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BACKGROUND: An appropriate prediction model for adverse prognosis before peritoneal dialysis (PD) is lacking. Thus, we retrospectively analysed patients who underwent PD to construct a predictive model for adverse prognoses using machine learning (ML). METHODS: A retrospective analysis was conducted on 873 patients who underwent PD from August 2007 to December 2020. A total of 824 patients who met the inclusion criteria were included in the analysis. Five commonly used ML algorithms were used for the initial model training. By using the area under the curve (AUC) and accuracy (ACC), we ranked the indicators with the highest impact and displayed them using the values of Shapley additive explanation (SHAP) version 0.41.0. The top 20 indicators were selected to build a compact model that is conducive to clinical application. All model-building steps were implemented in Python 3.8.3. RESULTS: At the end of follow-up, 353 patients withdrew from PD (converted to haemodialysis or died), and 471 patients continued receiving PD. In the complete model, the categorical boosting classifier (CatBoost) model exhibited the strongest performance (AUC = 0.80, 95% confidence interval [CI] = 0.76-0.83; ACC: 0.78, 95% CI = 0.72-0.83) and was selected for subsequent analysis. We reconstructed a compression model by extracting 20 key features ranked by the SHAP values, and the CatBoost model still showed the strongest performance (AUC = 0.79, ACC = 0.74). CONCLUSIONS: The CatBoost model, which was built using the intelligent analysis technology of ML, demonstrated the best predictive performance. Therefore, our developed prediction model has potential value in patient screening before PD and hierarchical management after PD.
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Diálise Peritoneal , Humanos , Estudos Retrospectivos , Algoritmos , Prognóstico , Aprendizado de MáquinaRESUMO
Non-small cell lung cancer (NSCLC) is one of the leading causes of death worldwide. Brain metastases are a common complication of a wide range of human malignancies, particularly lung adenocarcinoma (LUAD). Brain-derived neurotrophic factor (BDNF), a member of the neurotrophin family, has been linked to several human malignancies and has been shown to promote LUAD tumorigenesis. However, its function in the tumour immune microenvironment (TIME) remains largely unexplored, especially in complex brain tissue environments. In this study, BDNF was found to be particularly increased in patients with advanced tumour stage, lymphatic metastasis, and distant metastasis, indicating a correlation with LUAD progression. We characterized the prognostic value of BDNF and defined BDNF as an unfavourable prognostic indicator through a common driver gene-independent mechanism in LUAD. Furthermore, patients with increased BDNF levels in primary LUAD might have a higher risk of developing brain metastasis (BM), and central nervous system (CNS) metastasis showed an elevated expression of BDNF compared to their matched primary lesions. Additionally, we investigated the interaction between BDNF and infiltrating immune cells in both primary lesions and paired BM using multiplex immunostaining. The results showed that BDNF might drive an immunosuppressive tumour microenvironment (TME) by re-education of tumour-associated macrophages (TAMs) toward a pro-tumorigenic M2 phenotype, particularly in BM. Our findings demonstrate that BDNF serves as an independent potential prognostic marker and correlates with BM in LUAD. As it is closely related to TAM polarization, BDNF may be a promising immune-related biomarker and molecular target in patients with LUAD.
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Adenocarcinoma de Pulmão , Neoplasias Encefálicas , Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Humanos , Fator Neurotrófico Derivado do Encéfalo , Prognóstico , Carcinogênese , Microambiente TumoralRESUMO
The mechanisms underlying acute kidney injury (AKI) and chronic kidney disease (CKD) progression include interstitial inflammation, cellular senescence, and oxidative stress (OS). Although vanin-1 (VNN1) plays an important role in OS, its contribution to the AKI-CKD transition remains unknown. Here, we explored the roles and mechanisms of VNN1 in the progression of the AKI-CKD transition. We observed that VNN1 expression was upregulated after ischemia/reperfusion (I/R) injury and high VNN1 expression levels were associated with poor renal repair after I/R injury. In VNN1 knockout (KO) mice, recovery of serum creatinine and blood urea nitrogen levels after I/R injury was accelerated and renal fibrosis was inhibited after severe I/R injury. Furthermore, in VNN1 KO mice, senescence of renal tubular cells was inhibited after severe I/R injury, as assessed by P16 expression and SA-ß-Gal assays. However, our results also revealed that VNN1 KO renal tubular cells did not resist senescence when OS was blocked. To elucidate the mechanism underlying VNN1-mediated regulation of senescence during the AKI-CKD transition, retinoblastoma 1 (RB1) was identified as a potential target. Our results suggest that the reduced senescence in VNN1 KO renal tubular cells was caused by suppressed RB1 expression and phosphorylation. Collectively, our results unveil a novel molecular mechanism by which VNN1 promotes AKI-CKD transition via inducing senescence of renal tubular cells by activating RB1 expression and phosphorylation after severe renal injury. The present study proposes a new strategy for designing therapies wherein VNN1 can be targeted to obstruct the AKI-CKD transition.
