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Objective To explore the mechanism of miR-141-5p and its effect on Imatinib(IM) resistance in CML.Methods qRT-PCR was used to detect miR-141-5p mRNA levels in IM resistant and sensitive patients.Western blot was used to detect the expression of proteins such as MMP-3,MMP-9,and Bcl-2 before and after transfection in K562 and K562/G01 cells.CCK-8 was used to detect of K562 and K562/G01 cell activity;Flow cytometry assay was used to detect the binding of miR-141-5p with ABCG1;Nude mice were used to validate the effect of miR-141-5p on tumors in vivo.Results The results showed that miR-141-5p was downregulated in IMresistant CML patients and IM-resistant CML cells and overexpression of miR-141-5p could inhibit the growth of IMresistant CML cells and promote their apoptosis.Research on tumor bearing mice had shown that miR-141-5p inhibits tumor growth in vivo.Finally,it was found that miR-141-5p could directly target ABCG1 in IM-resistant CML cells to regulate CML occurrence.Conclusion miR-141-5p and ABCG1 form a competing endogenous RNA(ceRNA) network to function in IM resistance,thus facilitating CML progression.

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Objective To investigate the effect of G protein-coupled receptor 108(GPR108) gene knockout on systemic inflammation in lipopolysaccharide(LPS)-induced sepsis mice. Methods Male C57BL/6 mice and GPR108 gene knockout mice were randomly divided into 4 groups: WT group, WT-LPS group, KO group, KO-LPS group. The physiological characteristics of mice in different groups were observed, and the morphological changes of liver and lung tissues were observed. Macrophages were extracted from bone marrow and subjected to flow cytometry to detect their M1 polarization status. The expression levels of IL-6 in liver and lung tissues, macrophages, and serum were also measured. Results KO-LPS group mice showed significant liver and lung tissue damage, with a significantly greater number of bone marrow-derived macrophages polarizing towards M1 in the KO-LPS group compared to the WT-LPS group. Additionally, at the tissue, cellular, and serum levels, the expression of IL-6 in the KO-LPS group mice was significantly higher than that in the WT-LPS group mice(P<0.05). Conclusion During the systemic inflammatory infection induced by LPS in mice, the lack of GPR108 exacerbates the systemic inflammatory response. GPR108 has an inhibitory effect on the inflammatory response in mice with LPS-induced sepsis.

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Objective To investigate the inhibitory effect of curcumin(Cur) on IL-1β-induced cartilage damage and to study the relationship between the regulatory mechanisms of the DUSP1/p38 MAPK signalling pathway in the above process. Methods Chondrocytes(C28/I2) and postoperative primary chondrocytes from osteoarthritis patients were divided into control and experimental groups, and the experimental group was treated with different concentrations of Cur(0, 10, 20, 40, 60, 80 μmol/L) after applying the inflammatory induction treatment with IL-1β(10 μg/L). The cell proliferation inhibition rate was determined by cell viability assay(CCK-8), the apoptosis rate was detected by flow cytometry assay. Real-time fluorescence quantitative PCR(qRT-PCR), Western blot, and immunofluorescence assay were used to detect type II collagen α1 chain(Collagen Ⅱ), matrix metallopeptidase 13(MMP13), interleukin-1β(IL-1β), BCL2-related X protein(Bax), B lymphocytoma-2(Bcl-2), dual-specificity phosphatase 1(DUSP1), p38 mitogen-activated protein kinase(p38), and phosphorylated p38 mitogen-activated protein kinase(p-p38) RNA and protein expression levels. The role of the DUSP1/p38 MAPK axis in the inhibition of chondrocyte oxidative stress, apoptosis and inflammation by Cur was further validated using DUSP1 interfering RNA and p38 MAPK pathway inhibitor(SB). Results Cur significantly inhibited the IL-1β-induced decrease in chondrocyte viability and significantly reduced the levels of oxidative stress, apoptosis, and inflammation in chondrocytes; Cur inhibited the expression of MMP13, IL-1β, Bax, and p-p38 proteins, while the expression of Collagen II, Bcl-2, and DUSP1 proteins significantly increased; IL-1β and interfering RNA silencing DUSP1 activated the p38 pathway, while Cur inhibited the activation of the p38 pathway; the use of p38 MAPK pathway inhibitors reduced cellular inflammation. Conclusion Cur attenuates IL-1β-induced oxidative stress, apoptosis and inflammation in chondrocytes by promoting the expression of DUSP1 protein and inhibiting the activation of p38 MAPK pathway.

