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ObjectiveTo investigate the causes of false negative results in the detection of Streptococcus agalactiaeusing fluorescent quantitative PCR (qPCR) targeting the CAMP factor gene ( cfb), and to perform a comprehensive analysis of the associated molecular mechanisms. MethodsA total of 76 vaginal secretion samples were evaluated using both qPCR based on cfb gene and bacterial culture methods. Four suspicious strains exhibiting negative qPCR results but positive culture findings were identified using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), latex agglutination antigen detection, and the CAMP test. Whole genome analysis was conducted utilizing the MGI DNBSEQ-T7 and Nanopore-PromethION 48 sequencing platforms. Phylogenetic and circular evolutionary trees were constructed using the 16S rRNA gene for strain verification. Multilocus sequence typing (MLST) was conducted, and cfb sequences were aligned and analyzed based on spliced sequences and original data. Specific primers targeting the cfbgene were designed for full-length amplification, followed by verification through agarose gel electrophoresis. ResultsThe four strains identified as suspicious were classified as S.agalactiae through MALDI-TOF MS, antigen detection, and 16S rRNA gene analysis, with MLST typing indicating ST862. Phenotypic analysis revealed a negative CAMP test. Whole genome sequence alignment failed to detect the cfb gene or any homologous sequences, and molecular testing confirmed the absence of cfb gene PCR amplification products, thereby confirming its complete deletion. Conclusion This deletion is identified as the molecular mechanism responsible for the false negative qPCR detection of S. agalactiae when targeting this specific gene. It is recommended that the qPCR detection targeting a single cfb gene has limitation, and this may impact clinical diagnosis and treatment decisions. This limitation warrants careful consideration.

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ObjectiveTo investigate the impact of mandibular retrognathism on brain function in rats and its potential mechanisms. Method Male SD rats aged 3 to 4 weeks (w) were randomly divided into control group ( n=10) and mandible retrusion (MR) group ( n=30), with the MR group further divided into three subgroups: 1, 2, or 4 weeks, with 10 rats in each subgroup. The MR group used modified metal tubes, which were bonded to the lingual side of the maxillary incisors in rats, forcing the mandible to retract by 2 mm-2.5 mm, thereby achieving the goal of constructing a mandibular retrognathism model; the control group received no interventions. The effects of mandibular retraction on the spatial learning ability and memory ability of rats were explored by behavioral experiments such as Morris water maze, Y maze and new object recognition 1, 2, and 4 weeks after modeling, respectively. Hematoxylin-eosin (HE) and Nissl staining were employed to observe the histomorphological changes in CA1, CA3, and DG regions of hippocampal tissues; Immunohistochemistry was employed to detect the protein expression levels of tropomyosin receptor kinase B (TrkB) in the CA1, CA3, and dentate gyrus (DG) regions of the hippocampus. Western blot was used to assess the expression changes in the TrkB/phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) pathway in hippocampal tissue. Quantitative analysis of apoptosis-related proteins in the hippocampus, including Bcl-2-associated X protein (Bax), B-cell lymphoma-2 protein (Bcl-2), and Caspase-3, was conducted using qRT-PCR and Western blot. Results Behavioral experiments showed that rats in the MR group, especially the MR4 w group, were significantly weaker in learning and spatial memory than the control group, MR1 w, and MR2 w（ P＜0.05）. HE staining showed that pyramidal cells in the control group were arranged orderly with regular morphology. In contrast, the MR group exhibited a reduction in the number of cells in the hippocampal CA1, CA3, and dentate gyrus (DG) regions. The cells were sparsely and irregularly arranged, with nuclear condensation and enlarged cellular spaces. Nissl staining further demonstrated damage to the Nissl bodies in the MR group. Meanwhile, the TrkB/PI3K/AKT signaling pathway was activated in the MR group, and the expression of p-TrkB, PI3K, and p-AKT proteins was up-regulated ( P<0.05); Western blot and qRT-PCR indicated that MR up-regulated the expression of pro-apoptotic protein Bax, apoptotic effector protein Caspase-3 ( P<0.05), and inhibited the expression of anti-apoptotic protein Bcl-2 ( P<0.05). Conclusion Mandibular retraction impairs brain function and affects cognition and learning memory in rats. This may be achieved by regulating the TrkB/PI3K/AKT signaling pathway to induce apoptosis in hippocampal neurons.