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Injúria Renal Aguda , Insuficiência Renal Crônica , Traumatismo por Reperfusão , Injúria Renal Aguda/genética , Injúria Renal Aguda/metabolismo , Animais , Senescência Celular , Fibrose , Rim/metabolismo , Camundongos , Camundongos Knockout , Insuficiência Renal Crônica/metabolismo , Traumatismo por Reperfusão/metabolismoRESUMO
BACKGROUND: The study aims to explore the incidence and risk factors of cognitive dysfunction in hemodialysis patients. METHODS: PubMed, Embase, Cochrane Library, and Web of Science databases were searched for clinical studies on the association between hemodialysis and cognitive dysfunction from the database's inception to 1 December 2022. Two researchers independently completed data extraction and risk of bias assessments for the included studies. All statistical analyses were performed using STATA15.0 software. RESULTS: Ten studies were included in this meta-analysis, with a total of 5535 hemodialysis patients, that is, 2033 patients with cognitive dysfunction and 3502 patients with normal cognitive function. The Newcastle-Ottawa Scale scores of the included studies were greater than 5. Meta-analysis results suggested that the incidence of cognitive dysfunction in hemodialysis patients was (effect size = 51%, 95% confidence interval [CI] [0.33, 0.69]), and hemodialysis patients with cognitive dysfunction were often older than those with normal cognition (standard mean difference [SMD] = 0.49, 95% CI [0.31, 0.68]). Female gender was a risk factor for cognitive dysfunction in hemodialysis patients (relative risk [RR] = 1.21, 95% CI [1.04, 1.41]); diabetes (RR = 1.33, 95% CI [1.04, 1.71]) and stroke (RR = 1.66, 95% CI [1.08, 2.55]) increased the incidence of cognitive dysfunction in hemodialysis patients. CONCLUSIONS: The most important risk factors for cognitive dysfunction associated with hemodialysis might be female gender, old age, diabetes, and stroke. Close attention should be paid to such patients for early prevention.
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Disfunção Cognitiva , Diabetes Mellitus , Acidente Vascular Cerebral , Humanos , Feminino , Incidência , Diálise Renal/efeitos adversos , Disfunção Cognitiva/epidemiologia , Disfunção Cognitiva/etiologia , Fatores de RiscoRESUMO
Phagocytic clearance of apoptotic cells by the macrophages (efferocytosis) is impaired in sepsis, but its mechanism is poorly understood. Extracellular cold-inducible RNA-binding protein (eCIRP) is a novel damage-associated molecular pattern that fuels inflammation. We identify that eCIRP-induced neutrophil extracellular traps (NETs) impair efferocytosis through a novel mechanism. Coculture of macrophages and apoptotic thymocytes in the presence of recombinant murine CIRP (rmCIRP)-induced NETs significantly inhibited efferocytosis. Efferocytosis was significantly inhibited in the presence of rmCIRP-treated wild-type (WT), but not PAD4-/- neutrophils. Efferocytosis in the peritoneal cavity of rmCIRP-injected PAD4-/- mice was higher than WT mice. Milk fat globule-EGF-factor VIII (MFG-E8), an opsonin, increased macrophage efferocytosis, whereas the inhibition of efferocytosis by NETs was not rescued upon addition of MFG-E8, indicating disruption of MFG-E8's receptor(s) αvß3 or αvß5 integrin by the NETs. We identified neutrophil elastase in the NETs significantly inhibited efferocytosis by cleaving macrophage surface integrins αvß3 and αvß5 Using a preclinical model of sepsis, we found that CIRP-/- mice exhibited significantly increased rate of efferocytosis in the peritoneal cavity compared with WT mice. We discovered a novel role of eCIRP-induced NETs to inhibit efferocytosis by the neutrophil elastase-dependent decrease of αvß3/αvß5 integrins in macrophages. Targeting eCIRP ameliorates sepsis by enhancing efferocytosis.
