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Objective To investigate the effects of 3-phosphoinositide-dependent protein kinase-1(PDK1) on the purkinje cells(PCs) in aging mice. Methods The footprint test, elevated beam-walk test and rotarod test were used to measure the gait traits, motor balance and motor coordination of mice. Immunoflourescence was used to detect the structure of the cerebellum and the morphology of cells in the cerebellum. The expression level of PDK1, protein kinase C gamma(PKCγ) and protein kinase B(also known as AKT) and the phosphorylation of ribosomal protein S6(rpS6) in the cerebellum were detected by Western blot. Results The mice showed motor defects in the overlap between forelimb and hindlimb, motor balance and motor coordination ability in an age-dependent manner. Furthermore, the cerebellum and the soma size of the PCs decreased in an age-dependent manner. However, the density of the PCs was not affected by age. Additionally, the expression level of the PDK1, PKCγ, AKT and the phosphorylation of rpS6 in the cerebellum were reduced with age. Conclusion The cerebellar size and the cell soma size of PCs decrease in the old mice. These effects may contribute to the reduction of the expression of PKCγ and AKT and the phosphorylation of rpS6 in the old mice, ultimately affecting the cerebellar motor coordination.

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Objective To observe the effects of apocynum venetum leaves extract(AVLE) on anxiety-like behavior caused by chronic restraint stress(CRS). Methods Male C57BL/6 mice were randomly divided into 4 groups, 5 per group: control group(equivalent physiological saline 0.2 ml/10 g), CRS model group(equivalent physiological saline 0.2 ml/10 g), AVLE treatment group(AVLE 120 mg/kg) and positive control group(paroxetine hydrochloride 3 mg/kg). The anxiety mouse model of chronic restraint stress was established, and the anxiety level of mice treated with apocynum venetum leaf extract was evaluated by open field test and elevated cross maze test. Immunofluorescence staining was used to detect the changes of serotonin receptor in hypothalamic paraventricular nucleus. Chemogenetics combined with adeno-associated virus was used to detect the effect of hypothalamic oxytocin neurons on the anxiety regulation of AVLE. Results Compared with the model group, apocynum venetum leaves extract could increase the total distance of open field exercise, the activity time, the time of mice entering the central area(P<0.05), and the activity distance and time of mouse elevated maze(P<0.05). The expression of serotonin 1A receptor in the paraventricular nucleus of hypothalamus was enhanced after treatment with AVLE, and inhibition of serotonin 1A receptor and oxytocin neurons in hypothalamic paraventricular nucleus could decrease activity level in mice(P<0.05). Conclusion AVLE can improve the anxiety state and activity ability of CRS mice, and its effect is related to serotonin receptor and oxytocin neurons in hypothalamic paraventricular nucleus.

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Objective To investigate the effects and underlying mechanisms of the macrophage migration inhibitory factor(MIF) inhibitor(S,R)-3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazoleacetic acid methyl ester(ISO-1) on sepsis-induced acute kidney injury(AKI). Methods Human renal tubular epithelial HK-2 cells were divided into Con group(without any treatment), ISO-1 group(10 μg/ml ISO-1 treatment for 24 h) and LPS group(10 μg/ml LPS treatment for 24 h), LPS+ISO-1 group(10 μg/ml LPS treatment for 24 h followed by 10 μg/ml ISO-1 treatment for 24 h). ELISA was used to measure the levels of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), and interleukin-6(IL-6) in the cell supernatants. Reactive oxygen species(ROS) levels were assessed using the 6-carboxyl-2 ′,7′-dichlorodihydrofluorescein diacetate fluorescent indicator(DCFH-DA) method. Apoptosis levels were detected by TUNEL staining, and Western blot was employed to analyze the expression of proteins of Kelch like ECH associated protein 1(Keap1), NFE2 like bZIP transcription factor 2(Nrf2), heme oxygenase-1(HO-1), as well as apoptosis-related proteins Bcl-2, Bax, and cleaved Caspase-3(c-Caspase-3). A sepsis mouse model was established using the cecal ligation and puncture(CLP) method, and the mice were divided into four groups: sham-operated(Sham), ISO-1 control(ISO-1), CLP, and ISO-1 treatment(CLP+ISO-1). After the experiment, mouse kidney tissues were collected for HE staining to observe pathological changes. Blood urea nitrogen(BUN), serum creatinine(Scr), myeloperoxidase(MPO) levels in kidney tissues, glutathione(GSH) and superoxide dismutase(SOD) activities were measured. Western blot was also used to detect the expression of MIF and proteins in the Nrf2/Keap1 signaling pathway and apoptosis-related proteins in kidney tissues. Results Compared to the Con group, the LPS and LPS+ISO-1 groups showed significantly increased levels of TNF-α, IL-1β, IL-6, TUNEL-positive rates, ROS levels, and protein expressions of Keap1, Bax, and c-Caspase-3 in HK-2 cells(P<0.05), while the expressions of Nrf2, HO-1, and Bcl-2 significantly decreased(P<0.05). The ISO-1 group showed no significant changes(P>0.05). Compared to the LPS group, the LPS+ISO-1 group exhibited significantly decreased levels of TNF-α, IL-1β, IL-6, TUNEL-positive rates, ROS levels, and protein expressions of Keap1, Bax, and c-Caspase-3, while the expressions of Nrf2, HO-1, and Bcl-2 significantly increased(P<0.05). In the mouse experiments, compared to the Sham group, the CLP and CLP+ISO-1 groups showed severe kidney tissue damage, increased levels of serum BUN, Scr, and kidney MIF, Keap1, Bax, and c-Caspase-3 protein expressions(P<0.05), while GSH, SOD activities, and protein expressions of Nrf2, HO-1, and Bcl-2 significantly decreased(P<0.05). The ISO-1 group showed no significant changes(P>0.05). Compared to the CLP group, the CLP+ISO-1 group showed significant improvements in the aforementioned indicators(P<0.05). Conclusion The specific MIF inhibitor ISO-1 can ameliorate sepsis-induced AKI by inhibiting oxidative stress, inflammatory response, and apoptosis bothin vitroandin vivo. The mechanism may be through Nrf2/Keap1 signaling pathway.

