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Objective To investigate the effects and mechanisms of Astragalus Polysacharin (APS) on the proliferation and metastasis of breast cancer cells by regulating miR-107 and miR-346-mediated macrophage polarization in breast cancer-derived exosomes. Methods Forty 8-week-old female BALB/c mice were selected and breast cancer xenograft models and 4T1 transplanted tumor models were established. The mice were divided into the control group and the APS group. The APS group mice received daily intragastric administration of APS for 25 days, while the control group mice were given the same amount of normal saline. After all treatments were completed, the mice were euthanized, and tumor tissues were isolated. Western blot and flow cytometry were used to detect the expressions of proliferating cell nuclear antigen (PCNA), Ki-67, CD206, CD163, inducible nitric-oxide synthase (iNOS), and CD86. The apoptosis of single-cell suspensions in tumor tissues was analyzed. Human breast cancer cell line MDA-MB-231 was cultured and stimulated with APS, and exosomes from the cell culture medium were collected. The proliferation, migration, and invasion of cells were detected by CCK-8 assay, scratch assay, permeability chamber cell invasion assay, and qRT-PCR. Differentially expressed genes were screened by bioinformatics. ResultsBy measuring the expressions of molecules related to breast cancer cell proliferation and metastasis, it was shown that APS treatment reduced the expressions of proliferation-related proteins (PCNA and Ki-67) and metastasis-related proteins (Vimentin) in MDA-MB-231 xenograft tumor tissues; and the polarization of tumor-associated macrophages was observed. APS treatment of 4T1 transplanted tumor tissues could reduce the number of M2 macrophages and increase the number of M1 macrophages, resulting in an increase in the ratio of M1/M2 macrophages and an increase in cell apoptosis in 4T1 transplanted tumor tissues. The expressions of related proteins iNOS and CD86 increased, and CD206 and CD163 decreased. After APS treatment, the exosomes produced by MDA-MB-231 reduced the polarization of M2 macrophages and affected the expressions of miR-107 and miR-346. Conclusion APS inhibits the polarization of M2 macrophages by regulating the expression of miR-107 or miR-346 in breast cancer cell-derived exosomes, ultimately inhibiting the proliferation and metastasis of breast cancer cells.

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Objective To investigate the effects of polydatin on a high-fat diet-induced non-alcoholic fatty liver disease (NAFLD) mouse model and hepatoma G2 (HepG2) cell model, and to reveal its potential molecular mechanisms. MethodsThirty 6-week-old male SPF C57BL/6J mice were randomly divided into a normal diet group and a high-fat diet group. After the NAFLD mouse model was established in the high-fat diet group, they were further divided into a model group and a polydatin treatment group. The polydatin treatment group was administered polydatin by gavage at a dose of 250 mg/(kg·d) for 10 weeks, during which body weight was monitored and oral glucose and insulin tolerance tests were performed. At the end of the experiment, a series of tests to evaluate the effects of polydatin on mouse liver weight, blood lipids, liver lipid accumulation, and liver injury markers were performed. The expression of G 0/G 1 switch gene 2(G0S2) and adipose triglyceride lipase (ATGL) was measured by qRT-PCR and Western blot, and gene expression was further verified using immunohistochemical staining. The effects of polydatin on HepG2 cell activity was assessed by CCK-8 assay, lipid accumulation was observed by oil red O staining, and the expression of G0S2 and ATGL was detected by qRT-PCR and Western blot. Results Polydatin significantly reduced the body weight, liver weight, and serum and liver tissue levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), triglyceride (TG), and total cholesterol (TC) in mice ( P<0.05), alleviated pathological liver damage, decreased G0S2 expression ( P<0.05), and increased ATGL expression ( P<0.05). At the cellular level, polydatin reduced lipid droplet accumulation, improved lipid metabolism, decreased G0S2 expression ( P<0.05), and increased ATGL expression ( P<0.05). Even in cells with knockdown of G0S2, polydatin still promoted fat decomposition ( P<0.01). Conclusion Polydatin promotes hepatic fat breakdown by regulating the expression of G0S2 and ATGL, helping to alleviate metabolic disorders and liver damage in the NAFLD mouse model caused by a high-fat diet, offering a new strategy for treating NAFLD.

