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BACKGROUND: A decreased autophagic capacity of bone marrow mesenchymal stromal cells (BMSCs) has been suggested to be an important cause of decreased osteogenic differentiation. A pharmacological increase in autophagy of BMSCs is a potential therapeutic option to increase osteoblast viability and ameliorate osteoporosis. AIM: To explore the effects of sinomenine (SIN) on the osteogenic differentiation of BMSCs and the underlying mechanisms. METHODS: For in vitro experiments, BMSCs were extracted from sham-treated mice and ovariectomized mice, and the levels of autophagy markers and osteogenic differentiation were examined after treatment with the appropriate concentrations of SIN and the autophagy inhibitor 3-methyladenine. In vivo, the therapeutic effect of SIN was verified by establishing an ovariectomy-induced mouse model and by morphological and histological assays of the mouse femur. RESULTS: SIN reduced the levels of AKT and mammalian target of the rapamycin (mTOR) phosphorylation in the phosphatidylinositol 3-kinase (PI3K)/AKT/mTOR signaling pathway, inhibited mTOR activity, and increased autophagy ability of BMSCs, thereby promoting the osteogenic differentiation of BMSCs and effectively alleviating bone loss in ovariectomized mice in vivo. CONCLUSION: The Chinese medicine SIN has potential for the treatment of various types of osteoporosis, bone homeostasis disorders, and autophagy-related diseases.

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Objective:To investigate the safety and efficacy of intravascular intervention in ruptured intracranial vertebral artery dissecting aneurysm (IVADA).Methods:A retrospective analysis was performed; 25 patients with ruptured IVADA (25 aneurysms) admitted to Department of Neurosurgery, First Affiliated Hospital of University of Science and Technology of China from January 2020 to June 2023, were chosen. Aneurysm and parent artery occlusion or stent-assisted spring coil embolization were performed according to location of the aneurysms, degrees of aneurysm immediate embolization were evaluated by Raymond grading, and perioperative adverse events were recorded. The patients were followed up for 6-48 months, and aneurysm recurrence was determined according to DSA results; prognoses were assessed by modified Rankin Scale (mRS), with scores of 0-2 as good prognosis and scores of 3-6 as poor prognosis.Results:All 25 patients had unilateral ruptured IVADA, 10 (40%) received aneurysm and parent artery occlusion (occlusion of dissection segment) and 15 (60%) received stent-assisted embolization. Immediately after surgery, 19 patients (76%) had grading I embolization, 4 (16%) grading II embolization, and 2 (8%) grading III embolization. No aneurysm rupture or stent related thrombosis was observed during procedure; 3 patients (12%) died after procedure, with postoperative rebleeding in 1, postoperative cerebellar infarction with respiratory failure in 1, and severe pneumonia in 1. In the 22 survivals, 18 had good prognosis and 4 had poor prognosis. In the 5 relapsed patients (all accepted stent-assisted embolization), 4 underwent re-intervention, and one with visualization at aneurysm neck was relatively stable on re-examination and accepted regular follow up.Conclusion:Aneurysm and parent artery occlusion can be used for non-dominant vertebral artery aneurysms not involving posterior inferior cerebellar artery, whose recurrence rate is lower than that of stent-assisted coil embolization.

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Ions play a vital role in regulating various biological processes, including metabolic and immune homeostasis, which involves tumorigenesis and therapy. Thus, the perturbation of ion homeostasis can induce tumor cell death and evoke immune responses, providing specific antitumor effects. However, antitumor strategies that exploit the effects of multiion perturbation are rare. We herein prepared a pH-responsive nanomodulator by coloading curcumin (CU, a Ca2+ enhancer) with CaCO3 and MnO2 into nanoparticles coated with a cancer cell membrane. This nanoplatform was aimed at reprogramming the tumor microenvironment (TME) and providing an antitumor treatment through ion fluctuation. The obtained nanoplatform, called CM NPs, could neutralize protons by decomposing CaCO3 and attenuating cellular acidity, they could generate Ca2+ and release CU, elevating Ca2+ levels and promoting ROS generation in the mitochondria and endoplasmic reticulum, thus, inducing immunogenic cell death. Mn2+ could decompose the endogenous H2O2 into O2 to relieve hypoxia and enhance the sensitivity of cGAS, activating the cGAS-STING signaling pathway. In addition, this strategy allowed the reprogramming of the immune TME, inducing macrophage polarization and dendritic cell maturation via antigen cross-presentation, thereby increasing the immune system's ability to combat the tumor effectively. Moreover, the as-prepared nanoparticles enhanced the antitumor responses of the αPD1 treatment. This study proposes an effective strategy to combat tumors via the reprogramming of the tumor TME and the alteration of essential ions concentrations. Thus, it shows great potential for future clinical applications as a complementary approach along with other multimodal treatment strategies.

