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1.
Cell Mol Life Sci ; 81(1): 114, 2024 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-38436813

RESUMO

Hyperuricemia is an independent risk factor for chronic kidney disease (CKD) and promotes renal fibrosis, but the underlying mechanism remains largely unknown. Unresolved inflammation is strongly associated with renal fibrosis and is a well-known significant contributor to the progression of CKD, including hyperuricemia nephropathy. In the current study, we elucidated the impact of Caspase-11/Gasdermin D (GSDMD)-dependent neutrophil extracellular traps (NETs) on progressive hyperuricemic nephropathy. We found that the Caspase-11/GSDMD signaling were markedly activated in the kidneys of hyperuricemic nephropathy. Deletion of Gsdmd or Caspase-11 protects against the progression of hyperuricemic nephropathy by reducing kidney inflammation, proinflammatory and profibrogenic factors expression, NETs generation, α-smooth muscle actin expression, and fibrosis. Furthermore, specific deletion of Gsdmd or Caspase-11 in hematopoietic cells showed a protective effect on renal fibrosis in hyperuricemic nephropathy. Additionally, in vitro studies unveiled the capability of uric acid in inducing Caspase-11/GSDMD-dependent NETs formation, consequently enhancing α-smooth muscle actin production in macrophages. In summary, this study demonstrated the contributory role of Caspase-11/GSDMD in the progression of hyperuricemic nephropathy by promoting NETs formation, which may shed new light on the therapeutic approach to treating and reversing hyperuricemic nephropathy.


Assuntos
Armadilhas Extracelulares , Hiperuricemia , Insuficiência Renal Crônica , Humanos , Hiperuricemia/complicações , Actinas , Ácido Úrico , Caspases , Inflamação , Fibrose , Gasderminas , Proteínas de Ligação a Fosfato
2.
Nucleic Acids Res ; 51(21): e108, 2023 Nov 27.
Artigo em Inglês | MEDLINE | ID: mdl-37870443

RESUMO

DNA methylation is essential for a wide variety of biological processes, yet the development of a highly efficient and robust technology remains a challenge for routine single-cell analysis. We developed a multiplex scalable single-cell reduced representation bisulfite sequencing (msRRBS) technology. It allows cell-specific barcoded DNA fragments of individual cells to be pooled before bisulfite conversion, free of enzymatic modification or physical capture of the DNA ends, and achieves read mapping rates of 62.5 ± 3.9%, covering 60.0 ± 1.4% of CpG islands and 71.6 ± 1.6% of promoters in K562 cells. Its reproducibility is shown in duplicates of bulk cells with close to perfect correlation (R = 0.97-0.99). At a low 1 Mb of clean reads, msRRBS provides highly consistent coverage of CpG islands and promoters, outperforming the conventional methods with orders of magnitude reduction in cost. Here, we use this method to characterize the distinct methylation patterns and cellular heterogeneity of six cell lines, plus leukemia and hepatocellular carcinoma models. Taking 4 h of hands-on time, msRRBS offers a unique, highly efficient approach for dissecting methylation heterogeneity in a variety of multicellular systems.


Assuntos
Metilação de DNA , DNA , Humanos , Ilhas de CpG/genética , Metilação de DNA/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Células K562 , Reprodutibilidade dos Testes , Análise de Sequência de DNA/métodos , Linhagem Celular Tumoral
3.
Artigo em Inglês | MEDLINE | ID: mdl-38244230

RESUMO

BACKGROUND AND HYPOTHESIS: Acute kidney injury (AKI) could progress to chronic kidney disease (CKD) and the AKI-CKD transition has major clinical significance. A growing body of evidence has unveiled the role of pyroptosis in kidney injury. We postulate that GSDMD and GSDME exert cumulative effects on the AKI-CKD transition by modulating different cellular responses. METHODS: We established an AKI-CKD transition model induced by folic acid in wildtype (WT), Gsdmd-/-, Gsdme-/-, and Gsdmd-/-Gsdme-/- mice. Tubular injury, renal fibrosis and inflammatory responses were evaluated. In vitro studies were conducted to investigate the interplay among tubular cells, neutrophils, and macrophages. RESULTS: Double deletion of Gsdmd and Gsdme conferred heightened protection against AKI, mitigating inflammatory responses, including the formation of neutrophil extracellular traps (NETs), macrophage polarization and differentiation, and ultimately renal fibrosis, compared with wildtype mice and mice with single deletion of either Gsdmd or Gsdme. Gsdme, but not Gsdmd deficiency, shielded tubular cells from pyroptosis. GSDME-dependent tubular cell death stimulated NETs formation and prompted macrophage polarization towards a pro-inflammatory phenotype. Gsdmd deficiency suppressed NETs formation and subsequently hindered NETs-induced macrophage-to-myofibroblast transition (MMT). CONCLUSION: GSDMD and GSDME collaborate to contribute to AKI and subsequent renal fibrosis induced by folic acid. Synchronous inhibition of GSDMD and GSDME could be an innovative therapeutic strategy for mitigating the AKI-CKD transition.

