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Primary hyperoxaluria type 1 (PH1) is a childhood-onset autosomal recessive disease, characterized by nephrocalcinosis, multiple recurrent urinary calcium oxalate stones, and a high risk of progressive kidney damage. PH1 is caused by inherent genetic defects of the alanine glyoxylate aminotransferase (AGXT) gene. The in vivo repair of disease-causing genes was exceedingly inefficient before the invention of base editors which can efficiently introduce precisely targeted base alterations without double-strand DNA breaks. Adenine base editor (ABE) can precisely convert A·T to G·C with the assistance of specific guide RNA. Here, we demonstrated that systemic delivery of dual adeno-associated virus encoding a split-ABE8e could artificially repair 13% of the pathogenic allele in AgxtQ84X rats, a model of PH1, alleviating the disease phenotype. Specifically, ABE treatment partially restored the expression of alanine-glyoxylate-aminotransferase (AGT), reduced endogenous oxalate synthesis and alleviated calcium oxalate crystal deposition. Western blot and immunohistochemistry confirmed that ABE8e treatment restored AGT protein expression in hepatocytes. Moreover, the precise editing efficiency in the liver remained stable six months after treatment. Thus, our findings provided a prospect of in vivo base editing as a personalized and precise medicine for PH1 by directly correcting the mutant Agxt gene.
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Hiperoxalúria Primária , Hiperoxalúria , Humanos , Ratos , Animais , Criança , Oxalato de Cálcio , Edição de Genes , RNA Guia de Sistemas CRISPR-Cas , Hiperoxalúria Primária/genética , Hiperoxalúria Primária/terapia , Transaminases/genética , Transaminases/química , Transaminases/metabolismo , Alanina , MutaçãoRESUMO
The COVID-19 caused by SARS-CoV-2 has led to a global pandemic that continues to impact societies and economies worldwide. The main protease (Mpro) plays a crucial role in SARS-CoV-2 replication and is an attractive target for anti-SARS-CoV-2 drug discovery. Herein, we report a series of 3-oxo-1,2,3,4-tetrahydropyrido[1,2-a]pyrazin derivatives as non-peptidomimetic inhibitors targeting SARS-CoV-2 Mpro through structure-based virtual screening and biological evaluation. Further similarity search and structure-activity relationship study led to the identification of compound M56-S2 with the enzymatic IC50 value of 4.0 µM. Moreover, the molecular simulation and predicted ADMET properties, indicated that non-peptidomimetic inhibitor M56-S2 might serve as a useful starting point for the further discovery of highly potent inhibitors targeting SARS-CoV-2 Mpro.
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COVID-19 , Pirazinas , SARS-CoV-2 , Humanos , Antivirais/farmacologia , COVID-19/prevenção & controle , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Inibidores de Proteases/farmacologia , SARS-CoV-2/efeitos dos fármacos , Proteínas não Estruturais Virais , Pirazinas/química , Pirazinas/farmacologia , Tratamento Farmacológico da COVID-19RESUMO
KIAA1429 is an important 'writer' of the N6-methyladenine (m6A) modification, which is involved in tumour progression. This study was conducted to explore the mechanism of action of KIAA1429 in colon adenocarcinoma (COAD). KIAA1429-silenced COAD cell and xenograft tumour models were constructed, and the function of KIAA1429 was explored through a series of in vivo and in vitro assays. The downstream mechanisms of KIAA1429 were explored using transcriptome sequencing. Dimethyloxalylglycine (DMOG), an activator of HIF-1α, was used for feedback verification. The expression of KIAA1429 in COAD tumour tissues and cells was elevated, and KIAA1429 exhibited differential expression at different stages of the tumour. Silencing of KIAA1429 inhibited the proliferation, migration, and invasion of HT29 and HCT116 cells. The expression levels of NLRP3, GSDMD and Caspase-1 were decreased in KIAA1429-silenced HT29 cells, indicating the pyroptotic activity was inhibited. Additionally, KIAA1429 silencing inhibited the growth of tumour xenograft. Transcriptome sequencing and reverse transcription quantitative polymerase chain reaction revealed that after KIAA1429 silencing, the expression of AKR1C1, AKR1C2, AKR1C3 and RDH8 was elevated, and the expression of VIRMA, GINS1, VBP1 and ARF3 was decreased. In HT29 cells, KIAA1429 silencing blocked the HIF-1 signalling pathway, accompanied by the decrease in AKT1 and HIF-1α protein levels. The activation of HIF-1 signalling pathway, mediated by DMOG, reversed the antitumour role of KIAA1429 silencing. KIAA1429 silencing inhibits COAD development by blocking the HIF-1 signalling pathway.
