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The role of Sidt2 in the process of glucose and lipid metabolism has been recently reported. However, whether Sidt2 is involved in the metabolic regulation in skeletal muscle remains unknown. In this study, for the first time, using skeletal muscle-selective Sidt2 knockout mice, we found that Sidt2 was vital for the quality control of mitochondria in mouse skeletal muscle. These mice showed significantly reduced muscle tolerance and structurally abnormal mitochondria. Deletion of the Sidt2 gene resulted in decreased expression of mitochondrial fusion protein 2 (Mfn2) and Dynamin-related protein 1 (Drp1), as well as peroxisome proliferator-activated receptor γ coactivator-1 (PGC1-α). In addition, the clearance of damaged mitochondria in skeletal muscle was inhibited upon Sidt2 deletion, which was caused by blockade of autophagy flow. Mechanistically, the fusion of autophagosomes and lysosomes was compromised in Sidt2 knockout skeletal muscle cells. In summary, the deletion of the Sidt2 gene not only interfered with the quality control of mitochondria, but also inhibited the clearance of mitochondria and caused the accumulation of a large number of damaged mitochondria, ultimately leading to the abnormal structure and function of skeletal muscle.
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Membrana Celular , Lisossomos , Músculo Esquelético/metabolismo , Proteínas de Transporte de Nucleotídeos/metabolismo , Animais , Autofagia/fisiologia , Linhagem Celular , Regulação da Expressão Gênica , Predisposição Genética para Doença , Camundongos , Camundongos Knockout , Microscopia Eletrônica de Transmissão , Mitocôndrias Musculares/metabolismo , Músculo Esquelético/citologia , Doenças Musculares/genética , Proteínas de Transporte de Nucleotídeos/genéticaRESUMO
The autophagy, which can be regulated by lysosomal membrane proteins, plays a critical role in maintaining normal podocyte function. TM7SF1 is a novel lysosomal membrane protein, but its effect on autophagy is still unknown. This study aimed to identify the role of TM7SF1 in mouse podocyte (MPC5) autophagy. Interestingly, we detected an increase in LC3BII and SQSTM1/P62 in MPC5 through inhibiting TM7SF1, and which can be completely corrected after blocking the autolysosome degradation with chloroquine (CQ). Moreover, inhibition of TM7SF1 expression did not increase the mRNA level of SQSTM1/P62. Theses results suggested that inhibition of TM7SF1 led to impaired degradation of autophagy products, which manifest as an abnormal accumulation of LC3BII and SQSTM1/P62. Further studies showed that the downregulation of TM7SF1 resulted in a significant decrease in the number of acid lysosomes, which directly led to decreases in the number and function of autolysosomes. In conclusion, TM7SF1 is therefore essential for autolysosomes degradation pathway at the end of autophagy flow, and for the maintenance of podocyte function.
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Autofagia , Podócitos/citologia , Podócitos/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Animais , Autofagia/efeitos dos fármacos , Linhagem Celular , Cloroquina/farmacologia , Regulação para Baixo/efeitos dos fármacos , Lisossomos/efeitos dos fármacos , Lisossomos/metabolismo , Camundongos , Proteínas Associadas aos Microtúbulos/metabolismo , Podócitos/efeitos dos fármacos , Proteína Sequestossoma-1/metabolismoRESUMO
In patients with chronic kidney disease, the abnormal activation of inflammatory pathways is usually an important factor leading to renal fibrosis and further deterioration of renal function. Finding effective intervention targets of the inflammatory signaling pathway is an important way to treat chronic kidney disease. As a newly discovered lysosomal membrane protein, the correlation between SID1 transmembrane family member 2 (Sidt2) and the inflammatory signaling pathway has not been reported. The aim of this study was to investigate the effect of Sidt2 on inflammation by inhibiting the expression of the Sidt2 gene in a mouse mesangial cell line mediated by a lentiviral CRISPR/Cas9 vector. Hematoxylin and eosin staining