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Objective To investigate the ameliorative effect of vitamin D on sciatic nerve chronic compression injury (CCI) rats and its regulatory effect on nuclear factor κB (NF-κB)/cystathione-β-synthase (CBS)-hydrogen sulfide (H 2S) system. Methods Fifty rats were randomly divided into five groups: control group, model group, Vitamin D group, activator group, and activator + vitamin D group, with 10 rats in each group. Except for the control group, the rats in the other groups underwent sciatic nerve ligation to establish the CCI model. The vitamin D group received intraperitoneal injection of 500 mg/kg Vitamin D, the activator group received intraperitoneal injection of 10 mg/kg NF-κB activator lipopolysaccharide, and the activator + vitamin D group received intraperitoneal injection of 500 mg/kg vitamin D along with 10 mg/kg lipopolysaccharide. The control and model groups received intraperitoneal injection of an equal volume of saline, once per day for 2 weeks. The following parameters were compared among the groups: paw withdraw thermal latency (PWTL), mechanical withdrawal threshold (MWT), serum 25-hydroxyvitamin D 3 [25(OH)D 3] levels, H2S content, calcitonin gene-related peptide (CGRP), prostaglandin E2 (PGE2) levels, and the expression of vitamin D receptor (VDR), NF-κBp65, and CBS proteins in the L4-L6 dorsal root ganglia of rats. Results Compared with the control group, the model group showed decreased PWTL, MWT, 25(OH)D 3 levels and VDR protein expression, while H 2S content, CGRP and PGE2 levels, and NF-κBp65 and CBS protein expression significantly increased ( P<0.05). Compared with the model group, the vitamin D group exhibited increased PWTL, MWT, 25(OH)D 3 levels and VDR protein expression ( P<0.05), while H 2S content, CGRP and PGE2 levels, and NF-κBp65 and CBS protein expression significantly decreased ( P<0.05). Compared with the activator group, the activator + Vitamin D group showed increased PWTL, MWT, 25(OH)D 3 levels and VDR protein expression, and decreased H 2S content, CGRP and PGE2 levels and decreased NF-κBp65, CBS protein expression ( P<0.05). Compared with the vitamin D group, the activator + vitamin D group showed decreased PWTL, MWT, 25(OH)D 3 levels and VDR protein expression, while H 2S content, CGRP and PGE2 levels, and NF-κBp65 and CBS protein expression significantly increased ( P<0.05). ConclusionExogenous supplementation of vitamin D can relieve nerve pain and reduce pain sensitivity in CCI rats, possibly by inhibiting NF-κB/CBS-H 2S signaling pathway.

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Objective To explore the effect of interferon regulatory factor 2 binding protein ( IRF2BP2) on the proliferation of AML cells and its molecular mechanism. Methods The CRISPR-Cas9 gene editing technology was used to knock out IRF2BP2 in human AML cell lines Kasumi-1 and U937, and Western blot was performed to detect the knockout efficiency of IRF2BP2 protein. Cell morphology was observed using a microscope. Cell phenotypes were analyzed by CCK-8 assay, colony formation experiments, and flow cytometry. RNA-Seq was performed to identify differentially expressed genes between the IRF2BP2 knockout group and the control group in the U937 cell line. Gene Set Enrichment Analysis (GSEA) was conducted to explore the downstream molecular mechanisms. Western blot was used to detect the expression of downstream differentially expressed genes. The Cleavage Under Targets and Tagmentation (CUT&Tag) technique was applied to identify the direct targets of the IRF2BP2 protein, and the corresponding binding signals were visualized using the Integrated Genomics Viewer (IGV). ResultsCompared with the control group, after knocking out IRF2BP2, the CCK-8 experiment showed that AML cell proliferation was inhibited ( P<0.05); the number of colonies in the IRF2BP2 knockout group decreased ( P<0.05), and the proportion of G1 phase was prolonged ( P<0.05); in U937 cell lines, knocking out IRF2BP2resulted in significant enrichment of differential genes in MYC-related signaling pathways, and the protein expression levels of pathway molecules MYC, CDK4, and CDK2 decreased with the downregulation of IRF2BP2; using IRF2BP2 antibodies in U937 cell lines for CUT&Tag experiments, IGV visualization analysis showed a significant increase in signal peaks in the MYC promoter region. Conclusion IRF2BP2 affects the cell cycle and proliferation of AML cells by targeting and regulating MYC.

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Objective To investigate the effect of receptor of activated protein kinase C 1 (RACK1) on lipopolysaccharide (LPS)-induced barrier function in rat pulmonary microvascular endothelial cells (RPMVEC) and its interplay with the Sonic hedgehog (SHH) signaling pathway. Methods RPMVEC were cultured in vitroand randomly divided into si-NC, si-NC+LPS, si-RACK1, si-RACK1+LPS, si-RACK1+LPS+Vismodegib and Vismodegib+SAG groups. The RACK1 of RPMVEC was silenced by small interfering RNA (siRNA) technology and the cells were treated with LPS(10 mg/L), SHH signaling pathway inhibitor (Vismodegib) (20 μmol/L) and SHH signaling pathway agonist (SAG) (1 μmol/L). Following the intervention, the expressions of RACK1 and caveolin-1 in RPMVEC were detected by immunofluorescence, while the transendothelial electrical resistance (TEER) was evaluated using the method of Transwell, and the expression levels of RACK1, glioma-associated oncogene homolog 1 (Gli-1) and caveolin-1 were detected by Western blot. Results Silencing RACK1 significantly increased the TEER value of RPMVEC induced by LPS ( P<0.05). The expression of caveolin-1 decreased ( P<0.05), and the expression of Gli-1 increased ( P<0.05). Inhibiting the SHH signaling pathway could reverse the increased TEER value of LPS-induced RPMVEC caused by silencing RACK1 ( P<0.05), and the expressions of RACK1 and caveolin-1 increased ( P<0.05). Activation of the SHH signaling pathway increased the TEER value of LPS-induced RPMVEC caused by silencing RACK1 (P<0.05), and the expressions of RACK1 and caveolin-1 decreased ( P<0.05). Conclusion RACK1 plays a role in LPS-induced hyperpermeability of RPMVEC, and its effect may be achieved by modulating the SHH signaling pathway and caveolin-1.

