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Objective To evaluate the change of energy metabolism during cisplatin-induced acute kidney injury. Methods Adult CD-1 male mice were intraperitoneally injected with a single dose of cisplatin(20 mg/kg), and renal function and renal tissue pathology were tested; gene expression was analyzed and signaling pathways were enriched in cisplatin-treated renal tubular epithelial cells using transcriptome; the contents of renal glycolysis and amino acid metabolites were analyzed using liquid chromatography-tandem mass spectrometry(LC-MS/MS). Results Serum urea nitrogen and blood creatinine significantly increased in cisplatin-treated mice. Pathological histology observed swelling and shedding of renal tubular epithelial cells. Transcriptome analysis revealed that 2 632genes were upregulated and 2 799 genes were downregulated in cisplatin-treated HK-2 cells. GO and KEGG analysis showed that differential genes were enriched in energy metabolism. The GSEA analysis results showed that cisplatin caused an upregulation of the oxidative phosphorylation pathway and a downregulation of the glycolysis pathway in renal tubular epithelial cells, further KEGG analysis demonstrated that cisplatin caused changes in the expression of amino acid genes in renal cells. Metabolomics showed that the contents of glycolytic intermediates and several amino acids were altered in the kidney of cisplatin-treated mice. Conclusion Cisplatin-induced acute renal injury is accompanied by modification in renal tubular cell glycolysis and amino acid metabolism.

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Objective To investigate the relationship between circular RNA homeodomain interacting protein kinase 3(circHIPK3) and the activation of rat microglia(RM) cells. Methods In vitro, RM cells were cultured and randomized into normal and oxygen-glucose deprivation/reperfusion(OGD/R) groups, and the expression level of circHIPK3 in each group was detected by RT-qPCR. The circHIPK3 lentiviral vector with puromycin resistance was constructed, and the overexpression(OE) group and negative control(NC) group were set up. The optimal multiplicity of infection(MOI) for RM cells was determined based on fluorescence expression, and puromycin was used to screen RM cells stably expressing circHIPK3. The cells of OE and NC groups were treated with OGD/R, and the expression levels of ionized calcium binding adaptor molecule 1(Iba-1) and eukaryotic tumor necrosis factor receptor superfamily(CD40) were detected by Western blot. The circHIPK3 translational protein potential was analyzed by the circRNAdb database, while the potential binding microRNAs on circHIPK3 were predicted by circBank and Starbase databases. Results OGD/R down-regulated circHIPK3 in RM cells(P<0.000 1). The sequencing results were accurate and the lentiviral vector of circHIPK3 was constructed successfully. The optimal MOI of RM cells was 80, puromycin at a concentration of 2 μg/ml was used to screen RM cell lines stably expressing circHIPK3. RT-qPCR results showed that the expression level of circHIPK3 was significantly higher in the OE group compared with the NC group(P<0.01). Western blot results revealed that the expression levels of Iba-1 and CD40 in the OE group were markedly lower than those in the NC group(P<0.05). Protein translation analysis showed that circHIPK3 encoded a polypeptide of 404 amino acids with two internal ribosome entry sites(IRES) and an open reading frame(ORF). Database analysis uncovered that circHIPK3 could target eight specific miRNAs, namely hsa-miR-3529-5p, hsa-miR-379-5p, hsa-miR-506-3p, hsa-miR-33, hsa-miR-450b-5p, hsa-miR-551b-3p, hsa-miR-193, and hsa-miR-508-3p. Conclusion The overexpression of circHIPK3 effectively suppresses OGD/R-induced activation of RM cells. It has the potential to encode peptides and may act as a miRNA sponge. These findings provide a foundation for further study of circHIPK3 functions.

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Objective To investigate the effects of DNA demethylation drugs combined with histone deacetylase inhibitors on fragile X mental retardation 1 neighbor protein(FMR1NB) expression and its promoter methylation in human oral cancer cells and try to find a strategy of weakening the heterogeneity of FMR1NB expression. Methods Human oral cancer cell lines Cal27 and SCC-9 were treated with decitabine(DAC), an inhibitor of DNA methyltransferase, combined with trichostatin A(TSA) and valproic acid(VPA), inhibitors of histone deacetylase. Then reverse transcription-polymerase chain reaction(RT-PCR), quantitative real-time PCR(qRT-PCR)and Western blot were used to detect the expression of FMR1NB and pyrosequencing was used to detect the methylation of FMR1NB promoter. Results Compared with the blank control group, DAC and its combination with TSA and VPA significantly induced the expression of FMR1NB mRNA and protein in Cal27 and SCC-9 cells. Compared with DAC alone group, FMR1NB mRNA expression of each DAC-combined drug groups significantly increased, but FMR1NB protein did not significantly change in Cal27 cells; for SCC-9 cells, except for DAC+TSA group, the mRNA and protein levels of FMR1NB significantly increased in all other groups. In addition, there was no significant difference in the expression of FMR1NB mRNA and protein between the three-combined drugs group and two-combined drugs groups. Further methylation assay showed that the methylation level of the overall FMR1NB promoter and its each CpG site measured were reduced to varying degrees in all treatment groups except for three-combination drug group of SCC-9. Conclusion DAC and its combination with TSA and VPA can enhance the expression of FMR1NB by mediating the demethylation of FMR1NB promoter, wherein the enhanced expression effect of the combination of the two drugs is stronger, suggesting that they have the potential to weaken the heterogeneity of FMR1NB expression and improve the immunotherapy effect of oral cancer.

