RESUMO
BACKGROUND: Epithelial-to-mesenchymal transition (EMT) is a critical event contributing to more aggressive phenotypes in cancer cells. EMT is frequently activated in radiation-targeted cells during the course of radiotherapy, which often endows cancers with acquired radioresistance. However, the upstream molecules driving the signaling pathways of radiation-induced EMT have not been fully delineated. METHODS: In this study, RNA-seq-based transcriptome analysis was performed to identify the early responsive genes of HeLa cells to γ-ray irradiation. EMT-associated genes were knocked down by siRNA technology or overexpressed in HeLa cells and A549 cells, and the resulting changes in phenotypes of EMT and radiosensitivity were assessed using qPCR and Western blotting analyses, migration assays, colony-forming ability and apoptosis of flow cytometer assays. RESULTS: Through RNA-seq-based transcriptome analysis, we found that LPAR5 is downregulated in the early response of HeLa cells to γ-ray irradiation. Radiation-induced alterations in LPAR5 expression were further revealed to be a bidirectional dynamic process in HeLa and A549 cells, i.e., the early downregulating phase at 2 ~ 4 h and the late upregulating phase at 24 h post-irradiation. Overexpression of LPAR5 prompts EMT programing and migration of cancer cells. Moreover, increased expression of LPAR5 is significantly associated with IR-induced EMT and confers radioresistance to cancer cells. Knockdown of LPAR5 suppressed IR-induced EMT by attenuating the activation of ERK signaling and downstream Snail, MMP1, and MMP9 expression. CONCLUSIONS: LPAR5 is an important upstream regulator of IR-induced EMT that modulates the ERK/Snail pathway. This study provides further insights into understanding the mechanism of radiation-induced EMT and identifies promising targets for improving the effectiveness of cancer radiation therapy.
Assuntos
Metaloproteinase 1 da Matriz , Neoplasias , Linhagem Celular Tumoral , Transição Epitelial-Mesenquimal/genética , Células HeLa , Humanos , Metaloproteinase 9 da Matriz , RNA Interferente Pequeno , Receptores de Ácidos LisofosfatídicosRESUMO
BACKGROUND: Preoperative prediction of microvascular invasion (MVI) is critical for treatment strategy making in patients with hepatocellular carcinoma (HCC). We aimed to develop a deep learning (DL) model based on preoperative dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) to predict the MVI status and clinical outcomes in patients with HCC. METHODS: We retrospectively included a total of 321 HCC patients with pathologically confirmed MVI status. Preoperative DCE-MRI of these patients were collected, annotated, and further analyzed by DL in this study. A predictive model for MVI integrating DL-predicted MVI status (DL-MVI) and clinical parameters was constructed with multivariate logistic regression. RESULTS: Of 321 HCC patients, 136 patients were pathologically MVI absent and 185 patients were MVI present. Recurrence-free survival (RFS) and overall survival (OS) were significantly different between the DL-predicted MVI-absent and MVI-present. Among all clinical variables, only DL-predicted MVI status and a-fetoprotein (AFP) were independently associated with MVI: DL-MVI (odds ratio [OR] = 35.738; 95% confidence interval [CI] 14.027-91.056; p < 0.001), AFP (OR = 4.634, 95% CI 2.576-8.336; p < 0.001). To predict the presence of MVI, DL-MVI combined with AFP achieved an area under the curve (AUC) of 0.824. CONCLUSIONS: Our predictive model combining DL-MVI and AFP achieved good performance for predicting MVI and clinical outcomes in patients with HCC.