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ObjectiveTo explore the value of nomogrambased on DCE-MRI radiomics combined with clinical-radiological featuresin predicting HR status in breast cancer with Her-2 low expression. MethodsA total of 198 patients of Her-2 low expression breast cancerwho underwent standardized breast MRI in our hospital from January 2019 to February 2025 were retrospectively analyzed. Patients were divided into HR (+) group ( n=137) and HR (-) group ( n=61). The cases were divided into a training set (138 cases) and a testing set (50 cases) in a 7:3 ratio.Clinical-radiological model was based on clinical and traditional radiological features,radiomics modelwas based on DCE-MRI, and combined modelwas constructed, respectively. The nomogram was drawn, andreceiver operating characteristic curve was used to compare the performance of different models in predicting HR status. Results The Radiomics score (Radscore) between the HR (+) group and the HR (-) group showed statistical differences inboth the training and testing sets (both P<0.001). The AUC of the clinical-radiological model based on lesion mobility, Ki67, TIC type, enhancement pattern and maximum diameterfor predicting HR status in the training set and testing set were 0.643 and 0.616, respectively. The AUC of the radiomics model in the training set and testing set were 0.897 and 0.860, respectively. The nomogram drawn by combining clinical-radiological features and Radscore showed better predictive performance in both the training set (AUC=0.913) and testing set (AUC=0.898) than the clinical-radiological model (all P<0.05). ConclusionThe nomogramcombinedby DCE-MRIradiomics and clinical-radiological features can effectively predict HR status of breast cancer with low Her-2 expression, which is helpful to the building of individualized treatment plan for breast cancer patients.

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Objective To investigate the mechanism by which icariin (ICA) inhibits the invasion and metastasis of human triple-negative breast cancer (TNBC) cells via downregulation of the transforming growth factor-β/Smad (TGF-β/Smad) signaling pathway. M ethodsTNBC cells (MDA-MB-231 and MDA-MB-468) were cultured in vitro and divided into four groups: an experimental group treated with 15 μmol/L ICA; a model group treated with 10 μmol/L TGF-β receptor inhibitor LY2109761; a combination group (LY2109761+ICA) treated with both 15 μmol/L ICA and 10 μmol/L LY2109761; and a control group. Cell proliferation, migration, and invasion were assessed using CCK-8, colony formation, 5-ethynyl-2′-deoxyuridine (EdU), wound healing, and Transwell assays. The expression levels of epithelial-mesenchymal transition (EMT)-related proteins, as well as TGF-β1, Smad2, and phosphorylated Smad2 (P-Smad2) were detected by immunofluorescence and Western blot. ResultsCCK-8 results showed that cell proliferation decreased gradually with increasing concentrations of ICA ( P<0.05). Colony formation and EdU assays indicated significantly inhibited proliferation in the ICA-treated group compared to the control ( P<0.05). Wound healing and Transwell assays demonstrated reduced migration and invasion capabilities in the experimental group relative to the control ( P<0.05). Compared to the model group, the LY2109761+ICA group exhibited further suppression of invasion ( P<0.05). Immunofluorescence revealed decreased Vimentin expression in the experimental group ( P<0.05), with an even more pronounced reduction in the LY2109761+ICA group ( P<0.01). Western blot analysis showed that the protein levels of N-cadherin, matrix metalloproteinase-9 (MMP9), Vimentin, TGF-β1, Smad2, and P-Smad2 were downregulated in the experimental group compared to the control ( P<0.05). These proteins were further suppressed in the LY2109761+ICA group compared to the model group ( P<0.05). ConclusionsICA inhibits TNBC cells proliferation, invasion, metastasis, and EMT by downregulating the TGF-β/Smad signaling pathway.

