ObjectiveTo investigate the impact of melanoma-associated antigen D4 （MAGED4） on the malignant biological behavior of hepatocellular carcinoma （HCC）， providing experimental evidence for its evaluation as a potential therapeutic target in HCC. MethodsGenes positively correlated with MAGED4 expression in HCC were screened using the LinkedOmics database， followed by GO functional and KEGG pathway enrichment analyses. Endogenously low-expressing SMMC-7721 and Huh7 cells were selected， with normal hepatic LO2 cells serving as controls. Stable overexpression cell lines were established via lentiviral infection （MAGED4 overexpression group）， with RT-PCR and Western blot validating overexpression efficiency. The effects of MAGED4 overexpression on cell proliferation， cell cycle distribution， migration， and adhesion were systematically analyzed using CCK-8 assays， flow cytometry， scratch healing assays， Transwell migration assays， and cell adhesion assays. ResultsBioinformatics analysis indicated that MAGED4 co-expressed genes were significantly enriched in biological processes closely associated with tumor proliferation and metastasis， including cell-cell adhesion， extracellular matrix organization， and the PI3K-Akt signaling pathway. A stable MAGED4 overexpression cell model was successfully established. In vitro functional assays demonstrated that MAGED4-overexpressing SMMC-7721 and Huh7 cells exhibited a mesenchymal-like morphology， whereas LO2 cells maintained an epithelial-like morphology. Compared with the empty-virus control groups， the MAGED4-overexpressing groups showed significantly increased proliferative activity （48 h： t=3.887， 22.070， 8.866， all P<0.05； 72 h： t=3.231， 23.840， 9.848， all P<0.05； 96 h： t=5.123， 49.800， 5.909， P<0.05， P<0.01， P<0.05）， wound healing rates （12 h： F=40.90， 44.00， 200.50， all P<0.001； 24 h： F=112.00， 34.89， 249.20， all P<0.001）， and numbers of migrated cells （F=239.20， 59.79， 132.80， all P<0.001）， along with significantly decreased numbers of adherent cells （F=73.65， 85.91， 138.00， all P<0.001）， while no significant differences were observed in cell cycle phase distribution in all three cell lines. ConclusionMAGED4 enhances proliferation and migration while suppressing adhesion in both HCC and normal hepatocytes， however， induces mesenchymal-like morphological changes exclusively in HCC cells， suggesting that MAGED4 is capable of driving HCC progression and may serve as a potential therapeutic target for HCC.

Objective: To determine the expression of melanoma-associated antigens D4(MAGED4) and SIRT7 in human glioma, and to analyze the potential effects of MAGED4 and SIRT7 on glioma cell proliferation. Methods: The MAGED4 and SIRT7 expression levels and their correlation were compared by the China glioma genome atlas(CGGA), human protein atlas(HPA), and UALCAN databases. Survival analysis, ROC curve analysis, and Cox regression analysis were used to predict the outcome of MAGED4 and SIRT 7 in glioma patients. Gene ontology(GO) and Kyoto encyclopedia of genes and genomes(KEGG) signaling pathway enrichment analysis were used to explore the biological functions of MAGED4 and SIRT7 in glioma. Western blot experiment was used to investigate whether MAGED4 protein exerted its regulatory effects on the activity of the PI3K/AKT signaling pathway via SIRT7. The effect of MAGED4 on cell proliferation in glioma through SIRT7 was explored by CCK-8. Results: The analysis results of CGGA, UALCAN, and HPA databases showed that the expression levels of MAGED4 and SIRT7 in glioma tissues were higher than those in normal brain tissue, and the expression were positively correlated. Results of survival, ROC, and Cox analysis showed that high expression of MAGED4 and SIRT7 mRNA were risk factors for poor prognosis in glioma. Results of KEGG enrichment analysis showed that MAGED4 and SIRT7 were associated with the PI3K/AKT signaling in glioma, and Western blot results showed that MAGED4 activated the PI3K/AKT signaling pathway by regulating SIRT7. The CCK-8 results showed that MAGED4 promotes the proliferation of glioma cells through SIRT7. Conclusion MAGED4 and SIRT7 are highly expressed in glioma and associated with poor prognosis, and MAGED4 promotes glioma cell proliferation through activation of the PI3K/AKT signaling pathway by SIRT7.