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Objective To synthesis a Gd-labeled molecular probe capable of responding to fibroblast activation protein(FAP) highly expressed in tumor-associated fibroblasts for magnetic resonance imaging studies of pancreatic cancer.Methods The chemical structure of Z-Gly-Pro-Cys-Lys(Gd-DOTA)-CBT(contrast agent 1) was characterized through mass spectrometry analysis.High-performance liquid chromatography(HPLC) was used to verify the contrast agent 1 formed a dimer under the cleavage of fibroblast activation protease(FAP) in vitro,and further self-assembled to form Gd-nanoparticles.The cell counting Kit-8(CCK-8) assay was used for biological safety analysis.BxPc-3 tumor-bearing mice were established and randomly divided into three groups,3 mice intravenously(i.v.) injected with 0.08 mmol/kg contrast agent 1 through tail vein were designated as the experimental group,3mice i.v.injected with 0.08 mmol/kg Gd-DTPA were designated as the control group,and 3 mice i.v.injected with normal saline were designated as the blank control group(n=3).Dynamic MR scanning from 0 to 3 hours was performed for them,respectively.The enhancement effects of T1-and T2-weighted imaging were observed and the rate of change in the tumor-to-muscle ratio(T/M) over time was analyzed.Results Contrast agent 1 efficiently responded to FAP in vitro,forming dimers under the action of FAP,and further self-assembled to form nanoparticle structures.Contrast agent 1 has good biocompatibility.The BxPc-3 tumor-bearing mice showed that the T1-weighted contrast of the tumor reached its highest level at 2 h post injection of contrast agent 1 or Gd-DTPA.In T1-weighted imaging,the T/M at 2 h in the experimental group was 135.20%±0.06% of 0 h,the T/M at 2 h in the Gd-DTPA group was 115.70%±0.05% of 0 h and the T/M at 2 h in the saline group was 113.5%±0.02% of 0h.Compared with Gd-DTPA control group,the experimental group had significantly enhanced T1-weighted MR contrast,and the difference was statistically significant(P<0.01).In tumor T2-weighted imaging,the T/M quantitative analysis showed that the tumor T2-weighted contrast enhancement of the three groups was relatively small,and the T/M of the experimental group and the control group at 0.5 h was 94.60±0.03% and 106.30±0.04% of the 0 h,respectively.In the normal saline group,T/M at 0.5 h was 102.20±0.002% of that at 0 h,and no significant enhancement of the T2-weighted MR contrast of tumor was observed.Conclusion Successfully construct a FAP-responsive molecular probe which can self-assemble to form a Gd-nanoparticle structure in tumor cells under the cleavage of FAP,effectively improving the enrichment of Gd in the target region,and enhancing T1-weighted magnetic resonance imaging of pancreatic cancer,which has potential clinical application value.

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Objective To investigate the relationship between tropomodulin 3(TMOD3) and the malignant biological characteristics of hepatocellular carcinoma, and the predictive potential of TMOD3 as a biomarker for the recurrence and metastasis of hepatocellular carcinoma. Methods Firstly, the structure of TMOD3 and its subcellular localization in cells and tissues were analyzed using database of Human Protein Atlas. Then explored the differential expression of TMOD3 in hepatocellular carcinoma tissues and normal liver tissues and its impact on clinical pathological characteristics and prognosis using TCGA and GEO datasets. Subsequently, the STRING database was utilized to explore the interacting proteins of TMOD3, followed by enrichment analysis conducted using the Metascape database. Finally, Logistic regression was used to analyze the independent risk factors for metastasis of hepatocellular carcinoma and evaluated the importance of predictive variables using ROC curves and Wald tests. Survival analysis was conducted using the Kaplan-Meier curve and the Log-rank test. Results TMOD3 was localized to actin filaments in cells, and compared with normal tissues, the expression level of TMOD3 in liver cancer tissues is higher(P<0.05), and high expression of TMOD3 was closely related to lymph node metastasis and distant metastasis of hepatocellular carcinoma patients(P<0.05). Enrichment analysis results revealed that TMOD3 and its interacting proteins mainly function and signaling pathways related to tumor invasion and migration. Logistic regression found that TMOD3 was an independent risk factor for recurrence and metastasis of hepatocellular carcinoma(OR: 4.359, 95%CI: 1.235-15.384,P=0.022). Survival analysis revealed that high expression of TMOD3 was associated with poor OS, DFS, and RFS of hepatocellular carcinoma patients(P<0.05). Both ROC analysis and Wald test indicated that TMOD3 has good predictive characteristics for recurrence and metastasis of hepatocellular carcinoma. Conclusion TMOD3 is closely associated with the invasion and metastasis of hepatocellular carcinoma and is an independent risk factor for the recurrence and metastasis of liver cancer. TMOD3 performs well in predicting the recurrence and metastasis of hepatocellular carcinoma and has the potential to become a biomarker for predicting the recurrence and metastasis of hepatocellular carcinoma.