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Objective To explore the correlation between small intestinal bacterial overgrowth (SIBO) and cognitive impairment in patients undergoing peritoneal dialysis (PD). Methods60 PD patients and 46 non-dialysis patients with end-stage renal disease (ESRD) were included in this study. SIBO was detected by lactulose hydrogen methane breath test (LHMBT). The cognitive function levels of the subjects were evaluated using the Montreal cognitive assessment (MoCA) and mini-mental state examination (MMSE). The emotional states of the patients were assessed by self-rating anxiety scale (SAS) and self-rating depression scale (SDS). The gastrointestinal symptoms were evaluated using gastrointestinal symptom rating scale (GSRS). According to the results of LHMBT, the PD patients were further divided into two subgroups: SIBO-positive and SIBO-negative. The general conditions, laboratory data, and scores of each scale were compared between the two groups. Furthermore, according to the MoCA scores, the PD patients were divided into two groups: those with normal cognitive function and those with cognitive dysfunction. The positive rates of SIBO between the two groups were compared. Binary Logistic stepwise regression was used to explore whether SIBO was an independent risk factor for cognitive impairment in PD patients. At the same time, other independent influencing factors for cognitive impairment in PD patients were analyzed. Result s The positive rate of SIBO in PD patients was 40%. Among them, the positive rate of SIBO in the group with normal cognitive function during peritoneal dialysis was 27.27%, and the positive rate of SIBO in the group with cognitive dysfunction during peritoneal dialysis was 55.56%. There was a statistically significant difference in the positive rate of SIBO between the two groups ( P =0.026). Further analysis by multivariate Logistic stepwise regression showed that in addition to age ( OR=1.118, 95% CI 1.007-1.242, P =0.037), educational attainment（ OR=0.655，95% CI 0.442-0.972， P =0.036）, and hemoglobin （ OR=0.946，95% CI0.895-0.998， P =0.044）, SIBO （ OR=7.613，95% CI1.160-49.979， P =0.034）was also an independent risk factor for cognitive impairment in PD patients. ConclusionThe incidence of SIBO in PD patients is relatively high and is associated with multiple factors, among which SIBO may be closely related to cognitive dysfunction in PD patients.

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Objective To establish an in vitrocell model of Cobalt dichloride (CoCl 2)-induced epithelial-mesenchymal transition (EMT) in rat renal tubular epithelial cells (NRK-52E). MethodsNRK-52E cells were cultured in vitroand randomly divided into a blank control group (NC group) and a CoCl 2 treatment group. The CoCl 2 treatment group underwent 1, 2, 3, and 4 cycles of treatment, with each cycle consisting of CoCl 2 treatment for 9 h followed by recovery in CoCl 2-free medium for 3 h. The optimal concentration and time of CoCl 2-induced EMT were screened using the CCK-8 assay. Morphological changes in cells were observed using light microscopy and phalloidin staining. The expression levels of EMT marker proteins were detected by Western blot and immunofluorescence. ResultsCompared with the NC group, stimulation by 100 μmol/L CoCl 2 for 48 h significantly induced apoptosis ( P<0.01), meeting the requirements for subsequent experiments. Western blot results showed that the expression of hypoxia-inducible factor-1α (HIF-1α) and α-smooth muscle actin (α-SMA) significantly increased in the 3-cycle group treated with 100 μmol/L CoCl 2 for 9 h followed by recovery for 3 h ( P<0.001), indicating the most pronounced fibrotic response. Observations under light microscopy and rhodamine-labeled phalloidin staining revealed that the morphological changes and cytoskeletal rearrangement of NRK-52E cells were most significant in the 3-cycle group treated with 100 μmol/L CoCl 2 for 9 h followed by recovery for 3 h, demonstrating the best model stability. Immunofluorescence results showed that compared with the NC group, the fluorescence intensity of the fibrous matrix protein Collagen I significantly increased in the 3-cycle group treated with 100 μmol/L CoCl 2 for 9 h followed by recovery for 3 h ( P<0.001). ConclusionThe protocol involves treating NRK-52E cells with 100 μmol/L CoCl₂ for 9 h, following 3 h of recovery in CoCl₂-free medium. Repeating this cycle three times can establish an in vitro EMT model.