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Adenocarcinoma , Neoplasias do Colo , Metiltransferases , Transdução de Sinais , Humanos , Neoplasias do Colo/metabolismo , Neoplasias do Colo/genética , Neoplasias do Colo/patologia , Animais , Camundongos , Adenocarcinoma/genética , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Metiltransferases/metabolismo , Metiltransferases/genética , Células HT29 , Camundongos Nus , Inativação Gênica , Masculino , Células HCT116 , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Camundongos Endogâmicos BALB C , Feminino , Progressão da Doença , Ensaios Antitumorais Modelo de Xenoenxerto , Regulação Neoplásica da Expressão Gênica , Fator 1 Induzível por Hipóxia/metabolismo , Fator 1 Induzível por Hipóxia/genética , HialuronoglucosaminidaseRESUMO
Epidemiological and clinical data have demonstrated that exposure to cadmium (Cd), a toxic heavy metal, is associated with an increased risk of female infertility. Granulosa cells, the main somatic cells comprising ovarian follicles, are one of the main targets of Cd in the ovaries. However, the mechanism by which Cd induces cytotoxicity in granulosa cells has not been fully elucidated. In this study, we exposed human ovarian granulosa cells (KGN cells) to Cd and conducted in vitro cell experiments and multi-omics (metabolomics and transcriptomics) methods to elucidate these mechanisms. Cd exposure was found to not only induce the apoptosis of the KGN cells but also further reduced mitochondrial function by decreasing mitochondrial membrane potential, ATP production, and respiratory chain complex activity as well as increasing mitochondrial reactive oxygen species (ROS) production. A total of 443 differentially expressed metabolites (160 upregulated and 283 downregulated) and 5200 differentially expressed genes (4634 upregulated and 566 downregulated) were observed in the Cd exposed-cells. The multi-omics data showed that Cd interfered with citric acid cycle (TCA cycle), amino acid (including alanine, glycine, serine, threonine, arginine, and proline) metabolism, and calcium signaling. These findings help to better elucidate the potential toxicity mechanisms of Cd on granulosa cells and the ovary.
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Cádmio , Multiômica , Humanos , Feminino , Cádmio/toxicidade , Cádmio/metabolismo , Células da Granulosa/metabolismo , Folículo Ovariano , Ovário/metabolismo , ApoptoseRESUMO
Strain sensors that can rapidly and efficiently detect strain distribution and magnitude are crucial for structural health monitoring and human-computer interactions. However, traditional electrical and optical strain sensors make access to structural health information challenging because data conversion is required, and they have intricate, delicate designs. Drawing inspiration from the moisture-responsive coloration of beetle wing sheaths, we propose using Ecoflex as a flexible substrate. This substrate is coated with a Fabry-Perot (F-P) optical structure, comprising a "reflective layer/stretchable interference cavity/reflective layer", creating a dynamic color-changing visual strain sensor. Upon the application of external stress, the flexible interference chamber of the sensor stretches and contracts, prompting a blue-shift in the structural reflection curve and displaying varying colors that correlate with the applied strain. The innovative flexible sensor can be attached to complex-shaped components, enabling the visual detection of structural integrity. This biomimetic visual strain sensor holds significant promise for real-time structural health monitoring applications.