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Objective To investigate whether norepinephrine(NE) regulates the oxidative stress in human endometrial epithelial cells(hEECs) by activating nuclear factor E2-related factor 2(Nrf2)/heme oxygenase-1(HO-1) signal pathway. Methods Cultured hEECs were used. The expression of α and β adrenergic receptors was detected by reverse transcription-polymerase chain reaction(RT-PCR). Cell counting kit-8(CCK-8) assay was applied to test the effect of NE on cell viability, then the cells were divided into Control group and NE treatment group, and the appropriate concentrations were chosen. The expression of tight junction proteins Occludin and zona occludens-1(ZO-1), apoptosis-related proteins apoptosis-related protein B-cell lymphoma-2 protein(Bcl-2) and Bcl-2 associated X protein(Bax), antioxidant proteins Nrf2 and HO-1 were examined by Western blot. The apoptosis was detected by flow cytometry. The malonaldehyde(MDA) and superoxide dismutase(SOD) in the cell culture medium were detected by enzyme-linked immunosorbent assays kit(ELISA). Results The mRNA expression of α1 a, α1 b, α2 a, α2 b, α2 c, β1, β3 was detected in the hEECs. After the NE treatment, no significant change in cell viability was observed in low concentration(5 μmol/L and 10 μmol/L) groups, while 15 μmol/L and 20 μmol/L NE treatments for 6 h or 24 h promoted the cell viability significantly. The expression of ZO-1 and Occludin increased significantly in 15 μmol/L group after 24 h treatment, the expression of ZO-1 decreased in 6 h treatment group, significant down regulation was observed after 15 μmol/L NE application, the expression of Occludin increased in 6 h group. The cell apoptosis increased compared with the control group after NE stimulation, especially a significantly increase in 6 h 15 μmol/L group was detected, while the fall in increased apoptosis was observed after 24 h treatment. The ration of Bcl-2/Bax>1. The expression of Nrf2 and HO-1 was elevated by NE. There was no obvious change in MDA level while significant elevation in SOD was detected in cell culture medium. Conclusion Nrf2/HO-1 signal is activated after application of NE to the hEECs, which may responsible for the upregulation of SOD, antioxidant and anti-apoptotic effect in the hEECs.

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Objective To explore and optimize thein vitroprimary culture method of astrocytes in neonatal mouse cerebral cortex, which provides a better solution for thein vitroculture of astrocytes. Methods In order to optimize thein vitroculture method of mouse cerebral cortex astrocytes, 3-day-old C57BL/6J mouse cerebral cortex tissues were taken, meninges and blood vessels were removed, digested by pancreatic enzymes and centrifuged, and high-glucose dulbecco′s modified eagle medium(DMEM) was added to form cell suspension, which was purified by differential adhesion method, cross hand method and constant temperature shaking method. The cells were inoculated in poly-D-lysine-coated culture bottles with different culture densities, and the purity of astrocytes was determined by morphological observation and immunofluorescence staining. Results The cells were inoculated at a density of 5×106cells per bottle with good effect and high activity. The purity of astrocytes reached 99% by using high sugar DMEM medium combined with differential adhesion method, cross hand method and constant temperature shaking method. Conclusion The primary culture method of astrocytes in mouse cerebral cortex is successfully established and optimized.

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Objective To investigate the effects of cynomorium songaricum extract on cognitive dysfunction of Alzheimer disease(AD) model mice based on network pharmacology and animal experiments. Methods Network pharmacology was used to predict the related targets and signal pathways of the extract of cynomorium songaricum to improve AD. Senescence accelerated mice P8(SAMP8) were selected as the model of AD. Based on the results of the preliminary experiment, 0.17 g/(kg·d) was selected as the optimal dosage for the extract of cynomorium songaricum. The extract of cynomorium songaricum [0.17 g/(kg·d), Donepezil hydrochloride [2.0 mg/(kg·d)] and normal saline were given orally for 28 days according to the groups. Morris water maze evaluated the learning and cognitive function of animals. The number of neurons in cornu ammonis 1(CA1) of hippocampus was observed by Nissl staining. The expression of recombinant Beclin 1(Beclin-1), Sequestosome 1(p62), light chain 3(LC-3) protein was detected by immunohistochemical method. The protein expression levels of phosphoinositide 3-kinase(PI3K), protein kinase B(AKT) and glycogen synthase kinase3β(GSK-3β) in the hippocampus of mice in each group were detected by Western blot. Results Based on the network pharmacology study, it was predicted that the biological mechanism of cynomorium songaricum to improve AD might be the regulation of autophagy, and the possible signaling pathway was PI3K/AKT/GSK-3β. The results of animal experiments showed that the extract of cynomorium songaricum could improve the spatial memory learning ability of AD model mice, improve the damage of hippocampal neurons, significantly increase the number of neurons, and increase the expression levels of PI3K, p-AKT/AKT, p-GSK-3β/GSK-3β, Beclin-1 and LC3 in the hippocampus of mice. The expression level of p62 decreased. There was no significant difference between male and female mice during the experiment. Conclusion The extract may improve the cognitive dysfunction of male and female AD models by activating autophagy mediated by PI3K-AKT-GSK-3β signaling pathway, and there is no significant gender difference in the effect.

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Objective To search for new biomarkers to predict prognosis in colorectal cancer(CRC) patients. Methods A prognostic model was developed for colorectal cancer with immune-related genes from the cancer genome atlas(TCGA) database using one-way Cox regression analysis and least absolute shrinkage and selection operator(LASSO) regression analysis. Moreover, the immune infiltration characteristics of patients in high and low risk groups was compared by sstimation of stromal and immune cells in malignant tumor tissues using expression data(ESTIMATE) and cell-type identification by estimating relative subsets of RNA transcripts(CIBERSORT). In addition, the expression levels of immune checkpoints were analyzed in patients from different risk groups. The sensitivity of patients in the two risk groups to chemotherapeutic agents was also compared based on genomics of drug sensitivity in cancer(GDSC). Results It was found that the prognostic model constructed based on immune genes could better predict the overall survival(OS) of CRC patients, and the results showed area under curve(AUC) values of 0.764(95%CI:0.751-0.793), 0.773(95%CI:0.761-0.779), and 0.760(95%CI:0.742-0.774) for 1-, 3-, and 5-year OS, respectively. Patients in the low-risk group had higher expression levels of immune checkpoints and more abundant immune cells such as T cells(P<0.001), dendritic cells(P<0.001), macrophages(P<0.001), neutrophils(P<0.001). Patients in the high-risk group might be more sensitive to some chemotherapeutic agents such as axitinib, imatinib, methotrexate, pazopanib, rapamycin, sunitinib and tasigarnib. Conclusion A prognostic model based on 19 immune genes was effective in predicting the prognosis of CRC patients. The number and activity of immune cells in the immune microenvironment in different patients may be an important factor influencing their response to immunocheck inhibitors and chemotherapeutic agents.