and microscopy found that the mesangial cells lost their normal morphology after inhibiting the expression of Sidt2, showing that the cell body became smaller, the edge between the cells was unclear, and part of the nucleus was pyknotic and fragmented, appearing blue-black. The expressions of IKK ß, p-IKK α/ß, NF-κB p65, p-NF-κB p65, p-IκBα, IκBα, and TNF-α in the NF-κB pathway of the Sidt2 -/- group were higher than those of the Sidt2 +/+ group. p-Jak2 and IL6 increased in the Jak/Stat pathway, and p-ERK and p-P38 increased in the MAPK pathway. The expressions of IKK ß, p-IKK α/ß, NF-κB p65, p-NF-κB p65, p-IκBα, IκBα, and TNF-α in the NF-κB pathway of the Sidt2 +/++LPS group were significantly higher than those in the Sidt2 +/+ group. The expressions of IKK ß, p-IKK α/ß, NF-κB p65, p-NF-κB p65, p-IκBα, IκBα, and TNF-α in the Sidt2 -/-+LPS group were higher than those in the Sidt2 -/- group. The expressions of p-IKK α/ß, NF-κB p65, p-NF-κB p65, p-IκBα, IκBα, and TNF-α in the Sidt2 -/-+LPS group were higher than those in the Sidt2 +/++LPS group. In the Jak/Stat pathway, the protein expressions of p-Jak2 and IL6 in the Sidt2 +/++LPS group were higher than those in the Sidt2 +/+ group. The expressions of p-Jak2 and IL6 in the Sidt2 -/-+LPS group were higher than those in the Sidt2 -/- group. The expressions of p-Jak2 and IL6 in the Sidt2 -/-+LPS group were higher than those in the Sidt2 +/++LPS group. The expressions of p-JNK, p-ERK, p-P38, and ERK in the MAPK pathway in the Sidt2 +/++LPS group were higher than those in the Sidt2 +/+ group. The expressions of p-JNK, p-ERK, p-P38, and ERK in the Sidt2 -/-+LPS group were higher than those in the Sidt2 -/- group. The expressions of p-JNK, p-ERK, p-P38, and ERK in the Sidt2 -/-+LPS group were higher than those in the Sidt2 +/++LPS group. These data suggested that deletion of the Sidt2 gene changed the three inflammatory signal pathways, eventually leading to the damage of glomerular mesangial cells in mice.
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Perfilação da Expressão Gênica , Inflamação/metabolismo , Células Mesangiais/metabolismo , Proteínas de Transporte de Nucleotídeos/metabolismo , Animais , Sistemas CRISPR-Cas , Citocinas/metabolismo , Regulação da Expressão Gênica , Taxa de Filtração Glomerular , Quinase I-kappa B/metabolismo , Lentivirus/genética , Lipopolissacarídeos/metabolismo , Lisossomos/metabolismo , Sistema de Sinalização das MAP Quinases , Camundongos , Camundongos Knockout , Inibidor de NF-kappaB alfa/metabolismo , Transdução de Sinais , Fator de Transcrição RelA/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Apolipoprotein M (ApoM) is involved in lipid metabolism, and especially is involved in reverse cholesterol transport. However, the relationship between ApoM and apoptosis has been rarely reported. This study aimed to investigate the effect of ApoM on apoptosis using an ApoM gene-deficient mice (ApoM-/-) model and a mouse mesangial cell model with suppressed ApoM gene expression. First, we observed by transmission electron microscopy that mitochondrial damage and endoplasmic reticulum stress were abnormally altered in the kidneys of ApoM-/- mice compared with wild-type mice, showing mitochondrial swelling, vacuolization, myeloid changes, and expansion of the rough endoplasmic reticulum. At the molecular level, the expression of pro-apoptotic related proteins such as AIF, Bax, chop, clever-caspase 3, clever-caspase 7, clever-caspase 9, and clever-caspase 12 increased, and the expression of anti-apoptotic protein Bcl-2 decreased. Secondly, by interfering with the expression of the ApoM gene in mouse mesangial cells, we found that, compared with the control group (NC-si), the cells of the experimental group (siApoM) showed decreased cell viability, nuclear chromatin condensation, nuclear lysis, and an increased proportion of early apoptotic cells. The results in cells at the molecular level were consistent with those at the tissue level. These data indicated that the deletion of the ApoM gene led to upregulation of apoptosis in mouse kidney tissues and mesangial cells through the mitochondrial and endoplasmic reticulum pathways.