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The study aimed to compare the impact of four chemical additives on fermentation characteristics, aerobic stability and in vitro digestibility of total mixed ration (TMR) silage based on soy sauce residue. The TMR (35% soy sauce residue + 45% Napier grass + 20% concentrate) was placed into silos (10 L). The experiment followed the completely random design, treated with different chemical additives: (1) distilled water (control); (2) 0.1% potassium sorbate (SP); (3) 0.1% sodium benzoate (SS); (4) 0.5% calcium propionate (SC); (5) 0.5% sodium diacetate (SD). Total of 100 silos (5 treatments × 4 aerobic exposure days × 5 replicates) were ensiled for 60 days. After exposure to the air, the samples were analyzed for the dynamic change of fermentation parameters at 4, 9 and 15 days, and the data was analyzed as repeated measures. The content of butyric acid and ammonia nitrogen was maintained at a low level. The highest (p < 0.05) lactic acid (LA) content and the lowest (p < 0.05) pH value were measured in SP. At the first 4 days of aerobic exposure, TMR silages treated with four chemical additives were more stable relative to the control, as indicated by the low pH value and yeast counts. Furthermore, the highest (p < 0.05) LA content and the lowest (p < 0.05) pH value indicated that SP performed superior aerobic stability compared with other chemical additives. The SP shows higher (p < 0.05) 72 h cumulative gas production (GP72) and in vitro neutral detergent fiber digestibility (IVNDFD) relative to the control. In conclusion, the SP performed superior in improving fermentation characteristics, aerobic stability and in vitro digestibility of TMR silages based on soy sauce residue.
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We demonstrate the effective establishment of long-range electrostatic interactions among colloidal silica nanospheres through acid treatment, enabling their assembly into colloidal crystals at remarkably low concentrations. This novel method overcomes the conventional limitation in colloidal silica assembly by removing entrapped NH4+ ions and enhancing the electrical double layer (EDL) thickness, offering a time-efficient alternative to increase electrostatic interactions compared with methods like dialysis. The increased EDL thickness facilitates the assembly of SiO2 nanospheres into a body-centered-cubic lattice structure at low particle concentrations, allowing for broad spectrum tunability and high tolerance to particle size polydispersity. Further, we uncover a disorder-order transition during colloidal crystallization at low particle concentrations, with the optimal concentration for crystal formation governed by both thermodynamic and kinetic factors. This work not only provides insights into assembly mechanisms but also paves the way for the design and functionalization of colloidal silica-based photonic crystals in diverse applications.
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Purpose: Endoplasmic reticulum (ER) stress regulates mucus hypersecretion, and may activate downstream factors via TBK1 signaling to induce gene expression. However, it remains unclear whether ER stress promotes airway mucus secretion through the TBK1 pathway. We aimed to investigate the role of the TBK1 pathway in the regulation of MUC5AC expression in a mouse model of house dust mite (HDM)-induced allergic asthma. Materials and Methods: Mice with HDM-induced asthma and human bronchial epithelial BEAS-2B cells were treated with amlexanox, an anti-allergy drug (25 µM), or 4-PBA (10 mM). Tissue and cell samples were collected. Tissue samples were stained with hematoxylin and eosin (H&E) or periodic acid Schiff (PAS) to evaluate pathology. Protein expression was analyzed by western blotting and immunofluorescence. Results: Mice exposed to HDM presented ER stress and hypersecretion of mucus Muc5ac from airway epithelial cells (p < 0.001). Similar results were observed in BEAS-2B cells following exposure to HDM. Both in vivo and in vitro studies revealed that HDM-induced ER stress induced MUC5AC overexpression via TBK1 signaling. Amlexanox and 4-PBA markedly reduced mucus production and weakened the TBK1 signal, which mediates MUC5AC hypersecretion. Conclusion: TBK1 plays a pivotal role in HDM-induced ER stress, leading to overproduction of MUC5AC in the asthmatic airway epithelium. The overproduction of MUC5AC can be significantly decreased by inhibiting TBK1 or ER stress using 4-PBA. These findings highlight potential target-specific therapies for patients with chronic allergic asthma.