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Objective To establish aNBR1-knockout lung cancer mouse model through CRISPR-Cas9 technology by using adeno-associated virus(AAV) as a vector to specifically inhibitNBR1expression and to investigate the impact ofNBR1knockout on tumor growth and immune cell infiltration and regulation. Methods sgRNAs targeting mouseNBR1(Gene ID: 17966) was designed using the online tool CRISPOR(http://crispor.tefor.net/crispor.py). AAV6 was utilized as the vector for sgRNA delivery, and the efficiency of gene knockout was confirmed using PCR and DNA sequencing methods. To determine the best AAV infection approach in mice, 6 C57BL/6J mice were randomly divided into intranasal and endotracheal groups. After 28 days, lung tissue sections were assessed for enhanced green fluorescent protein expression to identify the more efficient infection method for subsequent experiments. Lung tumor growth, as well as immune cell infiltration and activation status in tumor tissues, were detected using methods including HE staining, immunohistochemistry, immunofluorescence, and flow cytometry. Results DNA sequencing and immunofluorescence results indicated successful construction of the AAV6-U6-sgNBR1-CAG-Cre-GFP vector with stable knockout efficiency. Fluorescence microscopy showed higher efficiency of lung infection in mice through intratracheal administration(P<0.05). HE staining revealed reduced tumor area in mouse lungs after targetedNBR1knockout compared to the control group(P<0.01). Immunofluorescence and flow cytometry results demonstrated enhanced functional activity of CD8+T lymphocytes in lung cancer tissues of mice with targetedNBR1knockout, characterized by increased effector T lymphocytes and decreased exhausted T lymphocytes(P<0.01). Conclusion Using CRISPR/Cas9 technology, we construct a lung cancer mouse model with targetedNBR1knockout. We verify that targeted inhibition of NBR1 expression significantly enhances the functional activity of CD8+T lymphocytes in lung tissues, resulting in suppressed tumor growth, reduced tumor burden, and extended survival in lung cancer mice. This study lays an experimental foundation for investigations into the mechanisms and functions ofNBR1and other genes in lung adenocarcinoma cells.

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Objective To explore the damaging effect of the IP3-protein kinase C pathway on human retinal vascular endothelial cells under high glucose conditions. Methods Human retinal vascular endothelial cells in the logarithmic growth phase were taken and divided into the experimental group(10 mmol/L glucose and 20 mmol/L glucose) and the control group(5 mmol/L glucose). The morphological changes of cells were observed under different concentrations of glucose culture medium. The apoptosis of human retinal vascular endothelial cells was detected by flow cytometry. Real-time fluorescent quantitative reverse transcription polymerase chain reaction was used to detect the RNA expression level of PKC in the IP3-protein kinase C(PKC) pathway, and Western blot was used to detect the protein expression level of PKC in cells. Results Under high glucose conditions, the volume of human retinal vascular endothelial cells increased, the degree of extension decreased, and the apoptosis rate increased. Compared with the control group, the apoptosis rate of human retinal vascular endothelial cells in the 20 mmol/L glucose group increased(P<0.05); the IP3 level of human retinal vascular endothelial cells in the 20 mmol/L glucose group(587.9±15.2) ng/ml was lower than that of the control group(738.9±1.0) ng/ml(P<0.05). Under high glucose treatment, the expression levels of PKC mRNA and protein increased(P<0.05). Conclusion The IP3-PKC pathway may be involved in the damage process of human retinal vascular endothelial cells under high glucose conditions, and thus may play a role in diabetic retinopathy.

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Objective To investigate theBMI1-NOTCH signaling pathway that regulates proliferation, migration, invasion, apoptosis, and cisplatin(CDDP) sensitivity in human oral squamous cell carcinoma(OSCC) cells. Methods Human OSCC cell line CAL27 was used to construct the CAL27 cell line with lentivirus, which knocked down or overexpressedBMI1gene. The knockdown and control groups included: shBMI1-1, shBMI1-2, shBMI1-3, and shNC group, while the overexpression and control groups were BMI1 and NC group. RT-qPCR and Western blot were employed to verify transfection efficiency and select the cell group with the most effective knockdown. Western blot was used to detect the expression of NOTCH signaling pathway proteins, including NOTCH1, Delta-like ligand 1(DLL1), Jagged1(JAG1), and Hairy/enhancer-of-split 1(HES1), in the transformed CAL27 cells. Subsequently, the cells in each group were cultured with the drug solvents dimethyl sulfoxide(DMSO), CDDP, NOTCH pathway inhibitor gamma-secretase inhibitor(DAPT), and CDDP+DAPT, respectively, and the cell phenotype in each group were detected using CCK-8 assay, Wound healing assay, Transwell invasion assay, colony formation assay, and flow cytometry. Results Overexpression ofBMI1increased the expression levels of NOTCH pathway gene proteinsNOTCH1,DLL1,JAG1, andHES1(P<0.05). Under CDDP intervention, cells in the BMI1 group exhibited increased viability, invasion, migration, and colony formation abilities, and decreased apoptosis compared to the shBMI1 group(P<0.05). When comparing the CDDP group with the CDDP+DAPT group, the combination of medications resulted in a significant increase in the apoptosis rate and an increase in the sensitivity of cancer cells to CDDP(P<0.05). Conclusion BMI1may increase the cellular malignancy of CAL27 cell line by activating the NOTCH signaling pathway, promoting cell viability, migration, and invasion, as well as decreasing the cellular drug sensitivity to CDDP. Inhibition of the NOTCH pathway increases the cisplatin sensitivity of CAL27, and CDDP+DAPT has a synergistic cytotoxic effect to promote OSCC cell apoptosis.