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ObjectiveTo investigate the role of STAT3 phosphorylation in ferroptosis regulation and its impact on cisplatin resistance mechanisms in human osteosarcoma cells. Methods Human osteosarcoma HOS cells and cisplatin-resistant HOS/DDP cells were treated with cisplatin (0.5, 1, 2, 4, 8, 16, 32 mg/L), the ferroptosis inducer Erastin (Era) (2 μmol/L), and/or the ferroptosis inhibitor Ferrostatin-1 (Fer1) (10 μmol/L). Cell viability and proliferation were assessed using the Cell Counting Kit-8 (CCK-8) and colony formation assays, and cell migration was evaluated via a scratch assay. Reactive oxygen species (ROS), intracellular ferrous iron levels, mitochondrial membrane potential, mitochondrial function, malondialdehyde (MDA) levels, and the reduced glutathione/oxidized glutathione (GSH/GSSG) ratio were measured using commercial kits. The mRNA expression of ferroptosis-related genes was analyzed by quantitative reverse transcription polymerase chain reaction (RT-qPCR). The protein levels of glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), phosphorylated STAT3 (p-STAT3), and total STAT3 were determined by Western blot. Results With cisplatin treatment, HOS cells exhibited decreased cell viability, mitochondrial membrane potential, and GSH/GSSG ratio ( P < 0.01), along with elevated levels of ROS, ferrous ion, and MDA content ( P < 0.01). The protein levels of GPX4 ( P < 0.01), SLC7A11, and p-STAT3 also decreased ( P < 0.05). Coadministration with the ferroptosis inhibitor Ferrostatin-1 (Fer-1) reversed these aforementioned effects ( P < 0.05). In HOS/DDP cells, the mRNA levels of ferroptosis-suppressive genes ( GPX4, FTH1, SLC7A11, and AIFM2) were significantly higher than those in HOS cells ( P < 0.05), whereas the expression of ferroptosis-promoting genes ( ACSL4 and PTGS2) was significantly lower ( P < 0.05). The cisplatin-induced reductions in cell viability and mitochondrial membrane potential, as well as the increases in ROS, ferrous ion, and MDA levels, were less pronounced in HOS/DDP cells than in HOS cells. The SLC7A11 protein level showed no significant change. However, combined treatment with the ferroptosis inducer Erastin (Era) resulted in significant decreases in viability, mitochondrial membrane potential, and the GSH/GSSG ratio in HOS/DDP cells ( P < 0.05). Furthermore, the protein levels of p-STAT3, GPX4, and SLC7A11 were also markedly reduced ( P < 0.05). Conclusion The activation of ferroptosis mediated by p-STAT3 enhances cisplatin sensitivity in HOS/DDP cells.

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ObjectiveTo explore the regulation of interleukin 8 (IL-8) secretion by peptidylarginine deiminase 4 (PADI4) and its effect on the pathogenesis of systemic lupus erythematosus (SLE). MethodsqRT-PCR was used to compare the expression levels of IL-8 in neutrophils between healthy controls (HC) and SLE patients. ELISA was used to detect IL-8 secretion levels in the serum of HC and SLE patients, the correlation between serum IL-8 and SLE related serological indicators in lupus patients was analyzed. ELISA was used to detect the effect of HC or SLE serum on IL-8 secretion by neutrophils. Using PADI4-specific inhibitor GSK484 in primary neutrophils, or in PADI4-knockdown neutrophil-like HL-60 cells (dHL-60), IL-8 stimulated by N-formyl-met-leu-phe (fMLP) or immune complexes (ICs) was detected. ResultsCompared with HC, IL-8 was significantly higher expressed in neutrophils of SLE patients. Serum IL-8 levels significantly increased in lupus patients and were positively correlated with serum IgM levels. Serum from SLE patients induced neutrophils to secrete more IL-8. PADI4 inhibitor could upregulate the production of IL-8 in neutrophils. In dHL-60 cells, knockdown of PADI4 led to a significant increase in IL-8 secretion. ConclusionThe proinflammatory cytokine IL-8 is highly expressed in neutrophils and serum of SLE patients, regulated by PADI4 and correlated with lupus serological indicators. IL-8 plays a role in the development of SLE through inflammatory responses, and PADI4/IL-8 provides new thinking for SLE monitoring and therapy.

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Objective To investigate the effect of aloperine (Alo) on the proliferation, migration and invasion of gastric cancer cells. Methods Human gastric cancer cell lines HGC-27 and AGS were treated with 0, 100, and 200 μmol/L Alo. CCK-8 was used to detect cell viability. Colony formation assay was used to detect cell proliferation. Flow cytometry was used to detect apoptosis. Scratch and Transwell migration assays were used to detect cell migration. Transwell invasion assay was used to detect cell invasion. Western blot was used to detect the expression of proteins related to proliferation, apoptosis, migration, invasion and Hippo pathway. Results Compared with the control group, the cell viability, number of colony formation, cell migration and invasion were significantly reduced after 100 and 200 μmol/L Alo treatment, and the apoptosis rate significantly increased ( P < 0.05 ). Additionally, the expressions of proliferating cell nuclear antigen (PCNA), B-cell lymphoma/leukemia-2 protein (Bcl-2), neural cadherin (N-cadherin), Vimentin, and transcriptional co-activator with PDZ-binding motif (TAZ) were significantly reduced, whereas the expressions of Bcl-2-associated X protein (Bax), phosphorylated yes-associated protein (p-YAP), and large tumor suppressor 1/2 (LATS1/2) significantly increased ( P < 0.05 ). Conclusion Alo inhibits the proliferation, migration and invasion of gastric cancer cells HGC-27 and AGS by regulating the Hippo signaling pathway.