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Armadilhas Extracelulares/imunologia , Macrófagos/imunologia , Neutrófilos/imunologia , Proteínas de Ligação a RNA/imunologia , Sepse/imunologia , Timócitos/imunologia , Animais , Técnicas de Cocultura , Armadilhas Extracelulares/genética , Masculino , Camundongos , Camundongos Knockout , Proteínas de Ligação a RNA/genética , Sepse/genéticaRESUMO
INTRODUCTION: The optimal dialysate bicarbonate concentration (DBIC) for hemodialysis (HD) remains controversial. Herein, we analyzed the effect of dialysate bicarbonate levels on mortality in HD patients. METHODS: Patients undergoing maintenance HD were recruited from the HD unit of the Daping Hospital. Patients were categorized into quartiles according to their DBIC level (quartile 1: <31.25 mmol/L, n = 77; quartile 2: 31.25-32.31 mmol/L, n = 76; quartile 3: 32.31-33.6 mmol/L; n = 81; quartile 4: ≥33.6 mmol/L, n = 79). Demographic and clinical data were collected. Survival curves were estimated using the Kaplan-Meier method. A Cox proportional hazards regression model was used to estimate the association between DBIC and all-cause mortality. RESULTS: We included 313 patients undergoing maintenance HD with a mean DBIC of 32.16 ± 1.59 mmol/L (range, 27.20-34.72 mmol/L). The patients in quartile 4 were more likely to have higher pre- and post-HD serum bicarbonate concentrations than those in other quartiles. The mortality rate was lowest in quartile 2 (10.53%). The survival time was significantly lower in the quartile 4 group than in the other quartiles (p = 0. 008, log-rank test). After full adjustment, the hazard ratio (per 3 mmol/L higher DBIC) for all-cause mortality was 4.29 (95% confidence interval, 2.11-8.47) in all patients, whereas no significant association was observed between DBIC and initial hospitalization. CONCLUSIONS: Our data indicate that DBIC is positively associated with all-cause mortality. A DBIC concentration of 31-32 mmol/L may benefit patient outcomes. This study provides an evidence-based medical basis for optimal dialysis prescription in the future.
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Soluções para Diálise , Falência Renal Crônica , Humanos , Bicarbonatos , Diálise Renal/métodos , Hospitalização , Modelos de Riscos Proporcionais , Falência Renal Crônica/complicaçõesRESUMO
Diabetic nephropathy (DN) is the leading cause of end-stage renal disease. Renal tubulointerstitial injury is an important pathophysiological basis that contributes to the progression of DN to end-stage renal disease. Stress-induced senescence of renal tubular epithelial cells (RTECs) forms a key link that causes tubulointerstitial injury. In recent years, it has been reported that organelles, such as endoplasmic reticulum, mitochondria, and lysosomes, in RTECs are damaged to varying degrees in DN, and that their functional imbalance may lead to stress-induced senescence of RTECs, thereby causing sustained cellular and tissue-organ damage, which in turn promotes the progression of the disease. However, the core mechanism underlying changes in the senescence microenvironment caused by stress-induced senescence of RTECs in DN is still not understood. In addition, the mechanism by which organelles lose homeostasis also needs to be further investigated. Herein, we described the specific pathophysiological mechanisms of renal tubular injury, stress-induced senescence of RTECs, and their association with organelles in the context of DN in order to provide reference for the next-step research, as well as the development of new therapeutic strategies.
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Diabetes Mellitus , Nefropatias Diabéticas , Falência Renal Crônica , Humanos , Nefropatias Diabéticas/etiologia , Rim , Túbulos Renais , Células EpiteliaisRESUMO
BACKGROUND: Peritoneal fibrosis induced by various factors during peritoneal dialysis (PD) can eventually lead to ultrafiltration failure and termination of PD treatment. The existing animal models are caused by a single stimulus, and cannot accurately simulate complex pathogenesis of peritoneal injury and fibrosis. We aim to develop an efficient and realistic mouse model of PD-associated peritoneal injury using daily intraperitoneal injection (I.P.) of human peritonitis PD effluent. METHODS: Eight-week-old male C57BL/6 mice were classified into six groups: saline control; 2.5% PD fluid; 2.5% PD fluid + lipopolysaccharide (LPS); 4.25% PD fluid; 4.25% PD fluid + LPS; and peritonitis effluent. Mice received daily I.P. for 6 weeks, and were sacrificed to determine peritoneal structural and functional damage, inflammation, and fibrosis. RESULTS: Mice in the peritonitis effluent group had low mortality. The submesothelial thickness in the peritonitis effluent group was significantly greater than that in the 2.5% PD fluid group. The peritonitis effluent group had increased expression of fibrosis markers (α-SMA, Collagen I, etc.), neutrophil granulocytes (MPO), and macrophages (CD68, F4/80) in the peritoneum based on immunohistochemical staining; and significantly higher expression of inflammation markers (IL-1ß, IL-6, etc.) and fibrosis markers (TGF-ß1, α-SMA, etc.) based on real-time qPCR. Modified peritoneal equilibration tests (PET) demonstrated that I.P. of peritonitis effluent reduced peritoneal ultrafiltration. CONCLUSION: Our novel animal model of PD-associated peritoneal injury faithfully simulates the clinical pathophysiological process. This animal model may be useful for study of the pathogenesis of PD-associated peritoneal injury and identification of novel treatments.
