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The enzyme AKR1C3 plays a crucial role in hormone and drug metabolism and is associated with abnormal expression in liver cancer, leading to tumor progression and poor prognosis. Nanoparticles modified with HSA can modulate the tumor microenvironment by enhancing photodynamic therapy to induce apoptosis in tumor cells and alleviate hypoxia. Therefore, exploring the potential regulatory mechanisms of resveratrol on AKR1C3 through the construction of HSA-RSV NPs carriers holds significant theoretical and clinical implications for the treatment of liver cancer. The aim of this study is to investigate the targeted regulation of AKR1C3 expression through the loading of resveratrol (RSV) on nanomaterials HSA-RSV NPs (Nanoparticles) in order to alleviate tumor hypoxia and inhibit the progression of hepatocellular carcinoma (HCC), and to explore its molecular mechanism. PubChem database and PharmMapper server were used to screen the target genes of RSV. HCC-related differentially expressed genes (DEGs) were analyzed through the GEO dataset, and relevant genes were retrieved from the GeneCards database, resulting in the intersection of the three to obtain candidate DEGs. GO and KEGG enrichment analyses were performed on the candidate DEGs to analyze the potential cellular functions and molecular signaling pathways affected by the main target genes. The cytohubba plugin was used to screen the top 10 target genes ranked by Degree and further intersected the results of LASSO and Random Forest (RF) to obtain hub genes. The expression analysis of hub genes and the prediction of malignant tumor prognosis were conducted. Furthermore, a pharmacophore model was constructed using PharmMapper. Molecular docking simulations were performed using AutoDockTools 1.5.6 software, and ROC curve analysis was performed to determine the core target. In vitro cell experiments were carried out by selecting appropriate HCC cell lines, treating HCC cells with different concentrations of RSV, or silencing or overexpressing AKR1C3 using lentivirus. CCK-8, clone formation, flow cytometry, scratch experiment, and Transwell were used to measure cancer cell viability, proliferation, migration, invasion, and apoptosis, respectively. Cellular oxygen consumption rate was analyzed using the Seahorse XF24 analyzer. HSA-RSV NPs were prepared, and their characterization and cytotoxicity were evaluated. The biological functional changes of HCC cells after treatment were detected. An HCC subcutaneous xenograft model was established in mice using HepG2 cell lines. HSA-RSV NPs were injected via the tail vein, with a control group set, to observe changes in tumor growth, tumor targeting of NPs, and biological safety. TUNEL, Ki67, and APC-hypoxia probe staining were performed on excised tumor tissue to detect tumor cell proliferation, apoptosis, and hypoxia. Lentivirus was used to silence or overexpress AKR1C3 simultaneously with the injection of HSA-RSV NPs via the tail vein to assess the impact of AKR1C3 on the regulation of HSA-RSV NPs in HCC progression. Bioinformatics analysis revealed that AKR1C3 is an important target gene involved in the regulation of HCC by RSV, which is associated with the prognosis of HCC patients and upregulated in expression. In vitro cell experiments showed that RSV significantly inhibits the respiratory metabolism of HCC cells, suppressing their proliferation, migration, and invasion and promoting apoptosis. Silencing AKR1C3 further enhances the toxicity of RSV towards HCC cells. The characterization and cytotoxicity experiments of nanomaterials demonstrated the successful construction of HSA-RSV NPs, which exhibited stronger inhibitory effects on HCC cells. In vivo, animal experiments further confirmed that targeted downregulation of AKR1C3 by HSA-RSV NPs suppresses the progression of HCC and tumor hypoxia while exhibiting tumor targeting and biological safety. Targeted downregulation of AKR1C3 by HSA-RSV NPs can alleviate HCC tumor hypoxia and inhibit the progression of HCC.
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ABSTRACT: Background : This study aims to determine the impact and mechanism of miR-21-3p on intestinal injury and intestinal glycocalyx during fluid resuscitation in traumatic hemorrhagic shock (THS), and the different impacts of sodium lactate Ringer's solution (LRS) and sodium bicarbonate Ringer's solution (BRS) for resuscitation on intestinal damage. Methods : A rat model of THS was induced by hemorrhage from the left femur fracture. The pathological changes of intestinal tissues and glycocalyx structure were observed by hematoxylin-eosin staining and transmission electron microscope. MiR-21-3p expression in intestinal tissues was detected by real-time quantitative polymerase chain reaction. The expression of glycocalyx-, cell junction-, and PI3K/Akt/NF-κB signaling pathway-related proteins was analyzed by western blot. Results : MiR-21-3p expression was increased in THS rats, which was suppressed by resuscitation with BRS. BRS or LRS aggravated the intestinal injury and damaged intestinal glycocalyx in THS rats. The expression of SDC-1, HPA, ß-catenin, MMP2, and MMP9 was upregulated, the expression of E-cad was downregulated, and the PI3K/Akt/NF-κB signaling pathway was activated in THS rats, which were further aggravated by BRS or LRS. The adverse effect of LRS was more serious than BRS. MiR-21-3p overexpression deteriorated the injury of intestinal tissues and intestinal glycocalyx; increased the expression of SDC-1, HPA, ß-catenin, MMP2, and MMP9 while decreasing E-cad expression; and activated the PI3K/Akt/NF-κB signaling pathway in BRS-resuscitated THS rats. Conclusion : MiR-21-3p aggravated intestinal tissue injury and intestinal glycocalyx damage through activating PI3K/Akt/NF-κB signaling pathway in rats with THS resuscitated with BRS.