4.
J Intensive Care Med ; 39(5): 465-476, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-37964547

RESUMO

BACKGROUND: Sepsis-associated acute kidney injury (SA-AKI) is a critical condition with significant clinical implications, yet there is a need for a predictive model that can reliably assess the risk of its development. This study is undertaken to bridge a gap in healthcare by creating a predictive model for SA-AKI with the goal of empowering healthcare providers with a tool that can revolutionize patient care and ultimately lead to improved outcomes. METHODS: A cohort of 615 patients afflicted with sepsis, who were admitted to the intensive care unit, underwent random stratification into 2 groups: a training set (n = 435) and a validation set (n = 180). Subsequently, a multivariate logistic regression model, imbued with nonzero coefficients via LASSO regression, was meticulously devised for the prognostication of SA-AKI. This model was thoughtfully rendered in the form of a nomogram. The salience of individual risk factors was assessed and ranked employing Shapley Additive Interpretation (SHAP). Recursive partition analysis was performed to stratify the risk of patients with sepsis. RESULTS: Among the panoply of clinical variables examined, hypertension, diabetes mellitus, C-reactive protein, procalcitonin (PCT), activated partial thromboplastin time, and platelet count emerged as robust and independent determinants of SA-AKI. The receiver operating characteristic curve analysis for SA-AKI risk discrimination in both the training set and validation set yielded an area under the curve estimates of 0.843 (95% CI: 0.805 to 0.882) and 0.834 (95% CI: 0.775 to 0.893), respectively. Notably, PCT exhibited the most conspicuous influence on the model's predictive capacity. Furthermore, statistically significant disparities were observed in the incidence of SA-AKI and the 28-day survival rate across high-risk, medium-risk, and low-risk cohorts (P < .05). CONCLUSION: The composite predictive model, amalgamating the quintet of SA-AKI predictors, holds significant promise in facilitating the identification of high-risk patient subsets.


Assuntos
Injúria Renal Aguda , Sepse , Humanos , Curva ROC , Unidades de Terapia Intensiva , Modelos Logísticos , Pró-Calcitonina , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/epidemiologia , Sepse/complicações , Sepse/epidemiologia , Estudos Retrospectivos
5.
Ren Fail ; 45(1): 2194451, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36994793

RESUMO

AIM: Idiopathic membranous nephropathy (IMN) is a common type of nephrotic syndrome, and is associated with acute kidney injury (AKI). We investigated the association of multiple variables with AKI in patients with IMN. METHODS: The data of 187 patients with biopsy-proven IMN were examined. Renal outcome was defined as progression to end-stage renal disease (ESRD). Binary logistic regression and Kaplan-Meier's analysis were used for statistical analysis. RESULTS: During follow-up, 46 (24.6%) patients developed AKI. The incidence of AKI was greater in males than females (p < .01). The AKI group had higher uric acid, lower serum PLA2R antibody positive, and worse baseline kidney function (all p < .01). Most patients in the AKI group had stage I (71.74%) or stage II (21.74%). The AKI group had higher renal tubular injury score and chronicity index (both p < .05). Binary logistic regression indicated that uric acid and baseline estimated glomerular filtration rate (eGFR) were independent risk factors for AKI in patients with IMN (p < .05). The optimal cutoff value of serum uric acid for predicting AKI was 402.50 µmol/L and the baseline eGFR was 96.83 mL/min/1.73 m2. Kaplan-Meier's analysis showed that the cumulative renal survival rate was lower in the AKI group (p = .047). CONCLUSIONS: AKI increases the risk of poor prognosis in IMN patients and the high uric acid and low baseline eGFR were considered independent predictors for developing AKI in patients with IMN.