As a potential vectored vaccine,Newcastle disease virus(NDV)has been subject to various studies for vaccine development,while relatively little research has outlined the immunomodulatory effect of the virus in antigen presentation.To elucidate the key inhibitory factor in regulating the interaction of infected dendritic cells(DCs)and T cells,DCs were pretreated with the NDV vaccine strain LaSota as an inhibitor and stimulated with lipopolysaccharide(LPS)for further detection by enzyme-linked immunosorbent assay(ELISA),flow cytometry,immunoblotting,and quantitative real-time polymerase chain reaction(qRT-PCR).The results revealed that NDV infection resulted in the inhibition of interleukin(IL)-12p40 in DCs through a p38 mitogen-activated protein kinase(MAPK)-dependent manner,thus inhibiting the synthesis of IL-12p70,leading to the reduction in T cell proliferation and the secretion of interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),and IL-6 induced by DCs.Consequently,downregulated cytokines accelerated the infection and viral transmission from DCs to T cells.Furthermore,several other strains of NDV also exhibited inhibitory activity.The current study reveals that NDV can modulate the intensity of the innate?adaptive immune cell crosstalk critically toward viral invasion improvement,highlighting a novel mechanism of virus-induced immunosuppression and providing new perspectives on the improvement of NDV-vectored vaccine.

Objective:To systematically evaluate the risk of hypotension induced by sodium-glucose transporter 2 (SGLT2) inhibitors in patients with type 2 diabetes mellitus.Methods:Randomized controlled trials of SGLT2 inhibitors in the treatment of type 2 diabetes mellitus, in which hypotension were evaluated as an outcome indicator of adverse events, were collected by searching relevant databases at home and abroad (up to February 15, 2023). Cochrane risk of bias assessment tool was used to evaluate the quality of the included studies. Stata 15.1 software was used to conduct Bayesian network meta-analysis, including drawing the network evidence plot, the league map of pairwise comparison, and the surface under the cumulative ranking curve (SUCRA) for hypotension risk under different interventions of SGLT2 inhibitors, and ranking the risks of hypotension induced by different interventions of SGLT2 inhibitors. The effect sizes were expressed by relative risk ( RR) and its 95% confidence interval ( CI). Results:A total of 20 studies were included in the analysis, involving 22 525 patients with 15 260 in the trial group and 7 265 in the control group. Drugs that used in the trial group included dapagliflozin (in 1 517 patients), canagliflozin (in 6 053 patients), and ertugliflozin (in 7 690 patients); drugs that used in the control group included glimepiride (in 482 patients) and placebo (in 6 783 patients). The results of the network meta-analysis showed that the risk of hypotension was higher after treatment with 300 mg of canagliflozin, compared with those with 5 mg and 15 mg of ertugliflozin, and placebo ( RR=2.13, 95% CI: 1.31-3.47; RR=2.21, 95% CI: 1.35-3.61; RR=2.49, 95% CI: 1.62-3.82; all P<0.05); the risk of hypotension was higher after treatment with 100 mg of canagliflozin, compared with placebo ( RR=1.61, 95% CI: 1.04-2.50, P<0.05); the differences in comparison between any other 2 interventions with SGLT2 inhibitors were not statistically significant. According to the relative risks for hypotension of different interventions with SGLT2 inhibitors in the results of SUCRA, interventions were ranked as ertugliflozin 5 mg, placebo, dapagliflozin 2.5 mg, ertugliflozin 10 mg, ertugliflozin 15 mg, canagliflozin 50 mg, canagliflozin 100 mg, dapagliflozin 5 mg, dapagliflozin 10 mg, canagliflozin 150 mg, and canagliflozin 300 mg. Conclusions:Different treatment regimens with SGLT2 inhibitors had different risks of hypotension in patients with type 2 diabetes. The risk of hypotension caused by ertugliflozin is lower, especially at the dose of 5 mg. The risk of hypotension caused by canagliflozin is higher, especially at relatively high doses.