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Intestinos , MicroARNs , Solución de Ringer , Choque Hemorrágico , Animales , Masculino , Ratas , Glicocálix/efectos de los fármacos , Glicocálix/metabolismo , Glicocálix/patología , Mucosa Intestinal/metabolismo , Mucosa Intestinal/efectos de los fármacos , Intestinos/patología , Intestinos/efectos de los fármacos , Intestinos/lesiones , Soluciones Isotónicas/farmacología , Soluciones Isotónicas/uso terapéutico , MicroARNs/metabolismo , MicroARNs/genética , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Ratas Sprague-Dawley , Resucitación , Choque Hemorrágico/tratamiento farmacológico , Choque Hemorrágico/metabolismo , Choque Hemorrágico/complicaciones , Transducción de Señal/efectos de los fármacos , Bicarbonato de Sodio/uso terapéutico , Bicarbonato de Sodio/farmacología , Solución de Ringer/farmacología , Solución de Ringer/uso terapéuticoRESUMEN
Sepsis-associated encephalopathy (SAE) is a prevalent complication of sepsis, with hippocampal neuroinflammation playing a crucial role in SAE-induced cognitive impairment. Maresin1 (MaR1), a bioactive docosahexaenoic acid (DHA) metabolite, demonstrates comprehensive anti-inflammatory and neuroprotective attributes. Yet, its protective efficacy against SAE-induced cognitive decline remains unexplored. In this investigation, we implemented a rat SAE model via cecal ligation and puncture (CLP), while lipopolysaccharide (LPS) stimulation of HT22 cells simulated an in vitro SAE model; both models were pre-treated with MaR1. We evaluated rat learning and memory using a water maze, assessed hippocampal neuron damage via Nissl and FJC staining, and observed mitochondrial alterations through TEM. In vivo and in vitro assays gauged levels of Fe2+, MDA, GSH, and SOD. Additionally, Iba1 expression in the hippocampus was examined via immunofluorescence, while SLC7A11 and GPX4 protein expression levels were determined using western blot. Our findings indicated CLP-induced learning and memory impairment in rats, along with heightened ROS, Fe2+, and MDA levels in hippocampal neurons, diminished GSH and SOD levels, and down-regulated ferroptosis-related proteins (GPX4 and SLC7A11). Remarkably, MaR1 treatment attenuated these adverse effects. In LPS-stimulated HT22 cells, MaR1 lowered lipid ROS and bolstered mitochondrial membrane potential. Nonetheless, the ferroptosis inducer Erastin reversed MaR1's protective effects. Transwell experiments further showed MaR1's potential to inhibit microglia activation triggered by ferroptosis in HT22 cells. Consequently, MaR1 may mitigate hippocampal neuroinflammation via activating the SLC7A11/GPX4 ferroptosis signaling pathway, thus ameliorating SAE-related cognitive impairment.
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Disfunción Cognitiva , Ferroptosis , Encefalopatía Asociada a la Sepsis , Sepsis , Animales , Ratas , Cognición , Disfunción Cognitiva/tratamiento farmacológico , Ferroptosis/efectos de los fármacos , Hipocampo , Lipopolisacáridos , Enfermedades Neuroinflamatorias , Especies Reactivas de Oxígeno , Sepsis/tratamiento farmacológico , Encefalopatía Asociada a la Sepsis/tratamiento farmacológico , Transducción de Señal , Superóxido Dismutasa , Ácidos Docosahexaenoicos/administración & dosificaciónRESUMEN
BACKGROUND: Pre-operative and post-operative hypoxemia are frequent complications of Stanford type A aortic dissection (AAD). This study explored the effect of pre-operative hypoxemia on the occurrence and outcome of post-operative acute respiratory distress syndrome (ARDS) in AAD. METHOD: A total of 238 patients who underwent surgical treatment for AAD between 2016 and 2021 were enrolled. Logistic regression analysis was conducted to investigate the effect of pre-operative hypoxemia on post-operative simple hypoxemia and ARDS. Post-operative ARDS patients were divided into pre-operative normal oxygenation group and pre-operative hypoxemia group that were compared for clinical outcomes. Post-operative ARDS patients with pre-operative normal oxygenation were classified as the real ARDS group. Post-operative ARDS patients with pre-operative hypoxemia, post-operative simple hypoxemia, and post-operative normal oxygenation were classified as the non-ARDS group. Outcomes of real ARDS and non-ARDS groups were compared. RESULT: Logistic regression analysis showed that pre-operative hypoxemia was positively associated with the risk of post-operative simple hypoxemia (odds ratios (OR) = 4.81, 95% confidence interval (CI): 1.67-13.81) and post-operative ARDS (OR = 8.514, 95% CI: 2.64-27.47) after adjusting for the confounders. The post-operative ARDS with pre-operative normal oxygenation group had significantly higher lactate, APACHEII score and longer mechanical ventilation time than the post-operative ARDS with pre-operative hypoxemia group (P < 0.05). Pre-operative the risk of death within 30 days after discharge was slightly higher in ARDS patients with pre-operative normal oxygenation than in ARDS patients with pre-operative hypoxemia, but there was no statistical difference(log-rank test, P = 0.051). The incidence of AKI and cerebral infarction, lactate, APACHEII score, mechanical ventilation time, intensive care unit and post-operative hospital stay, and mortality with 30 days after discharge were significantly higher in the real ARDS group than in the non-ARDS group (P < 0.05). After adjusting for confounding factors in the Cox survival analysis, the risk of death within 30 days after discharge was significantly higher in the real ARDS group than in the non-ARDS group (hazard ratio(HR): 4.633, 95% CI: 1.012-21.202, P < 0.05). CONCLUSION: Preoperative hypoxemia is an independent risk factor for post-operative simple hypoxemia and ARDS. Post-operative ARDS with pre-operative normal oxygenation was the real ARDS, which was more severe and associated with a higher risk of death after surgery.