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Apolipoproteínas M/fisiologia , Apoptose , Estresse do Retículo Endoplasmático , Deleção de Genes , Rim/ultraestrutura , Mitocôndrias/metabolismo , Animais , Apolipoproteínas M/genética , Proteínas Reguladoras de Apoptose/metabolismo , Células Cultivadas , Rim/citologia , Rim/metabolismo , Células Mesangiais/citologia , CamundongosRESUMO
Primary liver cancer has consistently exhibited a high prevalence and fatality rate, necessitating the investigation of associated diagnostic markers and inhibition mechanisms to effectively mitigate its impact. The significance of apolipoprotein M (ApoM) in impeding the progression of neoplastic ailments is progressively gaining recognition. However, a comprehensive understanding of its underlying mechanism in liver cancer advancement remains to be elucidated. Recent evidence indicates a potential association between ApoM and polyunsaturated fatty acids (PUFAs), with the peroxidation of phospholipids (PLs) containing PUFAs being recognized as a crucial element in the occurrence of ferroptosis. This prompts us to investigate the impact of the APOM gene on the progression of liver cancer through the ferroptosis pathway and elucidate its underlying mechanisms. The findings of this study indicate that the liver cancer cell model, which was genetically modified to overexpress the APOM gene, demonstrated a heightened ferroptosis effect. Moreover, the observed inhibition of the GSH (Glutathione) - GPX4 (Glutathione Peroxidase 4) regulatory axis suggests that the role of this axis in inhibiting ferroptosis is weakened. Through intersection screening and validation, we found that Mucin 1,cell surface associated (MUC1) can inhibit ferroptosis and is regulated by the APOM gene. Bioinformatics analysis and screening identified miR-4489 as a mediator between the two. Experimental results using the dual luciferase reporter gene confirmed that has-miR-4489 targets MUC1's 3'-UTR and inhibits its expression. In conclusion, this study provides evidence that the APOM gene induces a down-regulation in the expression of the ferroptosis-inhibiting gene MUC1, mediated by miR-4489, thereby impeding the advancement of liver cancer cells through the facilitation of ferroptosis.
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Apolipoproteínas M , Carcinoma Hepatocelular , Ferroptose , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas , MicroRNAs , Ferroptose/genética , Humanos , Apolipoproteínas M/genética , Apolipoproteínas M/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/metabolismo , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/metabolismo , Linhagem Celular Tumoral , Células Hep G2RESUMO
The falling weight deflectometer (FWD) detection system benefits from its outstanding characteristics of no damage, fast speed, and high precision. The warping deformation of cement concrete pavement occurs due to the temperature difference along the depth of the slab, which makes FWD detect different results under different temperature fields. In this study, we systematically carried out the cement pavement's temperature field and deflection test. The experimental data were analyzed to obtain the temperature variation law of the top and bottom of the pavement slab every day. By establishing a three-dimensional finite element model of cement pavement with a multi-layer elastic foundation type, the influence of the temperature difference at the bottom of the slab on the deflection of the center point of the slab corner load under different working conditions, different seasons, different loads and whether there is polymer filling in the void area was studied. We summarize the correlation between the temperature difference and the influence coefficient and propose the cement pavement void identification and polymer grouting effect evaluation method considering the temperature effect.
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As a self-degrading and highly conserved survival mechanism, autophagy plays an important role in maintaining cell survival and recycling. The discovery of autophagy-related (ATG) genes has revolutionized our understanding of autophagy. Lysosomal membrane proteins (LMPs) are important executors of lysosomal function, and increasing evidence has demonstrated their role in the induction and regulation of autophagy. In addition, the functional dysregulation of the process mediated by LMPs at all stages of autophagy is closely related to neurodegenerative diseases and cancer. Here, we review the role of LMPs in autophagy, focusing on their roles in vesicle nucleation, vesicle elongation and completion, the fusion of autophagosomes and lysosomes, and degradation, as well as their broad association with related diseases.
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Sarcopenia is a syndrome of age-related loss of muscle mass and strength that seriously affects human health, and there are currently no effective drugs to treat the disease. Linolenic acid as a common n-3 polyunsaturated fatty acid (n-3 PUFA) is known to have many beneficial functions. Some studies have found that n-3 PUFA might have the potential to improve sarcopenia. In this study, Caenorhabditis elegans (C. elegans) was used as a model animal to investigate the effects of linolenic acid on C. elegans muscles. The results showed that 50 µg mL-1 linolenic acid significantly improved sarcopenia by repairing mitochondrial function by promoting mitophagy and fighting oxidative stress (p < 0.05). This included the increase of the expression of the mitophagy gene pink-1 and DAF-16/FOXO transcription factors, respectively, by linolenic acid. This study could provide some evidence for the application of n-3 PUFA in improving sarcopenia.