Assuntos
Injúria Renal Aguda , Glomerulonefrite Membranosa , Masculino , Feminino , Humanos , Glomerulonefrite Membranosa/complicações , Ácido Úrico , Rim , Prognóstico , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/complicações , Estudos Retrospectivos
6.
RNA Biol ; 18(sup1): 232-243, 2021 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-34586036

RESUMO

RNA-seq has been widely used to reveal the molecular mechanism of variants of life process. We have developed an alternative method, MustSeq, which generates multiple second strands along a single 1st strand cDNA by random-priming initiation, immediately after reverse transcription for each RNA extract using sample-barcoded poly-dT primers, then 3' ends-enriching PCR is applied to construct the library. Unlike the conventional RNA seq, MustSeq avoids procedures such as mRNA isolation, fragmentation and RNA 5'-end capture, enables early pooling of multiple samples, and requires only one twentieth of sequencing reads of full-length sequencing. We demonstrate the power and features of MustSeq comparing with TruSeq and NEBNext RNA-seq, two conventional full-length methods and QuantSeq, an industrial 3' end method. In cancer cell lines, the reads distribution of CDS-exon as well as genes, lncRNAs and GO terms detected by MustSeq are closer than QuantSeq to TruSeq. In mouse hepatocarcinoma and healthy livers, MustSeq enriches the same pathways as by NEBNext, and reveals the molecular profile of carcinogenesis. Overall MustSeq is a robust and accurate RNA-seq method allowing efficient library construction, sequencing and analysis, particularly valuable for analysis of differentially expressed genes with a large number of samples. MustSeq will greatly accelerate the application of bulk RNA-seq on different fields, and potentially applicable for single cell RNA-seq.


Assuntos
Regiões 3' não Traduzidas/genética , RNA Mensageiro/genética , RNA-Seq/métodos , Análise de Sequência de RNA/métodos , Transcriptoma , Animais , Biblioteca Gênica , Células HeLa , Humanos , Células Jurkat , Células K562 , Camundongos , Camundongos Endogâmicos C57BL , RNA Mensageiro/análise
7.
J Neurosci Res ; 98(6): 1198-1212, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32291804

RESUMO

Microglial polarization to the anti-inflammatory M2 phenotype is essential in resolving neuroinflammation, making it a promising therapeutic strategy for stroke intervention. The actin cytoskeleton is known to be important for the physiological functions of microglia, including migration and phagocytosis. Profilin 1 (PFN1), an actin-binding protein, is involved in the dynamic transformation and reorganization of actin. However, the role of PFN1 in microglial polarization and ischemia/reperfusion injury is unclear. The role of PFN1 on microglial polarization was examined in vitro in BV2 microglial cells subjected to oxygen-glucose deprivation/reoxygenation (OGDR) and in vivo in male mice after transient middle cerebral artery occlusion (MCAO). Knockdown of PFN1 inhibited M1 microglial polarization and promoted M2 microglia polarization 48 hr after OGDR stimulation in BV2 cells and 7 days after MCAO-induced injury in male mice. RhoA/ROCK pathway was involved in the regulation of PFN1 during microglial polarization. Knockdown of PFN1 also significantly attenuated brain infarcts and edema, improved cerebral blood flow and neurological deficits in MCAO-injured mice. Inhibition of PFN1 effectively protected the brain against ischemia/reperfusion injuries by promoting M2 microglial polarization in vitro and in vivo.


Assuntos
Isquemia Encefálica/metabolismo , Polaridade Celular/fisiologia , Microglia/metabolismo , Profilinas/metabolismo , Quinases Associadas a rho/metabolismo , Proteína rhoA de Ligação ao GTP/metabolismo , Animais , Isquemia Encefálica/genética , Técnicas de Silenciamento de Genes , Masculino , Camundongos , Profilinas/genética , Transdução de Sinais/fisiologia , Quinases Associadas a rho/genética , Proteína rhoA de Ligação ao GTP/genética
8.
Clin Nephrol ; 84(5): 262-9, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26396095