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Carcinoma Hepatocelular , Aprendizado Profundo , Neoplasias Hepáticas , Carcinoma Hepatocelular/diagnóstico por imagem , Carcinoma Hepatocelular/cirurgia , Humanos , Neoplasias Hepáticas/diagnóstico por imagem , Neoplasias Hepáticas/cirurgia , Imageamento por Ressonância Magnética/métodos , Microvasos/diagnóstico por imagem , Microvasos/patologia , Invasividade Neoplásica/patologia , Estudos Retrospectivos , alfa-FetoproteínasRESUMO
This study aimed to observe the inhibitory effect of icariin against oxidative stress-induced calcification in aortic vascular smooth muscle cells(VSMCs) and elucidate the molecular mechanism of icariin in inhibiting endoplasmic reticulum stress(ERS)-mediated atherosclerotic calcification, so as to provide new ideas for exploring the anti-atherosclerotic mechanism of Epimedii Folium. The VSMCs in rat thoracic aorta were subjected to adherent culture and then treated with the complete calcification DMEM containing high glucose and hydrogen peroxide(H_2O_2) for three weeks. The resulting calcified VSMCs were divided into different treatment groups. Icariin was added one week after calcification induction for protecting the VSMCs, whose viability was then detected using cell counting kit-8(CCK-8). Alizarin red-S staining was conducted to observe the calcification degree. The activity of alkaline phosphatase(ALP) in VSMCs was measured using the disodium phenyl phosphate substrate and the calcium content was measured by arsenazo â ¢ method. The mRNA expression levels of ossification-related factors including osteocalcin(OC), osteopontin(OPN), Runt-related transcription factor 2(Runx2), and type â collagen(Col â a) were detected by real-time PCR. Western blot was carried out to determine the protein expression levels of α-smooth muscle actin(α-SMA), Runx2, activating transcription factor 4(ATF4), and eukaryotic translation initiation factor(eIF)-2α. The results showed that H_2O_2 significantly induced the calcification of VSMCs, increased the ALP activity and calcium content in VSMCs, promoted OC, OPN, Runx2, and Col â a mRNA expression and Runx2 protein expression, and reduced α-SMA protein expression. The ATF4 protein expression and eIF2α phosphorylation were also elevated significantly. Icariin reversed the calcification of VSMCs induced by H_2O_2, inhibited ALP activity and calcium content in VSMCs, down-regulated the mRNA expression levels of OC, OPN, Runx2 and Col â a and Runx2 protein expression, and relatively up-regulated the expression of α-SMA. The expression of ATF4 and phosphorylation of eIF2α also declined significantly. All these have demonstrated that icariin inhibited VSMCs calcification by down-regulating the ossification-related factors and lowering ALP activity and calcium content in VSMCs. Besides, the down-regulation of Runx2 expression and the inhibition of ATF4 and eIF2α-mediated cellular calcification pathway in ERS might also be involved in such calcification-suppressing process.
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Músculo Liso Vascular , Miócitos de Músculo Liso , Animais , Células Cultivadas , Flavonoides/metabolismo , Flavonoides/farmacologia , Músculo Liso Vascular/metabolismo , Estresse Oxidativo , RatosRESUMO
Ionic liquids (ILs) as green alternatives for volatile organic solvents are increasingly used in commercial applications. It is necessary to explore the cytotoxic mechanism of ILs to reduce the risk to human health. For this purpose, cell viability, apoptosis, cytochrome P450 3A4 (CYP3A4), glucose transporter type 2 (GLUT2), and microRNA-122 (miR-122) gene expression in HepG2 cells was evaluated after IL exposure. The results showed that ILs reduced the viability of HepG2 cells through apoptotic cell death. Moreover, ILs markedly upregulated the transcription and protein levels of CYP3A4, but did not affect the expression of GLUT2 in either messenger RNA level or protein level. Finally, ILs increased the expression of miR-122 and inhibition of miR-122 with miR-122 inhibitor blocked ILs-induced apoptosis in HepG2 cells. This finding may contribute to an increased understanding of the in vitro molecular toxicity mechanism of ILs to further understand IL-related human health risks.
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Apoptose/efeitos dos fármacos , Brometos/farmacologia , Citocromo P-450 CYP3A/metabolismo , Imidazóis/farmacologia , MicroRNAs/metabolismo , Transportador de Glucose Tipo 2/metabolismo , Células Hep G2 , HumanosRESUMO
This study investigated the role of long non-coding RNAs (lncRNAs) in the development of the palatal tissues. Cleft palates in mice were induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Expression levels of long non-coding RNA H19 (lncRNA H19) and insulin-like growth factor 2 (IGF2) gene were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The rate of occurrence of cleft palate was found to be 100% by TCDD exposure, and TCDD could cause short upper limb, cerebral fissure, webbed neck, and short neck. The expression levels of lncRNA H19 and IGF2 gene specifically showed embryo age-related differences on E13, E14, and E15 in the palatal tissues. The expression levels of lncRNA H19 and IGF2 gene showed an inverse relationship on E13, E14, and E15. These findings demonstrated that lncRNA H19 and IGF2 can mediate the development of mouse cleft palate.
Assuntos
Fissura Palatina/induzido quimicamente , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Dibenzodioxinas Policloradas/toxicidade , RNA Longo não Codificante/genética , Animais , Fissura Palatina/genética , Fissura Palatina/patologia , Feminino , Regulação da Expressão Gênica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Palato/metabolismo , Reação em Cadeia da Polimerase em Tempo RealRESUMO
In order to reveal the properties of polar metabolome in inflammatory cells, we selected LPS-induced RAW264.7 inflammatory cell models as the carrier for the research of metabolic fingerprint analysis. In this study, an ultra performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS)-based metabolomics protocol was optimized for the extraction of polar metabolites from RAW264.7 cell line. Then orthogonal partial least squares discriminant analysis (OPLS-DA) was used to process the metabolic data, and finally, a total of 17 metabolites were selected and identified. The results showed that MeOH-CHCl3-H2O (8â¶1â¶1) was chosen as the optimal extraction solvent to achieve higher number of chromatographic peaks, with the best relative extraction efficiency and stability. Comparing with the normal cells, the inflammatory cells presented an abnormal metabolism in protein, carbohydrate, nucleotide and phospholipids. In this study, a UPLC-Q-TOF/MS-based metabolomics protocol for the polar metabolites from RAW264.7 cell line was developed, which may provide important information for the study of mechanism of inflammation and the anti-inflammatory drugs.