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ObjectiveTo discover molecular markers associated with lung adenocarcinoma diagnosis/prognosis and drug resistance through screening of differentially expressed genes based on published chip data in gene expression databases using bioinformatics methods. Methods Comprehensive analysis was performed in available mRNA microarray datasets including lung adenocarcinoma tissues dataset GSE32863 and lung adenocarcinoma taxane-platin resistance dataset GSE77209 from the gene expression omnibus (GEO) database. Gene ontology enrichment analysis, gene pathway enrichment analysis and protein interaction network analysis were performed based on significantly correlated genes. The expression level of genes was validated in the cancer genome atlas (TCGA) dataset. Survival differences were assessed by the log-rank test in TCGA lung adenocarcinoma dataset. Based on the publications genomics of drug sensitivity in cancer (GDSC) database in CellMiner cross database (CellMiner CDB), Pearson correlation analysis was used to analyze the correlation between differentially expressed genes and the half-maximal inhibitory concentration (IC 50) of anticancer drugs. ResultsThere were a total of 77 genes which had a different expression in resistance lung adenocarcinoma cells and lung adenocarcinoma cancer tissues. The functional enrichment analysis showed that these co-different expression genes were mainly enriched in microtubule, extracellular exosome, cell cycle and signaling by nuclear receptors. Protein–protein interactions (PPI) network screened 6 most connected genes as molecular complex (MCODE). Among the MCODE, overexpressed ubiquitin conjugating enzyme E2 T (UBE2T), kinesin family member 20A(KIF20A), PCNA clamp associated factor (KIAA0101), pituitary tumor-transforming gene 1 (PTTG1) and NIMA related kinase 2 (NEK2) were associated with poor outcomes. Survival analysis results showed that these five genes were upregulated in lung adenocarcinoma tissues and drug-resistant cells and were significantly associated with poor prognosis in lung adenocarcinoma patients. Drug sensitivity analysis results suggested that high expression of PTTG1 and UBE2T was significantly associated with sensitivity to multiple anticancer drugs, including paclitaxel and docetaxel. RT-PCR validation showed that PTTG1 and UBE2T were highly expressed in docetaxel-resistant cells A549-TXR and H358-TXR. ConclusionPTTG1 and UBE2T holds the potential to be chemoresistance markers in lung adenocarcinoma.

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ObjectiveTo investigate the regulatory role of Porphyromonas gingivalis (Pg) infection on the EGFR/GSK3β signaling axis, and its impact on epithelial-mesenchymal transition (EMT) and cetuximab (Ctx) resistance in esophageal squamous cell carcinoma (ESCC). Methods Single cell RNA sequencing was employed to perform differential analysis of cellular subpopulations, identifying differentially expressed genes in ESCC tissues infected and non-infected with Pg. IHC was conducted to assess the expression of Pg and epidermal growth factor receptor (EGFR) in ESCC tissues. Western blot, RT-PCR, and IF staining were performed to evaluate EGFR expression in Pg infected ESCC cell lines KYSE70 and TE1. ESCC cells were treated with Pg and EGFR inhibitor Ctx, and divided into four groups: negative control (NC) group, P. gingivalis group, Ctx group, Pg+Ctx group. Cell proliferation, migration and invasion abilities were evaluated using CCK-8, plate cloning, wound healing and Transwell assay. Western blot analysis was performed to detect the expression of EMT and EGFR/GSK3β signaling pathway-associated proteins and their phosphorylation levels. Transforming growth factor-β1 (TGF-β1) was used to induce EMT in ESCC cells, promoting a transition from the epithelial phenotype to mesenchymal-like phenotype. The differential effects of Ctx on these two phenotypic states were subsequently compared. ResultsEpithelial cells were predominantly enriched in Pg-positive tissues, and Pg infection promoted the upregulation of EGFR expression in ESCC cells. Compared to the NC group, Pg treatment significantly enhanced the proliferation, invasion and migration capabilities of ESCC cells, and also increased chemoresistance to Ctx and reduced its antitumor efficacy. Pg induced EMT in ESCC cells via the EGFR/GSK3β signaling pathway. Notably, Ctx exhibited markedly weaker inhibitory effects on mesenchymal-like cells compared to epithelial ESCC cells. Conclusion Pg promotes ESCC cells proliferation, invasion and migration by regulating EMT through the EGFR/GSK3β signaling pathway, and enhances chemoresistance to Ctx.