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Proteínas de Caenorhabditis elegans , Ácidos Graxos Ômega-3 , Sarcopenia , Animais , Humanos , Caenorhabditis elegans/genética , Sarcopenia/tratamento farmacológico , Sarcopenia/metabolismo , Ácido alfa-Linolênico/farmacologia , Ácido alfa-Linolênico/metabolismo , Proteínas de Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/metabolismo , Mitofagia , Estresse Oxidativo , Ácidos Graxos Ômega-3/farmacologia , Ácidos Graxos Ômega-3/metabolismo , Fatores de Transcrição Forkhead/metabolismo , LongevidadeRESUMO
Polycyclic aromatic hydrocarbons (PAHs) in oil are affected by many factors, including temperature, time, and PAHs precursors. Phenolic compounds, as beneficial endogenous components of oil, are often associated with the inhibition of PAHs. However, studies have found that the presence of phenols may lead to increased levels of PAHs. Therefore, this study took Camellia oleifera (C. oleifera) oil as the research object, in order to study the effect of catechin in the formation of PAHs under different heating conditions. The results showed that PAH4 were generated rapidly during the lipid oxidation induction period. When the addition of catechin was >0.02%, more free radicals were quenched than generated, thus inhibiting the generation of PAH4. ESR, FT-IR, and other technologies were employed to prove that when the catechin addition was <0.02%, more free radicals were produced than quenched, causing lipid damage and increasing PAHs intermediates. Moreover, the catechin itself would break and polymerize to form aromatic ring compounds, ultimately leading to the conclusion that phenolic compounds in oil may be involved in the formation of PAHs. This provides suggestions for the flexible processing of phenol-rich oil to balance the retention of beneficial substances, and for the safe control of hazardous substances in real-life applications.
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Sulfonamide antibiotics in the environment not only disrupt the ecological balance but can also enter the human or animal body in various forms and cause harm. Therefore, exploring efficient methods to degrade sulfonamide antibiotics is crucial. In this study, we prepared biochar (BC) using corn straw, and TiO2/BC was obtained by doping different proportions of TiO2 into biochar with varying carbonization temperatures using the sol-gel method. Next, we investigated the degradation of sulfamethoxazole (SMX) in solution using the generated TiO2/BC under ultraviolet irradiation and studied the effects of various experimental parameters, such as the type of composite material, composite material addition, solution pH, and initial antibiotic concentration on SMX degradation. Under an initial SMX concentration of 30 mg/L, the composite with the best photocatalytic degradation performance was TiO2/BC-5-300 (i.e., 5 mL of TiO2 doping; 300 °C calcination temperature), with an addition amount of 0.02 g and a solution pH of 3. The degradation efficiency increased from 22.3% to 89%, and the most significant degradation effect occurred during the initial stage of photocatalytic degradation. In the TiO2/BC-5-300 treated SMX solution, the average rhizome length of bean sprouts was significantly higher than that of the untreated SMX solution and slightly lower than that of the deionized aqueous solution (3.05 cm < 3.85 cm < 4.05 cm). This confirmed that the photocatalytic degradation of SMX by the composite was effective and could efficiently reduce its impact on the growth of bean sprouts. This study provides essential data and theoretical support for using TiO2/BC in the treatment of antibiotic-contaminated wastewater.
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Margarine is a typical water-in-oil (W/O) emulsion fat product. Due to the presence of a water-oil interface, the oil oxidation in the emulsion system is the interface reaction, which is much faster than that in bulk oil and shows different oxidation mechanisms. The analysis of Rancimat and electron spin resonance indicated that α-tocopherol and EGCG show synergistic antioxidant effects in the margarine. After 20 days of accelerated oxidation storage, the antioxidant effect of the compound antioxidant (50 mg/kg α-tocopherol + 350 mg/kg EGCG) on the margarine was significantly higher than that of the single antioxidant α-tocopherol and EGCG. Based on the results of antioxidants partitioning, electrochemistry, fluorescence spectroscopy, and the oxidative decomposition of antioxidants, the possible mechanisms of interaction were the promotion of α-tocopherol regeneration by EGCG, and the fact that α-tocopherol and EGCG could act at different stages and positions of oxidation. This work will contribute to studying antioxidant interactions and can provide valuable suggestions for practical production. PRACTICAL APPLICATION: This study aims to improve the oxidative stability of margarine by adding α-tocopherol and epigallocatechin-gallate (EGCG) individually and in blends. The mechanism of compound antioxidant synergistic inhibition of margarine oxidation was analyzed, providing theoretical basis and scientific basis for the research and practical application of natural antioxidant synergistic mechanism.