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Disección Aórtica , Síndrome de Dificultad Respiratoria , Humanos , Síndrome de Dificultad Respiratoria/diagnóstico , Síndrome de Dificultad Respiratoria/epidemiología , Síndrome de Dificultad Respiratoria/etiología , Disección Aórtica/cirugía , Respiración Artificial , Pulmón , Hipoxia/diagnóstico , Hipoxia/epidemiología , Hipoxia/etiología , Estudios RetrospectivosRESUMEN
Objective: Neuroinflammation is a major etiology of cognitive dysfunction due to sepsis. Maresin1 (MaR1), identified as a docosahexaenoic acid (DHA)-derived metabolite from macrophages, has been demonstrated to exhibit potent neuroprotective and anti-inflammatory effects. Nevertheless, detailed functions and molecular mechanism of MaR1 in sepsis-induced cognitive dysfunction has not been fully elucidated. Here, we aimed to investigate potential neuroprotective effects of MaR1 on microglia-induced neuroinflammation in sepsis-induced cognitive impairment and to explore its anti-inflammatory mechanism. Methods: Different doses of MaR1 were administered to septic rats by via tail vein injection. The optimal dose was determined based on the 7-day survival rate of rats from each group. derived from macrophages with both anti-inflammatory to observe the ameliorative effects of MaR1 at optimal doses on cognitive dysfunction in septic rats. The effects of MaR1 on neuroinflammation-mediated microglial activation, neuronal apoptosis, and pro-inflammatory cytokine productions were in vivo and in vitro assayed, using Western blot, ELISA, TUNEL staining, Nissl staining, and the immunofluorescence method. To further elucidate anti-inflammatory machinery of MaR1, protein expressions of NLRP3 inflammatory vesicles and TLR4-NF-κB pathway-related proteins were subjected to Western blot assay. Results: After tail vein injection of MaR1 with different doses (2 ng/g, 4 ng/g, 8 ng/g), the results showed that 4 ng/g MaR1 treatment significantly increased the rats' 7-day survival rate compared to the CLP controls. Therefore, subsequent experiments set 4 ng/g MaR1 as the optimal dose. Morris water maze experiments confirmed that MaR1 significantly reduced space memory dysfunction in rats. In addition, in CLP rats and LPS-stimulated BV2 microglia, MaR1 significantly reduced activated microglia and pro-inflammatory cytokines levels and neuronal apoptosis. Mechanically, MaR1 inhibits microglia-induced neuroinflammation through suppressing activations of NLRP3 inflammatory vesicles and TLR4-NF-κB signal pathway. Conclusion: Collectively, our findings suggested that MaR1 might be a prospective neuroprotective compound for prevention and treatment in the sepsis process.
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INTRODUCTION: Acute lung injury (ALI) is a major cause of death in sepsis patients. The Six-transmembrane protein of prostate 2 (STAMP2) is a key regulator of inflammation, while its role in septic ALI remains unclear. MATERIAL AND METHODS: Male C57BL/6 mice were subjected to cecal ligation puncture (CLP) to induce experimental sepsis whereas lipopolysaccharide (LPS)-stimulated RAW 264.7 cells were used as the models of septic ALI in vivo and in vitro, respectively. Overexpression of STAMP2 in mouse lungs and RAW264.7 cells was performed with an adenoviral vector. We measured histological lung injury, lung wet/dry weight (W/D) ratio, and pulmonary myeloperoxidase (MPO) activity to assess lung injury extent. Cell counts in bronchoalveolar lavage fluid (BALF) were measured using Giemsa staining. The concentration of inflammatory factors was detected by enzyme-linked immunosorbent assay. The polarization of macrophages was evaluated by inducible nitric oxide synthase (iNOS) and F4/80 staining. The activation of cell apoptosis and NF-κB pathway was evaluated using Western blot, TUNEL staining, immunofluorescence, and immunohistochemistry. RESULTS: Overexpression of STAMP2 alleviated CLP-induced lung injury of mice with decreased W/D ratio of the lung, and MPO activity in lung tissue. STAMP2 overexpression reduced the lung infiltration of inflammatory cells, and the levels of TNF-a, IL-6, and macrophage chemoattractant protein-1 (MCP-1) in BALF. Overexpressed STAMP2 inhibited macrophage M1 polarization in lung tissues as indicated by F4/80 and iNOS stainings in lung tissue. STAMP2 overexpression inhibited RAW 264.7 cell apoptosis by increasing Bcl-2 and decreasing Bax and cleaved-caspase 3 expression. Besides, STAMP2 overexpression suppressed nuclear factor κB (NF-κB) p65 pathway activation, as evidenced by reduced phosphorylation of IκBα, and phosphorylation and translocation of NF-κB p65. In vitro study further proved that STAMP2 overexpression suppressed the NF-κB pathway (IκBα/p65) in macrophages and decreased macrophage M1 polarization and M1-associated inflammatory factor production (TNF-a, IL-6, and MCP-1). CONCLUSIONS: Our study for the first time demonstrated that STAMP2 might be able to reduce inflammation in sepsis-induced ALI by inhibiting macrophage M1 polarization through repressing NF-κB signaling activation.
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Lesión Pulmonar Aguda , Sepsis , Masculino , Animales , Ratones , FN-kappa B/metabolismo , Inhibidor NF-kappaB alfa , Interleucina-6 , Próstata/metabolismo , Ratones Endogámicos C57BL , Lesión Pulmonar Aguda/etiología , Lesión Pulmonar Aguda/metabolismo , Pulmón/patología , Sepsis/metabolismo , Macrófagos/metabolismo , Inflamación/patología , LipopolisacáridosRESUMEN
ABSTRACT: Background: Acute lung injury (ALI) induced by sepsis is distinguished by an inflammatory progression. Herein, we investigated the action of circular RNA kelch like family member 2 (circKlhl2) in sepsis-induced ALI. Methods: The animal or cell model of sepsis ALI was established by LPS stimulation. The contents of circKlhl2, microRNA-29b-3p (miR-29b-3p), rho-associated coiled-coil containing protein kinase 1 (ROCK1), CyclinD1, B-cell lymphoma-2 (Bcl-2), and cleaved-caspase 3 (C-caspase-3) were detected by quantitative real-time polymerase chain reaction and western blot, respectively. Cell viability was assessed by cell counting kit 8 assay. Cell cycle and apoptosis were evaluated by flow cytometry. The abundances of proinflammatory cytokines were detected by enzyme-linked immunosorbent assay. Besides, the targeted relationship between miR-29b-3p and circKlhl2 or ROCK1 was verified by dual-luciferase reporter assay, RNA immunoprecipitation assay and RNA pull-down assay. Results: Loss of circKlhl2 mitigated lung injury and proinflammatory cytokine expression in sepsis-ALI mice model and alleviated LPS-induced apoptosis and inflammatory response in microvascular endothelial cell (MPVECs) in vitro . The abundances of circKlhl2 and ROCK1 were boosted, while the miR-29b-3p level was diminished in the animal or cell model of sepsis-ALI. MiR-29b-3p inhibition abrogated circKlhl2 knockdown-mediated effects on MPVECs injury. Moreover, miR-29b-3p overexpression promoted cell proliferation and inhibited apoptosis and inflammation in LPS-treated MPVECs, while ROCK1 enhancement reversed these effects. Conclusion: CircKlhl2 expedited the sepsis-induced ALI by adjusting miR-29b-3p/ROCK1 axis.