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Diálise Peritoneal , Fibrose Peritoneal , Peritonite , Animais , Soluções para Diálise/efeitos adversos , Modelos Animais de Doenças , Humanos , Inflamação/complicações , Lipopolissacarídeos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Diálise Peritoneal/efeitos adversos , Fibrose Peritoneal/metabolismo , Peritônio/metabolismo , Peritonite/etiologiaRESUMO
Renal tubular injury contributes to the progression of diabetic nephropathy (DN). This study explored the role and mechanisms of E3-ubiquitin ligase Parkin in the renal tubular injury of DN. We found that Parkin expression gradually decreased and was inversely associated with IL-6, TGF-ß1, and GATA4 expression in the kidney during the progression of DN. Parkin over-expression (OE) reduced inflammation, fibrosis, premature senescence of renal tubular epithelial cells (RTECs), and improved renal function while Parkin knockout (KO) had opposite effects in DN mice. Parkin-OE decreased GATA4 protein, but not its mRNA transcripts in the kidney of DN mice and high glucose (HG)-treated RTECs. Immunoprecipitation indicated that Parkin directly interacted with GATA4 in DN kidney. Parkin-OE enhanced GATA4 ubiquitination. Furthermore, Parkin-KO upregulated growth arrest-specific gene 1 (GAS1) expression in renal tubular tissues of DN mice and GATA4-OE enhanced the HG-upregulated GAS1 expression in RTECs. Conversely, GAS1-OE mitigated the effect of Parkin-OE on HG-induced P21, IL-6, and TGF-ß1 expression in RTECs. These results indicate that Parkin inhibits the progression of DN by promoting GATA4 ubiquitination and downregulating the GATA4/GAS1 signaling to inhibit premature senescence, inflammation, and fibrosis in DN mice. Thus, these findings uncover new mechanisms underlying the action of Parkin during the process of DN.
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Proteínas de Ciclo Celular/metabolismo , Diabetes Mellitus Experimental/complicações , Nefropatias Diabéticas/prevenção & controle , Fator de Transcrição GATA4/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitinação , Animais , Proteínas de Ciclo Celular/genética , Células Cultivadas , Nefropatias Diabéticas/etiologia , Nefropatias Diabéticas/patologia , Transição Epitelial-Mesenquimal , Feminino , Fator de Transcrição GATA4/genética , Proteínas Ligadas por GPI/genética , Proteínas Ligadas por GPI/metabolismo , Glucose/metabolismo , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Prognóstico , Ubiquitina-Proteína Ligases/genéticaRESUMO
Tubulointerstitial inflammation plays a critical role in the progression of diabetic nephropathy (DN), and nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasomes contribute to renal interstitial inflammation in DN. Decreased expression of optineurin (OPTN) is also associated with the progression of DN. We investigated the role of OPTN in activation of the NLRP3 inflammasome in DN. We initially examined the renal biopsy tissues of 172 patients with type 2 DN and 32 nondiabetic patients with renal hamartoma. Expression of renal OPTN was significantly lower in patients with DN and negatively correlated with urinary levels of IL-1ß and IL-18. Confocal microscopy analysis of the biopsies indicated no NLRP3 or IL-1ß staining in OPTN-positive renal tubular epithelial cells (RTECs), indicating a negative correlation of OPTN expression with the activation of NLRP3 inflammasome. In vitro studies of murine RTECs indicated the levels of OPTN mRNA and protein decreased significantly after stimulation by a high glucose (HG) treatment. Relative to RTECs given HG, RTECs overexpressing OPTN showed significantly lower levels of NLRP3 expression, cleavage of caspase-1 and IL-1ß, and release of IL-1 ß and IL-18. Overexpression of OPTN in the presence of HG significantly increased the costaining of microtubule-associated protein 1A/1B-light chain 3-II and translocase of outer mitochondrial membrane 20 in RTECs, suggesting that OPTN enhances mitophagy. In addition, mitochondrial division inhibitor 1 blocked the inhibitory effect of OPTN overexpression on the activation of NLRP3 inflammasome in the presence of HG, indicating that OPTN overexpression inhibited NLRP3 inflammasome activation by enhancement of mitophagy. OPTN gene silencing significantly enhanced production of mitochondrial reactive oxygen species (mtROS) in the presence of HG. Compared with HG+OPTN small interfering RNA (siRNA)-treated RTECs, HG+OPTN siRNA+MitoTempo-treated RTECs attenuated NLRP3 inflammasome activation, suggesting that OPTN gene silencing activates the NLRP3 inflammasome by increasing mtROS in HG-treated RTECs. Taken together, our results demonstrate that OPTN inhibits the activation of NLRP3 inflammasome by enhancing mitophagy.-Chen, K., Feng, L., Hu, W., Chen, J., Wang, X., Wang, L., He, Y. Optineurin inhibits NLRP3 inflammasome activation by enhancing mitophagy of renal tubular cells in diabetic nephropathy.