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Antioxidantes , Catequina , Antioxidantes/farmacologia , Antioxidantes/química , alfa-Tocoferol/química , Margarina , Emulsões/química , Oxirredução , Catequina/química , Água , Estresse OxidativoRESUMO
Apolipoprotein M (ApoM) is considered a protective factor that inhibits the occurrence and development of liver cancer, but the specific underlying mechanisms require further investigation. Previous studies have demonstrated that ApoM gene knockout promotes the expression of the transcription factor sterol regulatory element-binding protein 1 (SREBP1; also known as SREBF1) in the livers of mice. SREBF1 is closely associated with factors involved in fatty acid synthesis and has a role in the promotion of tumor progression. The present study initially confirmed that the expression levels of ApoM in cancer tissues were significantly decreased compared with those in normal tissue, while the expression levels of SREBF1 were significantly increased. In addition, ApoM gene knockout significantly increased the expression levels of SREBF1 and the key glycolytic enzyme ATP-dependent 6-phosphofructokinase, liver type (PFKL). Binding site prediction and a dual-luciferase reporter gene assay indicated that SREBF1 regulates the promoter region of PFKL. To the best of our knowledge, the present study was the first to propose the regulation of glycolytic enzyme transcription levels by SREBF1. Furthermore, cell proliferation and Transwell assays demonstrated that ApoM gene knockout increased the expression levels of SREBF1 and further enhanced the activity of the promoter region of PFKL, ultimately promoting the proliferation, migration and invasion of liver cancer cells.
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Disintegrin and metalloproteinase 12 (ADAM12) is thought to trigger the occurrence and development of numerous tumours, including colorectal, breast, and pancreatic cancers. On the basis of The Cancer Genome Atlas (TCGA) datasets, in this study, the relationship between ADAM12 gene expression and hepatocellular carcinoma (HCC), the prognostic value of this relationship, and the potential mechanisms influencing HCC development were evaluated. The results showed that the ADAM12 gene was significantly and highly expressed in liver cancer tissue. The high expression of the ADAM12 gene in liver cancer tissue significantly and positively correlated with T stage, pathological stage, and residual tumour. Kaplan-Meier and Cox regression analyses revealed that ADAM12 gene expression is an independent risk factor influencing the prognosis of patients with liver cancer. Pathway analyses of ADAM12 in HCC revealed ADAM12-correlated signalling pathways, and the expression level of ADAM12 was associated with immune cell infiltration. In vitro experiments demonstrated that the expression level of ADAM12 in Huh-7 and Hep3B cells was significantly higher than that in other HCC cells. ShRNA transfection experiments confirmed that the expression levels of TGF-ß and Notch pathway-related proteins were significantly decreased. An EdU cell proliferation assay showed that a low level of ADAM12 gene expression significantly inhibited the proliferative activity of HCC cells. Cell cycle experiments showed that low ADAM12 expression blocked the G1/S phase transition. Overall, this research revealed that high ADAM12 gene expression implies a poor prognosis for patients with primary liver cancer. In addition, it is a potential indicator for the diagnosis of liver cancer.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , Proteína ADAM12/genética , Biomarcadores Tumorais/genética , Carcinoma Hepatocelular/diagnóstico , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Ciclo Celular , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Hepáticas/patologia , PrognósticoRESUMO
At present, there is no noninvasive biomarker of renal fibrosis. The potential diagnostic value of urinary exosome-derived circRNAs from glomerular disease patients for renal fibrosis is still uncertain. Here, we first detected the expression of hsa_circ_0008925 in TGF-ß1-cultured HK-2 cell-derived exosomes. Secondly, we collected urine samples from 95 biopsy-proven glomerular disease patients and 34 healthy controls. The expression of hsa_circ_0008925 was analyzed, and the correlation with renal function and pathological changes was calculated. The receiver operating characteristic (ROC) curve for the diagnosis of renal fibrosis was performed. The results showed that in exosomes derived from TGF-ß1-cultured HK-2 cells, the expression of hsa_circ_0008925 was increased compared with normal cultured. Further, the expression level of hsa_circ_0008925 was increased in urinary exosomes from renal fibrosis patients and correlated with serum creatinine, blood urea nitrogen (BUN), estimated glomerular filtration rate, and cystatin C. The level of hsa_circ_0008925 was furthermore correlated with the score of tubulointerstitial fibrosis (TIF) and the score of glomerular sclerosis. The ROC curve showed that hsa_circ_0008925 can diagnose renal fibrosis at a cut-off value of 0.093 with a sensitivity of 52.2% and specificity of 96.4%. In summary, we indicated that urinary exosomal hsa_circ_0008925 could be acted as a noninvasive biomarker for renal fibrosis in glomerular diseases patients.