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Objective To investigate the protective effects and mechanisms of soybean phospholipid powder on nerve cellsin vitroand rats neural tissues. Methods In the cell experiments, the cytotoxicity of soybean phospholipid powder with different concentrations on mouse microglia cells(BV2) and rat adrenal pheochromocytoma(PC12) cells was observed by cell counting kit-8(CCK-8) assay. The effect of soybean phospholipid powder on the NO level of BV2 cells was analyzed by NO determination experiment, and the synaptic growth of PC12 cells was observed under the microscope. In the animal experiment, the cognitive dysfunction of rat was simulated by scopolamine rat model. Then the learning and memory abilities of rat were tested by Morris water maze experiment; hippocampal tissue morphology and nerve cell density of scopolamine model mice were observed by hematoxylin-eosin staining(HE) staining. Results Soybean phospholipid powder had no obvious cytotoxicity on BV2 cells and PC12 cells within the concentration of 1 000 μg/ml. Compared with the control group, the NO secretion of BV2 cells pretreated with soybean phospholipid powder significantly decreased(P<0.01), and the neuronal synapse growth of PC12 cells significantly increased(P<0.01). In comparison to the model group, soybean phospholipid powder significantly improved the learning and memory ability of scopolamine model rats(P<0.05), reduced the neuronal damage in dentate gyrus(DG), cornu ammonis3(CA3), cornu ammonis1(CA1) areas of hippocampus, and increased the density of nerve cells(P<0.001). Conclusion Soybean phospholipid powder can play a neuroprotective role by reducing neuroinflammation and promoting neuronal synapse growth at the cellular level, and improve the learning and memory ability of rats with cognitive impairment, reduce hippocampal tissue damage.

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Objective To investigate effect of salidroside on the function and activation of rheumatoid arthritis fibroblast-like synoviocyte(HFLS-RA) by regulating the miR-20a-5p/tissue inhibitor of metalloproteinase-2(TIMP2) axis. Methods HFLS-RA cells were used as the research object. HFLS-RA cells were separated into control group, tumor necrosis factor-α(TNF-α) group, salidroside group, inhibitor NC group, miR-20a-5p inhibitor group, salidroside+mimic NC group, and salidroside+miR-20a-5p mimic group. qRT-PCR was applied to detect the expression of miR-20a-5p in HFLS-RA cells; enzyme-linked immunosorbent assay(ELISA) was applied to detect the levels of interleukin-1β(IL-1β) and IL-6 in the supernatant of HFLS-RA cells; cell counting kit-8(CCK-8) method and 5-ethynyl-2 ′-deoxyuridine(EdU) staining were applied to detect HFLS-RA cell proliferation; scratch experiment was applied to detect HFLS-RA cell migration; Western blot was applied to detect the expression of TIMP2, CyclinD1, and matrix metalloproteinase(MMP)-9 proteins in HFLS-RA cells; double lucifer-ase was applied to verify the relationship between miR-20a-5p and TIMP2. Results Compared with the control group, the expression of miR-20a-5p, the levels of IL-1β and IL-6, OD450value, EdU positive cell rate, scratch healing rate, and the expression of CyclinD1 and MMP-9 proteins in the TNF-α group increased, the expression ofTIMP2 protein decreased(P<0. 05); compared with the TNF-α group, the expression of miR-20a-5p, the levelsof IL-1β and IL-6, OD450value, EdU positive cell rate, scratch healing rate, and CyclinD1 and MMP-9 proteins expression decreased, the expression of TIMP2 protein increased in salidroside group(P<0. 05); compared withthe TNF-α group and inhibitor NC group, the expression of miR-20a-5p, the levels of IL-1β and IL-6, OD450value, EdU positive cell rate, scratch healing rate, and the expression of CyclinD1 and MMP-9 proteins in the miR-20a-5p inhibitor group decreased, the expression of TIMP2 protein increased(P<0. 05); compared with the sali-droside group and the salidroside + mimic NC group, the expression of miR-20a-5p, the levels of IL-1β and IL-6,OD450value, EdU positive cell rate, scratch healing rate, and the expression of CyclinD1 and MMP-9 proteins in the salidroside + miR-20a-5p mimic group increased, the expression of TIMP2 protein decreased(P<0. 05).There was a targeted regulatory relationship between miR-20a-5p and TIMP2.Conclusion Salidroside may inhibit TNF-α-induced HFLS-RA cell proliferation, migration and inflammatory response by regulating miR-20a-5p/TIMP2.

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Objective To explore the role of secreted frizzled-related protein 3(sFRP3), a regulator of the Wnt signaling pathway, in the activation and proliferation of murine cardiac fibroblasts(CFs). Methods Neonatal mice aged 1-3 days were obtained for surgical procedures to collect heart tissues. After digestion, CFs were isolated and cultured. Transforming growth factor-beta 1(TGF-β1) stimulation was used to induce activation and proliferation in CFs after they adhered to the culture dish. Once the model was confirmed, experimental and control groups were transfected with sFRP3 overexpression plasmids and empty plasmids for 24-48 hours. Expression levels of sFRP3, Periostin(POSTN), Type Ⅰ collagen(Collagen Ⅰ), and proliferating cell nuclear antigen(PCNA) were assessed at the molecular level using Western blot and qRT-PCR. Changes in cell proliferation capacity were examined using MTT, CCK-8, and EdU staining methods. Results In the TGF-β1-induced activation and proliferation model of CFs, compared to the control group, the model group exhibited decreased expression of sFRP3 protein and mRNA, while the expression of activation and proliferation-related proteins PCNA, POSTN, and Collagen Ⅰ was upregulated. Furthermore, in CFs overexpressing sFRP3 through plasmid transfection, the protein and mRNA expression of PCNA, POSTN, and Collagen Ⅰ decreased compared to the empty vector group. MTT, CCK-8, and EdU experiments indicated a significant decrease in the proliferative activity of CFs in the sFRP3 overexpression group compared to the empty vector group. Conclusion Overexpression of sFRP3 markedly inhibits the activation and proliferation of CFs, suggesting that sFRP3 may be a key gene involved in the regulation of CF activation and proliferation.