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Objective To investigate the effect of lansoprazole(LPZ) on cisplatin-induced acute kidney injury in mice and renal tubular epithelial cell injury.Methods C57BL/6J mice and renal tubular epithelial cells were divided into normal control(NC) group,lansoprazole(LPZ) group,cisplatin(CIS) group and cisplatin+lansoprazole(CIS+LPZ) group.In animal experiments,LPZ(25 mg/kg) was dissolved in normal saline,and the mice were intraperitoneally injected with LPZ for three consecutive days followed by cisplatin(20 mg/kg).Mice were fed normally for 3 days,and serum samples were collected to detect serum creatinine(CRE) and blood urea nitrogen(BUN) levels.HE staining and PAS staining were used to observe renal pathology,and transmission electron microscopy was used to observe renal ultrastructure.Western blot and immunohistochemistry were used to detect the changes in the expression levels of KIM-1,NGAL and pyroptosis-related proteins.In the cell experiment,the cells were treated with cisplatin(20 μmol/L) and LPZ(5 μmol/L) for 24 hours,and the expressions of KIM-1,NGAL and pyroptosis-related factors were detected by Western blot and Real-time PCR.Results Compared with the NC group,CRE and BUN levels increased in the CIS group,and LPZ treatment aggravated the serological indicators of the mice(P<0.001).Histopathological examination showed that compared with the CIS group,the LPZ+CIS group had obvious renal tubular dilatation,inflammatory cell infiltration and glycogen deposition in the renal tissue(P<0.001).Electron microscopy showed swelling of mitochondria,increased membrane density and decreased or absent mitochondrial crista in CIS group,which were aggravated by LPZ+CIS group.In vivo and in vitro experiments confirmed that LPZ treatment promoted CIS-induced acute kidney injury in mice and increased the expression levels of kidney injury factors KIM-1,NGAL and key factors of renal pyroptosis(Caspase 1,GSDMD,NLRP3 and IL-18) in renal tubular epithelial cells(P<0.001).Conclusion Lansoprazole promotes cisplatin-induced acute kidney injury by enhancing pyroptosis of renal tubular epithelial cells.

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Objective To study the effect and regulatory mechanism of secreted proteins from PSC on pancreatic ductal adenocarcinoma cells(PANC-1). Methods Conditioned medium(CM) from pancreatic stellate cells(PSC) was collected through an indirect co-culture method, and PANC-1 cells were cultured separately with CM for 0, 2, and 24 h. The proliferation phenotype of PANC-1 cells under different stimulation periods was detected using the CCK-8 assay. Proteomic analysis was performed to analyze the changes in protein levels of PANC-1 cells, and the most significant protein changes were validated using Western blot. Results Compared with the control group, the proliferation rate of PANC-1 cells increased after being stimulated by PSC derived CM; The results of proteomic analysis showed that the protein expression of metabolic pathways in PANC-1 cells increased continuously after being cultured in PSC CM for 0, 2, and 24 h. Western blot analysis confirmed an increasing trend of PIK3C2A in PANC-1 cells, indicating that the CM from PSC might promote the proliferation of PANC-1 cells by upregulating the expression of PIK3C2A. Conclusion The CM of PSC may promote the proliferation of PANC-1 cells by upregulating the expression of PIK3C2A, which improves the understanding of the mechanism of interaction between PSCs and pancreatic cancer cells in the tumor microenvironment.

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Objective To explore the effect of different concentrations of relaxin-2(RLN-2) on the proliferation and migration abilities of human immortalized keratinocytes(HaCaT cells). Methods HaCaT cells were cultured in media with different concentrations of RLN-2, and the cells were cultured in media without RLN-2 as the control group.The effect on cell proliferation was assessed by using the CCK-8 reagent, the cell migration ability was evaluated throughin vitrocell scratch assay, the cell cycle was examined by flow cytometry, and the expression levels of cell cycle proteins Cyclin B1 and Cyclin A2 were detected by Western blot. Results After being cultured for 24 hours under RLN-2 concentration ranging from 10～100 ng/ml, HaCaT cells showed progressively increased proliferation and migration capabilities compared to the control group, with elevated expression levels of cell cycle proteins Cyclin B1 and Cyclin A2 and an increased proportion of cells in S and G2/M phases, peaking at 100 ng/ml. However, HaCaT cells cultured with 200 ng/ml of RLN-2 exhibited reduced proliferation and migration capabilities, decreased expression levels of Cyclin B1 and Cyclin A2, and a lower proportion of cells in S and G2/M phases compared to the 100 ng/ml group. Conclusion RLN-2 can enhance the migration ability of HaCaT cells within an appropriate concentration range and may also promote cell proliferation by increasing the expression of related cell cycle proteins and the proportion of cells in S and G2/M phases.