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We investigated whether FTY-720 might have an effect on bleomycin-induced pulmonary fibrosis through inhibiting TGF-β1 pathway, and up-regulating autophagy. The pulmonary fibrosis was induced by bleomycin. FTY-720 (1 mg/kg) drug was intraperitoneally injected into mice. Histological changes and inflammatory factors were observed, and EMT and autophagy protein markers were studied by immunohistochemistry and immunofluorescence. The effects of bleomycin on MLE-12 cells were detected by MTT assay and flow cytometry, and the related molecular mechanisms were studied by Western Blot. FTY-720 considerably attenuated bleomycin-induced disorganization of alveolar tissue, extracellular collagen deposition, and α-SMA and E-cadherin levels in mice. The levels of IL-1β, TNF-α, and IL-6 cytokines were attenuated in bronchoalveolar lavage fluid, as well as protein content and leukocyte count. COL1A1 and MMP9 protein expressions in lung tissue were significantly reduced. Additionally, FTY-720 treatment effectively inhibited the expressions of key proteins in TGF-β1/TAK1/P38MAPK pathway and regulated autophagy proteins. Similar results were additionally found in cellular assays with mouse alveolar epithelial cells. Our study provides proof for a new mechanism for FTY-720 to suppress pulmonary fibrosis. FTY-720 is also a target for treating pulmonary fibrosis.

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The global emergence of SARS-CoV-2 variants has led to increasing breakthrough infections in vaccinated populations, calling for an urgent need to develop more effective and broad-spectrum vaccines to combat COVID-19. Here we report the preclinical development of RQ3013, an mRNA vaccine candidate intended to bring broad protection against SARS-CoV-2 variants of concern (VOCs). RQ3013, which contains pseudouridine-modified mRNAs formulated in lipid nanoparticles, encodes the spike(S) protein harboring a combination of mutations responsible for immune evasion of VOCs. Here we characterized the expressed S immunogen and evaluated the immunogenicity, efficacy, and safety of RQ3013 in various animal models. RQ3013 elicited robust immune responses in mice, hamsters, and nonhuman primates (NHP). It can induce high titers of antibodies with broad cross-neutralizing ability against the Wild-type, B.1.1.7, B.1.351, B.1.617.2, and the omicron B.1.1.529 variants. In mice and NHP, two doses of RQ3013 protected the upper and lower respiratory tract against infection by SARS-CoV-2 and its variants. We also proved the safety of RQ3013 in NHP models. Our results provided key support for the evaluation of RQ3013 in clinical trials.

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Objective:To analyze the effect and influencing factors of embolotherapy on headache in elderly patients with unruptured intracranial aneurysms.Methods:A retrospective analysis of clinical data of elderly patients(aged≥61 years)with unruptured intracranial aneurysms admitted to our hospital from January 2018 to December 2020 was performed.Headache assessment was performed by a quantitative 11-point headache scale in all patients preoperatively and at 6 months after endovascular treatment, and the difference between them was analyzed.Univariate analysis was applied to test the association between headache outcomes and clinical variables.Results:A total of 73 patients(mean age: 68.4 years old; age range: 61-86 years; 47 women)fulfilled the inclusion criteria.There were 53 patients(72.6%)who presented with preoperative headache(headache score≥1). Among them, 39 cases(73.6%)had an improvement in headache, 11(20.8%)remained unchanged, and 3(5.7%)aggravated, after endovascular treatment.The average preoperative headache score was 5(4, 6) vs.postoperative 3(1, 4), with statistical significance( Z=-5.036, P=0.000). Only the preoperative headache score was associated with outcomes of headache, and a higher headache score predicted a lack of headache relief( Z=-2.819, P=0.005). Conclusions:Embolotherapy of unruptured intracranial aneurysms can relieve headache in most elderly patients.Preoperative headache severity is correlated with postoperative headache outcomes.