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Objective To investigate the improvement of endoplasmic reticulum stress mediated by microRNA(miR)-34a-5p/transmembrane fusion protein 1(Notch1) signaling axis on hypoxia/reoxygenation(H/R) human cardiomyocytes. Methods Human cardiomyocytes were randomly divided into Control group, H/R group, mimic NC group, mimic group, inhibitor NC group and inhibitor group. Except the Control group, H/R injury model was established in other groups. The expression levels of miR-34a-5p and Notch1 were detected by quantitative real-time polymerase chain reaction(qRT-PCR), cell survival rate was detected by thiazolyl blue(MTT), cell apoptosis rate was detected by flow cytometry, and the targeting relationship between miR-34a-5p and Notch1 was detected by dual luciferase gene reporting method. The expressions of transcriptional activator 6(ATF6), inositol demand enzyme 1(IRE 1), protein kinase-like endoplasmic reticulum kinase(PERK) and glucose regulatory protein 78(GRP78) were detected by Western blot.Results miR-34a-5p targeted Notch1(P<0. 05); compared with Control group, the expression level of miR-34a-5p, apoptosis rate and protein expressions of ATF6, IRE1, PERK and GRP78 in H/R group increased, while the cell survival rate and Notch1 mRNA and protein expressions decreased(P<0. 05). Compared with H/R group and mimic NC group, miR-34a-5p expression, apoptosis rate, and proteinexpressions of ATF6, IRE1, PERK and GRP78 in mimic group increased, while cell survival rate and Notch1 mRNA and protein expressions decreased(P<0. 05). Compared with H/R group and inhibitor NC group, the expression of miR-34a-5p, apoptosis rate and protein expressions of ATF6, IRE1, PERK and GRP78 decreased in inhibitor group, while cell survival rate and Notch1 mRNA and protein expressions increased(P<0. 05).Conclusion miR-34a-5p can inhibit the apoptosis of H/R human cardiomyocytes, possibly through the targeted inhibition of Notch1-mediated endoplasmic reticulum stress.

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Objective To investigate the mechanism by which Erastin affects ferroptosis in lung fibroblasts. Methods Mouse lung fibroblasts(C57/B6-L) were treated with varying concentrations of the iron death inducer Erastin. Cell viability was assessed using the cell counting Kit-8(CCK-8) assay. Oxidative stress levels were visualized using a fluorescence microscope, and the expression of proteins related to the mitogen-activated protein kinase(MAPK) signaling pathway was analyzed using Western blot. Additionally, the p38 and extracellular regulated protein kinase(ERK) inhibitors SB203580 and U0126 were employed to further elucidate the mechanism by which Erastin induces iron death in lung fibroblasts. Results At a concentration of 100 μmol/L, Erastin effectively induced ferroptosis in lung fibroblasts, leading to an upregulation of oxidative stress. Furthermore, the phosphorylation levels of p38 and ERK proteins in the MAPK pathway were elevated(P<0.05). The addition of SB203580 and U0126 inhibitors resulted in a significant reduction in oxidative stress levels and a notable increased in cell activity in lung fibroblasts(P<0.05). Conclusion It can be concluded that Erastin induces ferroptosis in lung fibroblasts, potentially through the mediation of oxidative stressviathe MAPK signaling pathway.

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Objective To explore the effect ofadeno-associated virussense transfection up-regulating the expression level of the growth and differential factor 11(GDF11)in vivoon aortic injury in type 2 diabetic mellitus rats(T2DM). Methods Nine-week-old male SD rats were randomly selected to establish a T2DM model by using high-sugar and high-fat chow plus small-dose streptozotocin(STZ) combined induction. Both normal rats and diabetic model rats were randomly divided into five groups: blank control group(Control group), negative virus control group(NC group), GDF11adeno-associated virusgroup(GDF11 group), diabetic group(DM group), and diabetic + GDF11adeno-associated virusgroup(DM+GDF11 group). After 8 weeks of feeding, the serum concentrations of insulin(INS), advanced glycosylation end products(AGES), recombinant growth transforming factor 11(GDF11), total cholesterol(T-CHO), triglycerides(TG), low-density lipoproteins(LDL-C), high-density lipoproteins(HDL-C), asymmetric dimethylarginine(ADMA), and malondialdehyde(MDA) were assayed in the rats; periodic acid-schiff stain(PAS stain) was used to observe the sites of glycogen deposition, and hematoxylin-eosin staining(HE) was used to observe vascular damage. Scanning electron microscopy was used to observe the damage of vascular endothelial cells and vascular elastic fibers, and protein blotting and immunohistochemistry were used to detect the expression levels of vascular injury-related proteins. Protein blotting and immunohistochemistry were used to detect the expression levels of vascular injury-related proteins. Results The biochemical indexes showed that the serum concentrations of AGES, T-CHO, TG, LDL-C and MDA were higher in the DM group than those in the Control group(P<0.05), the concentrations of INS, GDF11, HDL-C and ADMA were significantly lower than those in the Control group(P<0.05), and the concentrations of AGES and HDL-C were not significantly lower in the DM+GDF11 group compared with the DM group(P<0.05). HDL-C was not significantly different from the DM group, and several other data were improved(P<0.05). Pathological staining suggested that PAS staining in the DM group suggested that glycogen particles deposited in the endothelium and subendothelium of the aorta, HE staining observed thickening of the aortic mesentery, endothelial cells and elastic fibers broke off in an irregular alignment, and electron microscopy observed endothelial damage in the vasculature and elastic fibers broke off in the DM group, and these changes attenuated in the DM+GDF11 group. Protein blotting and immunohistochemistry indicated that the expression of endothelial cell-associated proteins decreased in the DM group(P<0.05), and mesenchymal markers elevated in the DM group(P<0.05), these proteins were regressed in the DM+GDF11 group, and the difference was statistically significant(P<0.05). Conclusion Increasing the expression level of GDF11in vivocan improve aortic vascular injury caused by diabetes mellitus, which is inferred that it may be related to the inhibition of endothelial mesenchymal transition to protect the function of vascular endothelial cells and thus improve vascular injury.