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Macrófagos/metabolismo , Metabolômica , Animais , Cromatografia Líquida de Alta Pressão , Espectrometria de Massas , Camundongos , Células RAW 264.7RESUMO
To systematically evaluate the safety of Kudiezi injection. Databases such as Cochrane library, Medline, EMbase, Web of Science, Clinical Trials, CBM, CNKI, VIP, Wanfang and Chinese Clinical Trial Register were searched to collect the literature on all the study types of Kudiezi injection. Two researchers screened literature, assessed quality and extracted data according to inclusion and exclusion criteria. All studies were assessed by using internationally recognized methodological quality assessment tools or reporting quality evaluation criteria; Meta-analysis of adverse drug reaction/adverse events (ADR/AE) of Kudiezi injection was performed by using Stata 12.0 software. There were 411 clinical studies included, out of which 315 studies were analyzed finally. 18 072 patients in total used kudiezi injection, and there were 330 cases with ADRs and 13 cases with AEs. The most common ADR related system was the central and peripheral nervous system, with a weighted incidence of 2.9% [95%CI(0.022, 0.036)]. From the current evidence, the overall safety of Kudiezi injection was acceptable. Although data could be collected from all kinds of published reports, there are lack of mechanism experiments or observational studies with large samples of Kudiezi injection. Therefore, it is necessary to carry out further research on the safety of Kudiezi injection. Meanwhile, off label use of Kudiezi injection is common, so it is urgent for relevant governmental departments to formulate drug use specifications and provide better guidance for clinical drug use.
Assuntos
Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Medicamentos de Ervas Chinesas/efeitos adversos , Humanos , Incidência , InjeçõesRESUMO
Sufentanil has been used broadly in cardiac surgery, but the mechanisms by which it modulates coronary vascular tone after ischemia-reperfusion injury are largely unknown. Effects of sufentanil on coronary tone and on the relaxation of rat coronary arteries (CAs) in response to endothelium-dependent (acetylcholine) and endothelium-independent (sodium nitroprusside) relaxing agents in the presence of hypoxia-reoxygenation (H/R) was studied in an in vitro organ chamber setup. Sufentanil (10-7-10-4 mol/L) relaxed rat CA rings in endothelium-dependent and endothelium-independent manners. In endothelium-intact rings, preincubation of H/R-treated CAs with sufentanil (10-5 mol/L) significantly increased the acetylcholine response, but did not augment sodium nitroprusside-induced relaxation. Sufentanil-mediated potentiation of acetylcholine-induced relaxation was not affected by a nitric oxide synthase inhibitor or by intermediate- or small-conductance Ca2+-activated K+ channel blockers. However, potentiation was abolished by iberiotoxin (100 nmol/L), a selective inhibitor of large-conductance Ca2+-activated K+ channels, as well as Rp-cAMPS (30 µmol/L), a cyclic AMP-dependent protein kinase (PKA) inhibitor. Sufentanil induced endothelium-dependent and endothelium-independent relaxation and attenuated H/R-induced impairment of endothelium-dependent vasodilation in the rat CAs. The potentiating effect of sufentanil may involve activation of large-conductance Ca2+-activated K+ channels via cAMP-dependent mechanisms.
Assuntos
Vasos Coronários/efeitos dos fármacos , Endotélio Vascular/efeitos dos fármacos , Sufentanil/farmacologia , Vasodilatação/efeitos dos fármacos , Animais , Hipóxia Celular/efeitos dos fármacos , Hipóxia Celular/fisiologia , Vasos Coronários/fisiologia , Relação Dose-Resposta a Droga , Endotélio Vascular/fisiologia , Canais de Potássio Ativados por Cálcio de Condutância Alta/fisiologia , Masculino , Técnicas de Cultura de Órgãos , Ratos , Ratos Sprague-Dawley , Vasodilatação/fisiologiaRESUMO
To investigate the effects and mechanisms of total flavones of Epimedium (TFE) on oxidative stress induced by myocardial ischemia/reperfusion injury in rats, forty male SD rats were randomly divided into sham operated group, model group, diltiazem group and flavonoids of Epimedium low and high doses groups with 8 rats in each. Myocardial ischemia/reperfusion injury model was induced by ligaturing the left anterior descending artery for 30 min followed reperfusion for 4 h after TFE was taken by intragastric administration for 4 days. The degree of myocardial infarct was observed by N-BT staining. The concentrations of MDA and activities of SOD and T-AOC in cardiac tissue were measured by colorimetry. Serum TnI concentrations were checked by ELISA. HE stain was used to observe myocardium structure under light microscope. Expressions of SIRT1 and Nrf2 in cardiac tissue were evaluated by immunohistochemistry method and Western blot, respectively. Compared with the model group, the degree of myocardial infarct, MDA concentration in cardiac tissue and the levels of TnI in serum significantly decreased in the diltiazem group and flavonoids of Epimedium low and high doses groups (P<0.05 or P<0.01); flavonoids of Epimedium low and high doses groups and the diltiazem group also showed improvements in myocardium structure under ischemia/reperfusion injury. TFE significantly increased the activity of SOD and T-AOC and the expression of SIRT1 and Nrf2 in cardiac tissue when compared with the model group (P<0.05 or P<0.01). Therefore, TFE can increase anti-peroxidant capacity of myocardium tissue by using intrinsically anti-oxidant signaling pathway of SIRT1 and Nrf2, which can inhibit irreversible damage of cardiomyocytes in myocardial ischemia/reperfusion injury and protect normal function of cardiac tissue.