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Objective To investigate the impact of tumor necrosis factor ligand superfamily member 13(TNFSF13) derived from M2 macrophages on temozolomide(TMZ) resistanceviaregulating interferon regulatory factor 8(IRF8) in glioblastoma(GBM) cells. Methods Immunohistochemistry(IHC) was used to detect the expression of TNFSF13 in normal brain tissues and GBM tissues. ELISA was used to measure the expression of TNFSF13 in the conditioned media(CM) of M0-type macrophages and M2-type macrophages. M0-CM and M2-CM were used to culture U251 sensitive(U251/S) and resistant(U251/R) cells. The TMZ treatment group was also treated with 800 μmol/L TMZ. The U251/R cells were divided into the following groups: con group, M2vector-CM group, M2vector-CM+TMZ group, M2TNFSF13-CM group, M2TNFSF13-CM+TMZ group, si-IRF8 group, and si-IRF8+M2TNFSF13-CM group. CCK-8 assay was used to detect cell viability and calculate the IC50value. Transwell assay was used to detect cell invasion. Flow cytometry was used to detect apoptosis. Western blot was used to detect the expression of IRF8. Nude mouse xenograft models were constructed and the nude mice were divided into the following groups: U251+M2si-NCgroup, U251+M2si-TNFSF13group, U251+M2si-NC+TMZ group, U251+M2si-TNFSF13+TMZ group. The tumor volume and mass of each group were measured, and IHC was used to detect the expression of TNFSF13 and CD206 in tumor tissues of each group. Results Compared with adjacent tissues and M0-CM, the expression of TNFSF13 was up-regulated in cancer tissues and M2-CM. Compared with the M0-CM group, the IC50value of TMZ and the number of cell invasions in U251/S and U251/R cells in the M2-CM group significantly increased(allP<0.05). Overexpression of TNFSF13 in M2 macrophages could promote the IC50value of TMZ in U251/R cells, promote cell invasion, and inhibit cell apoptosis(allP<0.05). Overexpression of TNFSF13 promoted the expression of IRF8, and knocking down IRF8 could attenuate the TMZ resistance of U251/R mediated by overexpression of TNFSF13.In vivostudies showed that knocking down TNFSF13 alone or combined with TMZ treatment significantly inhibited tumor growth and reduced the expression of TNFSF13 and CD206. Conclusion TNFSF13 derived from M2 macrophages promotes TMZ resistance in GBM cells by activating IRF8.

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Objective To investigate the differences in enolase and laminin levels in vaginal epithelial tissues between mice successfully infected withGardnerellaand mice not infected with Gardnerella, providing information for further exploration of the correlation between enolase and laminin levels and the incidence of bacterial vaginosis. Methods Gardnerella strains isolated, purified, and identified from vaginal secretions of patients with bacterial vaginosis were used to infect the vagina of mice and establish a mouse model of bacterial vaginosis. Successful and failed mice was defined as successful and failed groups, respectively. Differential expression of enolase and laminin in the vaginal epithelial tissue of two groups of mice was detected by Western blot. Modeling success rate was statistically analyzed, and the expression differences of enolase and laminin was compared between two groups. Results One strain of Gardnerella vaginalis infected 10 SPF grade KM mice, 7 mice met the diagnostic criteria for bacterial vaginosis, and 3 mice failed to model, with a success rate of 70%. Western blot was used to detect protein expression levels, and the levels of laminin and enolase in the successfully modeled mouse vaginal epithelial tissue were significantly higher than those in the failed modeling group, with statistical differences between the two groups(P<0.05). Conclusion Enolase and laminin may be involved in the occurrence of bacterial vaginosis, however, further research is needed to determine the mechanisms through which they trigger the occurrence and development of the disease.

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Objective To investigate the expression changes of cyclin-dependent kinase inhibitor 3(CDKN3) indifferent tumors and its effect on tumor staging, prognosis and immunotherapy. Methods The expression characteristics of CDKN3 in different cancers using data from TCGA, CCLE, ICGC, and GTEx databases were evaluated. The GEPIA2 platform and Kaplan-Meier analysis were utilized to assess the effect of CDKN3 on tumor pathological staging and survival prognosis. The TIMER platform was employed to explore the influence of CDKN3 on the tumor immune microenvironment and immunotherapy. Its effect on immune checkpoint and key immunotherapeutic predictors using bioinformatics methods was explored. The GeneMANIA tool was used to construct the protein-protein interaction(PPI) network of CDKN3. Kyoto Encyclopedia of Genes and Genomes(KEGG) and Gene Onotology(GO) enrichment analyses were conducted to explore the biological processes and signaling pathways associated with CDKN3. The effect of CDKN3 on HepG2 cell proliferation, invasion, migration, and apoptosis was validated through transfection with CDKN3 siRNA. Results CDKN3 was found to be widely overexpressed in tumors. High expression of CDKN3 was often associated with advanced pathological staging and poor survival prognosis. CDKN3 expression was negatively correlated with most immune checkpoints and positively correlated with tumor mutation burden(TMB), microsatellite instability(MSI), and mismatch repair(MMR) genes. CDKN3 was associated with cell cycle, cellular senescence, and the p53 signaling pathway. Furthermore, EdU staining, JC-1 staining, Transwell, and Wound Healing assays confirmed that CDKN3 promoted HCC cell proliferation, invasion, and migration while inhibiting apoptosis. Conclusion Abnormal expression of CDKN3 is closely related to tumor staging, prognosis, and immune microenvironment characteristics, making it a potential prognostic marker and immunotherapy adjuvant target in cancer.