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Asma , Pyroglyphidae , Humanos , Camundongos , Animais , Pyroglyphidae/metabolismo , Asma/metabolismo , Estresse do Retículo Endoplasmático , Epitélio/metabolismo , Proteínas Serina-Treonina Quinases , Mucina-5AC/genética , Mucina-5AC/metabolismoRESUMO
To evaluate the feasibility and accuracy of AR-assisted pedicle screw placement using a new intraoperative rapid registration method of combining preoperative CT scanning and intraoperative C-arm 2D fluoroscopy in cadavers. Five cadavers with intact thoracolumbar spines were employed in this study. Intraoperative registration was performed using anteroposterior and lateral views of preoperative CT scanning and intraoperative 2D fluoroscopic images. Patient-specific targeting guides were used for pedicle screw placement from Th1-L5, totaling 166 screws. Instrumentation for each side was randomized (augmented reality surgical navigation (ARSN) vs. C-arm) with an equal distribution of 83 screws in each group. CT was performed to evaluate the accuracy of both techniques by assessing the screw positions and the deviations between the inserted screws and planned trajectories. Postoperative CT showed that 98.80% (82/83) screws in ARSN group and 72.29% (60/83) screws in C-arm group were within the 2-mm safe zone (p < 0.001). The mean time for instrumentation per level in ARSN group was significantly shorter than that in C-arm group (56.17 ± 3.33 s vs. 99.22 ± 9.03 s, p < 0.001). The overall intraoperative registration time was 17.2 ± 3.5 s per segment. AR-based navigation technology can provide surgeons with accurate guidance of pedicle screw insertion and save the operation time by using the intraoperative rapid registration method of combining preoperative CT scanning and intraoperative C-arm 2D fluoroscopy.
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Realidade Aumentada , Parafusos Pediculares , Cirurgia Assistida por Computador , Humanos , Cadáver , Fluoroscopia/métodos , Vértebras Lombares/diagnóstico por imagem , Vértebras Lombares/cirurgia , Cirurgia Assistida por Computador/métodos , Sistemas de Navegação CirúrgicaRESUMO
The gait acquisition system can be used for gait analysis. The traditional wearable gait acquisition system will lead to large errors in gait parameters due to different wearing positions of sensors. The gait acquisition system based on marker method is expensive and needs to be used by combining with the force measurement system under the guidance of rehabilitation doctors. Due to the complex operation, it is inconvenient for clinical application. In this paper, a gait signal acquisition system that combines foot pressure detection and Azure Kinect system is designed. Fifteen subjects are organized to participate in gait test, and relevant data are collected. The calculation method of gait spatiotemporal parameters and joint angle parameters is proposed, and the consistency analysis and error analysis of the gait parameters of proposed system and camera marking method are carried out. The results show that the parameters obtained by the two systems have good consistency (Pearson correlation coefficient r ≥ 0.9, P < 0.05) and have small error (root mean square error of gait parameters is less than 0.1, root mean square error of joint angle parameters is less than 6). In conclusion, the gait acquisition system and its parameter extraction method proposed in this paper can provide reliable data acquisition results as a theoretical basis for gait feature analysis in clinical medicine.
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Marcha , Extremidade Inferior , Humanos , Fenômenos Biomecânicos , Pé , Análise da Marcha , Reprodutibilidade dos TestesRESUMO
Influenza A viruses cause severe respiratory illnesses in humans and animals. Overreaction of the innate immune response to influenza virus infection results in hypercytokinemia, which is responsible for mortality and morbidity. However, the mechanism by which influenza induces hypercytokinemia is not fully understood. In this study, we established a mouse-adapted H9N2 virus, MA01, to evaluate the innate immune response to influenza in the lung. MA01 infection caused high levels of cytokine release, enhanced pulmonary injury in mice, and upregulated CD83 protein in dendritic cells and macrophages in the lung. Influenza virus neuraminidase (NA) unmasked CD83 protein and contributed to high cytokine levels. Furthermore, we provide evidence that CD83 is a sialylated glycoprotein. Neuraminidase treatment enhanced lipopolysaccharide (LPS)-stimulated NF-κB activation in RAW264.7 cells. Anti-CD83 treatment alleviated influenza virus-induced lung injury in mice. Our study indicates that influenza virus neuraminidase modulates CD83 status and contributes to the "cytokine storm," which may suggest a new approach to curb this immune injury.IMPORTANCE The massive release of circulating mediators of inflammation is responsible for lung injury during influenza A virus infection. This phenomenon is referred to as the "cytokine storm." However, the mechanism by which influenza induces the cytokine storm is not fully understood. In this study, we have shown that neuraminidase unmasked CD83 protein in the lung and contributed to high cytokine levels. Anti-CD83 treatment could diminish immune damage to lung tissue. The NA-CD83 axis may represent a target for an interruption of influenza-induced lung damage.