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Proteínas de Ciclo Celular/fisiologia , Nefropatias Diabéticas/patologia , Inflamassomos/fisiologia , Túbulos Renais Proximais/patologia , Proteínas de Membrana Transportadoras/fisiologia , Mitofagia/fisiologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/fisiologia , Adulto , Idoso , Animais , Proteínas de Ciclo Celular/análise , Proteínas de Ciclo Celular/genética , Células Cultivadas , Diabetes Mellitus Tipo 2/complicações , Nefropatias Diabéticas/metabolismo , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Feminino , Glucose/farmacologia , Humanos , Túbulos Renais Proximais/metabolismo , Masculino , Potencial da Membrana Mitocondrial , Proteínas de Membrana Transportadoras/análise , Proteínas de Membrana Transportadoras/genética , Camundongos , Camundongos Endogâmicos C57BL , Microscopia Confocal , Pessoa de Meia-Idade , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/análise , RNA Mensageiro/biossíntese , Espécies Reativas de Oxigênio/metabolismo , Proteínas Recombinantes/metabolismoRESUMO
Through-metal transfer of energy and data using piezoelectric transduce can avoid the potential leakage problem caused by physical penetrations and wired feed-through. The through-metal transfer efficiency of energy or data is determined by the relative pressure on the receiving PZT (piezoelectric transducer). Hence, in this paper, we first propose the Spatial Equivalent Plane Acoustic Pressure (SEPAP), which is defined as the integration of the acoustic pressure over the receiving area, to model the pressure on the receiving PZT. Then we analyze the features of SEPAP and the factors impacting it by utilizing COMSOL. Furthermore, we propose a low-cost and small-size prototype for simultaneous transfer of energy and bidirectional communication through metal by using two pairs of PZTs working on different resonant frequencies. Extensive experiment has been done on evaluating the match between SEPAP transfer efficiency and the power transfer efficiency and analyzing the achievable data rate for bi-directional communication. Test through a 20 mm aluminum alloy plate shows that power transmission with efficiency 20.3% and data communication rate up to 38.4 Kbps can be achieved simultaneously.
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BACKGROUND/AIMS: Stress-induced cell senescence, which contributes to cell cycle arrest and is independent of age, plays an important role in chronic kidney disease (CKD) progression. DcR2, as a senescent marker, exclusively expressed in senescent tubular epithelia. The objective of this study was to examine whether urinary DcR2 (uDcR2) could be a potential biomarker for tubulointerstitial fibrosis (TIF) in patients with immunoglobulin A nephropathy (IgAN). METHODS: This study included 210 IgAN patients and 80 healthy volunteers, with uDcR2 levels measured using enzyme-linked immunosorbent assay. We examined the relationship among uDcR2/Cr levels, renal function, and pathological parameters, using regression analysis to identify risk factors for TIF and the area under the curve (AUC) approach to predict TIF. Renal DcR2 expression was quantified by immunohistochemistry. Co-expression of DcR2 with fibrotic markers (α-smooth muscle actin [α-SMA], collagen III) was analyzed by confocal microscopy. RESULTS: Levels of uDcR2/Cr were significantly higher in IgAN patients and in those with more severe TIF, compared with healthy controls. Serum DcR2 levels were similar across groups. The proportion of IgAN patients with stages 1-2 CKD and T0 was highest among those with uDcR2/Cr <130 ng/g. In contrast, the majority of those with uDcR2/Cr >201 ng/g had stages 4-5 CKD and T2. Levels of uDcR2/Cr were positively associated with urinary albumin to creatinine ratio (ACR), urinary N-acetyl-ß-D-glucosaminidase (uNAG)/Cr, and TIF scores and negatively associated with estimated glomerular filtration rate (eGFR). uDcR2/Cr, uNAG, ACR, and eGFR were independent predictors for TIF, with AUC of 0.907 for uDcR2/Cr. This AUC value was higher than that observed for eGFR, uNAG/Cr, or ACR. The sensitivity and specificity of uDcR2/Cr in predicting TIF were 87.0 and 80.5%, respectively. Moreover, uDcR2/Cr levels were positively associated with the percentage of renal DcR2 expression. Renal DcR2 co-localized with α-SMA and collagen III in the kidneys of IgAN patients. CONCLUSIONS: Levels of uDcR2/Cr were closely associated with the severity of TIF and renal function parameters. uDcR2/Cr represents a potential biomarker for predicting TIF in IgAN patients.