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Objective To screen for potential biomarkers of anal fistula(AF) by using weighted gene co-expression network analysis(WGCNA), machine learning, immune infiltration analysis, and animal experiments. Methods Download transcriptome data from the gene expression omnibus containing AF and peri-fistula tissue(PF) for differential analysis. Gene ontology(GO) and Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analyses on differentially expressed genes(DEGs) were performed. WGCNA results were integrated with DEGs to screen for genes related to AF. Machine learning methods such as the least absolute shrinkage and selection operator(LASSO), support vector machine recursive feature elimination(SVM-RFE), and random forest(RF) were utilized to screen potential biomarkers for AF. Immune infiltration analysis was conducted and the AF rat model was replicated for validation. Results A total of 377 DEGs were obtained, mainly enriched in pathways such as B cell receptor signaling and chemokine signaling. Machine learning algorithms identified matrix metalloproteinase 13(MMP13) as a potential biomarker for AF. In AF samples, memory B cells, plasma cells, M0 macrophages, and M1 macrophages were higher than in PF samples, while resting CD4 memory T cells and resting dendritic cells were lower than in PF samples. MMP13 showed a positive correlation with M0 macrophages, activated mast cells, and immature B cells, and a negative correlation with resting mast cells. Experimental results showed that MMP13 expression levels were higher in rat AF samples compared to the control group. Conclusion The onset of AF involves various immune cells and signaling pathways. MMP13 is significantly upregulated in AF tissue and correlates with multiple immune cells, making it a potential novel biomarker of AF.

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Objective To establish an optimized method for isolation and purification of microglia from aged rat brain tissue, and the phenotype of microglia was detected by flow cytometry. Methods With young rats(3 months old) as control, the brain tissues of aged rats were immediately processed into single cell suspensions by mechanical dissociation and enzymatic digestion using type IV collagenase. Microglia were isolated on Percoll gradients(30%-37%-70%). The cells were stained with fluorescence-labeled antibodies and the phenotype of microglia was detected by flow cytometry. Results This study developed a method that enzymatic digestion and mechanical dissociation combined with density gradient centrifugation. More single cells could be obtained by using this method. And the survival rate of cells was more than 90%. The flow cytometric analysis showed that the expression of M1 microglia marker CD86 and MHC Ⅱ increased(P<0.01), and the expression of M2 microglia marker CD200R increased(P<0.01) in aged rats compared with that in young rats. Conclusion The use of type IV collagenase and mechanical digestion combined with density gradient centrifugation is good for isolating and purifying microglia from adult and aged rat brain tissue.

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Objective To investigate the effect of activation of the stimulator of interferon genes(STING) pathway on macrophage polarization function and its role in T-cell response.Methods Mouse macrophage RAW264.7 cells were used.STING signaling related proteins in RAW264.7 macrophage treated with STING agonist diABZI were analyzed by Western blot,including TANK-binding kinase-1(TBK1),interferon regulatory factor-3(IRF3),STING,p-TBK1,p-IRF3,p-STING.The polarization of macrophage RAW264.7 cells treated with diABZI was analyzed by flow cytometry.Co-culture of diABZI-treated RAW264.7 macrophage and T cells was applied to evaluate the change of T cell response.Results STING signaling related proteins were upregulated in macrophage RAW264.7 cells treated with diABZI for 3 hours.The expression of CD86 was upregulated on the surface of macrophages after 12 hours of diABZI treatment,and the CD86/CD206 ratio was elevated,which presented the M1 polarization phenotype.When coculturing diABZI-treated macrophage RAW264.7 cells with T cells,the cytokine secretion ability of T cells including CD4+T and CD8+T cells was enhanced and the expression of CD107a in CD8+T cells was upregulated.Conclusion STING signaling induces M1 polarization of macrophages which enhance the function of T cells,especially CD8+T cell immune response.