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Objective To investigate the value of metagene next-generation sequencing (mNGS) in the detection of pathogens in patients with pulmonary infection. Methods A retrospective analysis was performed on clinical data from 434 patients with pulmonary infections admitted over the past four years. Based on the presence of underlying comorbidities, patients were divided into underlying disease group ( n=262) and non-underlying disease group ( n=172). Pathogen detection was conducted using both mNGS and conventional tests. Clinical and laboratory parameters, radiographic findings, and pathogen detection results were systematically analyzed. The diagnostic performance of the two methods in identifying causative pathogens of pulmonary infections was compared. Results The positive rate of mNGS in 434 patients was higher than that of conventional tests, and the difference was statistically significant ( P<0.05). The efficacy of mNGS in detecting bacteria and viruses was significantly higher than that of conventional tests, and the difference was statistically significant ( P<0.05). Although the fungal detection rate of mNGS was higher than that of conventional tests, the difference was not statistically significant. Among them, the detection rates of Mycobacterium tuberculosis, Mycoplasma pneumoniae, Haemophilus influenzae, Streptococcus pneumoniae, Streptococcus constellation, Staphylococcus aureusand Aspergillus fumigatus were significantly higher than those of conventional tests, and the difference was statistically significant ( P<0.05). Subgroup analysis showed that the proportion of males, hospital stay, smoking prevalence and average age in the underlying disease group were higher than those in the non-underlying disease group, and the difference was statistically significant ( P<0.05), while there were no significant differences in antibiotic use and endotracheal intubation rate between the two groups. The most common pathogens detected by mNGS in the underlying disease group were Mycobacterium tuberculosis, Haemophilus influenzae, Streptococcus pneumoniae, Pseudomonas aeruginosa, human herpesvirus type 4and Aspergillus fumigatus, while the most common pathogens in the non-underlying disease group were Mycobacterium tuberculosis, Haemophilus influenzae, Streptococcus pneumoniae, Mycoplasma pneumoniaeand Klebsiella pneumoniae.The positive rate of mNGS in the two groups was significantly higher than that of conventional tests, and the difference was statistically significant ( P<0.05), while the difference in the positive rate of mNGS between the two groups was not statistically significant. Conclusion mNGS has significant advantages over conventional tests of pathogen in lung infection, and is less affected by underlying diseases, which can provide an etiological basis for lung infection.

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Objective To explore the regulatory effects of ginkgo biloba extract on airway inflammatory injury and Toll-like receptor 4(TLR4)/nucleotide-binding oligomerization domain-containing 3(NLRP3) pathway in rats with chronic obstructive pulmonary disease (COPD). Methods Thirty-six male SD rats were selected and randomly divided into four groups: the normal control group, the model group, the prednisone treatment group, and the ginkgo biloba extract treatment group, with 9 rats in each group. Except for the normal control group, the COPD rat models in the other groups was constructed by intratracheal instillation of lipopolysaccharide (LPS) combined with cigarette smoke exposure. After successful modeling, the rats were continuously administered drugs for 12 weeks, followed by sampling. The general conditions and respiratory symptoms of the rats were observed. The pathological changes of lung tissues were observed by hematoxylin-eosin (HE) staining technique; the mRNA and protein expression levels of TLR4, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and NLRP3 in rat lung tissues were detected by real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot. Results Compared with the normal control group, the lung tissues of rats in the model group were significantly damaged, and the protein and mRNA expression of TLR4, TNF-α, IL-1β, and NLRP3 increased ( P < 0.05). Compared with the model group, lung tissue damage was reduced in the prednisone group and the ginkgo biloba extract group, and TLR4, TNF-α, IL-1β, NLRP3 protein and mRNA expression decreased ( P < 0.05). Conclusion Ginkgo biloba extract may reduce TLR4, TNF-α, IL-1β, and NLRP3 expression in lung tissues of COPD rats and alleviate their airway inflammatory response by inhibiting the TLR4/NLRP3 signaling pathway.