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Objective 　To investigate the effects of C1q tumor necrosis factor-related protein 3 (CTRP3)-enhanced Sendai virus (SeV) vector-overexpressing Gata4, Mef2c, and Tbx5 (SeVGMT) in the treatment of myocardial infarction (MI) mice and to analyze whether ubiquitin carboxyl-terminal hydrolase L1 (UCHL1) mediates this therapeutic pathway. Methods 　The mice were divided into 7 groups ( n=12): Sham group, MI group, SeVGMT group, CTRP3-Lv group, UCHL1-sh group, SeVGMT+CTRP3-Lv group, and SeVGMT+CTRP3-Lv+UCHL1-sh group. In the Sham group, only the skin was incised without ligation, while the coronary artery was ligated 2-3 mm below the left atrial appendage in mice in other groups. PBS was injected at three points in the myocardial infarction boundary in the Sham and MI groups 30 minutes after ligation. Mice in other groups were injected with SeVGMT, CTRP3-Lv, or UCHL1-sh according to their grouping. Four weeks after treatment, fractional shortening (FS), ejection fraction (EF), left ventricular end-diastolic diameter (LVIDd), left ventricular end-systolic diameter (LVIDs), heart rate (HR), mean arterial pressure (MAP), serum creatine kinase isoenzyme MB (CK-MB), myocardial troponin I (cTnI) and lactate dehydrogenase (LDH) levels, and myocardial tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) levels in mice were detected. The pathological changes of myocardial tissue were detected by 2,3,5-triphenyltetrazolium chloride (TTC), hematoxylin-eosin (HE), Masson trichrome and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. The mRNA expressions of CTRP3 and UCHL1 were detected by qRT-PCR. The protein expressions of CTRP3, UCHL1, collagen Ⅰ, collagen Ⅲ, Bcl-2-associated X (Bax) and B-cell lymphoma/leukemia-2 (Bcl-2) in myocardial tissue were detected by Western blot or immunohistochemical staining. Results 　Compared with SeVGMT group and CTRP3-Lv group, the levels of EF, FS, HR and MAP in SeVGMT+CTRP3-Lv group increased ( P<0.05). The levels of LVIDd, LVIDs, CK-MB, cTnI, LDH, TNF-α, IL-1β and IL-6 decreased ( P<0.05). MI area, fibrosis area and TUNEL positive rate decreased ( P<0.05), the protein levels collagen Ⅰ, collagen Ⅲ and Bax decreased ( P<0.05), and Bcl-2 protein levels increased ( P<0.05). The mRNA and protein levels and relative staining intensity of CTRP3 and UCHL1 increased ( P<0.05). Compared with SeVGMT+CTRP3-Lv group, the addition of UCHL1-sh treatment (SeVGMT+CTRP3-Lv+UCHL1-sh group) significantly weakened the influence of SeVGMT+CTRP3-Lv on the above indexes ( P<0.05). Conclusion 　 CTRP3 mediated UCHL1 enhances the therapeutic effect of SeVGMT reprogrammed CFs on MI in mice.