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Objective To investigate the effects of knockdown of E2F transcription factor 1(E2F1) in human tongue squamous cell carcinoma cells on proliferation, migration, and invasion of tongue squamous cell carcinoma cells, and to explore its possible mechanisms. Methods The expression ofE2F1in human tongue squamous cell carcinoma was detected by RT-PCR, and the background expression ofE2F1gene in HOEC, SCC-9, CAL-27, TSCCa and SCC-25 cells was detected by RT-PCR, and the cell lines with significantly high expression of E2F1 were screened for subsequent experiments. CCK-8, EdU, colony formation, cell scratch, Transwell, apoptosis and cell cycle experiments were used to detect the effects of knockdown ofE2F1on proliferation, migration, invasion and apoptosis of human tongue squamous cell carcinoma cells after knockdown ofE2F1gene in tongue squamous cell carcinoma candidate cells. Western blot was used to detect the expression of epithelial mesenchymal transition markers E-cadherin, N-cadherin and snail family transcription inhibitor 1(Snail) protein and WNT signaling pathway markers WNT family members 3A(WNT3A), β-catenin and cyclin D1(CCND1). Results RT-PCR results showed that the expression ofE2F1in cancer tissues was significantly higher than that in adjacent tissues(P<0.001). At the same time, compared with HOEC cells, the expression ofE2F1in SCC-9, CAL-27, TSCCa and SCC-25 cells significantly increased(P<0.05), and the high expression in SCC-25 cells was the most obvious. SCC-25 would be used as an experimental cell line in subsequent experiments. After knocking downE2F1in SCC-25 cells, the viability of CCK-8 cells was significantly inhibited(P<0.001). The number of EdU positive cells decreased significantly(P<0.001). The number of cell clones was significantly reduced(P<0.001). The proportion of total apoptosis significantly increased(P<0.001). The proportion of cells in G1phase increased(P<0.01). The proportion of cells in G2phase decreased(P<0.01). The cell migration and invasion ability were significantly inhibited(P<0.001). Western blot showed that the expression of WNT signaling pathway proteins WNT3A, β-catenin and CCND1 decreased(P<0.001), while the expression of EMT-related proteins N-cadherin and Snail decreased(P<0.001), while the expression of E-cadherin increased(P<0.001). Conclusion Knockdown ofE2F1inhibits the proliferation, migration, invasion and EMT process of human tongue squamous cell carcinoma SCC-25 cells, and promotes apoptosis. This anti-tumor effect may be achieved by blocking the activation of WNT signaling pathway.

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Objective To investigate the alterations in hippocampal neurons,nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3) inflammasome,microglia,and myelin in rats with post-traumatic stress disorder(PTSD).Methods A PTSD rat model was established using the single prolonged stress(SPS) paradigm.Anxiety and cognitive functions were evaluated through the open field test,elevated plus maze,and Morris water maze.Histopathological changes in hippocampal neurons were assessed using hematoxylin and eosin(HE)staining.The expression levels of NLRP3 and iba-1(a microglial marker) in the hippocampus were examined using immunofluorescence staining.Immunohistochemical staining and Luxol Fast Blue staining were performed to investigate alterations in hippocampal myelin.Results Hippocampal neurons in PTSD rats exhibited damage,with increased activation of the NLRP3 inflammasome and microglia,and elevated myelin content.Conclusion The pathogenesis of PTSD may be associated with hippocampal neuronal damage,inflammatory responses,and changes in myelin.

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Objective To investigate the function and potential mechanism of hepatocellular carcinoma(HCC) cells-derived extracellular nicotinamide phosphoribosyl transferase(eNAMPT) in the angiogenesis of human umbilical vein endothelial cells(HUVEC). Methods The eNAMPT secretion of HCC cells were stimulated by hypoxia treatment and investigated the angiogenesis of HUVEC was investigated by tube formation assays. Transcriptomic RNA sequencing(RNA-seq) analyses of HUVEC under eNAMPT treatment were used to explore the mechanism of the effect of eNAMPT-insulin receptor(INSR) axis on angiogenesis. Western blot was used to detect the effects of eNAMPT-INSR axis on protein expression of HUVEC cells. qRT-PCR was used to detect the influence of eNAMPT-INSR axis on the expression of HUVEC intracellular related target genes. Results ELISA and Western blot results showed that the protein level of eNAMPT in the culture supernatant of HCC cells increased significantly after hypoxia stimulation.In vitroangiogenesis assay results showed that eNAMPT significantly increased the nodules number and total length of tubes in HUVEC. The transcriptome sequencing results of HUVEC showed that eNAMPT, as an insulin analogue, participated in the regulation of biological processes such as cell migration and angiogenesis in HUVEC, and might promote angiogenesis by activating insulin-like signaling pathways. Western blot results showed that the eNAMPT-INSR axis affected p-AKT and p-ERK1/2 protein levels in PI3K-AKT and MAPK-ERK1/2 signaling pathways in HUVEC. qRT-PCR results showed that the eNAMPT-INSR axis affected the expression of angiogenesis related target genes MMP2, MMP9 and VEGF in HUVEC. Conclusion eNAMPT secreted by HCC cells promotes angiogenesis by binding INSR to activate the PI3K-AKT and MAPK-ERK1/2 signaling pathways.

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Objective To investigate the effect of hirsutine combined with oxaliplatin on the antitumor activity of HGC-27 mice with gastric cancer. Methods Gastric cancer HGC-27 tumor-bearing mice model was established. The successfully established tumor-bearing mice were randomly divided into normal control group, model group, oxaliplatin group(10 mg/kg), hirsutine group(10 mg/kg) and combination group(10 mg/kg hirsutine + oxaliplatin 10 mg/kg). The intervention lasted for 2 weeks. The tumor growth of mice in each group was recorded, and the tumor inhibition rate, thymus index and spleen index were calculated. The proportion of CD4+T cells and CD8+T cell subsets in peripheral blood was detected by flow cytometry, and the CD4+/CD8+ratio was calculated. ELISA was used to measure the serum levels of interleukin(IL)-1β, IL-6 and tumor necrosis factor(TNF)-α. HE staining was used to observe the pathological changes in tumor tissues. Western blot was used to detect the protein expression levels of cleaved Caspase-3, Bcl-2 associated X protein(Bax), B-cell lymphoma-2(Bcl-2)and tumor proliferation antigen(Ki67), proliferating cell nuclear antigen(PCNA) in tumor tissues. Results Compared with the control group, the spleen index, thymus index, CD4+T cell ratio and CD4+/CD8+ratio in peripheral blood of the model group decreased(P<0.05), while the proportion of CD8+T cells in peripheral blood and the levels of TNF-α, IL-1β and IL-6 in serum increased(P<0.05). Compared with the model group, the tumor growth in oxaliplatin group, hirsutine group and combined group was significantly inhibited, the volume of tumor cells decreased, the heteromorphism was smaller, and the nucleosomes decreased less. The inhibitory rate, spleen index, thymus index, CD4+T cell ratio and CD4+/CD8+ratio in peripheral blood of the model group increased(P<0.05), the proportion of CD8+T cells in peripheral blood and the levels of TNF-α, IL-1β and IL-6 in serum decreased(P<0.05), while the protein expression levels of Bcl-2, Ki67 and PCNA in tumor tissues decreased and the cleaved Caspase-3 and Bax levels increased(P<0.05). Among them, the combination group of rhynchophylline and oxaliplatin had the most obvious intervention effect. Conclusion Hirsutine can inhibit the proliferation of tumor cells in gastric cancer bearing mice and promote their apoptosis, and has a more significant effect when combined with oxaliplatin.