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Oral cavity cancer is the sixth-leading cancer worldwide, which has become one of the important factors affecting oral health. The morbidity of oral squamous cell carcinoma (OSCC) accounts for nearly 80% of oral cavity cancer. The pathogenesis and prevention of OSCC have become a hot topic in this field. Previous studies have shown that bone morphogenetic proteins (BMPs) participated in the development, invasion and metastasis of gastric and bone cancers, thereby impacting the prognosis of the patients. Recent studies have found that BMPs also play a key role in the development, progression, metastasis, and invasion of OSCC, This review therefore summarizes the correlation and molecular mechanism between BMPs and the biological

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Objective To assess the risk factors for symptomatic intracranial hemorrhage (sICH) after endovascular therapy (EVT) in patients with acute anterior circulation large vessel occlusion. Methods A retrospective analysis was used to analyze the clinical data of 273 AIS patients with EVT, of which 158 were male and 115 were female; ages ranged from 33 to 94 years old [71 (59, 78) years old], and sICH occurred in 61 cases. The patients' gender, age, history of smoking and alcohol consumption, history of illness, history of oral anticoagulant/anti-antibiotic administration, oral anticoagulant/antiplatelet medication status, intravenous thrombolytic therapy, admission blood pressure, hospitalization days, occlusion location, procedure duration, number of times of thrombus retrieval, cause of stroke (TOAST classification), revascularization status (mTICI ≥2b), and admission ASPECT score, admission APACHEII score, and admission NHISS score were collected. Univariate analysis was used to compare the differences in the above indicators between the non-SICH group and the sICH group. Multivariable Logistic regression analysis was used to evaluate the independent factors for the occurrence of sICH after EVT. Results The results of univariate analysis showed that mTICI ≥ 2b, admission ASPECT score, and post-EVT sICH were significantly negatively correlated ( P<0.05), while occlusion location (ICA), admission APACHE-II score, admission NHISS score > 20, and admission mRS score were significantly positively correlated with post-EVT sICH ( P<0.05). The results of the multivariable Logistic regression analysis indicated that mTICI≥2b ( OR = 0.314, 95% CI0.106-0.929) and admission ASPECT score ( OR=0.775, 95% CI0.604-0.995) are protective factors for sICH after EVT, while occlusion location (ICA) ( OR=2.047, 95% CI1.033-4.054) and admission APACHE-II score ( OR=1.236, 95% CI1.140-1.340) were risk factors for sICH after EVT. Conclusion Higher admission APACHEII score, lower admission ASPECT score, occlusion location (ICA), and mTICI<2b are independent risk factors for the development of sICH after EVT in patients with acute anterior circulation large vessel occlusion.

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ObjectiveTo investigate the methylation status of the RUNX family transcription factor 3 (RUNX3) promoter and its mRNA expression in sepsis patients, and to analyze their relationship with the prognosis of sepsis. MethodsDifferentially expressed genes related to sepsis, including RUNX3, were identified from multiple datasets obtained from the gene expression omnibus (GEO) database. The gene expression and methylation sites were validated. A total of 120 patients with sepsis were included. Clinical data were recorded, and blood samples were collected at enrollment. Relative expression levels of RUNX3 in blood samples and promoter methylation status were detected using qPCR and methylation-specific PCR (MSP), respectively. Pearson correlation coefficients were used to analyze the correlation between RUNX3 levels in patient blood and clinical indicators. Kaplan-Meier analysis was performed to plot survival curves, and Cox proportional hazards regression analysis was conducted to identify factors affecting the prognosis of sepsis patients. Results Data set analysis revealed that RUNX3 was a differentially methylated gene associated with the prognosis of sepsis. The mRNA expression level of RUNX3 was lower in the non-survivor group compared to the survivor group ( P < 0.05), and the methylation ratio of RUNX3 was higher in the non-survivor group than in the survivor group ( P < 0.05). In sepsis patients, RUNX3 mRNA expression levels were negatively correlated with Interleukin-6 (IL-6), Procalcitonin (PCT), C-reactive protein (CRP), Acute Physiology and Chronic Health Evaluation (APACHE II) score, and Sequential Organ Failure Assessment（SOFA）score. Kaplan-Meier analysis showed that the 28-day survival rate in the methylated group was lower than that in the unmethylated group ( P < 0.05). Cox regression analysis results indicated that RUNX3 promoter methylation was an independent risk factor for predicting the 28-day prognosis of sepsis patients. ConclusionIn sepsis patients, the mRNA levels of RUNX3 were reduced, and the degree of promoter methylation was higher. RUNX3 promoter methylation was an independent risk factor for the 28-day prognosis of sepsis patients and could serve as a prognostic biomarker for sepsis.