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The endoplasmic reticulum is one of the most prominent membranous organelles that plays a significant role in maintaining intracellular homeostasis. Specific internal or external stimulus leading to the accumulation of unfolded or misfolded proteins, alteration in intracellular calcium levels, or disruptions in redox homeostasis will trigger ER dysfunction and, ultimately, endoplasmic reticulum stress（ERS). Physiologically, ERS is involved in maintaining the homeostasis of the epidermal skin barrier. Ultraviolet B (UVB) induces epidermal ERS, following the activation of a variety of physiological and pathological processes. It participates in the physiological processes of epidermal barrier homeostasis and keratinocyte differentiation, and also contributes to pathological processes such as UVB-induced skin inflammation and keratinocyte apoptosis leading to skin damage. Therefore, ERS is expected to become a therapeutic target for UVB-mediated skin-related diseases. Currently, the clinical potential of modulating UVB-induced keratinocyte ERS through the use of antioxidants, NF-κB inhibitors, and ER Ca 2+ regulators has been increasingly recognized. This review focuses on the mechanisms underlying UVB-induced skin damage, the molecular mechanisms of UVB-induced keratinocyte ERS, and the current progress of ERS-targeting drugs in UVB-induced skin damage, providing new insights into ERS’s roles in UVB-induced skin damage and novel therapeutic strategies targeting ERS for the clinical treatment of UVB-induced skin damage.

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ObjectiveTo explore the role of N6-methyladenosine (m6A) modification of RNA and Methyltransferase like Protein 14 methyltransferase (METTL14) in the pathogenesis of early-onset preeclampsia (ePE). MethodsPlacental tissues of 15 pregnant women with early-onset preeclampsia and 15 normal pregnant women were collected. The level of m6A was determined by colorimetry, and the expression of METTL14 was determined by RT-qPCR, Western blot and immunohistochemistry (IHC) experiments. By transfecting siRNA and plasmid, METTL14 levels of trophoblast cells were knocked down and overexpressed, and cell phenotype experiments were carried out in vitro. The effects of METTL14 on the proliferation, migration and invasion of trophoblast cells were investigated by CCK-8, scratch assay, Transwell assay and invasion assay. Results The level of m6A in placental tissue of ePE was lower than that of normal pregnancy. METTL14 was mainly expressed in the nuclei of trophoblast cells. Compared with normal pregnancy, the expression of METTL14 in placental tissue of ePE decreased, and the level of METTL14 mRNA was positively correlated with the level of m6A in placental tissue. The results of the CCK-8 experiment showed that compared with the control group, knockdown of METTL14 expression in trophoblast cells significantly reduced the cell proliferation rate, while the proliferation ability of trophoblast cells with overexpressed METTL14 was enhanced. The results of the scratch test showed that compared with the control group, the relative healing rate of scratches was significantly reduced after METTL14 knockdown, while it increased after the overexpression of METTL14. The results of the Transwell assay and invasion assay showed that compared with the control group, after knockdown of METTL14, the number of trophoblast cells passing through the chamber was significantly reduced, while the number of trophoblast cells with overexpressed METTL14 passing through the chamber increased. ConclusionThe total RNA m6A modification level in placental tissue of ePE is lower than that in the normal pregnancy group. The down-regulation of methyltransferase METTL14 is involved in the regulation of the total RNA m6A modification level. The overexpression of METTL14 can enhance the proliferation, migration and invasion abilities of trophoblast cells. It provides a new perspective for exploring the pathogenesis of early-onset preeclampsia.

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Objective To explore the expression of collagen type 1 alpha 2 (COL1A2) in cervical cancer and its correlation with immune infiltration. Methods Bioinformatics techniques were used to analyze the expression of COL1A2 in cervical cancer. Western blot and RT-qPCR were used to detect the expression of COL1A2 in cervical cancer tissues and cell lines. The correlation between the expression of COL1A2 and tumor immune cell infiltration was analyzed by tumor immune estimation resource (TIMER2.0). Gene set enrichment analysis (GSEA) was used to analyze the possible mechanism of COL1A2 in cervical cancer. Jaspar database was used to predict the transcription factors of COL1A2. Western blot and RT-qPCR were used to detect the expression of transcription factors in cervical cancer tissues and cell lines. Results The expression of COL1A2 was down-regulated in cervical cancer ( P < 0.05). The expression of COL1A2 was positively correlated with the levels of macrophages and myeloid dendritic cells ( P < 0.01). The proportions of 22 types of immune cells were different in different cervical cancer patients. In addition, compared with the high expression group of COL1A2, the proportion of M0 macrophages, M2 macrophages and resting memory CD4 + T cells increased in the low expression group of COL1A2, while the proportion of CD8 + T cells, activated memory CD4 + T cells, follicular helper T cells, activated NK cells and activated myeloid dendritic cells decreased ( P < 0.05). GSEA analysis showed that COL1A2 was related to immune-related signaling pathways, including Notch signaling pathway, interleukin-6/janus kinase/signal transducer and activator of transcription 3 (IL6/JAK/STAT3), Wnt/β-catenin signaling pathway, etc. ( P < 0.01). Jaspar database predicted that the transcription factor of COL1A2 was paired box protein 5 (PAX5), and the expression of PAX5 decreased in cervical cancer ( P < 0.05). Conclusion COL1A2 is expected to become a potential diagnostic biomarker and immunotherapy target for cervical cancer.