Assuntos
Epimedium/química , Flavonas/farmacologia , Traumatismo por Reperfusão Miocárdica/tratamento farmacológico , Estresse Oxidativo , Animais , Masculino , Miocárdio , Fator 2 Relacionado a NF-E2/metabolismo , Ratos , Ratos Sprague-Dawley , Sirtuína 1/metabolismo , Superóxido Dismutase/metabolismoRESUMO
An in vitro anti-thrombin bioassay was developed to investigate the chemical constituents which have anti-thrombin effect from the water soluble components of Salvia miltiorrhiza. Using Chromozym TH as a probe combined with ethyl acetate Semi-micro extraction was applied to measure p-nitroaniline by HPLC. According to the results, the inactivationrate of thrombin by sodium danshensu, salvianolic acid A and salvianolic acid B under a given set of conditions were 3.06%, 77.77% and 2.35%, respectively. In the water-soluble components, salvianolic acid A has a direct inhibition of thrombin, while sodium danshensu and salvianolic acid B have no significant effect on thrombin. The method is sensitive and low consumption. It can eliminate the interference absorbed for the sample itself which can be used for screening single or multiple direct antithrombin active ingredient of herbal extract.
Assuntos
Medicamentos de Ervas Chinesas/química , Fibrinolíticos/química , Salvia miltiorrhiza/química , Compostos de Anilina/química , Benzofuranos/química , Ácidos Cafeicos/química , Cromatografia Líquida de Alta Pressão , Lactatos/química , TrombinaRESUMO
Armand clematis stem (Clematidis Armandii Caulis, Chuanmutong) is a widely used Chinese herb to disinhibit urine and relieve stranguria. It is difficult to be identified owing to its various macroscopic feature and unknown characteristic compounds. Thus, total of 24 Chuanmutong samples and 7 related herbs including four manshurian aristolochia stem (Aristolochiae Manshuriensis Caulis, Guanmutong) and three akebia stem (Akebiae Caulis, Mutong) samples were collected and analyzed in the range of 4 000 - 400 cm⻹ by Fourier Transform Infrared (FTIR) and two-dimensional infrared correlation spectroscopy (2D-FTIR) techniques. The FTIR spectra of 24 Chuanmutong samples are consistent in the spectrum profiles, position and intensity of characteristic peaks. 20 of the 24 Chuanmutong samples were randomly selected as calibration samples to calculate and simulate mean spectrum. This mean spectrum is named as FTIR fingerprint of Chuanmutong with characteristic peaks at 3 412, 2 932, 1 739, 1 639, 1 509, 1 456, 1 426, 1 376, 1 332, 1 261, 1 159, 1 035, 897 ,609 cm⻹. Meanwhile, the limited level (Mean-3σ=0.992 6) to identify true or false Chuanmutong by correlation coefficient of FTIR spectra was calculated based on the 20 Chuanmutong calibration samples. Then, the rest 4 Chuanmutong, 4 Guanmutong and 3 Mutong samples were used as validation samples to evaluate the identification efficacy. The result shows that the FTIR spectra of 4 Chuanmutong validation samples were similar to the fingerprint. Their correlation coefficients of FTIR spectra were over the limited level and accepted as Chuanmutong. However, the spectra of Guanmutong and Mutong were significantly different from Chuanmutong fingerprint. The correlation coefficients of Guanmutong (0.902 1-0.940 4, n=4) and Mutong (0.954 9-0.978 9, n=3) FTIR spectra were less than the limited level and rejected from Chuanmutong. Furthermore, the number, position and intensity of auto-peaks on the 2D-FTIR were drastically different among the three herbs. It is concluded that the developed FTIR fingerprinting can be rapidly and accurately identify Chuanmutong and differentiate from related herbs.