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Objective To investigate the biocompatibility of copper-gadolinium nanomaterials and their antibacterial efficacy againstEscherichia coli(E.coli). Methods Copper-gadolinium nanomaterials were prepared using a hydrothermal method, and the samples were characterized and analyzed using instruments such as transmission electron microscopy. The biocompatibility of copper-gadolinium nanomaterials was evaluated through CCK-8 experiments with mouse fibroblast cells(L929). Copper-gadolinium nanomaterials were co-cultured withE.coli, and their antibacterial performance was observed and assessed. Results Transmission electron microscopy and Fourier transform infrared spectroscopy analysis confirmed the synthesis of the copper-gadolinium nanomaterials. The CCK-8 experiment results showed that the material had reliable biocompatibility within a concentration range of 0-2 μg/ml. The antibacterial experiments verified the excellent anti-E.coliactivity of copper-gadolinium, with the antibacterial effect positively correlating with concentration and co-cultivation time. When copper-gadolinium nanomaterials at a concentration of 2 μg/ml were co-cultured withE.colifor 4 hours, the antibacterial rate reached 99.99%. Conclusion The 2 μg/ml copper-gadolinium nanomaterials exhibit reliable biocompatibility and excellent anti-E.colicapabilities.

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Objective To explore the expression and distribution of μ-opioid receptors(MOR) in intracardiac ganglia(ICG) and the effect of its agonists on atrial fibrillation(AF). Methods (1) The chemical anatomical characteristics of ICG in normal SD rats were studied by immunofluorescence single staining. The expression and distribution of MOR in ICG were detected by using immunofluorescence double staining, Western blot, and RT-PCR.(2) Forty male SD rats weighing 250-300 g were randomly divided into 5 groups: normal group, AF model group(AF group), solvent control group(AF-NS group), MOR-specific agonist endomorphin-2(EM2) drug group(AF-EM2 group) and DAMGO drug group(AF-DAMGO group). The AF model was established by tail vein injection of acetylcholine(Ach) and CaCl2, and drug intervention was given during modeling. The duration of AF before and after drug administration was monitored by collecting electrocardiograms. The protein and mRNA expression of MOR in ICG and connexin 43(CX43) in atrial tissue were detected by the method of Western blot and qRT-PCR. Results (1) Peptide nerve fibers positive for substance P and calcitonin gene-related peptide were found in the ICG, along with parasympathetic neurons positive for acetylcholine transferase and sympathetic neurons positive for tyrosine hydroxylase. The mRNA transcripts and protein of MOR were expressed in the atrial posterior wall tissue. MOR immunoreactive products were mainly distributed in the cell bodies of ICG neurons, primarily in parasympathetic and sympathetic neurons.(2) Compared with the Normal group, the expression ofMORmRNA and protein in ICG of the AF rat decreased(P<0.05). Compared with the AF-NS group, the duration of AF was shortened in the AF-EM2 group and AF-DAMGO group, and the expression of CX43 increased(P<0.05). Conclusion MOR is mainly expressed in sympathetic and parasympathetic neurons in ICG. The expression of MOR decreases in AF rats. MOR agonists alleviate AF.

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Objective To establish a method for detecting the duration of the synthesis-phase(Ts) and the total cell-cycle(Tc) based on the dual labeling of 5-ethynyl-2′deoxyuridine(EdU) and 5-Bromodeoxyuridinc(BrdU). Methods HEK293T cells were double labeled with EdU and BrdU and then subjected to immunofluorescence staining. The fluorescence signals were collected using a High-Content Imaging System. The numbers of EdU-positive and BrdU-positive cells in the entire cell population were counted. Based on the cell cycle calculation formula in a previously published article, it was improved to accurately calculate the duration of Ts and Tc of cells. The cell cycle time of human embryonic stem cells was calculated using the established method. Results Based on the kinetic parameters of the EdU/BrdU dual labeling, the Ts of human embryonic stem cells was 10.9 h, and the Tc was 15.3 h. Conclusion The EdU/BrdU dual labeling method can be used to detect the duration of the cell cycle, providing an accurate and efficient method for cell cycle analysis.

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Objective To analyze the spatio-temporal distribution characteristics of pulmonary tuberculosis incidence in Xinjiang, and to provide reference for the prevention and control of pulmonary tuberculosis. Methods The reported incidence numbers of tuberculosis and population data from various counties, cities(prefectures), and districts in Xinjiang from 2010 to 2022 were collected. Based on spatial-temporal scan statistics, standard deviational ellipse analysis, and centroid migration models, the clustering and migration trends of tuberculosis incidence were evaluated. Utilizing Kriging interpolation techniques, an interpolation analysis of the 2022 incidence rate was conducted on an annual scale, with the reported incidence rate in 2018 serving as a temporal control, to identify hotspots of the spatio-temporal distribution. ArcGIS software was employed to visualize the continuous spatial trends of incidence rate changes. Results The annual reported incidence rate of tuberculosis in Xinjiang from 2010 to 2022 varied year by year, with fluctuations and increases prior to 2018, peaking in 2018, and then declining annually thereafter. The spatial distribution of the incidence rate exhibited a trend of initial clustering followed by diffusion, with the centroid of incidence shifting towards the northeast, yet the epicenter of the epidemic remained in Aksu Prefecture. The results of spatiotemporal scan statistics analysis revealed that the three-level aggregated areas of the epidemic encompassed a total of 41 prefectures, counties, and cities, with the tuberculosis incidence risk in the primary and secondary aggregated areas being significantly higher than that in other regions(P<0.01). The Kriging interpolation prediction map suggested that the four prefectures in southern Xinjiang continued to be high-risk regions for tuberculosis(incidence rate>300/100 000). The overall incidence rate in the northern region was relatively low, with the lowest rate observed in Urumqi, radiating outwards. Conclusion The incidence rate of tuberculosis in Xinjiang shows an upward trend before 2018, followed by a year-on-year decrease. The centroid of the incidence rate shifts towards the northeast. From 2010 to 2022, the tuberculosis epidemic in Xinjiang exhibits a notable spatiotemporal clustering, particularly prominent in the southwestern region, where the four prefectures constitute high-risk areas for tuberculosis. The prevention and control efforts of tuberculosis in Xinjiang should prioritize the regions with high tuberculosis incidence, intensifying prevention and control measures as well as policy support.