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Lesión Pulmonar Aguda , MicroARNs , Sepsis , Animales , Ratones , Lesión Pulmonar Aguda/metabolismo , Apoptosis/genética , Regulación hacia Abajo , Lipopolisacáridos/farmacología , MicroARNs/genética , MicroARNs/metabolismo , Quinasas Asociadas a rho/genética , Quinasas Asociadas a rho/metabolismo , Sepsis/complicaciones , Sepsis/genéticaRESUMEN
OBJECTIVE: Sepsis is the most common cause of death in the intensive care unit. Moreover, sepsis is the leading cause of acute lung injury (ALI). Serine-arginine protein kinase 1 (SRPK1) was demonstrated to promote the development of ALI. However, the potentials of SRPK1 in sepsis-induced ALI are still unknown. This study aimed to investigate the potentials of SRPK1 in sepsis-induced ALI and the underlying mechanisms. METHODS: Cecal ligation and puncture (CLP) was performed to establish a sepsis-induced ALI model in vivo. Primary human pulmonary microvascular endothelial cells (HPMECs) were exposed to lipopolysaccharide (LPS) to construct a sepsis-induced ALI model in vitro. Gene expression was detected using western blot and qRT-PCR. The interaction between forkhead box O3 (FOXO3) and NOD-like receptor thermal protein domain associated protein 3 (NLRP3) was detected using luciferase and Chromatin immunoprecipitation (ChIP) assay. Cellular functions were CCK-8, colony formation, PI staining, and flow cytometry assay. RESULTS: SRPK1 was downregulated in patients with sepsis-induced ALI. Overexpression of SRPK1 suppressed the pyroptosis of HPMECs as well as promoted cell proliferation. Additionally, SRPK1 overexpression alleviated sepsis-induced ALI in vivo. SRPK1 activated phosphatidylinositol3-kinase (PI3K) signaling pathways. Blocking the activation of PI3K degraded the cellular functions of HPMECs. Moreover, FOXO3 transcriptionally inactivated NLRP3 and suppressed its mRNA and protein expression. CONCLUSION: Taken together, SRPK1 suppressed sepsis-induced ALI via regulating PI3K/AKT/FOXO3/NLRP3 signaling. SRPK1 may be the potential biomarker for sepsis-induced ALI.
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Lesión Pulmonar Aguda , Arginina Quinasa , Sepsis , Humanos , Lesión Pulmonar Aguda/etiología , Lesión Pulmonar Aguda/metabolismo , Células Endoteliales/metabolismo , Proteína Forkhead Box O3/genética , Lipopolisacáridos , Pulmón/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Fosfatidilinositol 3-Quinasas , Proteínas Serina-Treonina Quinasas , Proteínas Proto-Oncogénicas c-akt/metabolismo , Sepsis/complicaciones , Sepsis/metabolismoRESUMEN
BACKGROUND AND PURPOSE: Hyperoxia-induced acute lung injury (HALI) is a critical life-threatening disorder characterized by severe infiltration immune cells and death of type II alveolar epithelial cells (AECII). However, little is known about the relations between immune cells and AECII in HALI. IL-17A is a pro-inflammatory cytokine mainly secreted by Th17 cells, contributing to the pathogenesis of various inflammatory diseases. The present study investigated the role of IL-17A in cell-cell communication between immune cells and AECII in HALI, and explored the therapeutic effect of salidroside (Sal, a natural anti-inflammatory agents) on HALI. METHODS: Mice with HALI were induced by exposure to hyperoxia over 90% for 12 h, 24 h, 48 h or 72 h, and the optimal timing was detected by H&E and Masson staining. Ferroptosis was confirmed by detecting the levels of MDA, Fe2+ and GPX4, and the morphological alterations of AECII under transmission electron microscopy. The expression of pro-inflammatory cytokine, including IL-6, TGF-ß1, IL-17A and IL-17A receptor (IL-17RA) were measured by Western blotting and immunohistochemical stanning. The ferroptosis-related Act1/TRAF6/p38 MAPK pathway was detected by Western blotting. The role of pro-inflammatory cytokine IL-17A for AECII ferroptosis, and the effect of Sal on HALI were investigated by administration of Y-320 (IL-17 inhibitor) and Sal respectively 3 days before mice exposed to hyperoxia. RESULTS: Mice exposed to hyperoxia for 24 h suffered sufficient HALI with inflammatory cell infiltration and collagen deposition, and exhibited features of ferroptosis under TME. Meanwhile, compared with sham mice, mice exposed to hyperoxia showed down-regulation of GPX4, and up-regulation of IL-6, TGF-ß1, IL-17A, IL-17RA, Act1, TRAF6, p38 MAPK and p-p38 MAPK. Moreover, inhibition of IL-17A with Y-320 or administration with Sal could reverse the effect caused by hyperoxia respectively. CONCLUSIONS: IL-17A is associated with immune cells infiltration in HALI, and contributes to ferroptosis of AECII that related to Act1/TRAF6/p38 MAPK pathway. Additionally, Sal protects against HALI throughout the whole pathogenic process. Video Abstract.
Oxygen inhalation has been widely used in the treatment of some diseases caused by hypoxia. This often leads people to mistakenly believe that oxygen inhalation is beneficial without harm. However, long-term high concentration oxygen inhalation will cause serious harm to the human body, sometimes even fatal. Hyperoxia causes lung cells to secrete proinflammatory factors, which promote the differentiation of infiltrated immune cells. The differentiated immune cells in turn act on lung cells and lead to their death. In short, this process is a vicious circle. Our research explores this process and is committed to finding a drug to reduce the damage of hyperoxia to the lungs when oxygen must be inhaled.