Assuntos
Clematis/química , Medicamentos de Ervas Chinesas/química , Caules de Planta/química , Aristolochia/química , Asteraceae/química , Espectrofotometria Infravermelho , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
OBJECTIVE: To study the correlation between various complications of chronic heart failure (CHF) patients and Chinese medical syndromes, thus indicating distribution laws of Chinese medical syndromes in various complications of CHF patients. METHODS: Chinese medical syndrome typing was performed in 630 CHF patients by cross-sectional study of the demographic data, history of present diseases, related information on Chinese medical four diagnostic methods, and the distribution of complications. Logistic regression analysis was used to determine the correlation of various complications of CHF patients and Chinese medical syndromes. RESULTS: In this study, recruited were common complications such as hypertension, diabetes, arrhythmia, hyperlipemia, and cerebral vascular accident, and so on. Main syndromes were sequenced as qi deficiency syndrome, blood stasis syndrome, water retention syndrome, yin deficiency syndrome, phlegm turbid syndrome, yang deficiency syndrome. Results of Logistic regression analysis indicated that correlation existed between common complications and Chinese medical syndromes. In CHF complicated hypertension patients, Logistic regression analysis showed qi deficiency syndrome and yang deficiency syndrome were negatively correlated with hypertension (P < 0.05). In CHF complicated diabetes patients, Logistic regression analysis showed phlegm turbid syndrome and water retention syndrome were positively correlated with diabetes (P < 0.05). In CHF complicated arrhythmia patients, there was no statistical difference in the distribution of each syndrome (P > 0.05). In CHF complicated hyperlipemia patients, Logistic regression analysis showed qi deficiency syndrome and water retention syndrome were negatively correlated with hyperlipemia (P < 0.05), while blood stasis syndrome, yin deficiency syndrome, and phlegm turbid syndrome were positively correlated with hyperlipemia (P < 0.01). In CHF complicated cerebral vascular accident patients, Logistic regression analysis showed qi deficiency syndrome and yang deficiency syndrome were negatively correlated with cerebral vascular accident (P < 0.01, P < 0.05). CONCLUSIONS: There existed certain correlations between complications of CHF and the distribution of main Chinese medical syndromes. It could be used as guidance for treating CHF and its various complications by Chinese medicine and pharmacy.
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Insuficiência Cardíaca/complicações , Insuficiência Cardíaca/diagnóstico , Medicina Tradicional Chinesa , Idoso , Doença Crônica , Estudos Transversais , Feminino , Humanos , Modelos Logísticos , Masculino , Deficiência da Energia Yang/complicações , Deficiência da Energia Yang/diagnóstico , Deficiência da Energia Yin/complicações , Deficiência da Energia Yin/diagnósticoRESUMO
Danjiangkou Reservoir is a critical water source for the South-to-North Water Diversion Project, which harbors a diverse bacterioplankton community with varying depths, and the understanding of its nitrogen and phosphorus cycle and associated driving factors remains limited. In this study, we selected five ecological sites within Danjiangkou Reservoir and conducted metagenomics analysis to investigate the vertical distribution of bacterioplankton communities in the surface, middle, and bottom layers. Furthermore, we analyzed and predicted the function of nitrogen and phosphorus cycles, along with their driving factors. Our findings revealed the dominance of Proteobacteria, Actinobacteria, and Planctomycetes in the Danjiangkou Reservoir. Significant differences were observed in the structure of bacterioplankton communities across different depths, with temperature ï¼Tï¼, oxidation-reduction potential ï¼ORPï¼, dissolved oxygen ï¼DOï¼, and Chla identified as primary factors influencing the bacterioplankton composition. Analysis of nitrogen cycle functional genes identified 39 genes, including gltB, glnA, gltD, gdhA, NRT, etc., which were involved in seven main pathways, encompassing nitrogen fixation, nitrification, denitrification, and dissimilatory nitrate reduction. Phosphorus cycle function gene analysis identified 54 genes, including pstS, ppx-gppA, glpQ, ppk1, etc., primarily participating in six main pathways, including organic P mineralization, inorganic P solubilization, and regulatory. Cluster analysis indicated that different depths were significant factors influencing the composition and abundance of nitrogen and phosphorus cycle functional genes. The composition and abundance of nitrogen and phosphorus cycle functional genes in the surface and bottom layers differed and were generally higher than those in the middle layer. Deinococcus, Hydrogenophaga, Limnohabitans, Clavibacter, and others were identified as key species involved in the nitrogen and phosphorus cycle. Additionally, we found significant correlations between nitrogen and phosphorus cycle functional genes and environmental factors such as DO, pH, T, total dissolved solids ï¼TDSï¼, electrical conductivity ï¼ECï¼, and Chla. Furthermore, the content of these environmental factors exhibited depth-related changes in the Danjiangkou Reservoir, resulting in a distinct vertical distribution pattern of bacterioplankton nitrogen and phosphorus cycle functional genes. Overall, this study sheds light on the composition, function, and influencing factors of bacterioplankton communities across different layers of Danjiangkou Reservoir, offering valuable insights for the ecological function and diversity protection of bacterioplankton in this crucial reservoir ecosystem.