RESUMO

OBJECTIVE: To evaluate the efficacy and safety of calcitriol in the prevention and treatment of glucocorticoid-induced osteoporosis. METHODS: 66 patients treated with glucocorticoids (GC) for primary nephrotic syndrome (NS) were randomly assigned to 3 groups. Groups were designated as follows: calcitriol alone (n = 22), calcitriol plus calcium carbonate (n = 23), or calcium carbonate alone (n = 21). Serum markers of bone metabolism and bone mineral density (BMD) were tested at 3 different time points: the initiation of GC treatment (baseline), 12 weeks, and 24 weeks after the initiation of treatment. RESULTS: Levels of serum 25-hydroxy vitamin D, serum osteocalcin, and total serum collagen type N-terminal extension of the peptide were significantly decreased following GC therapy (p < 0.05). ß-collagen serum-specific sequences were significantly increased following GC therapy. The above-mentioned changes were less dramatic in patients treated with calcitriol, although the differences were significant (p < 0.05). Changes in serum levels of calcium, phosphorus, alkaline phosphatase, and parathyroid hormone (PTH) were not significant. 24 weeks after the initiation of treatment, BMD of the lumbar spine and femoral bone significantly decreased in all of 3 groups. However, patients who received calcitriol had significantly higher BMD of the lumbar spine than patients who received calcium carbonate alone (calcitriol plus calcium carbonate vs. calcium carbonate alone: 0.82 ± 0.19 g/cm2 vs. 0.62 ± 0.23 g/cm2 p < 0.05; calcitriol vs. calcium carbonate alone 0.805 ± 0.203 g/cm2 vs. 0.615 ± 0.225 g/cm2 p < 0.05), respectively. No serious adverse events were observed. CONCLUSION: Calcitriol may be more effective than calcium carbonate in preventing and treating GC-induced osteoporosis in patients with NS.


Assuntos
Calcitriol/uso terapêutico , Glucocorticoides/efeitos adversos , Síndrome Nefrótica/tratamento farmacológico , Osteoporose/tratamento farmacológico , Adulto , Fosfatase Alcalina/sangue , Densidade Óssea/efeitos dos fármacos , Calcitriol/efeitos adversos , Carbonato de Cálcio/uso terapêutico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Osteocalcina/sangue , Osteoporose/induzido quimicamente , Hormônio Paratireóideo/sangue , Vitamina D/análogos & derivados , Vitamina D/sangue
9.
PeerJ ; 12: e17266, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38650643

RESUMO

Objective: Whether serum uric acid (SUA) at baseline could been identiûed as a risk factor for progression in IgA nephropathy (IgAN) patients remains unclear, therefore, long- term SUA control levels must be monitored. We aimed to investigate the relevant factors affecting time-averaged SUA (TA-SUA) and to assess the prognostic value of TA-SUA in IgAN. Methods: This retrospective study included 152 patients with IgAN. The relationships between TA-SUA and clinicopathological features and renal outcomes (defined as the doubling of the baseline serum creatinine level or end-stage renal disease) were analyzed in groups divided by quartiles of TA-SUA levels, the presence of hyperuricemia, and sex. Results: Patients with high TA-SUA levels had higher levels of baseline SUA, blood urea nitrogen (BUN), triglycerides, serum C3 and serum C4 and were more likely to be male and have hypertension, proteinuria, poor renal function, and pathological injuries including high grades of tubular atrophy/interstitial fibrosis (T1-T2). These patients had a poorer prognosis compared with patients with low TA-SUA levels. The TA-SUA level was positively correlated with baseline age and BUN, triglycerides, serum C3, and serum C4 levels, and negatively correlated with baseline eGFR. Survival curve analysis indicated that persistent hyperuricemia was associated with significantly poorer renal outcomes than normo-uricemia in both men and women. The TA-SUA level also was an independent predictor of renal outcome in patients with IgAN, with optimal cutoû values of 451.38 µmol/L (area under the curve (AUC) = 0.934) for men and 492.83 µmol/L (AUC = 0.768) for women. Conclusions: The TA-SUA level is associated with triglyceride level, complement component levels, renal function, and pathological severity of IgAN, and it may be a prognostic indicator in male and female patients with IgAN.


Assuntos
Glomerulonefrite por IGA , Ácido Úrico , Humanos , Glomerulonefrite por IGA/sangue , Glomerulonefrite por IGA/patologia , Masculino , Feminino , Ácido Úrico/sangue , Estudos Retrospectivos , Adulto , Prognóstico , Hiperuricemia/sangue , Pessoa de Meia-Idade , Progressão da Doença , Fatores de Risco , Falência Renal Crônica/sangue
10.
Front Immunol ; 13: 950076, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36052061