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Lesión Pulmonar Aguda , Ferroptosis , Hiperoxia , Ratones , Animales , Interleucina-17 , Factor 6 Asociado a Receptor de TNF/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo , Células Epiteliales Alveolares/metabolismo , Hiperoxia/metabolismo , Factor de Crecimiento Transformador beta1/metabolismo , Interleucina-6/metabolismo , Lesión Pulmonar Aguda/tratamiento farmacológico , Citocinas/metabolismo , Proteínas PortadorasRESUMEN
Background: For patients with multiple organ dysfunction syndrome (MODS), timely assessment of the condition and real-time adjustment of the treatment plan are of critical importance. To this end, transthoracic echocardiography (TTE) is widely used in clinical practice, but whether TTE can improve the short-term prognosis of MODS patients is currently unclear. Methods: We extracted data of patients from the Medical Information Mart for Intensive Care III (MIMIC-III) database and included cases according to inclusion and exclusion criteria. The primary endpoint was the mortality within 28 days after admission to the intensive care unit (ICU), and multi-variate regression analysis was conducted to infer the risk factors associated death within 28 days after ICU admission. Double robust analysis was conducted to investigate the correlation between TTE and the endpoint. A gradient-boosted model (GBM) was constructed to calculate the propensity score (PS) of patients received TTE, so as to reduce the difference of variates between these two groups. Results: A total of 13,844 MODS cases were included and were divided into a TTE group and a non-TTE group. There were 5,022 cases (36.28%) in the TTE group, 2,416 (48.10%) of whom were female; and 8,822 (63.72%) in the non-TTE group, 4,129 of whom (46.80%) were female. The covariates that contributed significantly to PS included chronic heart failure (CHF), creatine kinase (CK), troponin, partial pressure of oxygen (PO2), and sequential organ failure assessment (SOFA) scores. Double robust analysis showed that within 28 days after ICU admission, the TTE group had lower risk of death when compared with the non-TTE group. The adjusted odds ratio (OR) value of TTE for 28 days death was 0.73 [95% confidence interval (CI): 0.65-0.82; P<0.001]. The other 3 models had similar results, suggesting that conduct TTE for patients with MODS in ICU was associated with lower risk of 28 days mortality. Conclusions: TTE can reduce the mortality of MODS patients in the ICU.
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Breast cancer is one of the most common causes of female death globally. Numerous clinical drugs for breast cancer have been developed, but the unsatisfactory, inevitable side effects and drug resistance are the emerging threatens. Therefore, it is necessary to investigate the comprehensive mechanism of breast cancer. Enhancer of zeste homolog 2 (EZH2) is a candidate oncogenic driver in diverse cancers, such as breast cancer. The canonical role of EZH2 has been vastly investigated, but the non-canonical function of EZH2 in breast cancer remains unclear. Here, we demonstrated that EZH2 exacerbated breast cancer in non-canonical manner by methylating STAT3. EZH2 over-expressed in breast cancer patients and regulated STAT3 post-transcriptionally according to TCGA datasets. Chemical and genetic inhibition of EZH2 impeded proliferation and migration of breast cancer cells, which may be partially rescued by STAT3 over-expression. EZH2 physically interacted with STAT3 and methylated STAT3 directly, resulting in increased nuclear localization and chromatin of STAT3. Furthermore, the mutation of STAT3 methylation site, targeted by EZH2, impeded the transcriptional activity of STAT3. Eventually, disturbed STAT3 methylation by EZH2 in animal model showed decreased breast cancer growth. These data confirm that EZH2 exacerbates breast cancer by methylating STAT3 directly, and thus providing a promising therapeutic target for breast cancer.
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BACKGROUND: Heart failure with midrange ejection fraction (HFmrEF) has been recently acknowledged as a separate phenotype, but metabolomics evaluation of this subtype remains largely unexamined. METHODS: A quantitative metabolomics study on amino acids and acylcarnitines was performed to characterize different states of heart failure (HF) in 628 participants. Both multivariate orthogonal partial least squares- discriminant analysis and univariate Mann-Whitney U test were used to explore reliable metabolic profiles associated with different HF states. The resulting metabolites were further refined to obtain diagnostic metabolite scores (DMSs) with the use of ordinal logistic regression. Lasso-penalized regression was applied to produce a survival-associated prognostic metabolite score (PMS). The Cox proportional hazards model, Kaplan-Meier curves, and time-dependent receiver operating characteristics were used for a comprehensive assessment of prognostic value using PMS versus traditional clinical biomarkers. RESULTS: The optimized models identified a panel of 15 differential metabolites that were shared across different HF states, whereas some metabolites were associated with a specific state. PMS consisting of 9 metabolites demonstrated an appreciably better prognostic value (hazard ratio [HR] 1.62, 95% confidence interval [CI] 1.25-2.1) vs the natural logarithm of N-terminal pro-B-type natriuretic peptide (Ln[NT-proBNP]; HR 1.23, 95% CI 0.94-1.61; P < 0.001). The overall area under the receiver operating characteristic curve value of PMS was superior to that of Ln(NT-proBNP) in risk prediction for patients with HFmrEF and HF with reduced ejection fraction (HFrEF) subtypes (P < 0.001). CONCLUSIONS: Targeted metabolomics has provided a novel understanding of the molecular mechanism underlying HF. Both DMS and PMS clearly demonstrated HFmrEF as a distinct phenotype between a mild HF with preserved ejection fraction state and a severe HFrEF state. PMS exhibited superior prognostic value than Ln(NT-proBNP). Further investigation is needed with independent large-scale validation.