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Objective To prepare a kind of baicalin-loaded gelatin/chitosan coaxial electrospinning membrane to solve the problem of diabetic inflammatory wound healing. Methods The membrane was prepared by coaxial electrospinning technology, and its physicochemical and biological properties were evaluated by tensile test, in vitro simulation and cell co-culture. Results Baicalin combined with gelatin/chitosan membrane was prepared, the fiber diameter was about 100 nm, the tensile strength after crosslinking was 13.38 MPa, the release of baicalin could reach 1 week, and the inflammatory environment was simulated in vitro. Baicalin had anti-inflammatory effect within a certain concentration range, and the survival rate of 5-day cells was nearly 100%. Conclusion The prepared baicalin combined with gelatin/chitosan membrane has good mechanical strength and excellent biocompatibility, providing a material basis for solving the problem of difficult wound healing.

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O bjective To explore the association between single nucleotide polymorphism (SNP) in the arachidonate 15-lipoxygenase (ALOX15) gene and Helicobacter pylori ( H. pylori) infection as well as the risk of non-cardia gastric cancer in Baotou Han population, and to provide experimental evidence and data support for the screening of susceptible population for non-cardia gastric cancer. MethodsA total of 458 cases with non-cardia gastric cancer and 460 healthy examination people were collected. The 14C urea breath test (UBT) and enzyme-linked immunosorbent assay (ELISA) were used to detect H. pylori infection in the 460 healthy individuals. The genotypes of ALOX15 rs2619112, rs2619118, rs2664593, rs7220870 were detected by polymerase chain reaction-restriction fragment length polymorphism, and the association of SNP with H. pyloriinfection as well as the risk of non-cardia gastric cancer was statistically analyzed. Results The positive rate of H. pyloriinfection was 42.3%. ALOX15 rs2619112, rs2619118, rs2664593, and rs7220870 had no association with H. pylori infection. ALOX15 rs2619112, rs2664593, and rs7220870 were not associated with the risk of non-cardia gastric cancer. Compared with the carriers of (CC + CT) genotype, the carriers of rs2619118 TT genotype had an increased onset risk of non-cardia gastric cancer [ OR (95% CI) = 1.512 (1.110 - 2.060)]. The haplotype ACCC constructed by ALOX15 rs2619112, rs2619118, rs2664593, and rs7220870 could reduce the onset risk of non-cardia gastric cancer. The second-order interaction of ALOX15 rs2619112 and rs2619118 was associated with the risk of non-cardia gastric cancer ( P＜0.05). ConclusionALOX15 rs2619112, rs2619118, rs2664593, rs7220870 may not play a major role in H. pylori infection. ALOX15 rs2619118 TT genotype is a risk factor for the development of non-cardia gastric cancer. The haplotype ACCC constructed by ALOX15 rs2619112, rs2619118, rs2664593, and rs7220870 reduces the onset risk of non-cardia gastric cancer. The interaction of ALOX15 rs2619112 and rs2619118 has a synergistic effect in the development of non-cardia gastric cancer.