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Glomerulonefrite por IGA/complicações , Testes de Função Renal/métodos , Nefrite Intersticial/diagnóstico , Receptores Chamariz do Fator de Necrose Tumoral/metabolismo , Adolescente , Adulto , Idoso , Feminino , Glomerulonefrite por IGA/patologia , Humanos , Rim/patologia , Masculino , Pessoa de Meia-Idade , Nefrite Intersticial/patologia , Adulto JovemRESUMO
The present study was aimed to establish a modified method for culturing mouse renal proximal tubular epithelial cells (TECs). Renal cortex was isolated from mouse kidney and scissored into pieces. TECs were separated by digesting scissored renal cortex in type II collagenase combined with strainer filtration, and then cultured in DMEM. The morphology of TECs was observed under inverted microscopy. The cell proliferative ability was assessed by flow cytometry, and cell viability was analyzed by CCK-8 assay. The purity of TECs was identified by immunofluorescence. Immunofluorescence observation showed that more than 95% cells were epithelial marker CK18 positive and more than 90% cells expressed renal proximal TECs marker proteins, Villin, AQP1, and SGLT2. The cells could be subcultured for about 5 times. The cell proliferative ability declined following the repeated passage. This study introduced a modified efficient method for culturing highly purified mouse renal proximal TECs.
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Células Epiteliais/citologia , Túbulos Renais Proximais/citologia , Cultura Primária de Células , Animais , Células Cultivadas , Citometria de Fluxo , CamundongosRESUMO
Tubulointerstitial injury (TII) plays a crucial role in the progression of diabetic nephropathy (DN), but lack of specific and sensitive biomarkers for monitoring TII in DN management. This study is to investigate whether urinary decoy receptor 2 (uDcR2) could serve as a novel noninvasive biomarker for assessing TII in DN. We recruited 311 type 2 diabetics and 139 DN patients who were diagnosed by renal biopsy. uDcR2 levels were measured by ELISA, and renal DcR2 expression was detected immunohistochemically. Associations between uDcR2 and renal DcR2 and renal functional parameters were evaluated. Receiver operating characteristics (ROC) curve analyzed area under the curve (AUC) of uDcR2 for assessing TII. Double staining was undertaken for renal DcR2 with proximal and distal tubular markers; senescent markers p16, p21, and senescence-associated ß-galactosidase (SA-ß-gal); and fibrotic markers collagen I and IV. We found DcR2 was primarily expressed in renal proximal tubules; uDcR2 levels were elevated per albuminuria stratum and correlated with renal functional parameters in diabetics and were associated with percentage of tubular DcR2 and TII score in DN. The uDcR2 had an AUC of 0.909 for assessing TII in DN by ROC analysis. Almost all tubular DcR2 was coexpressed with p16 and p21, and nearly more than one-half of tubular DcR2 was positive for SA-ß-gal, primarily in collagen I- and IV-positive regions of DN. Our results indicate uDcR2 could potentially serve as a novel biomarker for TII and may reflect senescence of renal proximal tubular cells in DN pathogenesis.
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Diabetes Mellitus Tipo 2/complicações , Nefropatias Diabéticas/urina , Túbulos Renais Proximais/química , Receptores Chamariz do Fator de Necrose Tumoral/urina , Idoso , Área Sob a Curva , Biomarcadores/urina , Biópsia , Estudos de Casos e Controles , Senescência Celular , Colágeno Tipo I/análise , Colágeno Tipo IV/análise , Estudos Transversais , Inibidor p16 de Quinase Dependente de Ciclina/análise , Inibidor de Quinase Dependente de Ciclina p21/análise , Nefropatias Diabéticas/diagnóstico , Nefropatias Diabéticas/etiologia , Ensaio de Imunoadsorção Enzimática , Feminino , Fibrose , Humanos , Imuno-Histoquímica , Túbulos Renais Proximais/patologia , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Curva ROC , Regulação para Cima , Urinálise , beta-Galactosidase/análiseRESUMO
Background: Radix Bupleuri, a kind of Chinese herbal medicine with great clinical use, is often confused with its adulterants, and it is difficult to identify it without certain knowledge. The existing identification methods have their own drawbacks, so a new method is needed to realize the identification of Radix Bupleuri and its adulterants. Methods: We used Micro Computed Tomography (Micro-CT) to perform tomography scans on Radix Bupleuri and its adulterants, performed data screening and data correction on the obtained DICOM images, and then applied 3D reconstruction, data augmentation, and ResNext deep learning model for the classification study. Results: The DICOM images after data screening, data correction, and 3D reconstruction can observe the differences in the microstructure of Radix Bupleuri and its adulterants, thus enabling effective classification and analysis. Meanwhile, the accuracy of classification using the ResNext model reached 75%. Conclusion: The results of this study showed that Micro-CT technology is feasible for the authentication of Radix Bupleuri. The pre-processed and 3D reconstructed tomographic images clearly show the microstructure and the difference between Radix Bupleuri and its adulterants without damaging the internal structure of the samples. This study concludes that Micro-CT technology provides important technical support for the reliable identification of Radix Bupleuri and its adulterants, which is expected to play an important role in the quality control and clinical application of herbs.