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Objective To study the molecular mechanism of fat mass and obesity-associated protein(FTO) regulating hepatocellular carcinoma(HCC). Methods HepG2 cells of knock-down FTO were constructed, HepG2 cells of knock-down FTO and HepG2 cells were collected, and high-throughput sequencing was performed using Illumina Hiseq platform to screen the gene expression differences between the two groups. Through GO and KEGG enrichment analysis of these differential genes, FTO regulatory pathways were studied and downstream target genes of FTO were screened. The role of FTO downstream target gene in HCC was revealed by bioinformatic analysis and cell experiments. Results Transcriptome sequencing showed that 386 genes were differentially expressed between HepG2 cells of knock-down FTO and HepG2 cells, and they were involved in biological processes such as response to interferon-gamma. The expression of IFIT2, one of the most responsive interferon-stimulating genes, was up-regulated after FTO knockdown. Potential m6A methylation occurred at multiple sites of IFIT2. The survival of HCC patients with high expression of IFIT2 was significantly prolonged, and knock-down of IFIT2 promoted the growth and migration of HepG2 cells. Conclusion FTO may regulate IFIT2 by mediating m6A, and further promote the occurrence and development of HCC.

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Objective To investigate the mechanism of micellar curcumol(MC) regulating the immune microenvironment of ovarian cancer by promoting the polarization of M2-type macrophages to M1-type in ovarian cancer ascites. Methods(1) After the mice were divided into groups, a mouse ovarian cancer ascites model was constructed by using the mouse ovarian cancer cell line ID8. Then weight changes were observed, tumor tissue and ascites were collected. The expression of CD86 and CD206 on macrophages of the tumor tissue and ascites was detected by flow cytometry. The expression of protein kinase B(PKB/Akt)/mammalian target of rapamycin(mTOR) was detected by Western blot.(2) A human monocytic leukemia cell line(THP-1) was induced to transform into M2 macrophage(THP-1 M2 macrophage) in vitro, and then treated with 10 μg/ml MC. The apoptosis was detected by flow cytometry. The mRNA levels of macrophage mannose receptor(CD206), transforming growth factor-β(TGF-β), interleukin(IL)-1β and tumor necrosis factor-α(TNF-α) were detected by qRT-PCR. The expression of CD86 and CD206 was detected by flow cytometry, and Akt/mTOR expression and phosphorylation was detected by Western blot.Results(1)In vitrostudy showed that the average body weight of the MC group was lower than that of the control group. Compared with the control group, CD206 expression of macrophages decreased in tumor tissue and ascites in the MC group, while the expression of CD86 increased. The Akt and mTOR phosphorylation level of macrophages in the MC group′s ascites was lower than that in control group.(2)In vivostudy showed that there was no difference in apoptosis rate among the groups detected by flow cytometry. The mRNA expression level of CD206,TGF-β and the protein expression level of CD206 in MC group were significantly lower than those in the control group, while the mRNA expression of IL-1β, TNF-α and the protein expression level of CD86 were significantly higher than those in the control group. Compared with the control group, the phosphorylation level of Akt and mTOR in the MC group decreased.Conclusion MC promotes M1 polarization of macrophages in ascites to regulate the immune microenvironment of ovarian cancer, which may be related to the Akt/mTOR pathway.

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Objective To investigate the pathological role of detect protein kinase C(PKC)/transient receptor potential vanilloid subtype 1(TRPV1) pathway in trigeminal neuralgia(TN) in rats. Methods The infraorbital nerve-chronic constriction injury(ION-CCI) was used to establish a rat model of TN. The rats were randomly divided into Sham group, CCI group, CCI+DMSO group and CCI+GF109203X(a PKC inhibitor) group. The mechanical pain threshold of the rats was measured using a Von Frey brush. qRT-PCR and Western blot were used to detect PKC and TRPV1 in the trigeminal ganglion(TG). HE staining was used to observe the pathological changes of TG. Results The mechanical pain threshold significantly decreased(P<0.05), and the expression of phosphorylated PKC(p-PKC) and TRPV1 in TG significantly increased in the CCI group(P<0.05). Histopathological results showed that compared with the Sham group, the CCI group observed significant changes in TG such as increased inflammatory cell infiltration and nerve cell swelling. Injection of GF109203X effectively reduced the phosphorylation of PKC and the expression of TRPV1 in the TG of rats, and the mechanical pain threshold of the rats increased(P<0.05). Under the light microscope, cell swelling and inflammatory cells in the TG were reduced. Conclusion PKC/TRPV1 pathway may be involved in trigeminal neuralgia in rats.

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Objective To investigate the effects of hederagenin(HDG) on proliferation, migration, invasion and apoptosis of glioblastoma(GBM) cells and involved mechanism. Methods Human GBM cell lines U87, U251 and human brain glial cell line(HEB) were selected as the study subjects, and HDG 0 μmol/L(or 0 mg/kg) was used as the control group. MTT, EdU staining and cell plate cloning were used to detect the effect of HDG on the proliferation of GBM cells. Trypan blue staining was used to detect GBM cell death affected by HDG. The effects of HDG on migration and invasion of GBM cells were detected by cell scratch and Transwell assay. To analyze the effects of HDG on apoptosis of GBM cells, apoptosis-related proteins Bcl-2, Bax, p53 and cleaved caspase-3 were detected by Western blot. Mitochondrial potential change was detected by JC-10 staining, and apoptotic cell count was displayed by Annexin V-FITC staining. The effect of HDG on tumor bearing in GBM was analyzed by xenotransplantation in BALB/C mice.Results Compared with the control group(HDG 0 μmol/L), HDG significantly inhibited the proliferation, migration and invasion of U87 and U251 cells, and they were dependent on the use dose of HDG. Trypan blue staining showed that HDG obviously increased death number of GBM cells. The mitochondrial potential of GBM cells was remarkedly decreased, the number of apoptotic GBM cells obviously increased, the expressions of apoptosis-related proteins p53, Bax, cleaved-caspase3 were up-regulated and Bcl-2 was down-regulated by HDG in U87 and U251 cells. HDG significantly inhibited the size of subcutaneous GBM, the Ki67 positive rate of GBM cells and caused a large number of GBM cells to die in BALB/C mice. HDG had no obvious toxic effect on human HEB cells and the liver of tumor-bearing mice.Conclusion HDG can significantly inhibit the proliferation, migration and invasion of GBM cells and induce the apoptosis of them. The mechanism of HDG induced apoptosis of GBM cells may be through mitochondrial damage and regulation of p53 and Bcl-2/Bax expression.