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Objective:To construct a scientific research evaluation model through principal component analysis, and to explore scientific research evaluation methods for hospitals.Methods:The professional title, educational background, positions and scientific research output information of the scientific research personnel in the First Hospital of Jilin University from 2019 to 2020 were collected. Delphi expert consultation was used to determine the assignment value of each variable, and use SPSS 21.0 software was used to build the principal component analysis model and conduct model verification.Results:The study collected a total of 1 882 researchers′ information. The KMO value of the validity test and the Bartlett sphere test meet the requirements of principal component analysis (KMO=0.731, P<0.05); the model obtained a total of 7 principal components. Among them, principal component 1 represents researchers who published SCI papers, applying for national, provincial and ministerial level scientific research projects, and their part-time positions in academic societies. The second principal component represents the status of applying for patents and publications, and the third principal component represents the status of the awards. The scores of scientific research output of researchers were summarized and sorted according to disciplines, according to which the neurology, endocrinology and metabolism, neurosurgery, general surgery and orthopedics ranked better. The model verification results found that researchers with senior professional titles and doctoral degrees had the highest median weighted comprehensive score( P<0.05), suggesting that scholars with higher professional title levels and higher education received higher comprehensive scientific research output scores. Conclusions:The scientific research evaluation model constructed by this study can provide scientific data reference for the hospital scientific research evaluation.

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Objective:To observe the short-term efficacy and complications of intracranial wide-necked aneurysms treated by Neuroform Atlas stent-assisted coil embolization, and preliminary explore its safety and effectiveness.Methods:A retrospective analysis was performed. The clinical data of 33 patients with intracranial wide-necked aneurysms, admitted to our hospital from September 2020 to August 2021, were collected. All patients underwent Neuroform Atlas stent -assisted coil embolization, including 28 patients with single stent assisted coil embolization and 5 with double stent assisted embolization. Raymond grading was used to evaluate the degrees of immediate postoperative aneurysm embolization. Modified Rankin scale (mRS) was used to assess the prognoses 3-6 months after surgery. DSA was performed to recheck the recurrence of aneurysms and the patency of the parent artery.Results:DSA immediately after surgery showed that all aneurysms were tightly embolized (Raymond grading I). One patient developed intraoperative thrombosis, and blood flow was restored after tirofiban thrombolytic therapy, without new neurological dysfunction after surgery. Three-6 months after surgery, all patients had a good prognosis; DSA recheck was completed in 21 patients (63.6%), and no stenosis or thrombosis were found in the parent artery and no aneurysm recurrence was noted.Conclusion:The Neuroform Atlas stent-assisted coil embolization in treatment of intracranial wide-necked aneurysms has good efficacy and high safety.

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Sphingosine-1-phosphate (SIP) is an important bio- active phospholipid molecule, which is involved in the occurrence and development of cardiovascular diseases such as atherosclerosis, myocardial ischemia and reperfusion injury, myocardial infarction, myocardial fibrosis and myocardial remodeling, and it plays a wide range of biological effects in human cardiovascular system.SIP acts mainly in the form of binding of sphingosine-1-phosphate receptor (SI Pits), selectively binding vascular endothelial cells and smooth muscle cells, which plays a role in the fight against cardiovascular diseases.This paper reviews the biological effects of SIP in cardiovascular system, i- dentifies effective targets in cardiovascular diseases, and alleviates the damage caused by SI P.aiming to provide new ideas for the study of SIP in cardiovascular direction.

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Objective To investigate the effects of Smad7 knock down by lentivirus on rat cardiac fibroblasts proliferation, migration, cell differentiation and collagen secretion in vitro. Methods The primary cardiac fibroblasts were separated from the hearts of ten SD rats and identified by immunohistochemical method. The lentivirus transfection knocked down the expresson of Smad7 in cardiac fibroblasts, Western blotting was used to detect the efficiency of Smad7 knock down by lentivirus. The proliferation of cardiac fibroblasts was quantified by real-time unlabeled cell analyzer. Cell migration was evaluted by cell wound scratch assay. Western blotting was used to detect expression of α-smooth muscle actin（α-SMA） and collagen Ⅰ（Col Ⅰ）. Results Myocardial fibroblasts were successfully cultured and identified by immunocytochemical methods. The multiplicity of infection（MOI） that lentivirus transduction of myocardial fibroblasts was 100. After lentivirus transduction, 88.33% myocardial fibroblasts expressed green fluorescent protein, showed that the lentivirus could significantly reduce the protein expression of Smad7. Smad7 deficiency decreased the proliferation and migration of cardiac fibroblasts, increased the protein expression of α-SMA and decreased collagen secretion. The results indicated that Smad7 deficiency significantly down-regulated the proliferation and migration of cardiac fibroblasts, increased α-SMA protein expression and reduced ColⅠ protein expression. Conclusion Smad7 deficiency can significantly change the cardiac fibroblasts function, that is related to the pathological mechanism that lead to myocardial fibrosis