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Antígenos CD/metabolismo , Interações Hospedeiro-Patógeno/imunologia , Imunidade Inata/imunologia , Imunoglobulinas/metabolismo , Vírus da Influenza A Subtipo H9N2/patogenicidade , Lesão Pulmonar/etiologia , Glicoproteínas de Membrana/metabolismo , Neuraminidase/metabolismo , Infecções por Orthomyxoviridae/complicações , Proteínas Virais/metabolismo , Animais , Antígenos CD/genética , Células Dendríticas/imunologia , Células Dendríticas/virologia , Feminino , Imunoglobulinas/genética , Vírus da Influenza A Subtipo H9N2/enzimologia , Lesão Pulmonar/patologia , Macrófagos/imunologia , Macrófagos/virologia , Glicoproteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Neuraminidase/genética , Infecções por Orthomyxoviridae/virologia , Transdução de Sinais , Proteínas Virais/genética , Virulência , Antígeno CD83RESUMO
BACKGROUND: Alteration of commensal microbiota is highly correlated with the prevalence of allergic reactions to food in the gastrointestinal tract. The mechanisms by which microbiota modulate food allergen sensitization in the mucosal site are not fully understood. METHODS: We generate DCs specific knockout of retinoic acid receptor α (Rara) gene mice (DC KO Rara) to evaluate food sensitization. The bile acid-activated retinoic acid response was evaluated by flow cytometry, real-time RT-PCR and Illumina transcriptome sequencing. The global effect of Abx treatment on BA profiles in the mucosal lymph tissue mLN in mice was examined by UPLC-MS analysis. RESULTS: In this study, we demonstrate that depletion of commensal gut bacteria leads to enhanced retinoic acid (RA) signaling in mucosal dendritic cells (DCs). RA signaling in DCs is required for the production of food allergen-specific IgE and IgG1. Antibiotics induced an enlarged bile acid (BA) pool, and dysregulated BA profiles contributed to enhanced RA signaling in mucosal DCs. BA-activated RA signaling promoted DC upregulation of interferon I signature, RA signature, OX40L, and PDL2, which may lead to T helper 2 differentiation of CD4+ T cells. BA-activated RA signaling involved the farnesoid X receptor and RA receptor α (RARa) interaction. Depletion of bile acid reduces food allergen specific IgE and IgG1 levels in mice. CONCLUSION: Our research unveils a mechanism of food sensitization modulated by BA-RA signaling in DCs, which suggests a potential new approach for the intervention of food allergic reactions.
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Hipersensibilidade Alimentar , Tretinoína , Alérgenos/farmacologia , Animais , Ácidos e Sais Biliares/farmacologia , Cromatografia Líquida , Células Dendríticas , Humanos , Imunoglobulina E , Imunoglobulina G , Camundongos , Espectrometria de Massas em Tandem , Tretinoína/farmacologiaRESUMO
PURPOSE: To compare the clinical characteristics of pediatric urolithiasis patients with positive and negative molecular diagnoses. METHODS: The clinical characteristics corresponding to pediatric urolithiasis patients that had undergone exome sequencing at our hospital between January 2016 and May 2021 were collected. Genetic analysis results were used to separate patients into positive and negative molecular diagnosis groups. Multivariate logistic regression analyses adjusted for visiting age, sex, ethnicity, province, and body mass index were used to compare differences in medical history, diagnostic imaging findings, and renal function between individuals with and without molecular diagnoses. RESULTS: In total, 194 patients with pediatric urolithiasis of unknown etiology underwent exome sequencing and were included in the present study, of whom 63 obtained urolithiasis-related molecular diagnoses. Relative to cases without a molecular diagnosis, those with a positive molecular diagnosis were more likely to be associated with a positive family history (OR 2.84, 95% CI 1.29-6.29, p = 0.008), consanguineous parents (OR 24.7, 95% CI 1.34-454, p = 0.002), early onset (OR 1.26, 95% CI 1.09-1.45, p < 0.001), nephrocalcinosis (OR 10.6, 95% CI 3.06-36.6, p < 0.001), cast stone (OR 18.9, 95% CI 4.40-81.1, p < 0.001), multiple stones (OR 13.9, 95% CI 6.39-30.2, p < 0.001), bilateral stones (OR 7.04, 95% CI 3.47-14.2, p < 0.001), a lower estimated glomerular filtration rate (OR 1.17, 95% CI 1.07-1.28, p < 0.001), and chronic kidney disease (OR 26.9, 95% CI 1.42-526, p < 0.001). CONCLUSION: A positive family history, consanguineous parents, early onset, nephrocalcinosis, severe stone burden, and impaired renal function are signals of concern that are suggestive of inherited urolithiasis.