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Objective To investigate the associations of the single nucleotide polymorphism(SNP) rs2304733 in TEA domain transcription factor 1(TEAD1), rs7135838 and rs1990330 in TEA domain transcription factor 4(TEAD4) genes with the risk of non-cardia gastric carcinogenesis. Methods Enzyme linked immunosorbent assay(ELISA) was used to detect specific antibodies againstHelicobacter pylori(Hp)in serum samples of the normal control group. 470 normal controls were divided into Hp infection negative group(n=223) and positive group(n=247) based on antibody titers. In the 450 non-cardia gastric cancer cases and 470 controls, polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) was used to genotype the each SNP locus. The unconditional Logistic regression method was used to evaluate the associations between each SNP locus and the risk of non-cardia gastric carcinogenesis. Results The SNPs of TEAD1 and TEAD4 were not associated with Hp infection. TEAD1 rs2304733 was associated with the risk of non-cardia gastric cancer. Compared with the carriers of TT genotype, the carries of CT and CC genotypes had an increased risk of non-cardia gastric cancer(CTvsTT:OR=2.321, 95%CI: 1.690-3.188; CCvsTT:OR=5.140, 95%CI: 1.080-24.463). TEAD4 rs1990330 was associated with the risk of non-cardia gastric cancer. Compared with the carriers of GG genotype, those with GT genotype had an increased risk of non-cardia gastric cancer(OR=2.405, 95%CI: 1.480-3.908). TEAD4 rs7135838 was not associated with the risk of non-cardia gastric cancer. TEAD1 rs2304733, TEAD4 rs7135838 and rs1990330 had interaction effects on the risk of non-cardia gastric cancer(P<0.05). Conclusion In Baotou Han population, TEAD1 rs2304733 and TEAD4 rs1990330 do not play a major role in Hp infection, but may play a role in the risk of non-cardia gastric cancer. TEAD4 rs7135838 may not play a major role in the risk of Hp infection and non-cardia gastric cancer. TEAD1 rs2304733 and TEAD4 rs1990330 have the strongest synergistic effect on the risk of non-cardia gastric cancer, which is the best interaction model.

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Objective To explore the value of18F-FDG PET/CT dynamic imaging in the diagnosis of primary liver cancer and liver metastases. Methods In this study, the data of 94 patients with hepatic malignant lesions [hepatocellular carcinoma group(25 cases), cholangiocellular carcinoma group(27 cases), and liver metastases group(42 cases)] imaged by whole-body dynamic18F-FDG PET/CT were used as the research subjects, and the methods of(ANOVA) and subjects′ working characteristic curves(ROC) were applied to examine the patients with liver malignant tumours of different pathological types SUVmax, MRFDG, TBR and dmaxwere statistically analysed as imaging features. Results Among 94 patients, the differences in SUVmax, MRFDG, TBRSUV (max ) and TBRMRFDGwere statistically significant in the hepatocellular carcinoma, cholangiocellular carcinoma and hepatic metastasis groups(SUVmax: F=48.773, P<0.001; MRFDG: F=26.334, P<0.001; TBRSUV (max ): F=41.314, P<0.001; TBRMRFDG: F=20.821, P<0.001).The AUCs for the differential diagnosis of primary hepatocellular carcinoma and hepatic metastases for SUVmax, MRFDG, TBRSUV (max ), and TBRMRFDGwere 0.836, 0.851, 0.827, and 0.847, respectively.There was a positive correlation between SUVmax, MRFDG, and the hepatic malignant lesions′ There was a positive correlation between SUVmax, MRFDGand the maximum diameter dmax(SUVmax: r=0.4, P<0.05; MRFDG: r=0.2, P<0.05), and the difference was statistically significant. Conclusion18F-FDG PET/CT dynamic imaging has good differential diagnostic efficacy in different pathological types of liver malignant tumors. There is a positive correlation between SUVmax, MRFDGand dmaxin different pathological types of liver malignant tumors.

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Objective To explore the correlation between serum fibroblast growth factor-23(FGF23) concentration and heart failure and all-cause death in patients with end-stage renal disease(ESRD).Methods The prospective cohort study design was used in the present study. The ESRD patients who were admitted to the department of nephropathy in the Hospital and without heart failure symptoms were recruited in this study. The data of patients was collected through baseline questionnaires, physical examinations, echocardiography, and laboratory examinations.The serum FGF23 levels were measured by enzyme-linked immunosorbent assay(ELISA). The follow-up time was2 years. The onset of heart failure(ACC/AHA stage C-D) and all-cause death were composite endpoint events.The Cox proportional risk model was used to explore the risk factors of outcome events. Through subgroup analyses and interaction analyses, further exploration was conducted to determine whether there was heterogeneity in the association between FGF23 and outcome events in different subgroups.Results Ultimately, 107 ESRD patients were included in this study, with an average age of(52. 00 ± 12. 51) years. There were 39 males(36. 45%), and the median follow-up time was 23 months(21, 25 months). There were 32(29. 9%) outcome events, of which 22(20. 6%) onset of heart failure and 10(9. 3%) all-cause of deaths. The results of this study showed that the concentration of FGF23 in the outcome event group was significantly higher than that in the non-event group [(4. 40 ±1. 16) pmol/mlvs(3. 85 ± 0. 82) pmol/ml,P<0. 05]. The Cox proportional risk model showed that the elevated FGF23 was associated with increased risk of the composite endpoint events in ESRD patients(HR= 1. 730, 95%CI: 1. 164-2. 570,P= 0. 007). Subgroup analyses showed that there was an interactive effect between FGF23levels and gender on the risk of cardiovascular outcome events. Especially in male ESRD patients, the increased FGF23 level was correlated with a higher risk of cardiovascular events(P-interaction<0. 05).Conclusion Elevated serum FGF23 is an independent risk factor for the onset of heart failure and all-cause of mortality in ESRD patients,especially in male patients.