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Cellular senescence represents an irreversible state of cell-cycle arrest during which cells secrete senescence-associated secretory phenotypes, including inflammatory factors and chemokines. Additionally, these cells exhibit an apoptotic resistance phenotype. Cellular senescence serves a pivotal role not only in embryonic development, tissue regeneration, and tumor suppression but also in the pathogenesis of age-related degenerative diseases, malignancies, metabolic diseases, and kidney diseases. The senescence of renal tubular epithelial cells (RTEC) constitutes a critical cellular event in the progression of acute kidney injury (AKI). RTEC senescence inhibits renal regeneration and repair processes and, concurrently, promotes the transition of AKI to chronic kidney disease via the senescence-associated secretory phenotype. The mechanisms underlying cellular senescence are multifaceted and include telomere shortening or damage, DNA damage, mitochondrial autophagy deficiency, cellular metabolic disorders, endoplasmic reticulum stress, and epigenetic regulation. Strategies aimed at inhibiting RTEC senescence, targeting the clearance of senescent RTEC, or promoting the apoptosis of senescent RTEC hold promise for enhancing the renal prognosis of AKI. This review primarily focuses on the characteristics and mechanisms of RTEC senescence, and the impact of intervening RTEC senescence on the prognosis of AKI, aiming to provide a foundation for understanding the pathogenesis and providing potentially effective approaches for AKI treatment.
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Peritoneal fibrosis is a critical sequela that limits the application of peritoneal dialysis (PD). This study explored the role and mechanism of bone marrow mesenchymal stem cell-derived exosomes (BMSC-Exos) in preventing PD-associated peritoneal injury. C57BL/6 mice were randomized into three groups: a control (saline), peritoneal injury [2.5% glucose peritoneal dialysate + lipopolysaccharide (LPS)], and peritoneal injury + exosome group. After 6 weeks, mice were dissected, and the parietal peritoneum was collected. The level of peritoneal structural and functional damage was assessed. Additionally, transcriptome analysis of the peritoneum and miRNA sequencing on BMSC-Exos were performed. The parietal peritoneum had significantly thickened, and peritoneal function was impaired in the peritoneal injury group. Peritoneal structural and functional damage was significantly reduced after exosome treatment, while peritoneal inflammation, fibrosis, angiogenesis, and mesothelial damage significantly increased. Transcriptomic analysis showed that the BMSC-Exos affected the cell cycle process, cell differentiation, and inflammatory response regulation. Significant pathways in the exosome group were enriched by inflammation, immune response, and cell differentiation, which constitute a molecular network that regulates the peritoneal protective mechanism. Additionally, inflammatory factors (TNF-α, IL-1ß), fibrosis markers (α-SMA, collagen-III, fibronectin), profibrotic cytokines (TGF-ß1), and angiogenesis-related factor (VEGF) were downregulated at the mRNA and protein levels through BMSC-Exos treatment. BMSC-Exos treatment can prevent peritoneal injury by inhibiting peritoneal fibrosis, inflammation, and angiogenesis, showing a multitarget regulatory effect. Therefore, BMSC-Exos therapy might be a new therapeutic strategy for treating peritoneal injury.