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Nitrogênio , Fósforo , Plâncton , Fósforo/metabolismo , China , Nitrogênio/metabolismo , Plâncton/genética , Plâncton/metabolismo , Bactérias/genética , Bactérias/metabolismo , Bactérias/classificação , Proteobactérias/genética , Ciclo do Nitrogênio , Actinobacteria/genética , Actinobacteria/metabolismo , Genes BacterianosRESUMO
BACKGROUND: Atherosclerosis (AS) is the most prevalent cardiovascular disease, with an exceptionally high burden. High-fat diet (HFD) is a popular diet behavior, whereas low-dose radiation (LDR) is an environmental physical factor. There is evidence to suggest that an HFD may exacerbate the onset of atherosclerosis. Whether the combination effect of HFD and LDR would have potential on atherosclerosis development remains incompletely unclear. METHODS: In this study, ApoE-/- mice were used as atherosclerosis model animals to investigate the combination effects of HFD and LDR (10 × 0.01Gy, or 20 × 0.01Gy) on vascular lesions. Doppler ultrasound imaging, H&E staining, oil red O staining, western blotting, and immunohistochemistry (IHC) were used to assess the pro-atherosclerotic effects. LC-MS was used to detect the non-targeted lipidomic. RESULTS: Long-term exposure of low-dose radiation at an accumulated dose of 0.2Gy significantly increased the occurrence of vascular stiffness and the aortic lesion in ApoE-/- mice. The synergistic effect of HFD and LDR was observed in the development of atherosclerosis, which might be linked to both the dysbiosis of lipid metabolism and the stimulation of the inflammatory signaling system. Moreover, LDR but not HFD can activate the cGAS-STING signaling through increasing the yield of cytosolic mitochondrial DNAs as well as the expression of cGAS protein. The activation of cGAS-STING signal triggers the release of IFN-α/-ß, which functions as an inflammatory amplifier in the formation of atherosclerotic plaque. CONCLUSION: The current study offers fresh insights into the risks and mechanism that underlie the development of atherosclerosis by LDR, and there is a combination effect of LDR and HFD with the involvement of cGAS-STING signal pathway.
Assuntos
Aterosclerose , Dieta Hiperlipídica , Nucleotidiltransferases , Transdução de Sinais , Animais , Masculino , Camundongos , Apolipoproteínas E/genética , Apolipoproteínas E/metabolismo , Aterosclerose/metabolismo , Aterosclerose/etiologia , Aterosclerose/patologia , Dieta Hiperlipídica/efeitos adversos , Modelos Animais de Doenças , Proteínas de Membrana/metabolismo , Proteínas de Membrana/genética , Camundongos Knockout para ApoE , Nucleotidiltransferases/metabolismo , Nucleotidiltransferases/genética , Transdução de Sinais/efeitos da radiaçãoRESUMO
The green and efficient remediation of soil cadmium (Cd) is an urgent task, and plant-microbial joint remediation has become a research hotspot due to its advantages. High-throughput sequencing and metabolomics have technical advantages in analyzing the microbiological mechanism of plant growth-promoting bacteria in improving phytoremediation of soil heavy metal pollution. In this experiment, a pot trial was conducted to investigate the effects of inoculating the plant growth-promoting bacterium Enterobacter sp. VY on the growth and Cd remediation efficiency of the energy plant Hybrid pennisetum. The test strain VY-1 was analyzed using high-throughput sequencing and metabolomics to assess its effects on microbial community composition and metabolic function. The results demonstrated that Enterobacter sp. VY-1 effectively mitigated Cd stress on Hybrid pennisetum, resulting in increased plant biomass, Cd accumulation, and translocation factor, thereby enhancing phytoremediation efficiency. Analysis of soil physical-chemical properties revealed that strain VY-1 could increase soil total nitrogen, total phosphorus, available phosphorus, and available potassium content. Principal coordinate analysis (PCoA) indicated that strain VY-1 significantly influenced bacterial community composition, with Proteobacteria, Firmicutes, Chloroflexi, among others, being the main differential taxa. Redundancy analysis (RDA) revealed that available phosphorus, available potassium, and pH were the primary factors affecting bacterial communities. Partial Least Squares Discriminant Analysis (PLS-DA) demonstrated that strain VY-1 modulated the metabolite profile of Hybrid pennisetum rhizosphere soil, with 27 differential metabolites showing significant differences, including 19 up-regulated and eight down-regulated expressions. These differentially expressed metabolites were primarily involved in metabolism and environmental information processing, encompassing pathways such as glutamine and glutamate metabolism, α-linolenic acid metabolism, pyrimidine metabolism, and purine metabolism. This study utilized 16S rRNA high-throughput sequencing and metabolomics technology to investigate the impact of the plant growth-promoting bacterium Enterobacter sp. VY-1 on the growth and Cd enrichment of Hybrid pennisetum, providing insights into the regulatory role of plant growth-promoting bacteria in microbial community structure and metabolic function, thereby improving the microbiological mechanisms of phytoremediation.