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Objective To investigate the protective effect of neurotrophic factor(MANF) derived from midbrain astrocytes on myocardial injury induced by sepsis by activating SIRT1/AMPK signaling pathway. Methods 48 mice were randomly divided into 4 groups: control group, recombinant mouse MANF(rmMANF) group, cecal ligation and puncture(CLP) group and CLP+rmMANF group, with 12 mice in each group.The survival rate, sepsis score, anal temperature, blood biochemical indexes, pathological indexes of myocardial injury and the expression of endoplasmic reticulum stress(ERS) related proteins were detected 8 h after CLP.H9C2 cells were divided into control group(Con),LPS group, LPS+rmMANF group, LPS+rmMANF+EX527 group and LPS+rmMANF+Cpd C group.The cells were collected after 24 h treatment with LPS,and the expression of ERS protein and apoptosis in cells were analyzed. Results Compared with CLP group, the sepsis score and serum Lactate dehydrogenase(LDH),creatine kinase(CK),aspartateaminotransferase(AST) and blood urea nitrogen(BUN) levels in CLP+rmMANF group decreased significantly(P<0.01),and the anal temperature and serum albumin(ALB) levels increased significantly(P<0.05).Compared with CLP group, the expression of MANF in CLP+rmMANF group increased significantly(P<0.01),and the expression of glucose-regulated protein 78(GRP78),C/EBP homologous protein(CHOP) and the percentage of TUNEL positive cells decreased significantly(P<0.05).In vitro, LPS stimulation down-regulated the expression of SIRT1 and AMPK in H9C2 cells, while rmMANF further increased the expression level of SIRT1 and AMPK.Compared with LPS+rmMANF group, the expression of GRP78 and CHOP protein and the apoptosis rate of H9C2 cells in LPS+rmMANF+EX527 group and LPS+rmMANF+Cpd C group increased significantly(P<0.05). Conclusion rmMANF inhibits ERS related to sepsis-induced myocardial injury by activating SIRT1/AMPK signaling pathway, thereby protecting myocardial injury.

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Objective To explore the effects of high-altitude hypoxia on thyroid hormone(TH) synthesis by quasi-targeted metabolomics technology. Methods Twenty SPF male SD rats were randomly divided into Control group and Hypoxia group. An acute hypoxia injury model was established in SD rats by simulating hypoxia stress in a hypobaric oxygen chamber at an altitude of 6 km for 48 hours. The body weight, arterial oxygen partial pressure(PaO2) and blood oxygen saturation(SaO2) were detected. KEGG enrichment analysis was performed after the metabolites in the blood of two groups were detected by quasi-targeted metabolomics technology. The expression levels of sodium iodide symporter(NIS), thyroid peroxidase(TPO) and thyroglobulin(TG) were detected by RT-PCR and Western blot in TH synthesis pathway. The content of serum thyroxine(T4) and the expression levels of glutathione peroxidase(GSH-Px), superoxide dismutase(SOD) and malondialdehyde(MDA) were detected by ELISA. The expression levels of toll-like receptors-4(TLR-4), interleukin-6(IL-6), nuclear factor-κB/p65(NF-κB/p65) and tumor necrosis factor-α(TNF-α) were detected by RT-PCR, Western blot and ELISA. The expression levels of Pro apoptotic protein Bcl-2 associated X protein(Bax) and inhibitor of apoptosis protein B-cell lymphoma/leukemia-2(Bcl-2) were detected by RT-PCR and Western blot in rats thyroid tissue. Results Compared with the Control group, the body weight, PaO2and SaO2of rats in the Hypoxia group significantly decreased(P<0.01). The differential metabolites in arterial serum of hypoxia group rats were significantly enriched in the TH synthesis pathway, and the content of the pathway end product T4decreased significantly(P<0.01). In addition, the mRNA and protein expression levels of NIS, TPO, TG in rats thyroid tissue significantly decreased(P<0.05). The ELISA validation results showed that the changes of T4content were completely consistent with the above results. Compared with the Control group, the enzyme activities of SOD and GSH-Px in the serum of rats in the hypoxia group decreased, while the content of MDA increased(P<0.01); the mRNA, protein expression levels and contents of TLR-4, IL-6, NF-κ B/p65, TNF-α significantly increased(P<0.05), while the mRNA and protein expression levels of Bax in thyroid tissue significantly increased, Bcl-2 significantly decreased(P<0.05). Conclusion Hypoxia stress at high altitude leads to apoptosis of thyroid follicular epithelial cells by promoting oxidative stress and inflammatory response, which effects thyroid function and ultimately reduces thyroid hormone synthesis.