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O bjective To elucidate the molecular mechanism by which prohibitin 2 (PHB2) mediates periodontitis-induced bone tissue inflammation through regulating the nuclear factor kappa B (NF-κB) signaling pathway and its role in irreversible alveolar bone resorption. Methods Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC) were used to detect the expression differences of inflammatory factors and PHB2 in healthy and inflamed alveolar bone tissues of mice i n vivo. In vitro, an inflammatory model was established using lipopolysaccharide (LPS)-induced a mouse calvaria-derived preosteoblastic cell line, subclone E1 (MC3T3-E1) cells. Western blot and qRT-PCR were used to clarify the regulatory relationship between PHB2 and inflammatory factors, and immunofluorescence staining was performed to observe changes in PHB2 subcellular localization. PHB2 overexpression plasmids were constructed using molecular cloning, and RNA interference was employed to knock down PHB2 expression to assess its regulatory role in inflammation. Based on RNA-seq data, differential expression analysis based on the negative binomial distribution, version 2 (DESeq2) was used for differential expression analysis, and kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment along with gene ontology (GO) functional annotation were performed to identify key signaling pathways and differentially expressed genes. Results In the mouse periodontitis model, PHB2 expression was significantly upregulated in alveolar bone tissues. In the in vitro inflammatory cell model, PHB2 levels positively correlated with interleukin (IL)-6, IL-1β, and tumor necrosis factor-alpha (TNF-α) levels, and its subcellular localization shifted during inflammation. RNA-seq data and the detection of the level of phosphorylation of p65 protein (p-p65) demonstrated that PHB2 exacerbated inflammatory responses through the NF-κB signaling pathway and was mechanistically linked to upregulation of the upstream chemokine C-X-C motif chemokine ligand 10 (CXCL10). C onclusion PHB2 aggravates LPS-induced periodontitis inflammation via the NF-κB signaling pathway, providing new insights into the molecular mechanisms underlying the development of periodontitis.

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Objective To explore the therapeutic mechanism of puerarin in treating rheumatoid arthritis (RA) rats based on the serine/tyrosine protein kinase B (AKT)-phosphorylated forkhead box protein O1 (FOXO1)-interleukin-9 (AKT-FOXO1-IL-9) signaling pathway. Methods 36 rats were randomly divided into a blank group, a model group, a positive control group, and low, medium, and high dose groups of puerarin. Except for the blank group, the other groups were induced with type II collagen to establish a RA rat model. After successful modeling, different doses of puerarin and methotrexate were given to treat the rats. The body mass and toe thickness of the rats were measured, and biochemical indicators of rat blood rheology were detected. X-ray was used to observe changes in rat joint morphology. Safranin green staining and Alixin blue staining were used to observe the pathology of rat joint tissue. ELISA was used to detect the levels of IL-9 and rheumatoid factors in rat serum, and Western blot was used to detect changes in levels of AKT and FOXO1. ResultsCompared with the blank group, the model group had the lowest toe thickness, and X-ray images showed more obvious segmental stenosis and more severe marginal bone invasion; scaly like changes appeared at the edges of joints stained with safranin green, accompanied by the exudation of inflammatory cells and increased proliferation and secretion of chondrocytes; the expression levels of inflammatory factors IL-9 and rheumatoid factors were the highest, and the expression levels of AKT and FOXO1 proteins were the highest ( P<0.05) . Compared with the model group, the toe thickness of rats treated with different doses of puerarin decreased; X-ray images showed that the puerarin treatment group of rats showed improvement in plantar joint stenosis and marginal bone invasion; the results of safranin green staining showed that after treatment with different doses of puerarin, the infiltration of inflammatory cells decreased, and the expression levels of inflammatory factor IL-9, rheumatoid factors, AKT, and FOXO1 proteins decreased significantly ( P<0.05), with the high-dose puerarin group showing the most significant difference. Compared with the high-dose puerarin group, the positive control group showed a significant decrease in the above results and statistical differences ( P<0.05). ConclusionPuerarin has a good therapeutic effect on rats with RA by inhibiting the AKT-FOXO1-IL-9 pathway. The high-dose puerarin group (60 mg/kg) has the best therapeutic effect and the results show a dose-response relationship.

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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.

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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 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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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 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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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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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 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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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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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 behavior of OSCC.

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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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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 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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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.