RESUMO

Renal injury secondary to COVID-19 is an important factor for the poor prognosis of COVID-19 patients. The pathogenesis of renal injury caused by aberrant immune inflammatory of COVID-19 remains unclear. In this study, a total of 166 samples from 4 peripheral blood transcriptomic datasets of COVID-19 patients were integrated. By using the weighted gene co-expression network (WGCNA) algorithm, we identified key genes for mild, moderate, and severe COVID-19. Subsequently, taking these genes as input genes, we performed Short Time-series Expression Miner (STEM) analysis in a time consecutive ischemia-reperfusion injury (IRI) -kidney dataset to identify genes associated with renal injury in COVID-19. The results showed that only in severe COVID-19 there exist a small group of genes associated with the progression of renal injury. Gene enrichment analysis revealed that these genes are involved in extensive immune inflammation and cell death-related pathways. A further protein-protein interaction (PPI) network analysis screened 15 PPI-hub genes: ALOX5, CD38, GSF3R, LGR, RPR1, HCK, ITGAX, LYN, MAPK3, NCF4, SELP, SPI1, WAS, TLR2 and TLR4. Single-cell sequencing analysis indicated that PPI-hub genes were mainly distributed in neutrophils, macrophages, and dendritic cells. Intercellular ligand-receptor analysis characterized the activated ligand-receptors between these immune cells and parenchyma cells in depth. And KEGG enrichment analysis revealed that viral protein interaction with cytokine and cytokine receptor, necroptosis, and Toll-like receptor signaling pathway may be potentially essential for immune cell infiltration leading to COVID-19 renal injury. Finally, we validated the expression pattern of PPI-hub genes in an independent data set by random forest. In addition, we found that the high expression of these genes was correlated with a low glomerular filtration rate. Including them as risk genes in lasso regression, we constructed a Nomogram model for predicting severe COVID-19. In conclusion, our study explores the pathogenesis of renal injury promoted by immunoinflammatory in severe COVID-19 and extends the clinical utility of its key genes.


Assuntos
COVID-19 , Biologia Computacional , Biomarcadores Tumorais/genética , COVID-19/genética , Biologia Computacional/métodos , Humanos , Rim/patologia , Ligantes
11.
J Clin Med ; 11(22)2022 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-36431262

RESUMO

We aim to investigate the association of time-averaged hematuria (TA-hematuria) with the progression of IgA nephropathy (IgAN). Based on TA-hematuria during follow-up, 152 patients with IgAN were divided into a hematuria remission group (≤28 red blood cells [RBCs]/µL) and a persistent hematuria group (>28 RBCs/µL). The persistent hematuria group had a higher percentage of patients with macroscopic hematuria, lower levels of hemoglobin and TA-serum albumin, and more severe renal pathologic lesions. The composite endpoint is defined as a doubling of the baseline SCr level (D-SCr), or the presence of ESRD. During the mean follow-up of 58.08 ± 23.51 months, 15 patients (9.9%) reached the primary outcome of ESRD and 19 patients (12.5%) reached the combined renal endpoint. Kaplan-Meier analysis showed that the persistent hematuria group had a lower renal survival rate. The persistent hematuria patients who were incorporated with proteinuria (≥1.0 g/day) and low TA-serum albumin (<40 g/L) had the worst renal outcomes. Multivariate Cox regression indicated that TA-hematuria (hazard ratio [HR] = 0.004, 95% CI: 0.001, 0.008; p = 0.010) was independently associated with the progression of IgAN. Receiver operating characteristic analysis indicated the optimal TA-hematuria cutoff value for predicting the progression of IgAN was 201.21 RBCs/µL in females and 37.25 RBCs/µL in males.