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Objective To investigate the effects of long-term hyperglycemia on renal pathology of type 1 diabetic mellitus(T1DM) cynomolgus monkeys by establishing streptozotocin(STZ)-induced T1DM. Methods Eight 4-year-old male cynomolgus monkeys were randomly divided into control and model groups. Four cynomolgus monkeys were used in the control group, and four cynomolgus monkeys were injected with streptozotocin to create a T1DM model. Hematoxylin-eosin(HE) staining and periodic acid-Schiff(PAS) staining were used to observe renal pathological changes.The histological characteristics and changes were observed under transmission electron microscope. Morphometric measurements were used to analyze the glomerular area, the thickness of the glomerular basement membrane, the proportion of glomerular podocyte foot processes and the average area of foot processes. Results In comparison to the control group, the model group showed pathological changes in the kidneys, including increased glomerular area(P<0.01), thickened basement membrane, capillary loop compression, disordered arrangement of endothelial cell fenestrations, proliferation of mesangial cells, mesangial expansion, accumulation of mesangial matrix glycoproteins, a decrease in the number of foot processes(P<0.001), a decrease in the average size of the foot processes(P<0.05), widened podocyte slit diaphragm, and an increase in the number of fused foot processes(P<0.001). Conclusion Cynomolgus monkeys with T1DM suffer from renal pathological changes due to long-term hyperglycemia.

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Objective To simulate the tumor microenvironment though the 3D microtumor model which was constructed using droplet microfluidics.To explore its feasibility as a model for in vitro breast cancer research through3D microtumour fabrication,characterisation and sensitivity testing to chemotherapeutic drugs.Methods Breast cancer cells were encapsulated with hydrogel shells in collagen-rich microencapsulated cores to obtain breast cancer microtumours in vitro;breast cancer microtumours were co-cultured with 3D microencapsulated endothelial cells by Transwell system.The structure and growth characteristics of the microtumours were directly observed by microscopy;the CCK-8 assay was used to detect the proliferation of the cells under different culture models and the drug sensitivity to doxorubicin;flow cytometry was used to compare the differences in apoptosis during the proliferation process;and the differences in the migratory and invasive abilities of the cells were assessed by scratch assay and Transwell assay;the expression of epithelial-mesenchymal transition-related proteins was detected by Western blot.Results Breast cancer cells grew well in hydrogel nucleus-shell microcapsules;cell proliferation assays showed that 3D culture and 3D co-culture cells proliferated at a significantly lower rate than 2D culture;3D culture and 3D co-culture cells had enhanced migration and invasion ability and showed higher expression of EMT-related proteins compared to 2D culture;3D culture and 3D co-culture cells were significantly less sensitive to chemotherapeutic drugs compared to 2D culture.The sensitivity of 3D and 3D co-cultured cells to chemotherapeutic drugs was significantly reduced compared to 2D culture.Conclusion 3D cultures show similar morphology and biology to in vivo tumours and are more resistant to chemotherapeutic agents.

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Objective To construct and validate a prognosis model related to ferroptosis in urinary tract tumors using bioinformatics methods. Methods RNA-seq and clinical data from TCGA′s BLCA and KIRC datasets were analyzed to establish the prognostic model, and then were validated using ICGC and GEO data. Prognostic genes associated with ferroptosis were identified through univariate Cox, LASSO-Cox, and multivariate Cox regression analyses. Co-expression and protein-protein interaction(PPI) network analyses determined the relationships among these genes. Immune infiltration analysis explored the association between ferroptosis-related prognostic genes and the immune microenvironment. Functional enrichment analysis of differentially expressed genes between high and low-risk groups in BLCA and KIRC prognostic models was conducted to investigate potential mechanisms by which ferroptosis-related genes regulate BLCA and KIRC prognosis. Results Significant prognostic gene signatures associated with ferroptosis were identified in BLCA and KIRC. For BLCA, the genes EGR1, ZEB1, P4HB, WWTR1, JUN, CDO1,SCD,SREBF1,CAV1, and GALNT14 were significant. For KIRC, the genes ASMTL-AS1, CHAC1,MT1G, RRM2, TIMP1, DPEP1, GLRX5, and NDRG1 were significant. Ferroptosis-related miRNAs linked to the prognosis of both cancers were also identified. The constructed risk models based on these genes and miRNAs predicted patient prognosis in TCGA-BLCA and KIRC, with low-risk groups showing significantly higher overall survival(P<0.05). The hazard ratios for these models ranged from 2.54(95%CI: 1.73-3.74) to 4.74(95%CI: 3.47-6.47), with AUC values above 0.60. Co-expression analysis and PPI networks revealed high correlation levels between JUN and EGR1 in BLAC and between SCD and SREBF1. Immune infiltration analysis indicated positive correlations between EGR1, CAV1, JUN, and immune scores, while SREBF1 showed a negative correlation. Conclusion The prognosis model based on ferroptosis-related genes effectively predicts patient outcomes in BLCA and KIRC. This model can serve as a reference for targeting ferroptosis to assess the prognosis of BLCA and KIRC patients.