Assuntos
Exossomos , Células-Tronco Mesenquimais , Diálise Peritoneal , Fibrose Peritoneal , Camundongos , Animais , Fibrose Peritoneal/genética , Fibrose Peritoneal/metabolismo , Exossomos/metabolismo , Camundongos Endogâmicos C57BL , Diálise Peritoneal/efeitos adversos , Inflamação/metabolismoRESUMO
Acute kidney injury (AKI) is characterized by a rapid decrease in renal function with high mortality and risk of progression to chronic kidney disease (CKD). Ischemia and reperfusion injury (IRI) is one of the major causes of AKI. However, the cellular and molecular responses of the kidney to IRI are complex and not fully understood. Herein, we conducted unbiased proteomics and bioinformatics analyses in an IRI mouse model on days 3, 7, and 21, and validated the results using IRI, unilateral ureteral obstruction (UUO), and biopsies from patients with AKI or CKD. The results indicated an obvious temporal expression profile of differentially expressed proteins and highlighted impaired lipid metabolism during the progression of AKI to CKD. Acyl-coenzyme A oxidase 1 (Acox1), the first rate-limiting enzyme of peroxisomal fatty acid beta-oxidation, was then selected, and its disturbed expression in the two murine models validated the proteomic findings. Accordingly, Acox1 expression was significantly downregulated in renal biopsies from patients with AKI or CKD, and its expression was negatively correlated with kidney injury score. Furthermore, in contrast to the decreased Acox1 expression, lipid droplet accumulation was remarkably increased in these renal tissues, suggesting dysregulation of fatty acid oxidation. In conclusion, our results suggest that defective peroxisomal fatty acid oxidation might be a common pathological feature in the transition from AKI to CKD, and that Acox1 is a promising intervention target for kidney injury and repair.
RESUMO
Lung adenocarcinoma (LUAD) is a public enemy with a very high incidence and mortality rate, for which there is no specific detectable biomarker. Pregnancy zone protein (PZP) is an immune-related protein; however, the functions of PZP in LUAD are unclear. In this study, a series of bioinformatics methods, combined with immunohistochemistry (IHC), four-color multiplex fluorescence immunohistochemistry (mIHC), quantitative real-time PCR (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA), were utilized to explore the prognostic value and potential role of PZP in LUAD. Our data revealed that PZP expression was markedly reduced in LUAD tissues, tightly correlated with clinical stage and could be an independent unfavorable prognostic factor. In addition, pathway analysis revealed that high expression of PZP in LUAD was mainly involved in immune-related molecules. Tumor immune infiltration analysis by CIBERSORT showed a significant correlation between PZP expression and several immune cell infiltrations, and IHC further confirmed a positive correlation with CD4+ T-cell infiltration and a negative correlation with CD68+ M0 macrophage infiltration. Furthermore, mIHC demonstrated that PZP expression gave rise to an increase in CD86+ M1 macrophages and a decrease in CD206+ M2 macrophages. Therefore, PZP can be used as a new biomarker for the prediction of prognosis and may be a promising immune-related molecular target for LUAD.
RESUMO
BACKGROUND: Lung adenocarcinoma (LUAD) is a common subtype of non-small cell lung cancer with high morbidity and mortality rates and is usually detected at advanced stages because of the early onset of metastasis. Adenosine deaminase RNA-specific 1 (ADAR1) is an RNA editing enzyme that catalyzes the important physiological process of adenosine-to-inosine editing and has been shown to participate in the progression of LUAD. Increasing evidence has suggested that immune infiltration of the tumor immune microenvironment has prognostic value for most human solid organ malignancies; however, much is unknown about the functions of ADAR1. METHODS: The expression of ADAR1 was analyzed in The Cancer Genome Atlas -LUAD database and validated in our LUAD cohort. To assess the prognostic value of ADAR1, Kaplan-Meier survival analyses and Cox regression analyses were carried out in LUAD cohorts. The association between ADAR1 and LUAD immune infiltrates via analyses of cell-type identification by estimating relative subsets of known RNA transcripts. Furthermore, multiplex immunohistochemistry was used to confirm the relationship between ADAR1 expression and immune cells in the present cohort of patients with LUAD. RESULTS: ADAR1 was highly expressed in LUAD tissues and closely correlated with lymph node metastasis (LNM) (p < 0.01), advanced tumor stage (p < 0.05), and poor patient prognosis (p < 0.01), thus indicating that increased ADAR1 contributed to the progression of LUAD. LUAD with high ADAR1 expression can metastasize to lymph nodes that express more ADAR1 than the primary lesion. In addition, M0 macrophages and M2 macrophages increased and CD4+ T cells decreased in LUAD tissues with high ADAR1 expression. And the expression of ADAR1 in lymph node metastases was negatively correlated with the contents of CD4+ T cells (p = 0.0017) and M1 macrophages (p = 0.0037). CONCLUSION: The findings of our study suggested that ADAR1 may be useful in predicting prognosis and LNM in LUAD, and may serve as a promising immune-related molecular target for LUAD patients.