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Coronavirus disease 2019 (COVID-19), which is triggered by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, continues to threaten global public health. Developing a vaccine that only requires single immunization but provides long-term protection for the prevention and control of COVID-19 is important. Here, we developed an adeno-associated virus (AAV)-based vaccine expressing a stable receptor-binding domain (SRBD) protein. The vaccine requires only a single shot but provides effective neutralizing antibodies (NAbs) over 598 days in rhesus macaques (Macaca mulatta). Importantly, our results showed that the NAbs were kept in high level and long lasting against authentic wild-type SARS-CoV-2, Beta, Delta and Omicron variants using plaque reduction neutralization test. Of note, although we detected pre-existing AAV2/9 antibodies before immunization, the vaccine still induced high and effective NAbs against COVID-19 in rhesus macaques. AAV-SRBD immune serum also efficiently inhibited the binding of ACE2 with RBD in the SARS-CoV-2 B.1.1.7 (Alpha), B.1.351 (Beta), P.1/P.2 (Gamma), B.1.617.2 (Delta), B.1.617.1/3(Kappa), and C.37 (Lambda) variants. Thus, these data suggest that the vaccine has great potential to prevent the spread of SARS-CoV-2.

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Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), which causes coronavirus disease-2019 (COVID-19), interacts with the host cell receptor angiotensin-converting enzyme 2 (hACE2) via its spike 1 protein for infection. After the virus sequence was published, we identified two potent antibodies against SARS-CoV-2 RBD from antibody libraries using a phage-to-yeast (PtY) display platform in only 10 days. Our lead antibody JMB2002, now in a phase I clinical trial, showed broad-spectrum in vitro blocking activity against hACE2 binding to the RBD of multiple SARS-CoV-2 variants including B.1.351 that was reportedly much more resistant to neutralization by convalescent plasma, vaccine sera and some clinical stage neutralizing antibodies. Furthermore, JMB2002 has demonstrated complete prophylactic and potent therapeutic efficacy in a rhesus macaque disease model. Prophylactic and therapeutic countermeasure intervention of SARS-CoV-2 using JMB2002 would likely slow down the transmission of currently emerged SARS-CoV-2 variants and result in more efficient control of the COVID-19 pandemic.

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Depression is a serious mental illness with a high incidence. At present, we do not fully understand the specific pathological mechanisms of depression, and the efficacy of drug treatments is very limited. Recent studies have shown that epigenetic changes that occur in specific brain regions may be a key mechanism by which environmental factors to interact with individuals to influence the risk of depression. Therefore, drugs that target epigenetic regulation may become a new direction for the development of antidepressants. Histone deacetylase inhibitors (HDACi) are a class of compounds that inhibit histone deacetylase activity, which has been reported to be associated with depression; this article addresses the use of HDACi in preclinical studies, and their potential therapeutic role and limitations of use in depression.