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Objective To investigate the expression of transient receptor polycystic protein 2(TRPP2) in oral squamous epithelial cell and its effect on the invasion and migration of oral squamous cell carcinoma(OSCC), and to explore the potential signaling pathway of TRPP2 affecting OSCC metastasis. Methods The OSCC model with TRPP2 knockdown was constructed by CRISPR-Cas9 lentivirus plasmid transfection technique. The effect of TRPP2 protein knockdown was verified by Western blot. The effect of TRPP2 on OSCC proliferation was detected by CCK-8 assay and clone formation assay. RT-qPCR was used to detect the target genes associated with TRPP2 metastasis to OSCC. Western blot and RT-qPCR were used to detect the expression of EpCAM and its transcription factors associated with unfolded protein response(UPR). The effects of TRPP2 on the invasion and migration of OSCC were examined by invasion test and scratch test. Results Compared with HOK in oral epithelial cells, the expression of TRPP2 in OSCC was significantly higher. When TRPP2 was knocked down, OSCC proliferation and clonalformation were significantly enhanced. Compared with the control group, a total of 494 differential genes were significantly expressed in TRPP2 knockdown transcription profile, among which 234 genes were up-regulated and 260 genes were down-regulated. The expression of EpCAM gene, which is related to cell adhesion, was up-regulated. In addition, UPR related genes PERK, ATF6, GRP78 were up-regulated, while ATF6 and EpCAM were down-regulated in OSCC compared to HOK cells. The expression of ATF6 and EpCAM in oral squamous cell carcinoma cells was up-regulated by TRPP2 knockdown, and the cell migration and invasion ability decreased. The ATF6 inhibitor ceapin-A7(5 μmol/L) restored the OSCC migration and invasion ability of TRPP2 knockdown. Conclusion TRPP2 is highly expressed in OSCC. When TRPP2 is knocked down, OSCC proliferation ability is enhanced, migration and invasion ability are inhibited. TRPP2 mediates the expression of EpCAM through activation of UPR, thus affecting the invasion and migration of oral squamous cell carcinoma.

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Objective To analyze the clinical distribution and drug resistance trends of clinical isolates ofKlebsiella pneumoniaefrom a tertiary hospital in Henan Province from 2017 to 2021, and to provide recommendations for the rational use of antibiotics in the hospital. Methods A retrospective study was conducted to collect and organize the drug sensitivity test results and clinical data of clinical isolates ofKlebsiella pneumoniaein the hospital from 2017 to 2021. The data was analyzed using WHO NET 5.6 and SPSS 26.0 software. Results A total of 1 379 strains ofKlebsiella pneumoniaewere isolated in the hospital from 2017 to 2021, with an isolation rate of 17.87%. There was no significant difference in the isolation rates ofKlebsiella pneumoniaeover the entire five-year period.The top three sources of isolates were sputum(65.99%), secretion(12.98%), and urine(8.85%). The main departments for collecting isolates were the respiratory department(26.03%) and the ICU(24.58%), with the isolation rate in other departments being less than 5%. The drug sensitivity results showed that, except for the resistance rate of cefoperazone, the resistance rates of other antibacterial drugs showed statistically differences over the five years, generally showing a stable or decreasing trend. The carbapenem drugs imipenem and meropenem decreased from 33.1% and 33.5% in 2017 to 13.7% and 14.4% in 2021, respectively. In addition, the resistance rates of chloramphenicol and tetracycline class minocycline showed an increasing trend. Conclusion Klebsiella pneumoniaeis still the main pathogen isolated in the hospital, mainly in the respiratory department and ICU, with sputum, urine, and blood being the main specimens. The resistance rates have shown a stable or decreasing trend over the past five years, with a significant decrease in carbapenem resistance rates. However, attention should still be paid to the increasing resistance rates of chloramphenicol and minocycline. Clinically, antibiotics should be used rationally based on drug sensitivity results.