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Objective To compare the ultrasound signs of symptomatic joint lesions in rheumatoid arthritis(RA) and gouty arthritis(GA), musculoskeletal ultrasound(MSUS) was utilized. Methods A retrospective analysis was performed for 85 hospitalized patients with RA and 55 hospitalized patients with GA in the same period, and the differences in general data, diseased joints and ultrasound signs between the two groups were compared. Results The gender, age and diseased joints of the two groups were statistically significant(allP<0.05). The detection rate of knee joint lesions was the highest; the RA group had high sensitivity, high specificity of meniscal injury, and high diagnostic efficiency of bone erosion, while the diagnostic performance of the three combined ultrasound signs of punctate strong echo, double track sign and tophi in the GA group was higher than that of any individual diagnosis, and the sensitivity and specificity were also higher. The course of disease in the RA group was positively correlated with bone erosion(P<0.05), and the course in the GA group was positively correlated with tophi(P<0.05). Conclusion The ultrasound signs of RA and GA are different, and MSUS has good value in the diagnosis and differential diagnosis of the two.

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Objective To analyze the clinical characteristics of patients with adenomyosis(AM) and endometrial cancer(EC) and their role in preoperative diagnosis. Methods A total of 142 patients with AM complicated with EC or endometrial atypical hyperplasia(EAH) and AM with abnormal uterine bleeding(AUB) during the same period were collected from this hospital. They were divided into AM with EC or EAH group(71 cases) and control group(71 cases) according to whether they were complicated with EC or EAH. The clinical data of the patients were retrospectively analyzed, and the clinical characteristics and treatment strategies of the patients with adenomyosis complicated with endometrial cancer were discussed.Results Univariate analysis showed that age, increased proportion of CA125 and CA125, uterine volume, proportion of hypertension and abnormal endometrial ultrasonography in patients with AM with EC or EAH were significantly higher than those in the control group, and the proportion of dysmenorrhea was significantly lower than that in the control group, and the difference was statistically significant(P<0. 05). Multivariate Logistic regression analysis showed that endometrial thickening, endometrial echogenicity, and endometrial thickening combined with echogenicity were independent risk factors for AM with EC or EAH.Conclusion In clinical practice, the patients with AM with endometrial thickening or uneven echo-echo of ultrasound should be treated by hysteroscopy and curettage before further operation plan is formulated, so as to achieve the purpose of early diagnosis and treatment of EC.

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Objective To observe the expression of Galectin-3 in peritoneal dialysis(PD) fluid in patients with different dialysis ages, and to conduct correlation analysis with vascular endothelial growth factor(VEGF), fibronectin(FN) and related clinical indicators. Methods A total of 109 PD patients who were regularly followed up in the department of nephrology were divided into four groups according to different peritoneal dialysis ages. The concentrations of Galectin-3, VEGF and FN were determined by enzyme-linked immunosorbent assay. The expression of Galectin-3 in peritoneal dialysate of the 4 groups was compared, the correlation with VEGF, FN and clinical related indexes was analyzed, and the correlation was analyzed by Spearman test. Results The concentration of VEGF in peritoneal dialysis patients in group D significantly increased(P<0.05). Galectin-3 expression levels were positively correlated with VEGF(r=0.358,P=0.022), but not significantly correlated with FN(r=0.121,P=0.452). Galectin-3 was positively correlated with clinical indicators parathyroid hormone(PTH)(r=0.201,P=0.037), C-reactive protein(CRP)(r=0.357,P<0.001), left ventricular posterior wall dimensions(LVPWD)(r=0.213,P=0.026), and negatively correlated with clinical indicators total cholesterol(TC)(r=-0.316,P=0.001). Conclusion The concentration of Galectin-3 in the dialysate of long-term peritoneal dialysis patients is significantly elevated, indicating that the expression of galectin-3 increases with the extension of peritoneal dialysis time, suggesting that the detection of galectin-3 levels may be helpful for the evaluation of early peritoneal fibrosis. The positive correlation with VEGF may suggest its role in promoting peritoneal angiogenesis and fibrosis. Moreover, it is positively correlated with clinical indicators PTH, CRP and LVPWD, suggesting that it has certain clinical guiding significance on microinflammatory state and myocardial remodeling.