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Objective:To investigate the application value of electroencephalogram (EEG) combined with regional cerebral oxygen saturation (rSO 2) in monitoring cerebral perfusion during carotid endarterectomy (CEA). Methods:A retrospective analysis of clinical data of 42 patients with atherosclerotic carotid artery stenosis admitted to and accepted CEA in our hospital from January 2018 to December 2019 was performed. CEA was performed under EEG combined with rSO 2 monitoring. The efficacy and safety of EEG combined with rSO 2 in monitoring cerebral perfusion abnormalities during CEA were analyzed. Results:After carotid artery occlusion, 24 patients (57.1%) had normal EEG and rSO 2; 15 (35.7%) had abnormal changes of EEG, among whom 13 (31.0%) were accompanied by rSO 2 anomaly; 16 (38.1%) had abnormal rSO 2, among whom 13 (31.0%) were accompanied by EEG anomaly. Of these 18 patients with abnormal EEG and/or rSO 2 monitoring, 17 patients recovered after increasing their blood pressure and 1 patient recovered after diverter tube usage. Intraoperative EEG and rSO 2 monitoring results were consistent (Kappa=0.745, P=0.000). The positive rates of combined monitoring, EEG alone or rSO 2 alone were 42.9%, 35.7% and 38.1%, respectively. All patients were evaluated clinically and radiologically before discharge, and no new ischemic lesions or clinical symptoms were found. Conclusions:EEG and rSO 2 monitoring are well consistent in CEA; the combined monitoring can make up for the deficiency of single monitoring to increase surgical safety.

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Objective:To observe the short-term efficacy and complications of flow diverter device Tubridge in the treatment of complex intracranial aneurysms, and to summarize the indications and experience of Tubridge in treatment of complex intracranial aneurysms.Methods:Thirty-one patients with complex intracranial aneurysms accepted treatment with assistance of Tubridge in our hospital from January 2020 to December 2020 were enrolled. Raymond grading was used to evaluate the degrees of aneurysm embolization immediately after surgery. Modified Rankin scale (mRS) was used to evaluate the prognoses 6 months after surgery. The embolization of aneurysms was determined in these patients by DSA 6 and 12 months after surgery.Results:Thirty-seven aneurysms were noted in these 31 patients. A total of 32 diverters (Turbridge) were implanted into these 37 aneurysms, and all of which were successfully delivered and released. Thirteen aneurysms were filled with spring coils (8 with Raymond grading I, 2 with Raymond grading II, 3 with Raymond grading III by DSA immediately after surgery), and 24 aneurysms were not filled with spring coils. Postoperative ischemic complications occurred in 1 patient, and no hemorrhagic complications or death were noted. All 31 patients had good prognosis at follow-up. Six months after surgery, DSA re-examination in 24 patients (77.4%) showed that complete occlusion of aneurysms was achieved in 17 patients (70.8%). DSA re-examination 12 months after surgery showed complete occlusion of aneurysms in 19 of the 25 patients (76%).Conclusion:The new domestic flow diverter device Tubridge has a high success rate, less perioperative complications, high safety, and good recovery in the treatment of complex intracranial aneurysm.

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Aim To investigate the protective effect of 1, 8-Cineole on the injury of human aortic endothelial cells (HAECs) induced by high glucose (HG) via regulating autophagy. Methods Cells were incubated with different doses of 1, 8-Cineole followed by exposing to HG for 60 h, and MTT assay was used to analyse cell viability, lactate dehydrogenase (LDH) was used to detect cytotoxicity, and Western blot was used to detect Beclin1, LC3-II/I, p62, caspase-3 and caspase-9 expressions. Autophagy inhibitor (chloroquine, CQ) was treated on HAECs, and the expressions of Beclinl, LC3-II/I, p62, caspase-3 and caspase-9 were measured by Western blot. Results 1, 8-Cineole increased cell viability, reduced the content of LDH, activated autophagy and inhibited apoptosis. Compared with control group, the expression of Beclinl, LC3-II/I, p62, caspase-3 and caspase-9 in CQ group increased. Simultaneously, the expression of above-mentioned between CQ + HG group and CQ + HG + CH group. Conclusions 1, 8-Cineole has protective effect on the injury of HAECs induced by high glucose, and its effect is related to improving autophagy flux.