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ObjectiveTo investigate the role of transcription factor EB (TFEB) -autophagy pathway in rifampicin-induced liver injury and its possible mechanism. Methods Forty 6-8-week-old C57/BL6 mice were randomly divided into five groups: control group, model group, TFEB low-dose agonist group, TFEB high-dose agonist group, and autophagy agonist group, with 8 mice in each group. Except for the control group, the other four groups were given rifampicin 200 mg/（kg·d）by gavage daily. TFEB agonist was administered intraperitoneally at a low dose of 20 mg/kg and a high dose of 50 mg/kg for 7 days at 1 h after rifampicin administration. Autophagy agonist was administered by gavage at a dose of 10 mg/kg 6 h before rifampicin administration on day 1. The experiment was completed 7 days after modeling. The degree of liver injury was evaluated by detecting liver function indexes and liver pathological changes. Western blot was used to detect the protein expression total TFEB, chelator 1（p62）, microtubule-associated protein light chain 3（LC3）, benzyl chloride 1（Beclin-1）, sodium taurocholate co-transporting polypeptide（NTCP）and bile salt export pump（BSEP）levels in liver nucleus/liver tissue were quantified. Results Compared with the control group, the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), direct bilirubin (DBIL), and total bile acid (TBA) in the model group increased ( P < 0.05), and obvious pathological changes were observed in the liver. Compared with the model group, the high dose and low dose of TFEB agonist and autophagy agonist groups had reductions in the above indicators ( P < 0.05). Compared with the low-dose TFEB agonist group, the high-dose TFEB agonist group had reductions in the above indicators ( P < 0.05). The proportion of TFEB in the nucleus was (1.0±0.10) in the control group, (0.6±0.05) in the model group, (0.8±0.08) in the low-dose agonist group, and (0.9±0.07) in the high-dose agonist group ( P < 0.05). Autophagy agonist group (0.7±0.06) ( P < 0.05). Compared with the control group, the levels of NTCP and BSEP in the liver of the model group decreased ( P < 0.05), and the expression of NTCP and BSEP in the TFEB low-dose and high-dose agonist groups were restored, and the expression of NTCP and BSEP in the autophagy agonist group also increased ( P < 0.05). Compared with the control group, the protein expression levels of TFEB, LC3-Ⅱ/LC3-Ⅰand Beclin-1 in the liver tissue of the model group significantly decreased ( P < 0.05), while the protein expression level of p62 significantly increased ( P < 0.05). Compared with the model group, the protein expression levels of TFEB, LC3-Ⅱ/LC3-Ⅰand Beclin-1 in the liver tissue of the TFEB agonist high-dose group, low-dose group and autophagy agonist group increased ( P < 0.05), while the protein expression level of p62 decreased ( P < 0.05). Conclusion TFEB can improve rifampicin-induced liver injury by activating autophagy pathway, and the main mechanism may be related to the up-regulation of NTCP and BSEP expression.

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Respiratory diseases represent prevalent conditions that pose a significant threat to public health. The pathogenesis of these diseases is complex involving a variety of cells and cytokines. Cell culture technology serves as a fundamental tool for investigating pathological mechanisms and drug efficacy. However, traditional monolayer cell culture fail to mimic the intricate cell-cell interactions, limiting its applicability. In contrast, cell co-culture offers a more physiologically relevant approach by closely mimicking the multicellular microenvironment. This advancement provides a robust model for elucidating pathological pathways and evaluating pharmacological interventions in respiratory diseases. In this review, the applications of cell co-culture technologies in common respiratory diseases, including chronic obstructive pulmonary disease, idiopathic pulmonary fibrosis, lung cancer, asthma, and pneumonia are summarized, in order to propose strategies for the prevention and treatment of these diseases.

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ObjectiveTo evaluate the effects of different surface treatment methods and simulated aging on the roughness, microhardness, and color stability of lithium disilicate glass ceramics(LDC). Methods The LDC were grouped and performed 5 kinds of surface treatments:control group, polishing group, polishing + polishing paste group, glazing group, polishing + glazing group. The roughness and microhardness of the specimen were measured. Then the specimens were divided into two subgroupswhich were treated with water and staining solution. The specimens were measured color parameters before and after processing. The above data was analyzed. ResultsThe LDC had changes in surface roughness and microhardness after different surface treatments. The polishing + polishing paste group had the lowest surface roughness, and the surface microhardness of the LDC decreased after glazing. After simulating aging, the difference of color and transparency of the polishing + polishing paste group, glazing group, and polishing + glazing group were the smallest. Conclusion Different surface treatments and simulated aging have a certain impact on the roughness, microhardness, and color stability of LDC. Fine polishing with polishing paste has a comparable anti staining effect to glazing on LDC.

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ObjectiveTo investigate the relationship between the transgingival characteristics of implant restorations and peri-implant soft tissue with peri-implant mucositis and peri-implantitis using three-dimensional measurement techniques. MethodsA total of 121 bone-level implant restorations in posterior tooth positions from 73 patients were evaluated using three-dimensional data analysis software, based on clinical and radiographic examination data. The following characteristics were assessed: emergence angles, including the mucosal emergence angle (MEA), deep angle (DA), and total angle (TA); emergence profile; and peri-implant soft tissue. Generalized estimating equations were used to analyze the relationship between these characteristics and the occurrence of peri-implant mucositis and peri-implantitis. Results TA and MEA showed significant statistical relevance to peri-implant mucositis. Implant restorations with a TA greater than 30° had a 1.7 times higher risk of developing peri-implantitis compared to those with a TA of less than 30°. ConclusionMEA and TA of bone-level implant restorations in posterior tooth positions are significantly correlated with peri-implant mucositis. An excessive TA increases the likelihood of peri-implantitis. It is recommended that both the coronal and overall emergence angles of implant restorations should not exceed 30°.