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Nefrocalcinose , Insuficiência Renal Crônica , Urolitíase , Criança , Feminino , Humanos , Masculino , Estudos Retrospectivos , Urolitíase/diagnóstico , Urolitíase/genéticaRESUMO
Cystinuria is a genetic disorder of cystine transport, including defective protein b0,+AT (encoded by SLC7A9), and/or rBAT (encoded by SLC3A1). Patients present hyperexcretion of cystine in the urine, recurrent cystine lithiasis, and progressive decline in kidney function. Moreover, heterodimer transport is defective. To date, little omics data are accessible regarding this metabolic disease caused by membrane proteins. Since membrane function is closely related to changes in the lipidome, we decided to explore the changes in kidney tissue of a self-established cystinuria rat model by performing lipidomic analysis by LC-MS/MS. Our results demonstrated that Slc7a9 deficiency changed the lipid profile of the renal cortex and induced vital modifications in the lipidome, including major alterations in ChE, LPA, and PA. Among those alterations, this lipidomic study highlights the lipid changes that participate in inflammatory responses during cystinuria. As a result, lipid research, perhaps has great potential, for it may lead to the identification of novel therapeutic targets for the prevention and treatment of cystinuria.
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Cistinúria , Sistemas de Transporte de Aminoácidos Básicos/genética , Sistemas de Transporte de Aminoácidos Básicos/metabolismo , Animais , Cromatografia Líquida , Cistina/metabolismo , Cistinúria/genética , Cistinúria/metabolismo , Rim/metabolismo , Metabolismo dos Lipídeos , Lipidômica , Lipídeos , Ratos , Espectrometria de Massas em TandemRESUMO
The available findings on the association between iron status and risk of stroke remain controversial. We used multivariable logistic regression and restricted cubic spline models to explore the association between iron exposures and risk of stroke in the US National Health and Nutrition Examination Survey (NHANES 2007-2016, n = 24,627). A total of 941 (3.82%) stroke cases were identified in this study. In women, the ORs with 95% CIs of prevalence of stroke were 0.92 (0.65-1.28), 0.66 (0.44-0.98) and 0.72 (0.49-1.08) across quartiles 2-4 compared with quartile 1 of iron intake, respectively. An inverse and L-shaped association between iron intake and risk of stroke in women was observed, and the curve plateaued at 20 mg/day. However, neither serum iron concentrations nor iron intake were significantly associated with riskof stroke in men. Our study found that iron intake was inversely associated with risk of stroke in a sex-dependent fashion.
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Ferro , Acidente Vascular Cerebral , Adulto , Estudos Transversais , Dieta , Feminino , Humanos , Masculino , Inquéritos Nutricionais , Acidente Vascular Cerebral/epidemiologia , Estados Unidos/epidemiologiaRESUMO
BACKGROUND: Half of all apple production worldwide comes from China. However, the geographic authentication of Chinese apples has not been well studied. We highlight the multi-element-based geographical discrimination of apples from the southwest cold highlands (SCH) of China. 565 samples from the SCH (138) and others (427) were obtained, and the content of fifteen elements were applied to construct models for discrimination. RESULTS: The SCH apples from 2017 to 2019 had higher concentrations of Mn, Zn, Cr, Cd, Se, Pb, and Fe, but lower concentrations of Na, B, Ni, and P. With sufficient training, linear discriminant analysis (LDA) discriminated the SCH, and the testing accuracy averaged 92.5% and 92.2%. Nonlinear discrimination models were more suitable than the linear models. Optimized random forest analysis was the model with the best fit, and with averaged training and testing it obtained a level of accuracy of 98.2% and 98.5%. CONCLUSION: The multielement-based discrimination of SCH apples could aid further studies of geographical origins. © 2021 Society of Chemical Industry.