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Parkinson's disease (PD) is the second major neurodegenerative disease.The pathogenesis of PI) is still unclear.It is generally believed that neural damage, mitochondrial dysfunction, inflammation, oxidative stress and autophagy dysfunction caused by the transmission and aggregation of a- synuclein play an important role in the occurrence and development of PD.More and more research show- that metabolic disorder is one of the pathogenesis of PD.We examined whether overexpression of a- synuclein could induce metabolic disorder in mice and the possible mechanisms.Mice were divided into two groups: Thyl-aSYN transgenic mice (TG) and the control wild-type (WT) group.The rotarod test was used to analyze motor function in mice.We detected the body weight, plasma insulin content, glucose tolerance and insulin tolerance in the two group mice.The morphology of islets in the two groups were observed by hematoxylin eosin (HE) staining, and the islets were isolated to detect the glucose- stimulated insulin secretion (GSIS).The results showed that compared with the WT group, exercise tolerance of 12-month-old TG group decreased by 23.1% (P < 0.05) , body weight increased by 7% (P < 0.01), glucose tolerance decreased (P < 0.05), insulin tolerance decreased (P < 0.05), and insulin contents in the peripheral blood decreased by 20% (P < 0.05).Compared with the WT group, the levels of ce -syn proteins in the pancreas of the TG group increased by 1.32 times (P < 0.05) , the area of islets in the TG group decreased (P < 0.05 ) , the number of islets decreased (P < 0.01) , and the insulin secretion function decreased (P< 0.01).This study showed that the role of a-synuclein in PD is not limited to the damage of dopaminergic neurons, it also can affect metabolism and the morphology and function of peripheral organs, which provides a new theoretical basis for the pathogenesis of PD.

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Objective:Our study investigates the expression of Motor neuron and pancreatic homeobox 1 ( MNX1) in breast cancer tissues, and the effects of MNX1 on the proliferation, cell cycle, apoptosis, migration and invasion of MDA-MB-231 cells were studied by constructing the MNX1 knockdown MDA-MB-231 cell line. Methods:A retrospective study was conducted on 73 breast cancer tissues and adjacent normal tissues from 73 breast cancer patients in the First Affiliated Hospital of Guangxi Medical University. qRT-PCR was used to detect the relative expression of MNX1 in tissues and cells. Western blot detects the protein level of MNX1 in the tissue. We designed the shRNA MNX1 and constructed the MNX1 viral vector with low expression. The viral vector was further used to infect triple-negative breast cancer cells. The MNX1 with the best silencing effect was designed to silence the breast cancer cell line MDA-MB-231 as the silence group ( shMNX1), and the negative control group (Control) of lentivirus infection was designed to carry out follow-up cell function tests.CCK-8 method was used to detect cell proliferation ability. Transwell method was used to detect cell migration and invasion ability. Use flow cytometry to detect apoptosis. Each experiments at least 3 times independent experiments and measurement data with normal distribution were represented as the ( Mean± SD). The t-test was used for the comparison of two sample means. Results:The qRT-PCR showed that the expression level of MNX1 mRNA in breast cancer tissue was higher than that in adjacent tissues ( P<0.05). WB showed that the expression level of MNX1 protein in breast cancer tissue was significantly higher than that in adjacent tissues ( P<0.01) . CCK-8 experiment results showed that the OD (450 nm) of breast cancer cells in the silence group at 24, 48 and 72 h was lower than that of the negative control group ( P<0.05). The results showed that silencing the MNX1 gene can inhibit the proliferation of breast cancer cells MDA-MB-231. The results of Transwell migration experiment showed that the number of cells passing through the Transwell chamber in the silent group and the negative control group were 217.00±33.23 and 490.00±45.56, respectively, and the difference was significant ( P<0.05). The results of Transwell invasion experiment showed that the number of cells in the silent group and the negative control group passing through the Transwell chamber were (91.00±12.79)and (419.00±49.37), respectively, and the difference was statistically significant ( P<0.05). Our results show that silencing the MNX1 gene can inhibit the migration and invasion ability of breast cancer cell line MDA-MB-231. The results showed that the apoptosis rate of breast cancer cells in the silent group was (3.81±0.41)%, and the negative control group was (2.13±0.16)%. The apoptosis rate of breast cancer cells in the silent group was higher than that in the negative control group, and the result was statistically significant ( P<0.05). MNX1 promotes the apoptosis of triple-negative breast cancer cells. Conclusion:MNX1 is a new breast cancer gene, silencing the expression of MNX1 gene inhibits the proliferation, migration and invasion of breast cancer cells and promotes cell apoptosis, which provides a new regulatory mechanism and therapeutic target for breast cancer.