12.
Cell Death Dis ; 13(8): 693, 2022 08 08.
Artigo em Inglês | MEDLINE | ID: mdl-35941120

RESUMO

Renal fibrosis is a common consequence of various progressive nephropathies, including obstructive nephropathy, and ultimately leads to kidney failure. Infiltration of inflammatory cells is a prominent feature of renal injury after draining blockages from the kidney, and correlates closely with the development of renal fibrosis. However, the underlying molecular mechanism behind the promotion of renal fibrosis by inflammatory cells remains unclear. Herein, we showed that unilateral ureteral obstruction (UUO) induced Gasdermin D (GSDMD) activation in neutrophils, abundant neutrophil extracellular traps (NETs) formation and macrophage-to-myofibroblast transition (MMT) characterized by α-smooth muscle actin (α-SMA) expression in macrophages. Gsdmd deletion significantly reduced infiltration of inflammatory cells in the kidneys and inhibited NETs formation, MMT and thereby renal fibrosis. Chimera studies confirmed that Gsdmd deletion in bone marrow-derived cells, instead of renal parenchymal cells, provided protection against renal fibrosis. Further, specific deletion of Gsdmd in neutrophils instead of macrophages protected the kidney from undergoing fibrosis after UUO. Single-cell RNA sequencing identified robust crosstalk between neutrophils and macrophages. In vitro, GSDMD-dependent NETs triggered p65 translocation to the nucleus, which boosted the production of inflammatory cytokines and α-SMA expression in macrophages by activating TGF-ß1/Smad pathway. In addition, we demonstrated that caspase-11, that could cleave GSDMD, was required for NETs formation and renal fibrosis after UUO. Collectively, our findings demonstrate that caspase-11/GSDMD-dependent NETs promote renal fibrosis by facilitating inflammation and MMT, therefore highlighting the role and mechanisms of NETs in renal fibrosis.


Assuntos
Armadilhas Extracelulares , Nefropatias , Obstrução Ureteral , Caspases/metabolismo , Armadilhas Extracelulares/metabolismo , Fibrose , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Rim/patologia , Nefropatias/patologia , Macrófagos/metabolismo , Miofibroblastos/metabolismo , Proteínas de Ligação a Fosfato/metabolismo , Proteínas Citotóxicas Formadoras de Poros , Transdução de Sinais , Obstrução Ureteral/genética
13.
Ann Transl Med ; 10(2): 77, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-35282095

RESUMO

Background: Central venous stenosis (CVS) of radiocephalic arteriovenous fistula (RCAVF) affects RCAVF function and longevity. Ultrasound screening for CVS is limited by acoustic window. Herein, we analyzed the quantitative axillary venous (AxV) spectrum in hemodialysis patients via RCAVF, and constructed central venous stenosis index (CVSI) model based on the spectrum parameters to early detect resting asymptomatic CVS. Methods: From August 2017 to May 2021, stage 5 chronic kidney disease (CKD) patients dialysed via RCAVF at the First Affiliated Hospital of Fujian Medical University were included in this study. No CVS-related symptoms were found and the pulsation at the arteriovenous anastomosis was normal. However, the patients had the sensation of swelling in the ipsilateral upper limb during dialysis; the venous pressure advanced upon the completion of dialysis; or both (n=52). The inclusion criteria were as follows: (I) Ultrasound (US) showed that the temporal phases of the AxV spectrum were "normal"; and (II) CVS was confirmed by digital subtraction angiography (DSA). The exclusion criteria were as follows: (I) stent placement; (II) multiple stenosis; and (III) placement of central venous catheter. A total of 37 patients participated in the analysis. Eighteen patients were included in the CVS group, and 19 cases without CVS were included in the control group. Independent sample t-test was used to screen each parameter of the AxV spectrum, and a CVSI model was constructed by principal component analysis (PCA). The receiver operating characteristic curve (ROC) was applied to analyze the diagnostic value of CVSI. Results: According to the independent sample t-test, 9 parameters were found to have statistical significance (all P<0.05); they were analyzed by PCA, and the CVSI model was constructed. The ROC showed that CVSI had diagnostic value for CVS. When the cut-off value of CVSI was 7.13, the maximum value of the Youden index was 0.842, with a sensitivity of 100% and a specificity of 84.2%. Conclusions: The CVSI helps to early detect resting asymptomatic CVS and dramatically increases the detection rate of CVS.

14.
Front Bioeng Biotechnol ; 9: 717234, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34692653

RESUMO

Lupus nephritis (LN) is an important driver of end-stage renal disease (ESRD). However, few biomarkers are available for evaluating the diagnosis and prognosis of LN. For this study, we downloaded microarray data of multiple LN expression profiles from the GEO database. We used the WGCNA and R limma packages to identify LN hub genes and differentially-expressed genes (DEGs). We identified nine co-DEGs in the intersection with LN-related genes from the Genecards database. We found DEGs that are primarily associated with immune-related functions and pathways (including with the complement pathway, primary immunodeficiency markers, and MHC-like protein complexes) through our comprehensive GSEA, GO, and KEGG enrichment analyses. We used other LN and SLE validation datasets and discovered six explicitly expressed co-DEGs: HLA-DMA, HLA-DPA1, HLA-DPB1, HLA-DRA, IL10RA, and IRF8 in the LN set; ROC and Precision-Recall curve analyses revealed that these six genes have a good diagnostic efficacy. The correlation analysis with prognostic data from the Nephroseq database indicates that the differential expression of these co-DEGs is associated with a low glomerular filtration rate in that cohort. Additionally, we used a single-cell LN database of immune cells (for the first time) and discovered these co-DEGs to be predominantly distributed in different types of macrophages and B cells. In conclusion, by integrating multiple approaches for DEGs discovery, we identified six valuable biomarkers that are strongly correlated with the diagnosis and prognosis of LN. These markers can help clarify the pathogenesis and improve the clinical management of LN.