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Frutas/química , Malus/química , Oligoelementos/análise , China , Análise Discriminante , Frutas/classificação , Malus/classificaçãoRESUMO
Gastric carcinoma (GC) is a malignant tumor that has high mortality and morbidity worldwide. Although many efforts have been focused on the development and progression of GC, the underlying functional regulatory mechanism of GC needs more clarification. Metallothionein 1G (MT1G) is a member of the metallothionein family (MTs), and hypermethylation of MT1G occurred in a variety of cancers, including gastric cancer. However, the functional mechanism of MT1G in GC remains unclear. Here, we demonstrated that MT1G was down-regulated in GC tissues and cells. Overexpression of MT1G inhibited cell proliferation, foci formation and cell invasion, while knockdown of MT1G increased cell proliferation, foci formation and cell invasion. In addition, MT1G overexpression inhibited cell cycle progression and MT1G deficiency exerted opposite phenotype. p-AKT was negatively regulated by MT1G. In summary, our study reveals that MT1G exerts crucial role in regulating of cell proliferation and migration of gastric cancer, providing new insights for MT1G-related pathogenesis and a basis for developing new strategies for treatment of GC.
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The work aimed to investigate the effects of four organic acid salts on fermentation quality, aerobic stability, and in vitro rumen digestibility of total mixed ration (TMR) silage prepared with citric acid residue, wet brewers' grains, and Napier grass. The TMR was ensiled with the following: (1) no additives (control), (2) 0.1% sodium benzoate (SB), (3) 0.1% potassium sorbate (PS), (4) 0.5% sodium diacetate (SDA), (5) 0.5% calcium propionate (CAP) on a fresh weight basis. All silos (10 L) were opened after 60 days of ensiling to determine fermentation profiles and in vitro rumen digestibility, and then were subjected to a 9-day aerobic stability test. Four organic acid salts significantly (p < 0.05) increased dry matter contents, lactic acid bacteria count, and decreased ethanol content and yeast count compared with the control. The SDA and CAP significantly (p < 0.05) increased water-soluble carbohydrates, lactic acid, and crude protein contents, and decreased pH, ammonia nitrogen, neutral detergent fiber, and hemicellulose contents compared with other TMR silages after 60 days of ensiling. Organic acid salts significantly (p < 0.05) prolonged the hours of aerobic stability and significantly (p < 0.05) increased cumulative gas production and potential gas production compared with the control. The treatments of SDA and CAP significantly (p < 0.05) improved aerobic stability as indicated by higher (p < 0.05) lactic acid and water-soluble carbohydrates contents, and lower (p < 0.05) pH, ammonia nitrogen, ethanol contents, and yeast count compared with the control. The treatments of SDA and CAP significantly (p < 0.05) increased in vitro rumen parameters, as indicated by higher (p < 0.05) in vitro digestibility of dry matter, crude protein, and neutral detergent fiber after 60 days of ensiling. Overall, these results indicated that the addition of SDA and CAP could ensure the good fermentation quality and improve aerobic stability of TMR silages. By comprehensive consideration, CAP was recommended for improving fermentation quality, aerobic stability, and in vitro rumen digestibility of TMR silages prepared with wet brewers' grains, citric acid residue, and Napier grass.