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Objective To explore the correlation between Diabetes Complication Severity Index(DCSI), C-reactive protein/albumin ratio(CAR) and death in patients with diabetic foot ulcer(DFU) and to clarify their predictive value for all-cause death in DFU patients. Methods Retrospectively analyzed the clinical data of 354 DFU patients who were treated in the Endocrinology Department of the First Affiliated Hospital of Anhui Medical University from July 2019 to December 2022. Based on survival status during follow-up, patients were divided into a survival group(n=268) and a death group(n=86). Univariate and multivariate Cox regression analyses were used to identify risk factors for all-cause death in DFU patients. Receiver operating characteristic(ROC) curves were plotted to evaluate the predictive value of DCSI, CAR, and their combination for all-cause death in DFU patients. Kaplan-Meier curves were used to explore the impact of different DCSI and CAR levels on survival in DFU patients. Results Univariate Cox regression analysis showed that older age, history of hypertension, higher Wagner classification levels, and elevated levels of CRP, Scr, FDP, DCSI score, and CAR were associated with a higher risk of death in DFU patients(P<0.05). Higher levels of HGB, HCT, ALB, or eGFR were associated with a lower risk of death. Patients receiving combined insulin and oral hypoglycemic medication had a lower risk of death compared to those receiving only insulin therapy(P<0.05). Multivariate Cox regression analysis indicated that older age, higher levels of Scr, DCSI, and CAR were independent risk factors for all-cause death in DFU patients, while higher levels of ALB and combined insulin and oral hypoglycemic therapy were protective factors. ROC curve analysis showed that the AUC values for DCSI, CAR, and their combination were 0.652, 0.633, and 0.686, respectively. Kaplan-Meier curve analysis revealed that patients with high DCSI scores(≥4.5) had a lower survival rate compared to those with lower DCSI scores(<4.5). Similarly, patients with high CAR levels(≥0.124) had a lower survival rate compared to those with lower CAR levels(<0.124). Conclusion High levels of DCSI and CAR are independent risk factors for all-cause death in DFU patients. DCSI, CAR, and their combination have predictive value for all-cause mortality in DFU patients.

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Objective To analyze the safety and effectiveness of different types of stents with the treatment of vertebraebasilar artery dissection aneurysms(VBADA). Methods The clinical data of 80 patients with VBADA treated by intravascular intervention in the First Hospital of University of Science and Technology of China(Anhui Provincial Hospital) from February 2018 to November 2023 were retrospectively analyzed. Patients were divided into laser engraved stent group(n=34) and braided stent group(n=46) based on the type of stent used. O′Kelly-Marotta(OKM) grade was used to evaluate the embolization effect of aneurysms in DSA images, and a modified Rankin Scale(mRS) score was used to evaluate the clinical prognosis of patients. The baseline data, aneurysm characteristics, intraoperative and perioperative treatment details and postoperative follow-up details between the two groups were compared. Results There was no significant difference in baseline data, mRS score, ischemic and hemorrhage complications and mortality between the two groups(allP>0.05). Compared with the laser engraved stent group, the mean diameter of aneurysms was larger(P<0.000 1) and the proportion of ruptured aneurysm(P<0.01), parent artery stenosis and beaded vascular lesions(P<0.05) and branch artery dissecting aneurysm were higher(P<0.01) in the braided stent group. Conversely, the proportion of coil-assisted embolization(P<0.000 1) and the immediate aneurysm occlusion rate(OKM C and D)(P<0.000 1) were lower. Finally, 21 patients were obtained by controlling for maximum diameter of aneurysms and whether coil-assisted embolization, the aneurysm occlusion rate half a year later in the braided stent group was higher than that in the laser engraved stent group(P<0.05), but the recurrence rate of postoperative aneurysm was lower than that of laser engraved stent group(P<0.05). It was worth noting that the cure rate and vascular remodeling rate of middle-large size VBADA which the maximum diameter being over 15 mm in the braided stent group reached 72.2%, and the whole effect was ideal. Conclusion Braided stents are relatively safe and effective in the treatment of VBADA and are significantly better than laser engraved stents in reducing VBADA recurrence and remodeling lesion vessels without increasing postoperative complication.