15.
J Agric Food Chem ; 69(24): 6810-6819, 2021 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-34096300

RESUMO

In the dairy industry, glutamine (Gln) is often used as a feed additive to increase milk yield and quality; however, the molecular regulation underneath needs further clarification. Here, with bovine mammary epithelial cells (BMECs), the effects and mechanisms of Gln on cell growth and casein synthesis were assessed. When Gln was added or depleted from BMECs, both cell growth and ß-casein (CSN2) expression were increased or decreased, respectively. Overexpressing or inhibiting the mechanistic target of rapamycin (mTOR) revealed that Gln regulated cell growth and CSN2 synthesis through the mTORC1 pathway. A similar intervention of ADP-ribosylation factor 1 (Arf1) uncovered that Gln activated the mTORC1 pathway through Arf1. We next observed that both guanine nucleotide exchange factors, Cytohesin-1/2/3 (CYTH1/2/3, CYTHs) and ADP-ribosylation factor GTPase activating protein 1 (ARFGAP1), interacted with Arf1. Inhibiting CYTHs or ARFGAP1 showed that Gln supplement or depletion activated or inactivated Arf1 through CYTHs or ARFGAP1, respectively. Collectively, this study demonstrated that Gln positively regulated cell growth and casein synthesis in BMECs, which works through the CYTHs/ARFGAP1-Arf1-mTORC1 pathway. These results greatly enhanced current understanding regarding the regulation of the mTOR pathway and provided new insights for the processes of cell growth and casein synthesis by amino acids, particularly Gln.


Assuntos
Fator 1 de Ribosilação do ADP , Caseínas , Animais , Caseínas/metabolismo , Bovinos , Células Epiteliais/metabolismo , Glutamina , Glândulas Mamárias Animais/metabolismo , Alvo Mecanístico do Complexo 1 de Rapamicina/genética , Alvo Mecanístico do Complexo 1 de Rapamicina/metabolismo , Transdução de Sinais
16.
BMC Chem ; 13(1): 111, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31463480

RESUMO

BACKGROUND: In this work, we aim to develop and validate a fast, simple, and sensitive method for the quantitative determination of flibanserin and the exploration of its pharmacokinetics. METHODS: Ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was the method of choice for this investigation and carbamazepine was selected as an internal standard (IS). The plasma samples were processed by one-step protein precipitation using acetonitrile. The highly selective chromatographic separation of flibanserin and carbamazepine (IS) was realised using an Agilent RRHD Eclipse Plus C18 (2.1 × 50 mm, 1.8 µ) column with a gradient mobile phase consisting of 0.1% formic acid in water and acetonitrile. The analytes were detected using positive-ion electrospray ionization mass spectrometry via multiple reaction monitoring (MRM). The target fragment ions were m/z 391.3 → 161.3 for flibanserin and m/z 237.1 → 194 for carbamazepine (IS). The method was validated by linear calibration plots over the range of 100-120,000 ng/mL for flibanserin (R2 = 0.999) in rat plasma. RESULTS: The extraction recovery of flibanserin was in the range of 91.5-95.8%. The determined inter- and intra-day precision was below 12.0%, and the accuracy was from - 6.6 to 12.0%. No obvious matrix effect and astaticism was observed for flibanserin. The target analytes were long-lasting and stable in rat plasma for 12 h at room temperature, 48 h at 4 °C, 30 days at - 20 °C, as well as after three freeze-thaw cycles (from - 20 °C to room temperature). The proposed method has been fully validated and successfully applied to the pharmacokinetic study of flibanserin.