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Rúmen , Silagem , Aerobiose , Amônia/metabolismo , Animais , Carboidratos , Ácido Cítrico , Detergentes/metabolismo , Fibras na Dieta/metabolismo , Etanol/metabolismo , Fermentação , Ácido Láctico/metabolismo , Nitrogênio/metabolismo , Rúmen/metabolismo , Saccharomyces cerevisiae , Sais , Silagem/análise , ÁguaRESUMO
Primary hyperoxaluria type 1 (PH1) is a severe inherited disorder caused by a genetic defect in alanine-glyoxylate aminotransferase (AGXT), which results in recurrent urolithiasis and renal failure. Animal models that precisely reflect human PH1 phenotypes are lacking. We aimed to develop a novel PH1 rat model and study the mechanisms involved in PH1 deterioration. One cell stage Sprague-Dawley embryos were injected with the CRISPR/Cas9 system to introduce a Q84X mutation in Agxt. Liver tissues were harvested to determine Agxt expression. Urine oxalate, crystals, and electrolyte levels in AgxtQ84X and wild-type (WT) littermates were evaluated. Kidney tissues were used for Pizzolato staining and kidney injury evaluation. Data showed that Agxt mRNA and protein were absent in AgxtQ84X rats. At 4 and 24 wk, AgxtQ84X rats displayed 2.1- and 2.9-fold higher urinary oxalate levels, respectively, compared with WT littermates. As a result, calcium oxalate (CaOx) crystals in urine were revealed in all AgxtQ84X rats but in none of the WT rats. We also observed bladder stones in 36.4% of AgxtQ84X rats, of which 44.4% had renal CaOx deposition. Moreover, the elevated serum urea and creatinine levels indicated the impaired renal function in AgxtQ84X rats. Further investigation revealed significantly increased expression of inflammation-, necroptosis-, and fibrosis-related genes in the kidneys of AgxtQ84X rats with spontaneous renal CaOx deposition, indicating that these pathways are involved in PH1 deterioration. Collectively, these results suggest that this rat model has broad applicability in mechanistic studies and innovative therapeutics development for PH1 and other kidney stone diseases.NEW & NOTEWORTHY Primary hyperoxaluria type 1 is a severe inherited disorder that results in recurrent urolithiasis and renal failure. We generated an alanine-glyoxylate aminotransferase (Agxt)Q84X nonsense mutant rat model that displayed an early onset of hyperoxaluria, spontaneous renal CaOx precipitation, bladder stone, and kidney injuries. Our results suggest an interaction of renal CaOx crystals with the activation of inflammation-, fibrosis-, and necroptosis-related pathways. In all, the AgxtQ84X rat strain has broad applicability in mechanistic studies and the development of innovative therapeutics.
Assuntos
Hiperoxalúria/metabolismo , Rim/metabolismo , Nefrocalcinose/metabolismo , Transaminases/genética , Animais , Oxalato de Cálcio/metabolismo , Hiperoxalúria/genética , Cálculos Renais/sangue , Mutação/genética , Nefrocalcinose/genética , Oxalatos/metabolismo , Ratos , Insuficiência Renal/genética , Transaminases/metabolismoRESUMO
PURPOSE: To investigate the prevalence of inherited causes in an early onset urolithiasis cohort and each metabolic subgroup. METHODS: A retrospective analysis of both metabolic and genomic data was performed for the first 105 pediatric urolithiasis patients who underwent exome sequencing at our hospital from February 2016 to October 2018. Measurements included the diagnostic yield of exome sequencing in the entire cohort and each metabolic subgroup (hyperoxaluria, hypocitraturia, hypercalciuria, hyperuricosuria and cystine stone subgroups). The conformity between molecular diagnoses and metabolic evaluation was also evaluated. RESULTS: The present study involved a cohort of 105 pediatric patients with urolithiasis, from which diagnostic variants were identified in 38 patients (36%), including 27 primary hyperoxaluria and 11 cystinuria. In the metabolic subgroup analyses, 41% hyperoxaluria cases were primary hyperoxaluria caused by monogenic defects, and 100% of the causes of cystine stones could be explained by monogenic defects. However, no appropriate inherited causes were identified for hypocitraturia, hypercalciuria, or hyperuricosuria in the cohort. A high conformity (100%) was obtained between the molecular diagnoses and metabolic evaluation. CONCLUSION: Exome sequencing in a cohort of 105 pediatric patients with urolithiasis yielded a genetic diagnosis in 36% of cases and the molecular diagnostic yield varies substantially across different metabolic abnormalities.