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Aminoácidos/metabolismo , Carnitina/análogos & derivados , Insuficiencia Cardíaca , Metaboloma , Metabolómica/métodos , Volumen Sistólico , Análisis de Varianza , Biomarcadores/metabolismo , Carnitina/metabolismo , China/epidemiología , Femenino , Insuficiencia Cardíaca/diagnóstico , Insuficiencia Cardíaca/metabolismo , Insuficiencia Cardíaca/mortalidad , Insuficiencia Cardíaca/fisiopatología , Humanos , Estimación de Kaplan-Meier , Masculino , Persona de Mediana Edad , Gravedad del Paciente , Fenotipo , Valor Predictivo de las Pruebas , Pronóstico , Curva ROCRESUMEN
OBJECTIVE: To analyze the application of functional residual capacity (FRC)-guided optimal positive end-expiratory pressure (PEEP) in pulmonary retention in patients with acute respiratory distress syndrome (ARDS), and to explore the correlation between FRC and trans-pulmonary pressure and their predictive value for prognosis. METHODS: Seventy-eight ARDS patients on mechanical ventilation admitted to department of critical care medicine of the First Affiliated Hospital of Jinzhou Medical University from March 2018 to May 2019 were enrolled. According to random number table method, the patients were divided into experimental group and the control group. PEEP of all patients were gradually increased in recruitment after fully sedation and analgesia. The best PEEP was set by monitoring FRC in the experimental group, and by monitoring maximum oxygen in the control group set. The differences before and after 30 minutes and 2 hours recruitment manoeuvres in dynamic compliance (Cdyn), oxygenation index (PaO2/FiO2), and mechanical power (MP) were compared between the two groups. Pearson method was used to analyze the correlation between FRC and trans-pulmonary pressure. The predictive value of FRC and trans-pulmonary pressure for 28-day mortality in patients with ARDS was analyzed by receiver operating characteristic (ROC) curve. RESULTS: The optimal PEEP was (16.24±1.57) cmH2O (1 cmH2O = 0.098 kPa) in the experimental group and (14.11±1.15) cmH2O in the control group in recruitment maneuvres, with statistically significant difference between the two groups (t = 5.678, P = 0.000). Pearson correlation analysis showed that there was a significant correlation between FRC and trans-pulmonary pressure in ARDS patients (r = 0.759, P = 0.000). Cdyn and PaO2/FiO2 in the experimental group were higher than the control group at 30 minutes and 2 hours after recruitment maneuvres [Cdyn (mL/cmH2O): 61.16±3.55 vs. 58.54±5.25, 58.59±2.82 vs. 56.86±3.40; PaO2/FiO2 (mmHg, 1 mmHg = 0.133 kPa): 245.27±14.86 vs. 239.00±5.34, 192.25±5.11 vs. 188.86±5.07], MP was lower than the control group (J/min: 16.32±1.11 vs. 17.05±1.22, 15.22±1.25 vs. 17.03±1.50), the difference was statistically significant (all P < 0.05). The ROC curve analysis showed that both FRC and trans-pulmonary pressure had predictive value for the 28-day mortality of ARDS patients, and the area under the ROC curve (AUC) was 0.868, and 0.828 respectively (both P < 0.01). CONCLUSIONS: Measuring FRC in patients with ARDS during recruitment maneuvres can guide optimal PEEP. FRC was significantly correlated with trans-pulmonary pressure, and both of them had predictive value for 28-day mortality in ARDS patients.
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Respiración con Presión Positiva , Síndrome de Dificultad Respiratoria , Capacidad Residual Funcional , Humanos , Pulmón , PronósticoRESUMEN
OBJECTIVE: To verify the clinical safety and efficacy of new intelligent ventilation mode adaptive minute ventilation (AMV)+IntelliCycle ventilation in patients with mild-to-moderate acute respiratory distress syndrome (ARDS). METHODS: The patients with mild-to-moderate ARDS, admitted to intensive care unit (ICU) of the First Affiliated Hospital of Jinzhou Medical University from February 2018 to February 2019, were enrolled in the study. The patients were divided into synchronous intermittent mandatory ventilation+pressure support ventilation (SIMV+PSV) group and AMV+IntelliCycle group according to the random number table method. All patients were given mechanical ventilation, anti-infection, analgesia and sedation, nutritional support and symptomatic treatment of primary disease after admission. SV800 ventilator was used for mechanical ventilation. In the AMV+IntelliCycle group, after setting the minute ventilation volume (VE), inhaled oxygen concentration (FiO2) and positive end expiratory pressure (PEEP), the ventilator was turned on the full-automatic mode, and the preset value of VE percentage was 120%. In the SIMV+PSV group, the ventilator parameters were set as follows: the ventilation frequency was 12-20 times/min, the inspiratory expiratory ratio was 1:1-2, the peak inspiratory pressure (PIP) limit level was 35-45 cmH2O (1 cmH2O = 0.098 kPa), and the setting of FiO2 and PEEP was as the same as that of AMV+IntelliCycle group, the triggering flow was set to 2 L/min. All of the clinical parameters between the two groups were compared. The main outcomes were duration of mechanical ventilation, ventilator alarm times, manual operation times, and the mechanical power; the secondary outcomes were respiratory rate (RR), VE, tidal volume (VT), PIP, mouth occlusion pressure (P0.1), static compliance (Cst), work of breathing (WOB), and time constant at 0, 6, 12, 24, 48, 72, and 120 hours; and the blood gas analysis parameters of patients before and after ventilation were recorded. RESULTS: A total of 92 patients with mild-to-moderate ARDS were admitted during the study period, excluding those who quit the study due to death, abandonment of treatment, accidental extubation of tracheal intubation and so on. Eighty patients were finally enrolled in the analysis, with 40 patients in SIMV+PSV group and AMV+IntelliCycle group respectively. (1) Results of main outcomes: compared with the SIMV+PSV mode, AMV+IntelliCycle ventilation mode could shorten the duration of mechanical ventilation (hours: 106.35±55.03 vs. 136.50±73.78), reduce ventilator alarm times (times: 10.35±5.87 vs. 13.93±6.87) and the manual operations times (times: 4.25±2.01 vs. 6.83±3.75), and decrease the mechanical power (J/min: 12.88±4.67 vs. 16.35±5.04, all P < 0.05). But the arterial partial pressure of carbon dioxide (PaCO2) of AMV+IntelliCycle group was significantly higher than that of SIMV+PSV group [mmHg (1 mmHg = 0.133 kPa): 41.58±6.81 vs. 38.45±5.77, P < 0.05]. (2) Results of secondary outcomes: the RR of both groups was improved significantly with the prolongation of ventilation time which showed a time effect (F = 4.131, P = 0.005). Moreover, compared with SIMV+PSV mode, AMV+IntelliCycle mode could maintain a better level of RR, with intervention effect (F = 5.008, P = 0.031), but no interaction effect was found (F = 2.489, P = 0.055). There was no significant difference in VE, PIP, P0.1 or Cst between the two groups, without intervention effect (F values were 3.343, 2.047, 0.496, 1.456, respectively, all P > 0.05), but they were significantly improved with the prolongation of ventilation time in both groups, with time effect (F values were 2.923, 12.870, 23.120, 7.851, respectively, all P < 0.05), but no interaction effect was found (F values were 1.571, 1.291, 0.300, 0.354, respectively, all P > 0.05). The VT, WOB or time constant in both groups showed no significant changes with the prolongation of ventilation time, and no significant difference was found between the two groups, there was neither time effect (F values were 0.613, 1.049, 2.087, respectively, all P > 0.05) nor intervention effect (F values were 1.459, 0.514, 0.923, respectively, all P > 0.05). CONCLUSIONS: AMV+IntelliCycle ventilation mode can shorten the ventilation time of patients with mild-to-moderate ARDS, reduce mechanical power, and reduce the workload of medical care, but PaCO2 in the patients with AMV+IntelliCycle mode is higher than that in the patients with SIMV+PSV mode.