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Exossomos/metabolismo , Nefropatias/diagnóstico , Nefropatias/patologia , Glomérulos Renais/patologia , RNA Circular/urina , Adulto , Biomarcadores/urina , Células Cultivadas , Doença Crônica , Exossomos/genética , Feminino , Fibrose , Taxa de Filtração Glomerular , Humanos , Nefropatias/urina , Masculino , Pessoa de Meia-Idade , Fator de Crescimento Transformador beta1RESUMO
BACKGROUND: Chronic renal disease (CKD) is a common and irreversible loss of renal function. Renal fibrosis reflected the degree of renal dysfunction. However, the current biomarkers only characterize the renal function instead of indicating the fibrosis degree. The potential diagnostic value of urinary exosomes derived circRNAs for renal fibrosis needs to be further studied. METHODS: Urine exosomes from 3 chronic kidney disease (CKD) patients without renal fibrosis and 3 renal fibrotic patients were collected and human circRNAs microarray analysis were performed to detect the circRNAs expression profile. 110 biopsy-proven CKD patients and 54 healthy controls were enrolled and urine exosomes derived RNA was isolated. The expression of hsa_circ_0036649 was measured and the correlation with renal function parameter and pathological indicators was performed. The receiver operating characteristic (ROC) curve for the diagnosis of renal fibrosis was calculated. RESULTS: Human circRNAs microarray showed 365 circRNAs up expressed and 195 circRNAs down expressed in renal fibrotic patients compared to none fibrosis CKD patients. The expression of hsa_circ_0036649 was decreased in renal fibrotic patients according to RT-PCR and correlated with serum creatinine, blood urea nitrogen (BUN), estimated glomerular filtration rate and cystatin c. Further, the expression of hsa_circ_0036649 was correlated with the score of tubulointerstitial fibrosis (TIF) and the score of glomerular sclerosis. The ROC curve showed that hsa_circ_0036649 may predict renal fibrosis at a cut-off value of 0.597 with a sensitivity of 45.5% and specificity of 87.9%. CONCLUSION: Expression of urinary exosomes derived hsa_circ_0036649 associated with the degree of renal fibrosis. Its potential role as a biomarker in CKD remained to be supported by further follow-up studies.Key MessagescircRNAs profile in urine exosomes in renal fibrosis patients was revealed.The expression of urine exosomes derived hsa_circ_0036649 was correlated to renal function and fibrosis degree.circRNAs derived from urinary exosomes may become a new research direction for biomarkers of renal fibrosis.
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Exossomos , Insuficiência Renal Crônica , Biomarcadores/metabolismo , Exossomos/genética , Exossomos/metabolismo , Fibrose , Humanos , Rim/patologia , RNA Circular , Insuficiência Renal Crônica/diagnóstico , Insuficiência Renal Crônica/genética , Insuficiência Renal Crônica/patologiaRESUMO
BACKGROUND: Preeclampsia (PE) is a complex pregnancy-related disease that endangers the safety of maternal and fetal. The purpose of this study is to reveal the pathogenesis of preeclampsia and discover new predictors from the perspective of peptidomics. The umbilical cord blood of PE and control group was analyzed by peptidomics. A peptide named Regulation of Proliferation Process in Preeclampsia (ROPPIP) was screened out to explore its role in the proliferation, migration and apoptosis of trophoblast cells in preeclampsia. METHODS: We compared and analyzed the umbilical cord blood of patients with PE and normal pregnant women using liquid chromatography-tandem mass spectrometry (LC-MS). hTR-8/Svneo cells cultured in vitro were divided into ROPPIP group and a disordered peptide group as control. Cell Counting Kit-8 (CCK-8) assay, flow cytometry, Transwell chamber assays and western blot analysis were performed to detect cell proliferation, invasion, migration and apoptosis, in addition to the expression of Matrix metalloproteinase-2 (MMP2), nuclear associated antigen Ki67, B-cell lymphoma-2 (Bcl2), Caspase 3, and ß-actin protein. RESULTS: We identified 133 differential peptides. Of these, 51 were up-regulated while 82 were down-regulated. the polypeptide SFGVRMATASPTDGNV with low differential expression in the serum of PE patients was selected for the study, we named the polypeptide as Regulation of Proliferation Process in PE (ROPPIP). The experiment shows that ROPPIP can up-regulate the expression levels of MMP2, Ki67, and Bcl2 in HTR-8/Svneo cells, down-regulate the expression of caspase-3, promote the proliferation and migration of HTR-8/Svneo cells and inhibit the apoptosis induced by cisplatin, the activation of the phosphatidylinositol-3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) signaling pathway may be associated with the function of ROPPIP. CONCLUSIONS: ROPPIP promotes HTR-8/Svneo cells migration and proliferation, and inhibits apoptosis, by regulating the activation of the PI3K/AKT/mTOR signaling pathway.