17.
Int Immunopharmacol ; 70: 274-283, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30851708

RESUMO

Previous study has indicated that taraxasterol (TAR), one of bioactive pentacyclic triterpenes mainly isolated from Chinese medicine herb Taraxacum officinale, displays considerable anti-inflammatory effects in various kinds of models. However, its effects on rheumatoid arthritis (RA) have still not been elucidated. In this study, we aim to investigate its anti-inflammatory effects and underlying mechanisms of TAR against RA using both interleukin (IL)-1ß-stimulated human fibroblast-like synoviocytes rheumatoid arthritis (HFLS-RA) in vitro and collagen-induced arthritis (CIA) mice in vivo. Firstly, our results demonstrated that TRA significantly suppressed the IL-1ß-induced expressions of pro-inflammatory cytokines, including tumor necrosis factor-α (TNF-α), IL-6, and IL-8 and productions of matrix metalloproteinases (MMPs), like MMP-1 and MMP-3 in HFLS-RA in vitro. Moreover, TRA alleviated arthritis progressions and prevented inflammatory processes in the joint tissues of CIA mice in vivo. Further mechanism studies indicated that TRA blocked nuclear factor kappa B (NF-κB) activation via modulating inhibitor of kappa B (IκB), IκB kinase (IKK) and transforming growth factor-ß-activated kinase 1 (TAK1). Results also demonstrated that TRA suppressed the NOD-like receptor protein 3 (NLRP3) inflammasomes through blocking expressions of NLRP3, apoptosis-associated speck-like protein containing (ASC), and caspase-1 in both IL-1ß-induced HFLS-RA and CIA mice. In conclusions, current findings suggested that TRA might one of considerable therapeutic compounds for relieving rheumatoid arthritis progress via suppressing inflammations through modulating NF-κB and NLRP3 inflammasomes pathways.


Assuntos
Anti-Inflamatórios/uso terapêutico , Artrite Experimental/tratamento farmacológico , Artrite Reumatoide/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Fibroblastos/patologia , Esteróis/uso terapêutico , Sinoviócitos/efeitos dos fármacos , Triterpenos/uso terapêutico , Animais , Apoptose , Artrite Reumatoide/induzido quimicamente , Células Cultivadas , Modelos Animais de Doenças , Progressão da Doença , Humanos , Interleucina-1beta/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos DBA , NF-kappa B/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Transdução de Sinais , Sinoviócitos/patologia
18.
J Pharm Biomed Anal ; 162: 249-256, 2019 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-30268993

RESUMO

Idelalisib acts as a phosphoinositide 3 kinase inhibitor, which has been approved by the US FDA for the treatment of certain hematological malignancies. The aim of this study is to profile the metabolites of idelalisib in the liver microsomes of mouse, rat, rabbit, dog, monkey and human. Idelalisib at the concentration of 20 µM was incubated with the liver microsomes in the presence of NADPH, GSH and UDPGA. The incubation samples were analyzed by ultra-high performance liquid chromatography coupled with diode array detector and linear ion trap-orbitrap tandem mass spectrometer (UHPLC-DAD-LTQ-Orbitrap-MS), and the post-acquisition data was processed by Metworks software. Under the current experimental conditions, a total of 14 metabolites were detected. The structures of the metabolites were characterized based on their accurate masses, fragmental ions and retention times. Our results suggested the following: 1) idelalisib was prone to oxidative defluorination to give rise to desfluoroidelalisib (M13). This metabolite was reactive in nature as its corresponding GSH conjugate was detected (M4). Except GSH conjugation, this metabolite can further undergo oxygenation (M7 and M14), and glucuronidation (M3); 2) oxygenation was the major metabolic pathway in liver microsomes, leading to the metabolite M10 in all test species; 3) idelalisib can be directly conjugated with glucuronide to form N+-glucuronide (M1). Species-specific metabolic difference was observed between animals and human and rat and dog have closer metabolic profiles to human compared with other animal species.


Assuntos
Antineoplásicos/metabolismo , Microssomos Hepáticos/metabolismo , Inibidores de Proteínas Quinases/metabolismo , Purinas/metabolismo , Quinazolinonas/metabolismo , Animais , Cromatografia Líquida de Alta Pressão , Cães , Glucuronídeos/metabolismo , Glutationa/metabolismo , Haplorrinos , Humanos , Inativação Metabólica , Camundongos , Estrutura Molecular , Oxirredução , Coelhos , Ratos , Especificidade da Espécie , Espectrometria de Massas por Ionização por Electrospray , Espectrofotometria Ultravioleta , Espectrometria de Massas em Tandem
20.
Korean J Intern Med ; 38(6): 947-948, 2023 11.
Artigo em Inglês | MEDLINE | ID: mdl-37334509
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