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Objective: Pancreatic ductal adenocarcinoma (PDAC) is one of the most malignant digestive tract tumors with a poor prognosis and high recurrence rate. Recently, ferroptosis resistance has been found in PDAC. However, the underlying mechanism of ferroptosis resistance has not been fully elucidated. Cytochrome P4502J2 (CYP2J2) is the main enzyme which mediates arachidonic acid to produce epoxyeicosatrienoic acids (EETs) in human tissues. It has been reported that EETs involve in the development of cancer, while the roles of EETs in PDAC and ferroptosis remain unclear. This study aims to explore the effect of CYP2J2/EETs on ferroptosis of human pancreatic ductal adenocarcinoma cells PANC-1 cells and the underlying mechanisms.Methods: The tumor tissues and para-carcinoma tissues of 9 patients with PDAC were collected and the expression of CYP2J2 was detected with real-time PCR and Western blotting. Enzyme-linked immunosorbent assay (ELISA) was used to detect the level of 8,9-dihydroxyeicosatrienoic acid (8,9-DHET), and the degradation product of 8,9-epoxyeicosa-trienoic acid (8, 9-EET). PANC-1 cells were used in this study. The ferroptosis inducer erastin was used to induce ferroptosis. The intracellular long-chain acyl-CoA synthetase 4 (ACSL4) protein level, lactate dehydrogenase (LDH) activity, malondialdehyde (MDA) content, Fe2+concentration, and cell survival were detected. The 8, 9-EET was pretreated to observe its effect on erastin-induced ferroptosis in PANC-1 cells. Lentivirus was used to construct a CYP2J2 knockdown cell line to observe its effect on the ferroptosis of PANC-1 cells induced by erastin. A peroxisome proliferation-activated receptor γ (PPARγ) blocker was used to observe the effect of 8, 9-EET on erastin-induced glutathione peroxidase 4 (GPX4) and MDA content in PANC-1 cells.Results: High expression of CYP2J2 was found in PDAC, accompanied by an increased level of 8, 9-DHET. The 8, 9-EET pretreatment significantly attenuated the PANC-1 cell death induced by erastin. The 8, 9-EET reduced the Fe2+ concentration, LDH activity and MDA content, and ACSL4 protein expression in erastin-treated PANC-1 cells. The 8,9-EET also restored the ferroportin (FPN) and ferroptosis suppressor protein 1 (FSP1) mRNA expressions in erastin-treated PANC-1 cells. But CYP2J2 knockdown exacerbated the erastin-induced ferroptosis in PANC-1 cells. Besides, CYP2J2 knockdown furtherly down-regulated the gene expression of FPN and FSP1. The 8, 9-EET increased the expression of GPX4 in the erastin-treated PANC-1 cells, which was eliminated by a PPARγ blocker GW9662. And GW9662 abolished the anti-ferroptosis effects of 8,9-EET. Conclusion: CYP2J2/EETs are highly expressed in PDAC tissues. EETs inhibit the ferroptosis via up-regulation of GPX4 in a PPARγ-dependent manner, which contributes to the ferroptosis resistance of PDAC.

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Objective: Many studies have shown that respiratory syncytial virus persistent infection may be the main cause of chronic respiratory pathology. However, the mechanism is unclear. Cystic fibrosis transmembrane conduction regulator (CFTR) is an apical membrane chloride channel, which is very important for the regulation of epithelial fluid, chloride ion, and bicarbonate transport. CFTR dysfunction will lead to changes in bronchial secretions and impair mucus clearance, which is related to airway inflammation. In our previous study, we observed the down-regulation of CFTR in airway epithelial cells in respiratory syncytial virus (RSV) infected mouse model. In this study, we further investigated the expression and function of CFTR by constructing an airway epithelial cell model of RSV persistent infection. Methods: 16HBE14o- cells were infected with RSV at 0.01 multiplicity of infection (MOI). The expression of CFTR was detected by real-time RT-PCR, immunofluorescence, and Western blotting. The intracellular chloride concentration was measured by N-(ethoxycarbonylmethyl)-6-methoxyquinolium bromide (MQAE) and the chloride current was measured by whole-cell patch clamp recording. Results:16HBE14o-cells infected with RSV were survived to successive passages of the third generation (G3), while the expression and function of CFTR was progressively decreased upon RSV infection from the first generation (G1) to G3. Exposure of 16HBE14o-cells to RSV led to the gradual increase of TGF-β1 as well as phosphorylation of Smad2 following progressive RSV infection. Disruption of TGF-β1 signaling by SB431542 prevented Smad2 phosphorylation and rescued the expression of CFTR. Conclusion:RSV infection can lead to defective CFTR function in airway epithelial cells, which may be mediated via activation of TGF-β1 signaling pathway.