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In recent years, abnormal liver lipid metabolism has emerged as one of the important pathogenesis pathways of primary liver cancer. It is highly important to identify the mechanisms to explore potential prevention and treatment targets. Apolipoprotein M is specifically expressed in the liver and participates in liver lipid metabolism, but the evidence that ApoM affects primary liver cancer is insufficient. The Cancer Genome Atlas (TCGA) database and clinical case analysis, as well as animal level and cell level analysis suggest that the expression level of ApoM gene in cancer tissues is lower than that in paracarcinoma tissues. Further experimental research found that the deletion of ApoM significantly increased the proliferation of mouse liver cancer cells (Hepa1-6) and inhibited the level of apoptosis induced by cisplatin. In addition, mouse liver cancer cells lacking ApoM showed stronger migration and invasion capabilities in transwell experiments. In contrast, overexpression of ApoM in Hepa1-6 cells and Huh-7 cells showed an inhibition of proliferation, up-regulation apoptosis and reduced migration and invasion. In vivo, the deletion of the ApoM accelerated tumorigenesis in nude mice and allowed the mice to develop liver tumor mutations more quickly under the induction of N-nitrosodiethylamine and the survival time of mice was shorter than that control. Therefore, ApoM may be a potential protective factor to inhibit the occurrence and development of primary liver cancer.
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The aim of this study was to develop and evaluate a triptolide phospholipid complex (TPCX) for the treatment of rheumatoid arthritis (RA) by transdermal delivery. TPCX was prepared and characterized by differential scanning calorimetry (DSC), Fourier-transform infrared spectroscopy (FTIR) analysis, transmission electron microscope (TEM), and scanning electron microscope (SEM). The solubility of TPCX was determined. Then, a TPCX cream was prepared to evaluate its percutaneous permeability and the antiarthritis effect. The transdermal permeability was determined using the Franz method, and a microdialysis system was used for skin pharmacokinetic study. A rat model of RA was prepared to evaluate the pharmacological effects. TPCX increased the solubility of triptolide in water, and the percutaneous permeability of TPCX cream was greatly enhanced compared with triptolide cream. The skin pharmacokinetic study indicated that TPCX cream has a longer biological half-life (t1/2) and mean residence time (MRT), but it has a shorter Tmax than that of triptolide cream in vivo. The area under the curve (AUC0-t)/AUC0-∞) and the peak concentration (Cmax) of TPCX cream were obviously higher than those of triptolide cream. The TPCX-loaded cream alleviated paw swelling and slowed down the progression of arthritis by inhibiting the inflammatory response by down regulating the TNF-α, IL-1ß, and IL-6 levels, thus exhibiting excellent antiarthritic effects. In summary, the prepared TPCX effectively increases the hydrophilicity of triptolide, which is good for its percutaneous absorption and enhances its effect on RA rats. TPCX can be a good candidate for the transdermal delivery to treat RA.