Objective:To systematically evaluate the risk of hypotension induced by sodium-glucose transporter 2 (SGLT2) inhibitors in patients with type 2 diabetes mellitus.Methods:Randomized controlled trials of SGLT2 inhibitors in the treatment of type 2 diabetes mellitus, in which hypotension were evaluated as an outcome indicator of adverse events, were collected by searching relevant databases at home and abroad (up to February 15, 2023). Cochrane risk of bias assessment tool was used to evaluate the quality of the included studies. Stata 15.1 software was used to conduct Bayesian network meta-analysis, including drawing the network evidence plot, the league map of pairwise comparison, and the surface under the cumulative ranking curve (SUCRA) for hypotension risk under different interventions of SGLT2 inhibitors, and ranking the risks of hypotension induced by different interventions of SGLT2 inhibitors. The effect sizes were expressed by relative risk ( RR) and its 95% confidence interval ( CI). Results:A total of 20 studies were included in the analysis, involving 22 525 patients with 15 260 in the trial group and 7 265 in the control group. Drugs that used in the trial group included dapagliflozin (in 1 517 patients), canagliflozin (in 6 053 patients), and ertugliflozin (in 7 690 patients); drugs that used in the control group included glimepiride (in 482 patients) and placebo (in 6 783 patients). The results of the network meta-analysis showed that the risk of hypotension was higher after treatment with 300 mg of canagliflozin, compared with those with 5 mg and 15 mg of ertugliflozin, and placebo ( RR=2.13, 95% CI: 1.31-3.47; RR=2.21, 95% CI: 1.35-3.61; RR=2.49, 95% CI: 1.62-3.82; all P<0.05); the risk of hypotension was higher after treatment with 100 mg of canagliflozin, compared with placebo ( RR=1.61, 95% CI: 1.04-2.50, P<0.05); the differences in comparison between any other 2 interventions with SGLT2 inhibitors were not statistically significant. According to the relative risks for hypotension of different interventions with SGLT2 inhibitors in the results of SUCRA, interventions were ranked as ertugliflozin 5 mg, placebo, dapagliflozin 2.5 mg, ertugliflozin 10 mg, ertugliflozin 15 mg, canagliflozin 50 mg, canagliflozin 100 mg, dapagliflozin 5 mg, dapagliflozin 10 mg, canagliflozin 150 mg, and canagliflozin 300 mg. Conclusions:Different treatment regimens with SGLT2 inhibitors had different risks of hypotension in patients with type 2 diabetes. The risk of hypotension caused by ertugliflozin is lower, especially at the dose of 5 mg. The risk of hypotension caused by canagliflozin is higher, especially at relatively high doses.

Objective:To construct the content framework of the rehabilitation management application (APP) for patients with urinary incontinence after radical prostatectomy, so as to provide an effective network management platform for the rehabilitation management of patients wither urinary incontinence after radical prostatectomy.Methods:From November 2020 to May 2021, the content framework of rehabilitation management APP for patients with urinary incontinence after radical prostatectomy was initially constructed through literature review and group discussion. The objective sampling was used to select 15 experts from Beijing, Shanghai and other regions for two rounds of expert consultation, to construct the content framework of rehabilitation management APP for patients with urinary incontinence after radical prostatectomy.Results:Among two rounds of expert consultation, the effective recovery rates of the questionnaires were all 100.00% (15/15) , and the expert authority coefficients were 0.937 and 0.934, respectively. The Kendall harmony coefficients of all indicators in the second round of expert consultation ranged from 0.208 to 0.333, and the differences were all statistical (all P<0.01) . It was finally determined that the content framework of rehabilitation management APP for patients with urinary incontinence after radical prostatectomy included 3 first-level indicators, 10 second-level indicators, 43 third-level indicators, and 62 fourth-level indicators. Conclusions:The content framework of constructed rehabilitation management APP for patients with urinary incontinence after radical prostatectomy is highly scientific and practical, and can provide corresponding evidence.

【Objective】 To investigate the effect of transfusion-transmitted Zika virus (ZIKV) on the expression of non-coding circular RNA (hsa_circ_0001613) and the role of hsa_circ_0001613 in Zika virus replication. 【Methods】 Human adenocarcinomic alveolar basal epithelial cells (A549) were seeded on a 12-well plate at 1.8×10⁵/ well and infected with ZIKV at 0.05 MOI. The Total RNAs were isolated every day for 5 days after infection, and the relative expression level of hsa_circ_0001613 was detected by qRT-PCR. In addition, 10nM siRNA-hsa_circ_0001613 was transfected into 2×10⁵/ well A549 cells to specifically knock down the expression level of hsa_circ_0001613. 24h later, the cells were infected with ZIKV (MOI=0.05). Total RNAs were isolated at day 1-5 post-infection, proteins were extracted 96h post-infection. ZIKV replication, relative host antiviral gene expression, and interferon stimulated response element (ISRE) activity were tested using qRT-PCR, western blot and dual luciferase assay, respectively. 【Results】 The relative expression of hsa_circ_0001613 decreased significantly after 1-5 days of ZIKV infection. Knockdown of hsa_circ_0001613 inhibited ZIKV replication. Meanwhile, hsa_circ_0001613 knockdown significantly upregulated IFN-α/β and its downstream interferon-stimulated genes (ISGs) expression, also increased ISRE activity. 【Conclusion】 ZIKV infection significantly suppressed hsa_circ_0001613 expression in A4549 cells. Preliminary study indicated that hsa_circ_0001613 knockdown inhibited ZIKV replication possibly through activating type-Ⅰ IFN signaling pathway as showed by increased ISGs expression and ISRE activity.