Asunto(s)
Automatización , Respiración Artificial/métodos , Síndrome de Dificultad Respiratoria/terapia , Humanos , Respiración con Presión Positiva , Volumen de Ventilación Pulmonar , Ventiladores MecánicosRESUMEN
Although natural killer T cells (NKT cells) are thought to be generated from CD4+CD8+ (DP) thymocytes, the developmental origin of CD4-CD8- (DN) NKT cells has remained unclear. In this study, we found the level of NK1.1 expression was highest in DN cells, followed by CD4 and CD8 (SP) and DP cells. The level of NK1.1 expression was highest in CD44+CD25- (DN1) cells, after that CD44+CD25+ (DN2), finally, CD44-CD25- (DN3) and CD44- CD25+ (DN4) cells. Unexpectedly, cytoplasmic CD3 was not only expressed in SP and DP thymocytes but also in most DN thymocytes at various stages. The mean fluorescence of cytoplasmic and surface CD3 in DN cells was significantly lower than in mature (SP) T and NKT cells in the thymus and spleen. Interestingly, there were more NKT cells in DN-cytoplasmic CD3 expression cells was higher than in DN-surface CD3 expression cells. There were more CD3-NKT cells in DN1 thymocytes than in TCR-ß-NKT cells. NKT cells expressed higher levels of IL-7Rα which was correlated with CD44 expression in the thymus. Our data suggest that T cells and NKT cells follow similar patterns of expression with respect to cytoplasmic and surface CD3. Cytoplasmic CD3 could be used as a marker for early stage T cells. Both cytoplasmic CD3 and surface CD3 were expressed in mature T cells and immature T cells, including the immature cytoplasmic CD3+ surface CD3- and surface CD3+TCR-ß- cells in DN1-NKT thymocytes. CD44 could be used as an additional marker of NKT cells which may originate from cytoplasmic CD3-positive DN thymocytes that express CD44 and IL-7Rα in mice.
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Complejo CD3/metabolismo , Citoplasma/metabolismo , Receptores de Hialuranos/metabolismo , Células T Asesinas Naturales/citología , Receptores de Interleucina-7/metabolismo , Timocitos/citología , Animales , Antígenos Ly/metabolismo , Antígenos CD4/metabolismo , Antígenos CD8/metabolismo , Membrana Celular/metabolismo , Femenino , Ratones , Ratones Endogámicos C57BL , Subfamilia B de Receptores Similares a Lectina de Células NK/metabolismo , Bazo/citología , Timo/citologíaRESUMEN
BACKGROUND: TRIM32 is overexpressed in several human cancers. However, its expression pattern, biological characteristics and mechanisms in human non-small cell lung cancer (NSCLC) have not been reported. METHODS: We examined TRIM32 protein in 115 cases of NSCLC specimens. TRIM32 plasmid transfection and siRNA knockdown was carried out in NSCLC cell lines. AnnexinV/PI and JC-1 staining were performed to examine the change of apoptosis and mitochondrial membrane potential. Western blot was used to detect change of downstream proteins. RESULTS: We found that TRIM32 protein was upregulated in 69 cases and positively correlated with advanced TNM stage. TRIM32 overexpression also correlated with poor survival of NSCLC patients. Biological assays demonstrated that TRIM32 overexpression promoted while it depletion inhibited cell growth, colony formation and invasion. In addition, TRIM32 maintained NSCLC cell viability and reduced apoptosis when treated with cisplatin. JC-1 and CellRox staining demonstrated that TRIM32 could maintain mitochondrial membrane potential and reduce Reactive Oxygen Species (ROS) production after cisplatin treatment. Western blot analysis showed that TRIM32 overexpression downregulated caspase 3 cleavage and cytochrome c release. TRIM32 also positively regulated Bcl-2 protein expression and NF-κB signaling. Inhibition of NF-κB abolished the effects of TRIM32 on Bcl-2. CONCLUSION: Taken together, our results indicated that TRIM32 is overexpressed in NSCLC and regulates cisplatin resistance, possibly through NF-κB and Bcl-2.
RESUMEN
The purpose of this study was to investigate the protective effects of Saikosaponin a (SSa), a triterpene saponin derived from Radix bupleuri, on lipopolysaccharide (LPS)-induced acute lung injury (ALI) using a murine model. The mice were given SSa 1 h after intranasal instillation of LPS. Then, lung histopathological examination, the wet/dry (W/D) ratio, myeloperoxidase (MPO), and inflammatory cytokines in bronchoalveolar lavage fluid (BALF) were detected in this study. The results showed that SSa reduced lung pathological injury induced by LPS. Furthermore, LPS-induced lung W/D ratio, MPO activity, and inflammatory cytokines TNF-α and IL-1ß in BALF were significantly inhibited by SSa. In addition, SSa suppressed LPS-induced NF-κB activation and NLRP3 inflammasome expression. In conclusion, we found that SSa played a critical anti-inflammatory effect through inhibition of NF-κB and NLRP3 signaling pathways and protected against LPS-induced ALI.
