria of rat hippocampal tissues and mitochondria of HT-22 cells were detected by immunofluorescent staining and Western blotting. Results:(1) CCH rats had cognitive impairment: compared with the sham-operated group, CCH group had significantly increased frequency of entering the central area and reduced time in exploring new objects in open field experiment,and significantly decreased frequency of crossing the platform and prolonged escape latency in water maze experiment ( P<0.05). Compared with rats in the sham-operated group, rats in the CCH group had significantly decreased hippocampal ATP content ([18.820±1.177] nmol/L vs. [10.190±0.519] nmol/L) and increased ROS content ([4 488.00±255.70] AU vs. [11 644.00±530.20] AU, P<0.05). TUNEL results showed that the number of apoptotic neurons in the hippocampal CA1 area of CCH group was obviously increased than that in sham-operated group. Immunofluorescent staining results showed that 6mA and METTL4 mainly distributed in the mitochondria of hippocampal neurons in CCH group, and the 6mA and METTL4 expressions were obviously increased compared with those in the sham-operated group. Western blotting results showed that METTL4 expression in the hippocampal mitochondria of CCH group was significantly higher than that in the sham-operated group (1.729±0.168 vs. 1.000±0.000). (2) In vitro experiment: under transmission electron microscope, compared with the NC group, HT-22 cells in the OGD group showed obvious mitochondrial ridge disappearance, membrane rupture and vacuolation. Compared with the OGD group, the OGD+siMETTL4 group had significantly increased ATP production, decreased mtROS production, increased mitochondrial membrane potential, and increased mitochondrial complex I and III activities ( P<0.05). Immunofluorescent staining results showed that the mtDNA 6mA and METTL4 expressions in the OGD group were obviously higher than those in the NC group, and both mainly expressed in the mitochondria; mtDNA 6mA expression in the OGD+siMETTL4 group was obviously lower than that in OGD group. Western blotting results showed that METTL4 expression in the OGD+siMETTL4 group was significantly higher than that in the OGD group (1.578±0.261 vs. 2.970±0.280). Conclusion:Specific high expression of methylase METTL4 in hippocampal neurons of rats with cognitive impairment after CCH promotes the increased mtDNA 6mA expression and leads to mitochondrial energy metabolism disorders and increased ROS, which is speculated to be one of the mechanisms causing vascular cognitive impairment.