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The structural characteristics of the organic matter and biomarker distributions in Shengli lignite (SL) were comprehensively studied by combining a variety of modern analytical techniques and solvent extraction/thermal dissolution. Characterization of SL with Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, solid 13C nuclear magnetic resonance spectroscopy and thermogravimetry showed that organic matter in SL is rich in oxygen functional groups, such as C-O, >C=O, and -COOH, and hydrogen bonds. The hydrogen bonds mainly include -OH···π, self-associated -OH, -OH···ether O, tightly bound cyclic -OH, -OH···N, -COOH dimers, and -SH···N. The highest content of organic nitrogen and sulfur on SL surface are pyrrole nitrogen and aromatic sulfur, respectively. The proportions of aromatic and aliphatic carbons in SL are about 58% and 39%, respectively. The aromatic carbon is mainly composed of protonated aromatic and aromatic bridged carbons; methylene carbon has the highest content among the aliphatic carbons, with chains of average length of 1.43 carbon atoms. The average number of aromatic structural units in the carbon skeleton of SL is about 3, and each aromatic structural unit contains an average of 1-2 substituent groups. Thermogravimetric analysis clarified the distribution of the main types of covalent bonds in SL and their possible cracking temperatures during pyrolysis. The extracts and soluble portion of thermal dissolution from SL were analyzed by a gas chromatograph/mass spectrometer, and a series of biomarkers were identified, mainly concentrated in petroleum ether extract and cyclohexane thermal soluble portion. These included long-chain n-alkanes, isoprenoid alkanes, long-chain n-alkenes, terpenoids, n-alkan-2-ones, long-chain n-alkylbenzene, and long-chain n-alkyltoluene. The comprehensive characterization of the organic matter and the distribution of related biomarkers provided an important scientific basis for understanding the molecular structural characteristics and geochemical information on SL.
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OBJECTIVE: To develop a detection method for Cryptosporidium parvum oocysts from water samples, which combined immunomagnetic separation (IMS) with Taqman real-time PCR (qPCR). METHODS: Conditions of separation and enrichment of IMS method by using specific streptavidin magnetic beads coated with monoclonal antibodies Cp23 directed against C. parvum oocysts were optimized. Special primers of PCR and Taqman probes were designed referring to the 18S rDNA gene of C. parvum (GenBank Accession No. AB513881.1). The conserved genes were amplified from genomic DNA of C.parvum, and then cloned into Peasy-T1 vector. Tenfold dilutions of positive plasmids (10(4)-10(8) copy/microl) were used to construct a standard curves by Taqman qPCR. The specificity of the assay was determined using genomic DNA of C. baileyi, Toxoplasma gondii, C. canis and Escherichia coli. The sensitivity of this assay was evaluated by analyzing 10-fold serially dilutions of plasmids ranging from 10(0) to 10(8) copy/microl. Both IMS-qPCR assay and indirect immunofluorescent-antibody assay (IFA) were applied to detect 50 water samples collected from the dairy cattle farms in Hebei. RESULTS: The optimal incubation concentration and time of antibody Cp23 were 20 ng/ml and 30 min, respectively, and the catching time was 30 min, the recovery rate was more than 95%. PCR product was 272 bp, and identified by restriction enzyme digestion and nucleotide sequencing. There was a good linear relationship between the standard plasmids and Ct value (correlation r2 = 0.996 1) of the Taqman qPCR. No cross-reactivity was observed with C. baileyi, T. gondii, C. canis and E. coli. The sensitivity of C. parvum-specific assay was 10 copy/microl. Compared with IFA as golden standard method, the specificity and sensitivity of IMS-qPCR for 50 water samples was 100%(18/18) and 93.8% (30/32), respectively. CONCLUSION: The IMS-qPCR assay can be used to specifically detect C. parvum oocysts in water samples.