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Objective To explore the safety and effectiveness of transvaginal ischia spinous fascia fixation for pelvic organ prolapse. Methods The retrospective analysis of 124 patients who underwent surgical treatment for stage III-IV pelvic organ prolapse was conducted. Among them, 53 cases of transvaginal ischia spinous fascia fixation(ISFF) were performed as a study group(ISFF group) while 71 cases of transvaginal sacrospinous ligament fixation(SSLF) were performed as a control group(SSLF group). The operation time, postoperative hospitalization days, preoperative and postoperative hemoglobin values, indwelling urinary catheter time, postoperative pain scores, and the occurrence of complications were compared between the two groups, and the efficacy of the operation was objectively evaluated by using the staging method of pelvic organ prolapse(POP-Q). Also the scores of the pelvic floor impact questionnaire-7(PFIQ-7), the pelvic floor dysfunction questionnaire-20(PFDI-20), and the questionnaire of quality of life 12(PISQ-12) were used to evaluate the patients′ postoperative quality of life. Results The operation time and postoperative hospitalization days of patients in the ISFF group were less than those in the SSLF group, and the differences were statistically significant(P<0.05). The preoperative and postoperative hemoglobin values, retention time of urinary catheter, postoperative pain scores, and hospitalization costs of patients in the two groups were compared, and the differences were not statistically significant. At the 3-month postoperative outpatient follow-up, the objective success rate was 100% in two groups. The median follow-up time of patients in both groups was 24 months(12-41 months), and there were 2 cases of recurrence in the ISFF group, with a recurrence rate of 3.77% and a subjective success rate of 96.23%. While there were 3 cases of recurrence in the SSLF group and 2 cases of loss of visit, with a recurrence rate of 4.34% and a subjective success rate of 95.65%. 1 patient in the SSLF group presented with a pelvic hematoma with a diameter of about 5 cm after surgery. The hematoma disappeared after hemostasis and other symptomatic treatment. There was no organ injury or blood transfusion in both groups. Conclusion Transvaginal ischia spinous fascia fixation is a safe and effective treatment for pelvic organ prolapse, and it has the advantages of short operation time, fast postoperative recovery, fewer complications, and improvement of patients′ quality of life.

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Objective To identify possible associated genetic variants and characterise the clinical presentation of isolated ectopia lentis(IEL). Methods Forty-eight members with 5 generations of an IEL family were enrolled in this study. Peripheral blood samples of all members were collected, and clinical manifestations were observed through physical examination and routine ophthalmological examination. Whole-exome sequencing(WES) was performed for two patients to identify disease-causing variants. The target variants were verified by Sanger sequencing in family members and 200 normal controls. Then, candidate variants were verified using Sanger sequencing in family members and 200 healthy controls. SIFT, PolyPhen and MutationTester were used to predict the protein function. Results A total of 13 IEL patients in this family which inherited in an autosomal dominant pattern. The mean age at disease onset was 51.5 years. The main clinical phenotype of this ICE was characterised by ectopia lentis which anterior inclinated to the anterior chamber. As the anterior chamber became shallow, and the angle of the chamber became narrow, and eventually resulted in the secondary glaucoma. A heterozygous missense variant in the fibrillin gene-1(FBN1) gene(c. 3463G>A) was identified by WES, which was present in all patients but was absent in 200 healthy controls. SIFT, PolyPhen and MutationTester predicted that the variant affected protein function.Conclusion This IEL family is characterized by secondary glaucoma as the first symptom which is caused by ectopia lens with inclination. The c. 3463G>A of FBN1 gene may be the pathogenic mutation leading to IEL in this family.

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Objective To investigate the nutritional status and dietary structure of tuberculosis patients among different populations, analyze the factors influencing the nutritional status of tuberculosis patients, and provide theoretical basis for improving clinical nutrition and related issues in tuberculosis patients. Methods Tuberculosis patients, non-tuberculosis patients, and healthy individuals were randomly selected for a questionnaire survey. Descriptive analysis was conducted using SPSS 20.0 software. Statistical description was performed using rates and composition ratios, and qualitative data were described using relative numbers. Chi-square test was used to compare overall rates and composition ratios among different health conditions groups, with a significance level of α=0.05. Independent factors analysis of nutritional status body mass index(BMI) was conducted using multiple Logistic regression analysis for variables with statistically significant differences in the univariate analysis. Results Therewere differences in the nutritional status(χ2=62.184,P<0.05) and dietary diversity score(χ2=64.049,P<0.05) among tuberculosis patients, non-tuberculosis patients, and healthy individuals. Univariate analysis of nutritional status BMI showed statistically significant differences in gender, smoking, meat-based diet, vegetable-based diet, moderate diet diversity score, and 6 other variables for tuberculosis patients(P<0.05), and in gender, age, ethnicity, marital status, occupation, education level, smoking, drinking white wine, drinking beer, meat-based diet, moderate diet, and 11 other variables for healthy individuals(P<0.05). The variables with statistically significant differences in the univariate analysis were included in the multiple ordinal logistic regression analysis model for both tuberculosis patients and healthy individuals. The results showed that the level of education, vegetable intake, moderate food diversity score(DDS) of 4-6 were independent influencing factors of nutritional status BMI among tuberculosis patients(P<0.05); marital status was an independent influencing factor of nutritional status BMI among non-tuberculosis patients(P<0.05); while gender and occupation were independent influencing factors of nutritional status BMI among healthy individuals(P<0.05). Conclusion The dietary nutritional status of the three population groups varied. Targeted health education should be conducted, especially for tuberculosis patients, to address the issue of uneven dietary intake and promote good dietary habits among local tuberculosis patients.

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Objective To explore the related factors for the unilateral flap and bilateral flap by changing the original operation plan in the extraction of maxillary impacted mesiodens. Methods 81 patients with impacted mesiodens in the middle of the maxillary were retrospectively analyzed. The primary outcome variables were planned surgery(unilateral flap)and unplanned surgery(bilateral flap). The secondary outcome variables consisted of operation time and postoperative swelling. The predictive variables were as follows: the differential value of the shortest distance from the supernumerary tooth to the labial and palatal bone plates, which was divided into≥1.5 mm group and<1.5 mm group; the ratio of the distance from the adjacent tooth apex to the nasal floor, compared to the length of the supernumerary teeth, was recorded as≥1 and<1. A statistical software SPSS 20 was used to complete the statistical analysis. Results When the differential value was less than 1.5 mm, the possibility of unplanned surgery increased, and the probability of planned surgery was 0.085 times than that of unplanned surgery. With age growing each 1-year, the probability of planned surgery gradually decreased,HR=0.745. The postoperative swelling of the palatal approach was only 0.374 times than that of the labial approach. With age increasing, the operation time increased gradually,B=1.213. The ratio of the distance from the adjacent tooth apex to the nasal floor to the length of the supernumerary teeth did not affect the change of the surgical plan during the operation. Conclusion The shortest distance difference between the supernumerary teeth and the labial and palatal bone plates can be used as a reference for the selection of surgical approach for the extraction of maxillary impacted mesiodens.