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Objective To analyze the clinical characteristics of patients with abdominal type allergic purpura(HSP), to improve their diagnostic level, and to explore the risk factors for gastrointestinal bleeding in HSP patients. Methods A retrospective analysis was conducted on the clinical manifestations, laboratory data, imaging, endoscopic, and pathological characteristics of 98 patients with abdominal type HSP. Based on the occurrence of gastrointestinal bleeding, 98 patients were divided into a bleeding group and a non-bleeding group, and the risk factors for gastrointestinal bleeding in HSP patients were analyzed. Results Abdominal HSP often presented with abdominal pain, vomiting, vomiting blood, black stools, and bloody stools. Imaging often showed edema and thickening of the duodenum and jejunum, as well as enlargement of surrounding lymph nodes. Under endoscopy, the descending part of the duodenum and jejunum mucosa were commonly congested and edematous with erosion, and ulcers were seen in the distal ileum. Pathology commonly involved acute and chronic inflammation of the mucosa with congestion, edema, and local erosion. Patients with gastrointestinal bleeding had significantly higher levels of white blood cell count(WBC), neutrophil count(NEUT), C-reactive protein(CRP), D-dimer(D-D), and fibrinolytic products(FDP) compared to non-bleeding patients(P<0.05), while levels of red blood cell count(RBC), hemoglobin(HGB), and albumin(ALB) were significantly lower than those of non bleeding patients(P<0.05). Logistic regression analysis showed that decreased ALB and increased FDP were independent risk factors for gastrointestinal bleeding in patients with abdominal HSP(P<0.05). The areas under the ROC curves of ALB and FDP were(AUC=0.877, 95%CI:0.794-0.960,P<0.01) and(AUC=0.806, 95%CI:0.722-0.890,P<0.01), respectively. The maximum value of the Jordan index for ALB was 0.734, with sensitivity and specificity of 89.6% and 83.9%, respectively, and had a critical value of 38.2 g/L. The maximum value of the Jordan index for FDP was 0.577, with sensitivity and specificity of 64.2% and 93.5%, respectively, and had a critical value of 18.14 μg/ml. There was no statistically significant difference in the ROC curves between ALB and FDP. Conclusion For HSP with abdominal pain as the initial symptom, imaging and endoscopic examination are helpful for early diagnosis. Decreased ALB and elevated FDP are independent risk factors for gastrointestinal bleeding in adult patients with abdominal HSP.

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Objective To investigate the expression of the Y chromosome gene lysine-specific demethylase 5D(KDM5D) in male bladder cancer and its effects on cancer progression, thereby explaining gender-related prognostic differences in bladder cancer. Methods Bioinformatics analysis was conducted to identifyKDM5Das a differentially expressed Y chromosome gene with prognostic significance in bladder cancer based on data from the TCGA database. The expression ofKDM5Din bladder cancer cell lines, collected clinical bladder cancer tissues, and adjacent normal epithelial tissues were examined using quantitative Real-time PCR(qPCR), Western blot, immunofluorescence, tumor microenvironment analysis, and bioinformatics pathway analysis. The effects ofKDM5Don bladder cancer cell proliferation and migration were validated through cell experiments followingKDM5Dknockdown and overexpression. Additionally, clinical data onKDM5Dexpression in bladder cancer tissue arrays were statistically analyzed in relation to metastasis and recurrence. Results KDM5Dwas generally downregulated in male bladder cancer but exhibited variable expression in different bladder cancer cell lines and between recurrent and non-recurrent bladder cancers. Overexpression ofKDM5Dinhibited tumor cell proliferation and migration, and lowerKDM5Dexpression was associated with higher recurrence and metastasis rates. Bioinformatics analysis suggested thatKDM5Dmight influence tumor metastasis or recurrence by affecting immune cell aggregation and epithelial-mesenchymal transition. Conclusion KDM5Dis a protective gene specific to male bladder cancer and may be associated with the generally better prognosis observed in male compared to female bladder cancer patients.

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Objective To investigate the levels of cytidine/uridine monophosphate kinase 2(CMPK2) expression in CD4+T cells of systemic lupus erythematosus(SLE) patients and its correlation with clinical indicators. Additionally, to explore whether CMPK2 can induce pyroptosis in CD4+T cells of SLE patients through NLRP3, potentially providing a new target for the diagnosis and treatment of SLE. Methods RT-qPCR and Western blot analyses were used to assess the gene and protein expression levels of CMPK2 in SLE CD4+T cells and healthy controls(HC). Pearson or Spearman correlation analysis was performed to evaluate the relationship between CMPK2 mRNA expression levels and clinical indicators. Subsequently, the expression levels of pyroptosis-related proteins, including NLRP3, apoptosis-associated speck-like protein containing a CARD(ASC), caspase-1, gasdermin D(GSDMD), and the N-terminal domain of GSDMD(GSDMD-N), were examined in SLE CD4+T cells and HC. Furthermore, the protein expression levels of NLRP3, ASC, caspase-1, GSDMD, and GSDMD-N were detected after silencingCMPK2in SLE CD4+T cells. Results CMPK2 expression was significantly elevated in SLE CD4+T cells, exhibiting a positive correlation with SLE disease activity index(SLEDAI), anti-dsDNA antibody, anti-nucleosome antibody, anti-C1q antibody, and a negative correlation with complement C3 and C4 levels. Additionally, the expression levels of pyroptosis-related proteins, including NLRP3, ASC, caspase-1, GSDMD, and GSDMD-N significantly increased in SLE CD4+T cells(P<0.05), Moreover, the levels of cytokines IL-1β and IL-18 in the cell culture supernatants were elevated, and there was a notable increase in the rate of cellular pyroptosis(P<0.05). Silencing CMPK2 led to a reduction in the levels of these markers(P<0.05). Conclusion CMPK2 is highly expressed in SLE CD4+T cells and may serve as a diagnostic marker for SLE. Moreover, it is likely involved in the pathogenesis of SLE by promoting CD4+T cell pyroptosis through NLRP3.

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Objective To explore the clinical application value of neural network technology,and to screen the clinical early warning indicators of severe hand-foot-mouth disease(HFMD) by constructing a back propagation neural network(BPNN) model.Methods The clinical data of children with HFMD admitted to the Department of Infectious Diseases and Pediatrics of the First Affiliated Hospital of Xinxiang Medical University from January2019 to January 2023 were collected.The data were divided into 70% training samples and 30 % test samples using SPSS Modeler 18.0.The BPNN model and Logistic model were constructed to compare and evaluate the prediction accuracy and screening effect of the model.Results The clinical data of 589 children were collected and analyzed,including 324 cases in the mild group and 265 cases in the severe group.The prediction accuracy of the test set(n=178) of BP neural network model and Logistic regression model was 82.02% and 84.83%,respectively.The area under the ROC curve(95% CI) was 0.791(0.749-0.834) and 0.625(0.577-0.674),respectively.Among the predictive variables output by the BPNN model,the top five factors that had the greatest impact on the grouping were highest body temperature,duration of fever,glutamyl transpeptidase,aspartate aminotransferase,and globulin.There were 3 of the top 10 overlaps in the importance of the predictive variables output by the two models,which were the highest body temperature,duration of fever and limb shaking.Conelusion The BPNN model and Logistic regression model perform well in screening and verifying the risk factors of severe hand-foot-mouth disease,but the comprehensive prediction performance of BP neural network model is better.The top five influencing factors of severe HFMD screened by the BPNN model are the highest body temperature,duration of fever,glutamyl transpeptidase,aspartate aminotransferase and globulin.