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Cryptosporidium parvum/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Água/parasitologia , Cryptosporidium parvum/genética , DNA Ribossômico/isolamento & purificação , Separação Imunomagnética/métodosRESUMO
BACKGROUND AND AIM: Nonalcoholic fatty liver disease (NAFLD) is not only the top cause of liver diseases but also a hepatic-correlated metabolic syndrome. This study performed untargeted metabolomics analysis of NAFLD hamsters to identify the key metabolites to discriminate different stages of NAFLD. METHODS: Hamsters were fed a high-fat diet (HFD) to establish the NAFLD model with different stages (six weeks named as the NAFLD1 group and twelve weeks as the NAFLD2 group, respectively). Those liver samples were analyzed by untargeted metabolomics (UM) analysis to investigate metabolic changes and metabolites to discriminate different stages of NAFLD. RESULTS: The significant liver weight gain in NAFLD hamsters was observed, accompanied by significantly increased levels of serum triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), alanine aminotransferase (ALT), and aspartate aminotransferase (AST). Moreover, the levels of TG, LDL-C, ALT, and AST were significantly higher in the NAFLD2 group than in the NAFLD1 group. The UM analysis also revealed the metabolic changes; 27 differently expressed metabolites were detected between the NAFLD2 and NAFLD1 groups. More importantly, the levels of N-methylalanine, allantoin, glucose, and glutamylvaline were found to be significantly different between any two groups (control, NAFLD2 and NAFLD1). Receiver operating characteristic curve (ROC) curve results also showed that these four metabolites are able to distinguish control, NAFLD1 and NAFLD2 groups. CONCLUSION: This study indicated that the process of NAFLD in hamsters is accompanied by different metabolite changes, and these key differently expressed metabolites may be valuable diagnostic biomarkers and responses to therapeutic interventions.
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Background: Ganoderma lucidum is reportedly the best source of traditional natural bioactive constituents. Ganoderma triterpenoids (GTs) have been verified as an alternative adjuvant for treating leukemia, cancer, hepatitis and diabetes. One of the major triterpenoids, Resinacein S, has been found to regulate lipid metabolism and mitochondrial biogenesis. Nonalcoholic fatty liver disease (NAFLD) is a common chronic liver disease that has become a major public health problem. Given the regulatory effects on lipid metabolism of Resinacein S, we sought to explore potential protective effects against NAFLD. Methods: Resinacein S was extracted and isolated from G. lucidum. And mice were fed with high fat diet with or without Resinacein S to detect hepatic steatosis. According to Network Pharmacology and RNA-seq, we analyzed the hub genes of Resinacein S against NAFLD disease. Results: Our results can be summarized as follows: (1) The structure of Resinacein S was elucidated using NMR and MS methods. (2) Resinacein S treatment could significantly attenuate high-fat diet (HFD)-induced hepatic steatosis and hepatic lipid accumulation in mouse. (3) GO terms, KEGG pathways and the PPI network of Resinacein S induced Differentially Expressed Genes (DEGs) demonstrated the key target genes of Resinacein S against NAFLD. (4) The hub proteins in PPI network analysis could be used for NAFLD diagnosis and treatment as drug targets. Conclusion: Resinacein S can significantly change the lipid metabolism in liver cells and yield a protective effect against steatosis and liver injury. Intersected proteins between NAFLD related genes and Resinacein S-induced DEGs, especially the hub protein in PPI network analysis, can be used to characterize targets of Resinacein S against NAFLD.
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OBJECTIVE: To explore the brain effect mechanism and the correlation between brain functional imaging and cognitive function in treatment of depressive disorder (DD) with transcutaneous auricular vagus nerve stimulation (taVNS) based on the resting-state functional magenetic reasonance imaging (rs-fMRI). METHODS: Thirty-two DD patients were included in a depression group and 32 subjects of healthy condition were enrolled in a normal group. In the depression group, the taVNS was applied to bilateral Xin (CO15) and Shen (CO10), at disperse-dense wave, 4 Hz/20 Hz in frequency and current intensity ≤20 mA depending on patient's tolerance, 30 min each time, twice daily. The duration of treatment consisted of 8 weeks. The patients of two groups were undertaken rs-fMRI scanning. The scores of Hamilton depression scale (HAMD), Hamilton anxiety scale (HAMA) and Wisconsin card sorting test (WCST) were observed in the normal group at baseline and the depression group before and after treatment separately. The differential brain regions were observed before and after treatment in the two groups and the value of degree centrality (DC) of fMRI was obtained. Their correlation was analyzed in terms of HAMD, HAMA and WCST scores. RESULTS: The scores of HAMD and HAMA in the depression group were all higher than those in the normal group (P<0.05). After treatment, the scores of HAMD and HAMA were lower than those before treatment in the depression group; the scores of total responses, response errors and perseverative errors of WCST were all lower than those before treatment (P<0.05). The brain regions with significant differences included the left inferior temporal gyrus, the left cerebellar peduncles region 1, the left insula, the right putamen, the bilateral supplementary motor area and the right middle frontal gyrus. After treatment, the value of DC in left supplementary motor area was negatively correlated to HAMD and HAMA scores respectively (r=-0.324, P=0.012; r=-0.310, P=0.015); the value of DC in left cerebellar peduncles region 1 was negatively correlated to the total responses of WCST (r=-0.322, P=0.013), and the left insula was positively correlated to the total responses of WCST (r=0.271, P=0.036). CONCLUSION: The taVNS can modulate the intensity of the functional activities of some brain regions so as to relieve depressive symptoms and improve cognitive function.