【Objective】 To investigate the role of non-coding microRNA miR-16-5p in ZIKV replication and the underlying mechanism. 【Methods】 1×10⁵/mL HeLa cells were seeded in 24-well plate and infected with ZIKV(MOI=5). RNAs were harvested, and miR-16-5p expression levels were measured by qRT-PCR at 24, 48 and 72 hour post infection, respectively. HeLa cells were transfected with 20nM miR-16-5p mimic and infected with ZIKV(MOI=5) at 24h post transfection. RNAs were extracted and ZIKV RNA and several inflammation factors expression were tested using qRT-PCR at 48h post infection. HeLa cells were co-transfected with 1μg NFκB-luc and 10ng pRL-TK with 20nM miR-16-5p mimic, and then infected with ZIKV(MOI=5) for 24h before the luciferase expression was tested at 48h post infection. HeLa cells were transfected with 20nM miR-16-5p mimic and infected with ZIKV(MOI=5) at 24h post transfection, and cell apoptosis was assayed through flow cytometry. 【Results】 Compared with uninfected control, miR-16-5p expression was significantly decreased at 24h, 48h and 72h following ZIKV infection. MiR-16-5p over-expression inhibited ZIKV replication, while upregulated NFκB activity and inflammation factors expression compared with the negative mimic-transfected cells. MiR-16-5p overexpression also promoted HeLa cell apoptosis. 【Conclusion】 ZIKV infection downregulated intracellular miR-16-5p expression. Overexpression of miR-16-5p suppressed ZIKV infection. MiR-16-5p inhibited ZIKV replication and promoted cell apoptosis probably by activating NFκB pathway and stimulating inflammation factors expression.

Objective:To explore the application effect of pressure relief double knee pad in pressure injury in critically ill patients.Methods:A total of 635 patients who were admitted to Intensive Care Unit of Taizhou Hospital in Zhejiang Province from June 2020 to April 2021 were selected as the research objects by the convenient sampling method. They were divided into the control group ( n=315) and the experimental group ( n=320) by the random number table method. On the basis of the conventional lying position, patients in the control group placed the R-shaped pads on the back of the shoulder to the top of the pelvis, while patients in the experimental group slightly bent their lower limbs on the basis of the control group and placed it on the self-designed pressure relief double knee pad. The incidence of pressure injury, pressure index of sacrum and heel skin, the non-displacement rate of support appliances and the incidence of deep vein thrombosis (DVT) in the two groups were evaluated. Results:The incidence of pressure injury of patients in the experimental group was 0.63% (2/320) , which was lower than 3.17% (10/315) in the control group, and the difference was statistically significant ( P<0.05) . The pressure indexes of the sacrum and heel of the experimental group were respectively (66.10±13.43) and 0 mmHg, which were lower than (75.08±12.03) and (22.26±6.20) mmHg of the control group, and the differences were statistically significant ( P<0.05) . The non-displacement rate of support appliances in the experimental group was 89.62% (190/212) , which was higher than 52.63% (100/190) in the control group, and the difference was statistically significant ( P<0.05) . There was no statistically significant difference in the incidence of VTE between 2 groups ( P>0.05) . Conclusions:The use of pressure relief double knee pads can reduce the incidence of pressure injuries, the pressure of the skin of the sacrum and heel and increase the non-displacement rate of support appliances, which is worthy of clinical application.

Objective To investigate the variation characteristics of peripheral platelet parameters in children with dengue fever and to assess their use and value in diagnosis,prognosis and treatment for dengue fever.Methods Platelet parameters of 69 pediatric patients with dengue fever in acute febrile phase and remission period and 103 healthy controls were compared.The difference of platelet parameters between the liver function injury group and the normal liver function group,the coagulation dysfunction group and the coagulation function normal group,the severe group and the mild group with dengue fever was compared,the causes of thrombocytopenia in children with dengue was investigated so as to better understand the situation of bone marrow producing platelets.Results P-LCR,PDW,MPV were significantly higher in patients in acute febrile phase of dengue fever,compared with the healthy subjects(P＜0.01),while PCT,PLT were significantly lower in patients in acute febrile phase of dengue fever,compared with the healthy subjects(P＜0.01).MPV was significantly lower in patients with abnormal hepatic function when compared with patients with normal hepatic function(P＜0.05),while other platelet parameters did not show significant differences(P＞0.05).PLT and PCT were significantly lower in the severe patients than those in the mild patients(P＜0.05),while other platelet parameters did not show significant differences(P＞0.05).Peripheral platelet parameters did not show significant differences between patients with or without abnormal coagulation function (P＞0.05).Conclusion Peripheral platelet parameters may play a role in the diagnosis,treatment and prognosis of dengue fever in children.