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Tumor immune escape represents a pivotal determinant of cancer immunotherapy failure. It’s mechanistically linked to immunosuppressive tumor microenvironment (TME). The TME comprises tumor cells, immune cells, stromal components and extracellular matrix. These components interact synergistically to suppress antitumor immunity through multiple pathways, thereby promoting immune evasion. As crucial chromosomal stability regulators, centromere proteins (CENPs) remodel the TME via multifaceted mechanisms to potentiate immune evasion. This review synthesizes current knowledge on CENPs’ role in tumor immune evasion, offering novel insights for cancer diagnostics and immunotherapy.

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ObjectiveTo investigate the relationship between lower limb alignment and knee functional gait after total knee arthroplasty (TKA) for varus knee deformity. MethodsA total of 94 patients with knee osteoarthritis and varus deformity underwent mechanically aligned TKA. The knee society score (KSS), hip-knee-ankle angle (HKA), medial proximal tibial angle (mMPTA), and gait parameters (step length, gait speed, cadence, thigh jerk acceleration, and thigh swing work) were assessed preoperatively and one month postoperatively. The correlation between KSS scores and changes in lower limb alignment was also analyzed. ResultsThe KSS clinical score of the operated knee significantly improved from 39.19 ± 9.55 preoperatively to 73.01 ± 6.90 postoperatively ( P<0.001). The KSS functional score increased from 41.12±10.66 to 56.33±7.41( P<0.001). Postoperative gait analysis revealed significant improvements in step length, gait speed, cadence, and thigh swing work ( P<0.001), while thigh jerk acceleration showed no significant change ( P=0.525). The HKA angle of the affected limb increased from 170.61±4.39 preoperatively to 177.30±3.49 postoperatively ( P<0.001). Similarly, the mMPTA angle increased from 83.95±3.32 to 89.15±1.94 ( P<0.001). Correlation analysis indicated that The KSS clinical score was positively correlated with HKA ( P<0.001) and mMPTA ( P<0.05) of the lower limb force line. The KSS functional score was positively correlated with HKA ( P<0.05) but not with mMPTA ( P>0.05). ConclusionIn patients with severe knee osteoarthritis due to varus deformity, TKA significantly alleviates pain and improves both clinical and functional KSS scores within one month postoperatively. Restoring lower limb mechanical alignment is positively associated with joint functional recovery and enhances gait performance.

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Objective To treat malignant and non-malignant hematological diseases, allogeneic hematopoietic stem cell transplantation (alloSCT) is widely used. However, its application is limited by the risk of graft-versus-host disease (GVHD), difficulties in donor matching, and the high incidence of transplantation-related mortality due to myeloablative conditioning regimens. Therefore, there is an urgent need to develop new strategies that reduce the intensity of conditioning while enhancing donor engraftment efficiency. Methods In an alloSCT model, minor histocompatibility antigen-specific donor central memory T cells (miHA-TM) combined with a CD40 agonist were adoptively transferred to evaluate donor bone marrow engraftment under different irradiation doses. Results Minor histocompatibility antigen (miHA)-specific donor-derived central memory T cells (TM) facilitated recipient immune reconstitution without inducing GVHD. Donor-derived TM cells, however, underwent exhaustion during the process of recipient immune reconstitution, which compromised complete donor engraftment under reduced-intensity conditioning. At a low irradiation dose (4 Gy), the combination of miHA-TM and a CD40 agonist significantly promoted donor bone marrow engraftment. ConclusionmiHA-TM facilitates immune reconstitution in recipients without inducing graft-versus-host disease.

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ObjectiveTo investigate the expression level of signal transducer and activator of transcription 4(STAT4) in lung adenocarcinoma(LUAD), and to analyze its impact on the proliferation, invasion, and migration abilities of LUAD cells. MethodsGene expression omnibus data base（GEO），human protein atlas（HPA）and Kaplan-Meier Plotter databases were used to explore the expression of STAT4 in LUAD and its effect on the survival of patients with LUAD. Stable LUAD cell lines with STAT4 knockdown were constructed using lentiviral packaging technology, and the knockdown efficiency of STAT4 was assessed via RT-qPCR. The effects of STAT4 on LUAD cell proliferation were evaluated using CCK-8 assays and colony formation assays, while the impacts on cell migration and invasion were assessed through wound healing assays and Transwell assays. ResultsSTAT4 was lowly expressed in LUAD, and its low expression was unfavorable to the survival of LUAD patients. After STAT4 was knocked down in LUAD, the proliferation, invasion and migration abilities of lung adenocarcinoma cells were enhanced. ConclusionSTAT4 is a prognostic marker in LUAD which significantly inhibits the proliferation, invasion, and migration abilities of LUAD cells. STAT4 is expected to become a new therapeutic target for LUAD.