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Artrite Reumatoide/tratamento farmacológico , Diterpenos/farmacologia , Imunossupressores/farmacologia , Fenantrenos/farmacologia , Fosfolipídeos/química , Administração Cutânea , Animais , Área Sob a Curva , Química Farmacêutica , Diterpenos/administração & dosagem , Diterpenos/farmacocinética , Relação Dose-Resposta a Droga , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Compostos de Epóxi/administração & dosagem , Compostos de Epóxi/farmacocinética , Compostos de Epóxi/farmacologia , Meia-Vida , Imunossupressores/administração & dosagem , Imunossupressores/farmacocinética , Mediadores da Inflamação/metabolismo , Masculino , Fenantrenos/administração & dosagem , Fenantrenos/farmacocinética , Ratos , Ratos WistarRESUMO
Introduction. Chlamydia psittaci is an important cause of community-acquired pneumonia (CAP). The spectrum of CAP due to Chlamydia psittaci ranges from mild, self-limited to acute respiratory failure and the early identification of this disease can be challenging. Metagenomic next-generation sequencing (mNGS) of bronchoalveolar lavage fluid has the potential to improve the pathogen identification in severe CAP.Hypothesis/Gap Statement. Metagenomic next-generation sequencing (mNGS) of bronchoalveolar lavage fluid has the potential to rapidly identify pathogens in severe CAP. The early identification and appropriate use of antibiotics can improve the prognosis of severe CAP caused by Chlamydia psittaci.Aim. The aim of the study is to describe the clinical spectrum of severe psittacosis pneumonia to provide a better understanding of this disease and to demonstrate that mNGS is an effective method for pathogen detection.Methodology. Retrospective case analysis from November 2019 to November 2020 was performed. Sixteen cases of severe psittacosis pneumonia were diagnosed through mNGS. Clinical features, laboratory findings, imaging features, treatment and outcome were summarized.Results. Frequent symptoms included fever (16/16, 100%), dyspnoea (16/16, 100%), cough (12/16, 75%), sputum (11/16, 69%) and headache (9/16, 56%). The median leukocytosis was within the normal range, while C-reactive proteins, CK, LDH, AST, D-Dimer were significantly elevated. The feature of computed tomography included ground-glass opacity with consolidation and multiple lobar distributions. The total number of sequences of Chlamydia psittaci identified from bronchoalveolar lavage by mNGS varied from 58 to 57115. Five patients underwent noninvasive mechanical ventilation, four patients underwent high flow humidified oxygen therapy and one patient underwent invasive mechanical ventilation. Two patients had septic shock needing vasoactive medications. All of the sixteen patients experienced full recoveries.Conclusion. The symptoms of severe CAP caused by Chlamydia psittaci were not typical while laboratory results may have some clues. The mNGS technology can early detect of psittacosis, reduce unnecessary use of antibiotics and short the course of the disease.
Assuntos
Chlamydophila psittaci , Pneumonia Bacteriana , Psitacose , China/epidemiologia , Chlamydophila psittaci/genética , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Metagenômica , Pneumonia Bacteriana/epidemiologia , Psitacose/diagnóstico , Estudos RetrospectivosRESUMO
AIM: The metastasis of breast cancer is an important cause of tumor recurrence. This study highlights that tyrosine kinase inhibitors dasatinib (DAS) and rosiglitazone (ROZ) inhibit tumor growth and reduce the occurrence of tumor cell metastasis. Due to the poor water solubility, short half-time in the body of DAS and ROZ, which increases the difficulty of tumor treatment, as well as the demand for nano-drug delivery systems for organ-specific therapies. METHODS: Hyaluronic acid (HA) and DAS are bonded by a pH-sensitive ester bond to form an HA-DAS polymer. Then, ROZ was added as the core, D-A-tocopherol polydiethylene glycol isosuccinate (TPGS) and HA-DAS were used as carriers to form HA-DAS and TPGS mixed micelle system loaded with ROZ (THDR-NPs). The size and structure of THDR-NPs were characterized, the drug release, stability and biosafety of THDR-NPs were studied. In vitro, the cytotoxicity, targeting effect and tumor metastasis inhibition of THDR-NPs were evaluated in human breast cancer cell lines. In addition, the selective potency of designed THDR-NPs in depleting was further verified in vivo in the tumor-bearing nude mice model. RESULTS: The designed THDR-NPs have a particle size of less than 100 nm, good stability, biological safety and sustained release, and showed strong therapeutic effects on breast cancer models in vitro and in vivo. Moreover, it has been proved that THDR-NPs have the ability to inhibit tumor metastasis. CONCLUSION: DAS and ROZ were designed into micelles, the efficacy of THDR-NPs was higher than that of free drugs. These results indicate that nanoparticles have a good application prospect in the treatment of tumor metastasis.