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Objective To analyze the current situation, influencing factors and mechanism of knowledge-attitude-practice(KAP) regarding acquired immune deficiency syndrome(AIDS)/sexually transmitted diseases(STDs) among migrant workers in Chengdu's main urban area, so as to provide a basis for the development of effective prevention and control policies for this group. Methods Convenience sampling and systematic sampling were used to collect demographic information and data on knowledge, attitudes, and sexual behavior characteristics of AIDS/STDs of the participants. The collected data were organized and statistically analyzed by EpiData 3.1 and SPSS 26.0 software. Additionally, a KAP path analysis model was constructed by using AMOS 24.0 software. Results A total of 257 valid questionnaires were obtained. The AIDS awareness rate was 55.6%, with a mean scores of(5.59±1.61). The awareness rate of STDs was 37.4%, with a mean scores of(9.05±3.00). Discrimination attitudes towards AIDS and STDs were reported by 58 participants(22.6%) and 44 participants(17.1%) respectively. The prevalence of high-risk sexual behavior was 3.50%. Men(OR=0.500, 95%CI: 0.279-0.897) acted as deterrents to knowledge of AIDS. On the other hand, childlessness facilitated discrimination against AIDS(OR=2.748, 95%CI: 1.385-5.451) and STDs(OR=2.287, 95%CI: 1.084-4.825). There was lower likelihood of engaging in high-risk sexual behavior among migrant workers in Chengdu's main urban area who were older(OR=0.854, 95%CI: 0.785-0.929). The occurrence of high-risk sexual behaviors was influenced both directly and indirectly by attitudes towards AIDS and related knowledge. There was a positive correlation between knowledge about AIDS and STDs and attitudes towards them(r=0.15,0.24, bothP<0.05), as well as between attitudes towards AIDS and attitudes towards STDs(r=0.57,P<0.05). That is, the higher the scores of knowledge, the less likely one was to hold discrimination attitudes. Therefore, increasing the rate of knowledge awareness could reduce discrimination towards AIDS/STDs and the occurrence of high-risk sexual behaviors. Conclusion The level of AIDS/STDs knowledge among the migrant workers in Chengdu′s main urban area is concerning. Innovative interventions should be intensified in key areas and populations.

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Objective To explore the causal relationship between the abundance of intestinal flora(IF) and the risk of developing acute myeloid leukemia(AML) using two-sample bidirectional Mendelian randomization(MR) methods. Methods The MiBioGen Consortium and FinnGen databases were utilized for the IF abundance and AML genome-wide association datasets, respectively. The primary analysis method entailed variance inverse weighting, supplemented by weighted median test, while heterogeneity was assessed using Cochran′sQtest. Additionally, MR-PRESSO was employed to identify any outliers, while MR-Egger intercept was utilized to evaluate horizontal pleotropy. Results Higher abundance of genetically determinedDialister(P=0.034,OR=4.373,95%CI:1.329-14.392),Veillonaceae(P=0.009,OR=2.029,95%CI:1.839-3.866), andLachnospiraceae UCG008(P=0.034,OR=3.827,95%CI:1.107-13.228) were found to be linked to an increased risk of AML. In contrast,Ruminococcaceae(P=0.042,OR=0.176, 95%CI:0.033-0.939) andPeptococcaceae(P=0.025,OR=0.168, 95%CI:0.035-0.803) were associated with a reduced risk of this disease. Conclusion Higher levels of theDialister, theVeillonellaceae, and theLachnospiraceae UCG008are risk factors for AML, while the familiesRuminococcaceaeandPeptococcaceaeare identified as protective factors against AML.

abstract:
Glutamate receptors are widely distributed in the central nervous system(CNS) and participate in the delivery of excitatory neurotransmitters, including ionotropic glutamate receptors and metabotropic glutamate receptors. Metabotropic glutamate receptors(mGluRs) regulate neuronal excitability and synaptic plasticity, and are classified into group Ⅰ, Ⅱ and Ⅲ. The roles of group Ⅰ mGluRs in psychiatric and neurodegenerative diseases have been extensively studied, especially in the pathological progression of acute brain injury. Several studies have shown that group Ⅰ mGluRs can be involved in eliminating inflammatory damage, suppressing cell apoptosis, regulating neural network disorders, and promoting brain function recovery after acute brain injury. Therefore, group Ⅰ mGluRs have the potential to be effective targets for the treatment of acute brain injury. This paper reviews the distribution and function of group Ⅰ mGluRs in CNS, the pathological role in acute brain injury, and explore the potential of the development of drugs targeting group Ⅰ mGluRs.

abstract:
Pancreatic cancer is a highly malignant gastrointestinal cancer with a poor prognosis, and early diagnosis remains challenging. The use of reliable biomarkers can significantly enhance the early evaluation and management of this disease. Circulating tumor cells(CTCs) are released into the bloodstream and can be obtained easily through minimally invasive liquid-based biopsy, making them promising candidates for early tumor diagnosis, prognosis assessment, and monitoring therapeutic responses. This paper reviews the advancements in CTCs detection technology and their clinical applications in pancreatic cancer over the past decade, both domestically and internationally, which offer a new perspective on the early diagnosis and prognosis of pancreatic cancer.