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Objective To investigate the expression of human cysteine protease inhibitor S(CST4), Helicobacter pylori(HP) antibody typing and human epidermal growth factor receptor 2(HER2) in patients with gastric cancer and its predictive value for lymph node metastasis and TNM staging of gastric cancer. Methods A total of 150 gastric cancer patients admitted to Chaohu Hospital Affiliated to Anhui Medical University for surgery from December 2021 to December 2023 were selected as the study objects. Serum CST4 and HP antibody typing were performed before surgery, and HER2 expression levels in gastric cancer tissues were detected by immunohistochemistry after surgery. The clinicopathological data of gastric cancer patients with different serum CST4, HP antibody types and HER2 expression were compared and the correlation was analyzed. ROC curve was used to compare the predictive value of single and combined detection of serum CST4, HP antibody typing and HER2 expression in lymph node metastasis and TNM staging of gastric cancer. Results Patients with CST4, HPI and HER2-positive expression had deeper tumor invasion and were more likely to develop distant lymph node metastasis, and TNM stages were more likely to occur in Ⅲ and Ⅳ stages(P<0.05). Meanwhile, patients with CST4 positive expression were more likely to have vascular and nerve invasion(P<0.05). In patients with HPI positive expression, tumors were more likely to occur in the cardia and the bottom of the stomach, and the tumor diameter was larger and vascular invasion was more likely(P<0.05), while HER2 was negatively correlated with the degree of tumor differentiation, and the tumors were more likely to occur in high and secondary differentiation(P<0.05). ROC curve analysis showed that the AUC of single and combined detection of serum CST4, HPI and HER2 expression to predict lymph node metastasis of gastric cancer were 0.780, 0.676, 0.611 and 0.872, respectively(P<0.05). The AUC for TNM stage diagnosis of gastric cancer were 0.762, 0.635, 0.613 and 0.801(P<0.05), respectively. The diagnostic efficiency of combined detection was higher than that of single detection. Conclusion The expression of CST4, HPI and HER2 in serum is closely related to the depth of tumor invasion, lymph node metastasis and TNM stage. The expression of CST4, HPI and HER2 in serum can be used as an important predictor of lymph node metastasis and TNM stage in gastric cancer patients, providing a new idea for clinical diagnosis.

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Objective To investigate the changes of microorganisms and metabolites in joint effusion of patients with Rheumatoid arthritis(RA), Osteoarthritis(OA) and Urarthritis(UA). To provide new ideas for the study of the effect of microbiota on the pathogenesis of arthritis. Methods Joint effusion samples were collected from 20 patients with RA, 20 patients with OA, and 20 patients with UA. 16S rRNA gene sequencing and untargeted ultra-high performance Liquid chromatography-mass spectrometry(LC-MS) were used to explore the differences in microorganisms and metabolites among the three groups. Pearson correlation analysis was used to detect the correlation between effusion microbiota and metabolites. Results There were differences in microbial diversity and microbiota composition among the three groups. Combined with VIP>1 from OPLS-DA andP<0.05 from two-tailed Students t-test, 45 differential metabolites(Between RA and OA groups), 38 differential metabolites(Between UA and OA groups) and 16 differential metabolites(Between RA and UA groups), were identified. GO analysis and KEGG pathway analysis showed that the differential metabolic pathways among the three groups were mainly concentrated in citric acid cycle(TCA cycle), nucleotide metabolism, amino acid metabolism and glycolysis pathway. Correlation analysis of joint effusion microbiota and metabolites suggested that bacteria enriched in the three groups of joint effusion, such asPrevotella,Clostridium ruminosus,Prevotellaceae＿UCG-001, were related to many key metabolites such as lysozyme, uric acid, glucose, and L-glutamine. Conclusion This study shows that there are a variety of bacterial flora in joint cavity effusion of RA, OA, and UA patients, and the differential metabolites produced by them are involved in the pathogenesis of the three types of arthritis by affecting a variety of metabolic pathways.

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Dry eye disease is a chronic ocular surface inflammatory disease caused by abnormal tear quality or quantity and decreased tear film stability due to various reasons, and often accompanied by ocular discomfort such as itching, dryness, foreign body sensation, and visual dysfunction, which can seriously affect the patient′s quality of life and visual quality if not intervened in time. With the change of social lifestyles, the increase of environmental pollution and the trend of population aging, dry eye disease has become the most common ocular surface disease besides refractive error. Currently, dry eye disease is widely recognized as a non-infectious immune-related inflammatory disease, but the signaling pathways involved in dry eye disease are poorly understood. Whether dry eye disease is caused by excessive tear evaporation, insufficient tear production, or mucin deficiency, the ocular surface tissues(cornea/conjunctiva) inevitably undergo pathological processes such as aberrant proliferation, squamous epithelial hyperplasia, initiation of corneal damage repair mechanisms, and reduction in the number of conjunctival goblet cells, whereas the Wnt/β-catenin pathway is known to have a wide range of biological functions and plays an important role in cell proliferation, differentiation, and stemness maintenance. Therefore, this review describes the pathogenesis and potential experimental therapeutic options of the Wnt/β-catenin signaling pathway in dry eye disease from this perspective, aiming to provide new targets for the treatment of dry eye disease and achieve the goal of controlling the disease progression from the root.