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Lesión Pulmonar Aguda/prevención & control , Antiinflamatorios/farmacología , Lipopolisacáridos , Pulmón/efectos de los fármacos , Ácido Oleanólico/análogos & derivados , Saponinas/farmacología , Lesión Pulmonar Aguda/inducido químicamente , Lesión Pulmonar Aguda/metabolismo , Lesión Pulmonar Aguda/patología , Animales , Líquido del Lavado Bronquioalveolar/química , Caspasa 1/metabolismo , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Mediadores de Inflamación/metabolismo , Interleucina-1beta/metabolismo , Pulmón/metabolismo , Pulmón/patología , Masculino , Ratones Endogámicos BALB C , FN-kappa B/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Infiltración Neutrófila/efectos de los fármacos , Ácido Oleanólico/farmacología , Edema Pulmonar/metabolismo , Edema Pulmonar/patología , Edema Pulmonar/prevención & control , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
BACKGROUND: CD117 is expressed on double-negative (DN; CD4-CD8-) cells (Nat Rev Immunol 14:529-545; 2014), but whether it is expressed in other stages and its subsequent functions are unclear. We used an improved method of flow cytometry to analyze different populations of thymocytes (Sci Rep 4:5781; 2014). The expression of CD117 and CTLA-4 were directly assayed in the early stage of thymocytes. METHODS: Flow cytometry was used to analyze different populations of thymocytes, and T-cell proliferation assays, RT-PCR, and real-time RT-PCR were used to characterize the stem cells and examine the function of CD44+CD117+ cells. RESULTS: In DN cells, CD117 expression was greatest on CD44+CD25+ cells (DN2), followed by CD44+CD25- (DN1), CD44-CD25+ (DN3), and CD44-CD25- (DN4) cells. In thymocytes, CD117 expression was highest in DN cells, followed by single-positive (SP; CD4 or CD8) and double-positive (DP; CD4+CD8+) cells. Especially, CD117 expression was positively associated with CD44 and CTLA-4 expression. CTLA-4 expression was highest in DN cells, followed by SP and DP cells. CTLA-4 expression was positively associated with CD25, CD44, and Foxp3 expression. CD44+CD117+ T cells expressed more CTLA-4, which suppressed T-cell proliferation and blocked CTLA-4 to cause antibody-induced T-cell proliferation. CONCLUSION: These results suggest that CD44+CD117+ T cells are stem cells and a specific T-cell phenotype that initially develops in the thymus, but they do not progress through DN3 and DN4 stages, lack a DP stage, and potently suppress T-cell proliferation and modulate the CTLA-4 pathway.
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Antígeno CTLA-4/inmunología , Receptores de Hialuranos/inmunología , Proteínas Proto-Oncogénicas c-kit/inmunología , Células Madre/inmunología , Linfocitos T/inmunología , Animales , Antígeno CTLA-4/genética , Diferenciación Celular/genética , Proliferación Celular/genética , Citometría de Flujo , Humanos , Receptores de Hialuranos/genética , Activación de Linfocitos/inmunología , Ratones , Proteínas Proto-Oncogénicas c-kit/genética , Transducción de Señal , Células Madre/citología , Células Madre/metabolismo , Subgrupos de Linfocitos T/inmunología , Linfocitos T/citología , Linfocitos T/metabolismoRESUMEN
BACKGROUND: Heparin saline (HS) is theoretically superior to normal saline (NS) for maintaining the patency of central venous catheters (CVCs), but the comparative efficacy of them remains controversial. The aim of this systematic review and meta-analysis was to assess the efficacy of NS versus HS in the maintenance of the patency of CVCs in adult patients. METHODS: We searched PubMed, Embase and the Cochrane library databases. Randomized controlled trials (RCTs) evaluating the use of NS vs. HS to maintain the permeability of CVCs among adult patients were included in our meta-analysis. References of relevant papers were reviewed manually. No language restriction was applied. Non-human studies were excluded. Pooled relative risk (RR) was calculated using a Mantel-Haenszel random-effects model. We also performed subgroup analysis examining the effect of the duration of catheter placement on the outcome. All statistical tests were two-sided using a significance level of 0.05. RESULTS: Ten RCTs involving 7875 subjects (with analysis at patient, catheter, lumen and line access level) were included in this meta-analysis. Whether in terms of pooled or local analysis (RR with 95% confidence interval spans 1), NS can be equally, if not more effective, in keeping the CVCs open. Of studies reporting secondary outcomes (maneuver needed, heparin-induced thrombocytopenia, haemorrhage, central venous thrombosis and catheter-related bloodstream infection), heparinised saline was shown not to be superior to non-heparinised solution. Subgroup analysis in patients with short vs long term CVC placement was consistent with the main outcome partly and in particular for maintenance of catheter patency in patients with a long-term placement i.e. >30 days, the RR was 0.97 (n = 6589; 95% CI = 0.76 to 1.23; P = 0.796). However, for patients in whom the catheter was in place for <30 days, the RR was 1.52 (n = 1286; 95% CI = 1.02 to 2.27; P = 0.041). CONCLUSIONS: Based on the results of this meta-analysis, HS is not superior to NS in reducing CVCs occlusion. But in the short term, the use of HS is slightly superior to NS for flushing catheters from a statistical point of view.
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Catéteres Venosos Centrales , Heparina/administración & dosificación , Cloruro de Sodio/administración & dosificación , Irrigación Terapéutica/métodos , Irrigación Terapéutica/normas , Adulto , Obstrucción del Catéter/efectos adversos , Heparina/uso terapéutico , Humanos , Cloruro de Sodio/uso terapéutico , Trombosis/tratamiento farmacológico , Trombosis/prevención & controlRESUMEN
Tenuigenin (TNG) has been reported to have various pharmacological activities, such as anti-oxidative and anti-inflammatory activities. However, the protective effects of TNG on lipopolysaccharides (LPS)-induced acute kidney injury (AKI) are still not clear. The aim of this study was to investigate the protective effects and mechanism of TGN on LPS-induced AKI in mice. The kidney histological change, levels of blood urea nitrogen (BUN), and creatinine were measured to assess the protective effects of TNG on LPS-induced AKI. The levels of TNF-α, IL-1ß, and IL-6 in serum and kidney tissues were detected by ELISA. The extent of nuclear factor kappa-B (NF-κB) p65 and the expression of Toll-like receptor-4 (TLR4) were detected by western blot analysis. The results showed that TNG markedly attenuated the histological alterations, BUN and creatinine levels in kidney. TNG also suppressed LPS-induced TNF-α, IL-1ß, and IL-6 production. Furthermore, the expression of TLR4 and NF-κB activation induced by LPS were markedly inhibited by TNG. In conclusion, this study demonstrated that TNG protected against LPS-induced AKI by inhibiting TLR4/NF-κB signaling pathway.